Whole-Genome Sequencing and De Novo Assembly of Malassezia pachydermatis Isolated from the Ear Canal of a Dog with Otitis.
We have de novo assembled the genome sequence of Malassezia pachydermatis isolated from a canine otitis sample with Nanopore-only long reads. With 99× coverage and 8.23 Mbp, the genome sequence was assembled in 10 contigs, with 6 of them corresponding to chromosomes, improving the scaffolding of previous genome assemblies for the species.


Coronary collaterals: the role of MCP-1 during the early phase of acute myocardial infarction.
The collateral growth (arteriogenesis) of an individual may vary from complete to absent during the early phase of acute myocardial infarction (AMI). However, the mechanisms underlying the large differences in the extent and adequacy of collateralization remain unclear. We hypothesized that shear stress-induced activation of monocyte chemoattractant protein-1 could potently contribute to the development of coronary collaterals during the early phase of AMI. We enrolled forty patients with AMI who did not receive reperfusion therapy within 24 h after the onset of chest pain and who also underwent coronary angiography (CAG) from 1 to 7 days after admission (mean duration: 3.6+/-2.2 days). The grades of the collateral development were angiographically defined and grouped according to the grade of collaterals as absent (score 0, n=20) or well-developed (score 2, n=20) collateral circulation. The plasma concentrations of vascular endothelial growth factor (VEGF), endostatin, monocyte chemoattractant protein-1 (MCP-1), and stromal cell-derived factor-1 (SDF-1) were assessed by enzyme-linked immunosorbent assay and then these values were compared between the two groups. There were no differences in the demographic and angiographic characteristics except for the number of total occlusion in culprit lesion. The plasma MCP-1 levels were significantly higher in the group with well-developed collateral circulation compared to the group with absent collateral circulation (262+/-216 vs. 151+/-88 pg/ml, respectively, p=0.043). However, the plasma levels of VEGF, endostatin and SDF-1 were not different on comparisons between the groups (VEGF; 369+/-377 vs. 324+/-363 pg/ml, endostatin; 1.74+/-1.71 vs. 1.49+/-1.15 ng/ml, SDF-1; 1806+/-508 vs. 2091+/-772 pg/ml, respectively). During the early phase of AMI, the plasma levels of MCP-1 were significantly increased in the patients with well-developed collateral circulation as compared to those patients with absent collateral circulation. These findings suggested that the shear stress-induced overexpression of MCP-1 contributes significantly to the development of coronary collaterals during the early phase of AMI.


Conditioned medium derived from 3D tooth germs: A novel cocktail for stem cell priming and early in vivo pulp regeneration.
Conditioned medium (CM) from 2D cell culture can mitigate the weakened regenerative capacity of the implanted stem cells. However, the capacity of 3D CM to prime dental pulp stem cells (DPSCs) for pulp regeneration and its protein profile are still elusive. We aim to investigate the protein profile of CM derived from 3D tooth germs, and to unveil its potential for DPSCs-based pulp regeneration. We prepared CM of 3D ex vivo cultured tooth germ organs (3D TGO-CM) and CM of 2D cultured tooth germ cells (2D TGC-CM) and applied them to prime DPSCs. Influences on cell behaviours and protein profiles of CMs were compared. In vivo pulp regeneration of CMs-primed DPSCs was explored using a tooth root fragment model on nude mice. TGO-CM enhanced DPSCs proliferation, migration, in vitro mineralization, odontogenic differentiation, and angiogenesis performances. The TGO-CM group generated superior pulp structures, more odontogenic cells attachment, and enhanced vasculature at 4 weeks post-surgery, compared with the TGC-CM group. Secretome analysis revealed that TGO-CM contained more odontogenic and angiogenic growth factors and fewer pro-inflammatory cytokines. Mechanisms leading to the differential CM profiles may be attributed to the cytokine-cytokine receptor interaction and PI3K-Akt signalling pathway. The unique secretome profile of 3D TGO-CM made it a successful priming cocktail to enhance DPSCs-based early pulp regeneration.


Radiosynthesis and evaluation of [11C]AG-488, a dual anti-angiogenetic and anti-tubulin PET ligand.
Combinations of antiangiogenic and cytotoxic agents show promising results in the treatment of cancer. However, there is a lack of single agent with both antiangiogenic and cytotoxic activities for clinical application. AG-488 aka FLAG-003 is a novel ligand with established antiangiogenetic properties via activation of receptor thymidine kinase (RTK) and anti-tubulin properties in tumor cells. AG-488 is also reported to reduce tumor volume and prolong survival in preclinical animal models of glioblastoma multiforme, breast cancer and is in clinical stage. Higher expression of RTKs and tubulins is reported in various cancers. This study reveals the development of [11C]AG-488, a high affinity dual target inhibitor binding to RTK and anti-tubulin activities. We rationale that antiangiogenic RTK and anti-tubulin activity of [11C]AG-488 may enhance the tumor to tissue ratio, assisting in cancer drug development. [11C]AG-488 was synthesized in 35 ± 5 % radiochemical yield by radiomethylating the corresponding phenolate using [11C]CH3I. MicroPET studies in mice indicated blood-brain barrier penetration of [11C]AG-488 and retention in the brain. However, blocking studies with antitubulin and RTK agent HD-800 and microtubule depolymerizing agent MPC-6827 show increased binding of [11C]AG-488 in brain. The pattern of tracer binding in blocking conditions is similar to the baseline conditions. The higher binding may be due to the increased plasma uptake of radiotracer or the formation of more free tubulins due to microtubule dynamic instability during the blocking conditions.


[Argentine consensus on the diagnosis and therapeutics of treatment resistant schizophrenia].
Approximately 30% of people with schizophrenia fail to respond to first-line antipsychotic treatment which impacts the burden of the disease. Treatment-resistant schizophrenia (TRS) denotes patients with failure to respond to at least two adequate trials of different antipsychotics. Clozapine is a unique drug approved for treating treatment-resistant schizophrenia, however 1/3 of patients fail to respond to clozapine. Even though different strategies have been proposed for treating clozapine-resistant schizophrenia, the evidence is very limited, unclear, and of poor quality. A formal literature search was conducted and then, panel members were asked to complete 35 questions addressing different aspects of TRS. A modified Delphi method was used to unify expert opinion and achieve consensus. The expert consensus in diagnostic and treatment of TRS is the result of experts from the main national scientific societies under the organization of the Argentine Association of Biological Psychiatric (AAPB). The consensus statement aims to guide on diagnosis and treatment.


Congenital pyloric atresia, presentation, management, and outcome: a report of 20 cases.
Congenital pyloric atresia (CPA) is a very rare anomaly. It is usually seen as an isolated condition with excellent prognosis. Few cases are familial. These are usually associated with other hereditary conditions and have a poor prognosis. This is a review of our experience with 20 patients with CPA outlining aspects of diagnosis, associated anomalies and management. This is a retrospective analysis of 20 cases seen over a 22 year period (December 1990 to December 2012). Their records reviewed for: age, sex, presentation, prenatal history, associated anomalies, investigations, treatment, operative findings and the outcome. 20 cases (9 Males, 11 Females) were treated. 7 patients were full term and the remaining 13 were prematures. Their mean birth weight was 2.1 kg (1.1 kg to 3.9 kg). Polyhydramnios was seen in 13 patients (65%). Two were brothers and four were members of the same family. Isolated CPA was seen in 7 (35%); 13 had an associated conditions: epidermolysis bullosa (EB) in 8 (40%) and multiple intestinal atresias (MIA) in 5 (25%). Three patients had associated esophageal atresia. All were operated on except two who died early due to unrelenting sepsis. The variety of pyloric atresias encountered were as follows: pyloric diaphragm in 13 including double diaphragms in 2, pyloric atresia with a gap in 4 and pyloric atresia without gap in 3. Ten died postoperatively giving an overall survival of 40%. CPA is a very rare condition. Isolated CPA carries a good prognosis. Association of CPA with other familial and congenital anomalies like EB and MIA carries a poor prognosis.


Helicobacter pylori diversification during chronic infection within a single host generates sub-populations with distinct phenotypes.
Helicobacter pylori chronically infects the stomach of approximately half of the world's population. Manifestation of clinical diseases associated with H. pylori infection, including cancer, is driven by strain properties and host responses; and as chronic infection persists, both are subject to change. Previous studies have documented frequent and extensive within-host bacterial genetic variation. To define how within-host diversity contributes to phenotypes related to H. pylori pathogenesis, this project leverages a collection of 39 clinical isolates acquired prospectively from a single subject at two time points and from multiple gastric sites. During the six years separating collection of these isolates, this individual, initially harboring a duodenal ulcer, progressed to gastric atrophy and concomitant loss of acid secretion. Whole genome sequence analysis identified 1,767 unique single nucleotide polymorphisms (SNPs) across isolates and a nucleotide substitution rate of 1.3x10-4 substitutions/site/year. Gene ontology analysis identified cell envelope genes among the genes with excess accumulation of nonsynonymous SNPs (nSNPs). A maximum likelihood tree based on genetic similarity clusters isolates from each time point separately. Within time points, there is segregation of subgroups with phenotypic differences in bacterial morphology, ability to induce inflammatory cytokines, and mouse colonization. Higher inflammatory cytokine induction in recent isolates maps to shared polymorphisms in the Cag PAI protein, CagY, while rod morphology in a subgroup of recent isolates mapped to eight mutations in three distinct helical cell shape determining (csd) genes. The presence of subgroups with unique genetic and phenotypic properties suggest complex selective forces and multiple niches within the stomach during chronic infection.


Predicting event times in clinical trials when randomization is masked and blocked.
Timing for interim or final analysis of data in an event-based trial is often determined by the accrual of events during the study. Existing Bayesian methods may be used to predict the date of the landmark event using observed enrollment, event, and loss times when treatment arm information is masked. For event-based trials with a blocked randomization, knowledge of blocks in which patients are enrolled can provide additional information to improve predictions versus models that only assume a known treatment allocation proportion. We therefore propose to incorporate blocking information into existing methods for prediction. We derive a latent variable (LV) extension of a mixture model used in Donovan JM, Elliott MR, Heitjan DF. Predicting Event Times in Clinical Trials When Treatment Arm is Masked. J Biopharmaceut Stat 2006; 16:343-56 to incorporate block randomization (constrained LV) for predicting the landmark event and compare this model with (a) methods where blocking information is ignored (unconstrained LV), and (b) methods assuming a single population (SP). Comparison of the constrained and unconstrained LV models in our application shows that the constrained LV model has narrower prediction intervals. Simulation studies show that the constrained LV model can have better coverage probabilities for the prediction intervals than SP models if a treatment effect is present, and prediction intervals from the constrained LV model are narrower than those for the unconstrained LV model. These differences varied by block size and prediction time. We have limited focus to the exponential model for events. Coverage for the LV models may be somewhat reduced if no treatment effect is present. Extra information provided by knowledge of blocking can be used to decrease prediction interval width versus the unconstrained LV model, while providing better coverage properties than the SP model if a treatment effect is present.


Religion, spirituality, coping, and resilience among African Americans with diabetes.
Study aims were to explore how religion and spirituality impacted attitudes about self-management practices among African Americans with homelessness histories and to understand resilience in diabetes care practices. Qualitative semi-structured face-to-face interviews were conducted with 42 African Americans older than 18 years. All audio-taped interviews lasted between 1-1.5 hr, transcribed verbatim, and analyzed using ATLAS.ti (version 7.0). Five resilience themes emerged. While participants recognized diabetes as an illness requiring professional treatment, the context of balancing treatment with religion and spiritual practices mattered. The study findings highlight the importance of spirituality, religious beliefs, and coping strategies in diabetes self-care activities.


Forecasting the COVID-19 effects on energy poverty across EU member states.
The COVID-19 pandemic is manifesting its devastating effects in multiple directions, even indirectly affecting the health of citizens, for instance, by increasing the level of energy poverty (EP). As part of the studies that are trying to frame the consequences of the pandemic, this paper aims to identify the effects on EP in the coming years in a bid to identify the countries of the European Union most affected and the time span necessary to return to a path to reduce EP. For this purpose, an analysis based on the supervised learning algorithms of dynamic factor models is carried out. The outcomes of this investigation show that the negative effects of the pandemic on the level of EP will be reabsorbed very slowly, not before 2025, and in any case with substantial differences between countries, further widening the gap between countries with low levels of EP and those with greater EP levels.


Asymmetic division: dynastic intricacies of neuroblast division.
The cytoplasmic determinants Numb and Prospero are distributed asymmetrically into the daughter cells of Drosophila neuroblasts. The proteins encoded by the genes inscuteable, staufen and miranda are involved in the localisation of Prospero.


The production and characterization of SARS-CoV-2 RNA reference material.
SARS-CoV-2 in vitro transcribed RNA reference materials (RM), UME RM 2019 and UME RM 2020, were produced by Scientific and Technological Research Council of Turkey (TUBITAK), National Metrology Institute (UME), to be used as a quality control material for SARS-CoV-2 measurements, in liquid-frozen and lyophilized forms, respectively. These RNA RMs include ten internationally recommended SARS-CoV-2 target gene fragments (Pasteur-RdRp-IP2, Pasteur-RdRp-IP4, Charite-E, Charite-RdRp, CDC-N1, CDC-N2, China CDC-ORF1ab, China CDC-N, Hong Kong-ORF1b, and Hong Kong-N) for virus detection and one human gene fragment (RNase P) as an internal control. Two different platforms, RT-qPCR and RT-ddPCR, were used to characterize UME RM 2019 (UME RM 2020 was only characterized with RT-qPCR). The homogeneity studies were evaluated by RT-qPCR. According to these results, it has been shown that both reference materials are homogeneous for intended use. Short-term studies were also conducted similarly for mimicking transport conditions and UME RM 2020, which is produced in lyophilized form, unlike other reference materials available in the market, provides convenience for users by ensuring that the reference material remains stable for 17 days even at 45 °C temperature. The lyophilized formulation of the reference material had greater stability which would allow it to be shipped without cooling items. The development of such RNA reference materials provides quality control for existing and newly designed RNA-based virus detection tests and it helps the prevention and control of epidemics.


Linear solvation energy relationships of anionic dimeric surfactants in micellar electrokinetic chromatography II. Effect of the length of a hydrophilic spacer.
Anionic dimeric surfactants with hydrophilic spacers containing two to six oxygen atoms were synthesized and applied as pseudostationary phases in micellar electrokinetic chromatography. Their selectivity was determined via linear solvation energy relationships. There were no differences in cohesiveness, polarizability or dipolarity with increasing spacer length, but there was a clear trend in increasing hydrogen bond accepting ability, and a concomitant decrease in hydrogen bond donating ability. The different selectivity of these dimeric surfactants compared to sodium dodecylsulfate can be useful for optimizing separations of mixtures of solutes for which these types of interactions are important. Their critical micelle concentrations were in the range of 0.2-0.3mM, except for the surfactant with the shortest spacer (<0.03 mM), and are much lower than those of conventional surfactants used in micellar electrokinetic chromatography.


Combination antimicrobial therapy.
In spite of a large volume of data regarding the in vitro activity of single and combined antimicrobial activity, the clinical relevance of these studies is unclear. Few comparative trials of combined and single antibiotic therapy of human infection have been performed. Synergistic combination therapy has been shown to be beneficial in a few specific circumstances. Antagonistic combinations should be avoided in the treatment of meningitis, endocarditis, and infections of immunocompromised patients. The bactericidal titer of serum or spinal fluid should reflect adequacy of therapy of meningitis, endocarditis, and osteomyelitis, and adjustments can be made accordingly.


A novel digital arthrometer to measure anterior tibial translation.
Measurement of knee laxity after anterior cruciate ligament (ACL) injury is crucial for appropriate treatment and rehabilitation decision-making. This study examined the potential of a new digital arthrometer (Ligs, Innomotion, Shanghai, China) to quantify anterior tibial translation (ATT) in patients with ACL injuries and in healthy subjects. A total of 60 participants included 30 subjects with single-leg ACL injuries and 30 healthy subjects included as controls. The lower leg was immobilized. The thruster is positioned posterior to the lower leg and parallel to the tibial tuberosity in the sagittal plane. The load is applied vertically to the tibia under a dynamic load of 0-150 N, with continuous displacement recorded. The intrarater and interrater reliability will be examined. ATT and side-to-side differences (SSD) between the control and ACL injury groups were compared. Receiver operating characteristic (ROC) curves were analyzed, and the area under the curve (AUC) was calculated to determine the diagnostic accuracy of the Ligs. The interrater ICC was 0.909 and the intrarater ICC was 0.943. Significant differences in the SSD were observed between the control and ACL injury groups (for all P < 0.05), with the largest effect size (ES = 1.12) at 80 N. When comparing ATT at different loads between injured and healthy sides in the ACL injury group, displacement was statistically significant at different loads. At a load of 150 N, the AUC was the maximum (0.857) and the sensitivity and specificity were 0.87 and 0.73, respectively. A digital arthrometer can be used as a quantitative instrument to quantify knee laxity. Quantitative measurement of ATT and SSD under controlled loading can be an objective and effective tool for clinical practice.


How reliable is the manual correction of the autoscoring of a level IV sleep study (ApneaLink™) by an observer without experience in polysomnography?
This study aims to compare the manual correction of the automatic analysis of ApneaLink™ between a skilled observer in the interpretation of sleep studies and a subject trained only in the scoring of ApneaLink™ device. Ninety-six subjects performed the ApneaLink™ and polysomnography (PSG) simultaneously in the sleep laboratory. Two blind observers, who were independent from the results of the PSG, performed first the automatic scoring and then the hand correction from the ApneaLink™ device. The scorers of ApneaLink™ represented two physicians with different levels of training (scorer A: 20 years of experience in reading polysomnography plus 3 years of experience in the interpretation of ApneaLink™, scorer B: 1 year of experience in the analysis of ApneaLink™). The interobserver agreement was assessed with the intraclass correlation coefficient (ICC) and kappa statistics. The diagnostic accuracy of the manual analysis ApneaLink™ device was evaluated by the area under the receiver operator curve (AUC-ROC). Ninety patients were included (69 men; mean age, 49.6; median RDI, 13.9; median BMI, 29.3 Kg/m(2)). The ICC between the manual apnea/hypopnea index from ApneaLink™ and the respiratory disturbance index of the PSG for each observer was similar (scorer A, 0.902; CI 95% 0.80-0.95; vs. scorer B, 0.904; CI 95% 0.86-0.94; p = 0.9). The agreement between the observers on the presence or absence of obstructive sleep apnea syndrome (OSAS) was very good (kappa, 0.83; CI 95% 0.69-0.98). The AUC-ROC was similar between the observers (scorer A, 0.88; CI 95% 0.78-0.98; scorer B, 0.83; CI 95% 0.71-0.95; p = 0.5). The non-expert observer showed a very good agreement with the expert observer on the results of the manual correction of the ApneaLink™ autoscoring. Both observers had similar diagnostic accuracy to identify subjects with OSAS when compared with PSG.


The effects of limbic lesions on locomotion and stereotypy elicited by dopamine agonists in the rat.
The purpose of this experiment was to investigate the functional contributions of various limbic structures to locomotion and stereotypy induced by dopaminergic drugs. Female rats were randomly assigned to one of 5 groups (n = 10-14 rats/group) that received either a lesion of the hippocampus (colchicine + kainic acid), basolateral amygdala (quinolinic acid), frontal cortex (aspiration), nucleus accumbens (ibotenic acid), or served as unoperated controls. Beginning at least 2 weeks following surgery locomotion (measured as photocell beam breaks) elicited by D-amphetamine (0.0, 0.32, 1.0 and 3.2 mg/kg), SKF 82958 (0.0, 0.04, 0.08 and 0.16 mg/kg) or quinpirole (0.0, 0.25, 0.1 and 0.5 mg/kg) was determined. In agreement with previous results rats with hippocampal lesions were hyperactive in response to amphetamine. In comparison to these changes in drug-induced locomotion, lesions of the basolateral amygdala, and frontal cortex had only minor effects on drug-induced locomotion. Lesions of the nucleus accumbens produced consistent hyperactivity that was suppressed by doses of amphetamine or quinpirole that elicited behavioral stereotypy. These results provide evidence suggesting that, in comparison to other limbic structures that have substantial inputs to the nucleus accumbens, the hippocampus play a relatively prominent role in the modulation of drug-induced locomotion.


Tranexamic Acid in the Treatment of Melasma: A Review of the Literature.
Melasma is a common acquired pigmentary disorder marked by irregular hyperpigmented macules or patches and most commonly occurs in women of darker skin color. It is a chronic often-relapsing condition that causes negative psychosocial effects in those affected. Current treatments such as hydroquinone, kojic acid, and retinoids, among others, demonstrate variable efficacy and side-effect profiles. We conducted a comprehensive literature review examining the use of tranexamic acid (TA), a well-known anti-fibrinolytic agent, in the treatment of melasma. TA delivered orally, topically, and through physical methods works via the inhibition of ultraviolet (UV)-induced plasmin activity in keratinocytes. Predefined search terms were entered into PubMed. Articles were then independently screened by two authors to include only those written in the English language and relating to human subjects with at least mild melasma. The search identified 28 articles, 15 of which met the criteria for full review. The review revealed that TA treatment for melasma is equally effective or more effective than other standard therapies and may induce fewer side effects. Our comprehensive review suggests that TA may be a promising treatment option for melasma because of its demonstrated effectiveness alone and in combination with other modalities as well as its limited side-effect profile.


A scenario-based approach for urban water management in the context of the COVID-19 pandemic and a case study for the Tabriz metropolitan area, Iran.
The world's poorest countries were hit hardest by COVID-19 due to their limited capacities to combat the pandemic. The urban water supply and water consumption are affected by the pandemic because it intensified the existing deficits in the urban water supply and sanitation services. In this study, we develop an integrated spatial analysis approach to investigate the impacts of COVID-19 on multi-dimensional Urban Water Consumption Patterns (UWCPs) with the aim of forecasting the water demand. We selected the Tabriz metropolitan area as a case study area and applied an integrated approach of GIS spatial analysis and regression-based autocorrelation assessment to develop the UWCPs for 2018, 2019 and 2020. We then employed GIS-based multi-criteria decision analysis and a CA-Markov model to analyze the water demand under the impacts of COVID-19 and to forecast the UWCPs for 2021, 2022 and 2023. In addition, we tested the spatial uncertainty of the prediction maps using the Dempster Shafer Theory. The results show that the domestic water consumption increased by 17.57% during the year 2020 as a result of the COVID-19 pandemic. The maximum increase in water consumption was observed in spring 2020 (April-June) when strict quarantine regulations were in place. Based on our results, the annual water deficit in Tabriz has increased from ~18% to about 30% in 2020. In addition, our projections show that this may further increase to about 40-45% in 2021. Relevant stakeholders can use the findings to develop evidence-informed strategies for sustainable water resource management in the post-COVID era. This research also makes other significant contributions. From the environmental perspective, since COVID-19 has affected resource management in many parts of the world, the proposed method can be applied to similar contexts to mitigate the adverse impacts and developed better informed recovery plans.


Prophylactic use of antipyretic agents with childhood immunizations and antibody response: reason for concern?
In the pediatric primary care setting, well-child visits constitute over 50% of all encounters, treating over 24 million children annually. Anticipatory guidance topics vary based on different ages, but immunizations are a focal point of all well-child visits. This article addresses the prophylactic use of antipyretic agents with the administration of immunizations as a potential reason of concern. A literature review of the use of antipyretic agents in conjunction with immunizations and the effectiveness of treatment was performed. Based on several studies, the standard recommendation of administering antipyretic agents with immunization administration was a routine. Twenty years later, the scientific evidence was questioned. A pivotal study questioned these standards, noting no benefit and potential decreased immune response. Although the prophylactic use of antipyretics has been a standard in pediatrics, the lack of scientific support in the reduction of adverse effects of the vaccinations and the possibility of decreased immune response warrants further research.


Functional neural correlates of psychopathy: a meta-analysis of MRI data.
Neuroimaging studies over the last two decades have begun to specify the neurobiological correlates of psychopathy, a personality disorder that is strongly related to criminal offending and recidivism. Despite the accumulation of neuroimaging studies of psychopathy, a clear and comprehensive picture of the disorder's neural correlates has yet to emerge. The current study is a meta-analysis of functional MRI studies of psychopathy. Multilevel kernel density analysis was used to identify consistent findings across 25 studies (460 foci) of task-related brain activity. Psychopathy was associated with increased task-related activity predominantly in midline cortical regions overlapping with the default mode network (dorsomedial prefrontal cortex, posterior cingulate, and precuneus) as well as medial temporal lobe (including amygdala). Psychopathy was related to decreased task-related activity in a region of the dorsal anterior cingulate cortex overlapping with the salience network. These findings challenge predominant theories of amygdala hypoactivity and highlight the potential role of hyperactivity in medial default mode network regions and hypoactivity in a key node of the salience network during task performance in psychopathy.


UiO-66(Zr) coupled with Bi(2)MoO(6) as photocatalyst for visible-light promoted dye degradation.
This study provides the first attempt to combine UiO-66(Zr) with Bi2MoO6 for the preparation of composite via electrostatic interaction induced self-assembly process. Synthesized products were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared spectroscope (FT-IR), X-ray photoelectron spectroscopy (XPS), Brunauer-Emmett-Teller (BET) surface area analyse and UV-vis diffuse reflectance spectroscopy (UV-vis DRS). The photocatalytic activities of as-obtained Bi2MoO6/UiO-66(Zr) composites were assessed by degradation of Rhodamine B (Rh B) under the visible-light irradiation. Comparing to the pristine materials, the developed Bi2MoO6/UiO-66(Zr) composite with Bi:Zr molar ratio 2:1, labelled as BMUO-2, showed the best photocatalytic activity. It was supposed that the participation of Bi2MoO6 increased the use of solar energy and the existence of UiO-66(Zr) restrained the recombination rate of photo-generated electron-hole pairs. Moreover, Bi2MoO6/UiO-66(Zr) composite showed excellent stability and recyclability during cycling experiment. The mechanism and plausible pathway of Rh B degradation were also discussed in this paper.


Cortical bone failure mechanisms during screw pullout.
An experimental and computational study of screw pullout from cortical bone has been conducted. A novel modification of standard pullout tests providing real time image capture of damage mechanisms during screw pullout was developed. Pullout forces, measured using the novel test rig, have been validated against standard pullout tests. Pullout tests were conducted, considering osteon alignment, to investigate the effect of osteons aligned parallel to the axis of the orthopaedic screw (longitudinal pullout) as well as the effect of osteons aligned perpendicular to the axis of the screw (transverse pullout). Distinctive alternate failure mechanisms, for longitudinally and transversely orientated cortical bone during screw pullout, were uncovered. Vertical crack propagation, parallel to the axis of the screw, was observed for a longitudinal pullout. Horizontal crack propagation, perpendicular to the axis of the screw, was observed for a transverse pullout. Finite element simulation of screw pullout, incorporating material damage and crack propagation, was also performed. Simulations revealed that a homogenous material model for cortical bone predicts vertical crack propagation patterns for both longitudinal and transverse screw pullout. A bi-layered composite model representing cortical bone microstructure was developed. A unique set of material and damage properties was used for both transverse and longitudinal pullout simulations, with only layer orientations being changed. Simulations predicted: (i) higher pullout forces for transverse pullout; (ii) horizontal crack paths perpendicular to screw axis for transverse pullout, whereas vertical crack paths were computed for longitudinal pullout. Computed results agreed closely with experimental observations in terms of pullout force and crack propagation.


Detection of porcine circovirus type 2 (PCV2) in the Philippines and the complexity of PCV2-associated disease diagnosis.
Porcine circovirus type 2 (PCV2), an important pig viral pathogen, can cause porcine circovirus-associated disease (PCVAD), resulting in economic losses associated with decreased growth and mortalities. The diagnosis of PCVAD is complex requiring clinical, pathological and virological approaches. This study assessed PCV2 infection using histopathology and immunohistochemistry (IHC) on tissue samples and quantitative polymerase chain reaction (qPCR) on serum samples from 47 grower-finisher pigs allocated in three clinical groups in the Philippines. Typical PCV2 histopathological lesions were observed in mediastinal lymph nodes (MLN) of eight of 47 pigs. Lymphoid depletion was seen in all eight pigs and granulomatous inflammation in one of these pigs. Four of these eight pigs were PCV2 positive by both IHC and qPCR. IHC revealed PCV2 antigen in 8 pigs in at least one of the following tissues: MLN (5/8), spleen (3/8), tonsils (4/8) and lungs (5/8). PCV2 antigen was observed in 3/8 MLN with lymphoid depletion and in one MLN with depletion and granulomatous inflammation. The qPCR test showed that 33 sera had a non-detectable level, twelve had < 106 and two had > 106 PCV2 DNA copies/ml serum. One pig with lymphoid depletion had > 106 PCV2 DNA copies/ml serum, and another pig without MLN lesions also had > 106 PCV2 DNA copies/ml serum. These findings suggest that PCVAD is present in the Philippines and confirm the challenges of PCVAD diagnosis as different patterns of results were obtained from the different tests.


Transfemoral amputee intact limb loading and compensatory gait mechanics during down slope ambulation and the effect of prosthetic knee mechanisms.
Intact limb knee osteoarthritis is a prevalent secondary disability in transfemoral amputees. Walking down a ramp may increase this risk due to excessive limb loading. We sought to determine whether intact limb loading differed between transfemoral amputees and controls during down slope ambulation, and the compensatory strategies transfemoral amputees used to modify intact limb loading. Secondarily, we sought to determine the effect of prosthetic knee type. Five unilateral transfemoral amputees and five non-amputee controls walked down a ramp and the following outcome measures were compared between amputees and controls and across prosthetic knee type (C-leg versus Power Knee): step length, walking speed, leading limb ground reaction forces, and trailing and leading limb ankle and knee energy absorption. Linear mixed effects regression was used to test for association between gait variables and limb. There were no significant differences in intact limb loading between amputees and controls or between prosthetic knee types. Transfemoral amputees walked slower (C-leg - control = -0.29 m/s; P = 0.008, Power Knee - control = -0.38 m/s; P < 0.001) with a shorter intact limb step length (C-leg - control = -0.12 m/s; P < 0.001, Power Knee - control = -0.16 m/s; P < 0.001). The prosthetic trailing limb ankle absorbed less energy throughout stance than the trailing limb in controls (C-leg-control = -0.22 J/kg; P < 0.001, Power Knee - control = -0.22 J/kg; P < 0.001). Intact limb loading in transfemoral amputees is equivalent to controls during down ramp ambulation, in spite of reduced prosthetic trailing limb energy absorption. The primary compensatory strategies include a reduced ambulation speed and intact limb step length, which reduces center of mass velocity at heel contact.


A clinically relevant large-animal model for evaluation of tissue-engineered cardiac surgical patch materials.
Extracellular matrix (ECM) scaffolds may be useful as a tissue engineering approach toward myocardial regeneration in the infarcted heart. An appropriate large-animal model for testing the utility of biologically derived ECM in this application is needed. The purpose of this study was to develop such a model for optimal procedural success during and after patch implantation surgery. Myocardial infarction (MI) was created by embolization of the diagonal artery (DA) branch of the left anterior descending coronary artery with collagen suspension. After 4 to 6 weeks, 14 pigs received patch implant (ECM or expanded polytetrafluoroethylene). Six pigs were infarcted in the first DA and seven pigs in the second DA. Electrophysiology study was performed within 3 days before surgery. During surgery, the size and location of the infarct were measured. Infarcted myocardium (1.5-cm diameter) was transmurally excised under partial cardiopulmonary bypass. Patches (3-cm diameter) were sutured to the endomyocardial defect. Four pigs died postoperatively. After 1 month, 10 pigs were euthanized and the locations of patches were examined. Success rate of patch implant in the second DA (85.7%) was higher than the first DA (50%) group. Infarct size in the second DA was smaller than in the first DA (4.6+/-1.2 vs. 10.8+/-2.4 cm(2), P<.05). The second DA was more anteriorly positioned, which enabled easier access from the midsternal thoracotomy. However, the first DA was more laterally located requiring more manipulation of the heart during surgery. Electrophysiology revealed no ventricular tachyarrhythmia in the second DA but 33.3% in the first DA group (P<.05). At necropsy, the endocardial position of the first DA-infarct patches was anteroapical, whereas the second DA-infarct patches were more basolateral and often involved the anterior papillary muscle. The success rate of patch implant was associated with infarction size and location, and may be related to arrhythmic substrate. Experimental MI created by the second DA embolization is a feasible model for investigation of tissue-engineered cardiac patch implantation. This large-animal model is also suitable for study of cell therapy via endocardial catheter-based approaches or open surgical methods.


Multiplex PCR-based DNA array for simultaneous detection of three human herpesviruses, EVB, CMV and KSHV.
Human lymphotropic herpesviruses, Epstein-Barr virus (EBV), cytomegalovirus (CMV) and Kaposi's sarcoma-associated herpesvirus (KSHV) are responsible for a wide variety of human diseases. Due to an increase in diseased states associated with immunosuppression, more instances of co-morbid infections with these herpesviruses have resulted in viral reactivations that have caused numerous fatalities. Therefore, the development of rapid and accurate method to detect these viruses in immunocompromised patients is vital for immediate treatment with antiviral prophylactic drugs. In this study, we developed a new multiplex PCR method coupled to DNA array hybridization, which can simultaneously detect all three human herpesviruses in one single cell sample. Multiplex PCR primers were designed to amplify specific regions of the EBV (EBER1), CMV (IE) and KSHV (LANA) viral genomes. Pre-clinical application of this method revealed that this approach is capable of detecting as few as 1 copy of the viral genomes for KSHV and CMV and 100 copies of the genome for EBV. Furthermore, this highly sensitive test showed no cross-reactivity among the three viruses and is capable of detecting both KSHV and EBV viral genomes simultaneously in the lymphoblastoid cells that have been double infected with both viruses. Thus, this array-based approach serves as a rapid and reliable diagnostic tool for clinical applications.


Investigation of the potential effects of metformin on atherothrombotic risk factors in hyperlipidemic rats.
The increased mortality rate due to atherothrombotic events and related complications has necessitated the search for new pharmacological agents. Hyperlipidemia, thrombosis and oxidative stress are the primary underlying concerns in the pathogenesis of atherosclerosis. Metformin, although proved to be beneficial in micro and macrovascular complications of diabetes mellitus, its effects on pure cardiovascular subjects are still debatable. Hence, the aim of the present study was to investigate the effects of metformin on atherothrombotic risk factors in experimental hyperlipidemic rats. Hyperlipidemia was induced by an intra-peritoneal injection of criton X-100 (25 mg/kg). Assessment of the effects of metformin (300 mg/kg/day, 400 mg/kg/day and 500 mg/kg/day) on lipid profile, coagulation time (activated partial thromboplastin time and prothrombin time), fibrinogen level, thrombosis, lipid peroxidation, antioxidant enzymes level, plasma fluorescent oxidation products and aortic nitrite level revealed an overall improvement in the lipid profile at the dose of 400 mg/kg along with a significant reduction in oxidative stress as compared to criton X-100 treated control. Activated partial thromboplastin and prothrombin times were prolonged at all doses, while plasma fibrinogen level remained unaffected. Metformin pre-treatment also reduced endothelial cell damage in ferrous chloride induced thrombosis in carotid arteries. Thus, the results indicate a potential protective effect of metformin on atherothrombotic risk factors, as evident from an improvement in lipid profile, reduction in oxidative stress and thrombotic events.


Athlete Heart in Children and Young Athletes. Echocardiographic Findings in 331 Cases.
The changes of the athlete's heart are not well defined and characterized in children. We aimed to describe the morphological changes of the heart related to sport in young athletes. We evaluated a group of 331 young athletes under 18 years (mean 11.9 ± 3.2) who practice tennis: 58 (16.52%), football: 118 (33.62%), basketball: 16 (4.56%), athletics: 40 (11.4%), and swimming: 99 (28.21%). Type of sport, years of practice, and duration of the training were collected. All children underwent echocardiography with the following M-mode parameters: left atrium diameter (LAD), interventricular septum (IVS), and left ventricle posterior Wall (LVPW), diastolic diameter of the left ventricle (LVDD), and right ventricle outflow tract (RVOT). The major finding of our study was that 20% of the children had a Z score > 2 for the IVS and that increased to 30% for the children playing tennis or swimming. Also, other changes like LA and RVOT dilatation were observed in about 10 and 14% of the cases, respectively. Taken together, these figures indicate that cardiac remodeling is frequent in children. Further studies are needed to establish consensus-based criteria of athlete's heart in young children.


Serum M3/M21 in ovarian cancer patients.
Cytokeratins are polypeptides that constitute a subclass of intermediate filaments in epithelial cells. The aim of the present study was to evaluate the clinical usefulness of the serum evaluation of M3/M21 in patients with ovarian cancer. This retrospective study comprises 75 patients suffering from ovarian cancer FIGO stages Ia-III. M3/M21 reached a sensitivity of 78%, a specificity of 85%, a PPV of 89% and a NPV of 83% using a cut-off level of 45 U 1(-1). Forty-four women developed recurrent disease after complete remission during the observation period. M3/M21 showed lead time effects in 19 patients, ranging from 2 to 8 months (median 3.2 months). Elevated M3/M21 serum levels before therapy were associated with a poor overall survival (log-rank test, P = 0.02). Considering these preliminary results, the value of M3/M21 as a serum tumour marker, i.e. to evaluate the tumour burden, seems promising.


Reasons why children miss vaccinations in Western Kenya; A step in a five-point plan to improve routine immunization.
Global childhood vaccination coverage has stagnated over the past decade and raising coverage will require a collection of approaches since no single approach has been suitable for all countries or situations. The American Red Cross has developed a 5-Point Plana to geolocate under-vaccinated children and determine the reasons why they miss vaccination by capitalizing on the Red Cross Movement's large cadres of trusted community volunteers. The Plan was piloted in Bobasi sub-county in Western Kenya, with volunteers seeking to conduct a face-to-face interview in all households, visiting over 60,000 over 7 days. Six pockets of 233 children without a home-based vaccination record or missing an age-appropriate dose of Penta1, Penta3 or measles-containing vaccine were identified. Three activities were carried out to learn why these children were not vaccinated: 1) one-on-one interviews and 2) focus group discussions with the caregivers of the under-vaccinated children and 3) interviews with healthcare workers who vaccinate in Bobasi. Complacency was commonly reported by caregivers during one-on-one interviews while bad staff attitude or practice was most frequently reported in focus group discussions; health staff reported caregiver hesitency, not knowing vaccination due date and vaccine stock-outs as the most common reasons for caregivers to not have their child vaccinated. As reasons varied across the three different activities, the different perspectives and approaches helped characterize vaccination barriers. Civil society organizations working together with the Ministry of Health can provide valuable information for immunization managers to act on.


Protein tyrosine kinase activity as a prognostic parameter in breast cancer, a pilot study.
Protein tyrosine kinase (PTK) activity was assayed in cytosolic extracts from normal breast tissue, benign tumors, and 84 T1-T2, N0-N1 M0, breast carcinomas. Normal breast tissue extracts yielded an average value of 1.9 +/- 1.1 pmol 32P incorporated/min/mg protein, whereas a mean of 12.5 +/- 6.1 was obtained for cancer samples. With a median follow-up of 34 months, in the series of 40 patients classified N-, PTK positive patients presented a significantly smaller 3-year disease free survival than the PTK negative ones. Multivariate analysis shows that PTK activity emerges as a potential prognostic factor in breast cancer (p = 0.02). These preliminary results will be updated on a bigger cohort of patients.


A binational study of patient-initiated changes to antiretroviral therapy regimen among HIV-positive Latinos living in the Mexico-U.S. border region.
Research is lacking on factors associated with antiretroviral therapy (ART) sub-optimal adherence among U.S. Latinos, who are disproportionately affected by HIV and face substantial health care barriers. We examined self-reported, patient-initiated changes to ART (i.e., made small/major changes from the antiretroviral drugs prescribed) among HIV-positive Latinos. Trained interviewers administered surveys to 230 participants currently on ART in San Diego, U.S. and Tijuana, Mexico. We identified factors independently associated with ART changes. Participants were Spanish-language dominant (86%), mean age of 41 years, male (77%), and born in Mexico (93%). Patient-initiated changes to ART were reported in 43% of participants. Being female, having ≥1 sexual partner (past 3 months), ≤6 years since HIV diagnosis and poor health were associated with increased odds of ART changes. Findings raise concern about sub-optimal adherence among this binational population. Longitudinal studies are needed to further explore adherence barriers and avenues for intervention.


Primary care of patients with sexually transmitted diseases or genitourinary symptoms in Hong Kong.
To determine the adequacy of care received from general practitioners by patients with sexually transmitted diseases or genitourinary symptoms. Prospective study. Hong Kong. Diagnoses and drug data obtained from logbooks submitted by doctors studying for the Diploma in Family Medicine and candidates for Fellowship examinations between 1999 and 2002. Diagnosis or symptom of a sexually transmitted disease and prescribed treatment. Sexually transmitted diseases and genitourinary symptoms accounted for 1.1% of the workload of these community doctors in Hong Kong. The majority of patients were young adult males. The overall standard of treatment was inadequate: both multi-pharmacy and inappropriate treatment was common; in up to 30% of cases, doctors ignored local or international guidelines. Primary care doctors play an important role in the diagnosis and management of sexually transmitted diseases or genitourinary symptoms in Hong Kong. A high index of suspicion should be maintained and continuing education made available if doctors are to provide an equally high standard of care.


Salmonella surveillance and control at post-harvest in the Belgian pork meat chain.
Salmonella remains the primary cause of reported bacterial food borne disease outbreaks in Belgium. Pork and pork products are recognized as one of the major sources of human salmonellosis. In contrast with the primary production and slaughterhouse phases of the pork meat production chain, only a few studies have focussed on the post-harvest stages. The goal of this study was to evaluate Salmonella and Escherichia coli contamination at the Belgian post-harvest stages. E. coli counts were estimated in order to evaluate the levels of faecal contamination. The results of bacteriological analysis from seven cutting plants, four meat-mincing plants and the four largest Belgian retailers were collected from official and self-monitoring controls. The prevalence of Salmonella in the cutting plants and meat-mincing plants ranged from 0% to 50%. The most frequently isolated serotype was Salmonella typhimurium. The prevalence in minced meat at retail level ranged from 0.3% to 4.3%. The levels of Salmonella contamination estimated from semi-quantitative analysis of data relating to carcasses, cuts of meat and minced meat were equal to -3.40+/-2.04 log CFU/cm(2), -2.64+/-1.76 log CFU/g and -2.35+/-1.09 log CFU/g, respectively. The E. coli results in meat cuts and minced meat ranged from 0.21+/-0.50 to 1.23+/-0.89 log CFU/g and from 1.33+/-0.58 to 2.78+/-0.43 log CFU/g, respectively. The results showed that faecal contamination still needs to be reduced, especially in specific individual plants.


Substrate specificity of Staphylococcus aureus cysteine proteases--Staphopains A, B and C.
Human strains of Staphylococcus aureus secrete two papain-like proteases, staphopain A and B. Avian strains produce another homologous enzyme, staphopain C. Animal studies suggest that staphopains B and C contribute to bacterial virulence, in contrast to staphopain A, which seems to have a virulence unrelated function. Here we present a detailed study of substrate preferences of all three proteases. The specificity of staphopain A, B and C substrate-binding subsites was mapped using different synthetic substrate libraries, inhibitor libraries and a protein substrate combinatorial library. The analysis demonstrated that the most efficiently hydrolyzed sites, using Schechter and Berger nomenclature, comprise a P2-Gly↓Ala(Ser) sequence motif, where P2 distinguishes the specificity of staphopain A (Leu) from that of both staphopains B and C (Phe/Tyr). However, we show that at the same time the overall specificity of staphopains is relaxed, insofar as multiple substrates that diverge from the sequences described above are also efficiently hydrolyzed.


Metabolic control of the behavioural action of androgens in the dove brain: testosterone inactivation by 5 beta-reduction.
In vitro studies of androgen metabolism in the dove brain show that testosterone is rapidly converted to 5 beta-reduced metabolites (5 beta-dihydrotestosterone and the two corresponding 3 alpha/3 beta diols). The preoptic region and the anterior ventromedial and posterior hypothalamus which are target areas for androgen action on behaviour and the neuroendocrine system show significantly less 5 beta-reduction than adjacent parolfactory, septal and striatal areas not directly involved in androgen action. In contrast to 5 alpha-reduction and aromatization, which lead to the formation of active metabolites in the dove brain, 5 beta-reduction is likely to be part of a testosterone (T) inactivation mechanism because: (a) intrahypothalamic implants of 5 beta-dihydrotestosterone were ineffective for male behaviour in the dove; (b) 5 beta-reduced metabolites of T were not detected in hypothalamic cell nuclei following intramuscular injection of tritiated testosterone, and were rapidly eliminated from brain cells; (c) the 4 beta-diols were also found to be a major product of T catabolism in the liver, a site of androgen inactivation. Long-term androgen deficit induced by castration increased 5 beta DHT formation in the preoptic area, but not in an adjacent parolfactory area. The increase was reversed by exogenous testosterone and oestradiol-17 beta indicating that the 5 beta-reduction pathway in an androgen-sensitive brain area is influenced by hormonal conditions. It is suggested that 5 beta-reduction is a testosterone inactivation pathway involved in the regulation of brain sensitivity to androgens.


Experiences of role model instructors and nursing students about facilitator factors of role-modeling process: A qualitative research.
One of the key strategies in students' learning is the influence of models on them. Understanding the factors affecting the implementation of role-modeling process in education will help to make greater use of this training strategy. This study aimed to understand the experiences of role model instructors and nursing students about the facilitator factors in the role-modeling process. This qualitative study was carried out by using thematic analysis method and purposeful sampling. Data were collected until saturation by using three focus group discussions (n = 20) and two individual interviews with nursing instructors, as well as six semi-structured face-to-face interviews with role model instructors from five nursing faculties of Tehran universities in 2011. Six themes, "effort for humanistic and professional growth of students," "individual and managerial empowerment of instructor," "instructor and student's modeling," "motivation and effort of student," "strategies governing the education system," and "appropriate facilities and equipment," were extracted as the facilitating factors. For development of role-modeling process in nursing education, paying attention to personal and environmental factors, especially effort for professional and humanistic growth of nursing students is necessary.


Clinical relevance of adjustment disorder in DSM-III-4 and DSM-IV.
This study examines the diagnosis of adjustment disorder in DSM-III-R. In view of the methodological problems associated with this, we investigated clinical characteristics of patients to whom the diagnosis had been applied. The patients in question were treated at the University Psychiatric Outpatient Clinic in Lausanne, Switzerland (N = 1,308). Four groups of comparisons are made: between adjustment disorder, other diagnoses, and no diagnosis; between adjustment disorder with depressed mood and other mood disorders; between adjustment disorder with anxious mood and anxiety disorders; and between the clinical subdivisions of adjustment disorder. This diagnosis is characterized by a high number of marital stressors, low age, predominance of women, and short period of treatment. The clinical profile is marked by a predominance of depressive and mixed types, and the relationship between these is demonstrated. Finally, the course of a certain proportion of these disorders goes beyond the 6-month period stipulated by DSM-III-R. This result justifies the modifications introduced in DSM-IV.


Effect on intraocular pressure of extraocular muscle surgery for thyroid-associated ophthalmopathy.
To study the effect of extraocular muscle surgery on intraocular pressure (IOP) in patients with thyroid-associated ophthalmopathy. Retrospective, observational case series. The medical records of patients with restrictive myopathy secondary to thyroid-associated ophthalmopathy who underwent strabismus surgery from July 1, 1997 through July 31, 2003 were reviewed and analyzed retrospectively. Seventeen patients met the criteria and were included in this study. All patients were seen at the Thyroid Eye Center at the University of California, San Diego, a university-based tertiary referral center. The main outcome measure was IOP readings obtained before and after surgery in both primary gaze and upgaze. A statistically significant decrease in IOP in upgaze was noted after extraocular muscle recession. The mean IOP before surgery was 16.6 +/- 3.78 mm Hg in primary gaze and 23.2 +/- 7.27 mm Hg in upgaze. After strabismus surgery, the mean IOP after one month was 15.7 +/- 2.36 mm Hg (P = .215) in primary gaze and 18.9 +/- 2.96 mm Hg in upgaze (P = .001). Strabismus surgery resulted in a significant reduction in IOP in the early postoperative period in patients with restrictive myopathy secondary to thyroid-associated ophthalmopathy.


Establishment of appropriate glaucoma models using dexamethasone or TGFβ2 treated three-dimension (3D) cultured human trabecular meshwork (HTM) cells.
To establish appropriate ex vivo models for a glaucomatous trabecular meshwork (TM), two-dimensional (2D) and three-dimensional (3D) cultures of human trabecular meshwork cells (HTM) were prepared in the presence of 250 nM dexamethasone (DEX) or 5 ng/mL TGFβ2, and characterized by the following analyses; transendothelial electrical resistance (TEER) measurements, FITC dextran permeability, scanning electron microscopy and the expression of the extracellular matrix (ECM) including collagen (COL)1, 4 and 6, and fibronectin (FN), α-smooth muscle actin (α-SMA), tissue inhibitor of metalloproteinase (TIMP)1-4, and matrix metalloproteinase (MMP)2, 9 and 14. DEX and TGFβ2 both caused a significant increase or decrease in the TEER values and FITC dextran permeability. During the 3D spheroid culture, DEX or TGFβ2 induced a mild and significant down-sizing and an increase in stiffness, respectively. TGFβ2 induced a significant up-regulation of COL1 and 4, FN, α-SMA, and MMP 2 and 14 (2D) or COL1 and 6, and TIMP2 and 3 (3D), and DEX induced a significant up-regulation of FN (3D) and TIMP4 (2D and 3D). The findings presented herein indicate that DEX or TGFβ2 resulted in mild and severe down-sized and stiff 3D HTM spheroids, respectively, thus making them viable in vitro HTM models for steroid-induced and primary open angle glaucoma.


Characterization of a mutation that results in independence of oxidosqualene cyclase (Erg7) activity from the downstream 3-ketoreductase (Erg27) in the yeast ergosterol biosynthetic pathway.
In yeast, deletion of ERG27, which encodes the sterol biosynthetic enzyme, 3-keto-reductase, results in a concomitant loss of the upstream enzyme, Erg7p, an oxidosqualene cyclase (OSC). However, this phenomenon occurs only in fungi, as mammalian Erg27p orthologues are unable to rescue yeast Erg7p activity. In this study, an erg27 mutant containing the mouse ERG27 orthologue was isolated that was capable of growing without sterol supplementation (FGerg27). GC/MS analysis of this strain showed an accumulation of squalene epoxides, 3-ketosterones, and ergosterol. This strain which was crossed to a wildtype and daughter segregants showed an accumulation of squalene epoxides as well as ergosterol indicating that the mutation entailed a leaky block at ERG7. Upon sequencing the yeast ERG7 gene an A598S alteration was found in a conserved alpha helical region. We theorize that this mutation stabilizes Erg7p in a conformation that mimics Erg27p binding. This mutation, while decreasing OSC activity still retains sufficient residual OSC activity such that the strain in the presence of the mammalian 3-keto reductase enzyme functions and no longer requires the yeast Erg27p. Because sterol biosynthesis occurs in the ER, a fusion protein was synthesized combining Erg7p and Erg28p, a resident ER protein and scaffold of the C-4 demethyation complex. Both FGerg27 and erg27 strains containing this fusion plasmid and the mouse ERG27 orthologue showed restoration of ergosterol biosynthesis with minimal accumulation of squalene epoxides. These results indicate retention of Erg7p in the ER increases its activity and suggest a novel method of regulation of ergosterol biosynthesis.


Financing constraints and firm-level responses to the COVID-19 pandemic: International evidence.
We explore whether financing constraints affected the ways in which small and medium-sized enterprises navigated through the economic disruptions caused by the COVID-19 pandemic. We draw on data from a novel source, the COVID-19 Impact Follow-up Surveys conducted in 19 countries by the World Bank Enterprise Analysis Unit as a follow-up to enterprise surveys conducted in these countries prior to the COVID-19 outbreak. We find that previous bank-lending credit constraints magnified the effects of the pandemic. More specifically, credit-rationed firms were more likely to experience greater liquidity and cash flow problems and more likely than unconstrained firms to be delinquent in meeting their obligations to financial institutions during the economic crisis. Furthermore, these firms were less likely to have access to bank funding as a principal source of financing to address pandemic-induced cash flow and liquidity problems during the COVID-19 outbreak. We further find that credit-constrained firms were more likely to use trade credit, delay payments to suppliers or employees, and rely on government grants to cope with pandemic-related liquidity and cash flow problems. We find little evidence that credit-rationed firms were more likely to raise equity capital during this economic crisis. Finally, we find that financing constraints were more likely to hamper firms' ability to adjust business operations in response to exogenous shocks. This study contributes to the literature on the impact of credit constraints on firm behavior in times of crisis.


Properties and heterologous expression of the glucose transporter GHT1 from Schizosaccharomyces pombe.
Genomic DNA of the Schizosaccharomyces pombe glucose transporter, GHT1, was obtained by complementation of the glucose transport deficient Sz. pombe strain YGS-5. Here we describe the GHT1 gene that encodes a protein of 565 amino acids with a corresponding molecular mass of 62.5 kDa. This eukaryotic glucose transporter contains 12 putative transmembrane segments and is homologous to the HXT multigene family of S. cerevisiae with several amino acid motifs of this sugar transporter family. It is also homologous to other sugar carriers from human, mouse and Escherichia coli. The function of the Ght1 protein as a glucose transporter was proved both by homologous and heterologous expression in the Sz. pombe mutant YGS-5 and in the S. cerevisiae hxt mutant RE700A, respectively. Both transformed yeast strains transported D-glucose with substrate specificity similar to that in Sz. pombe wild-type cells. Moreover, the cells of the two transformed yeast strains accumulated 2-deoxy-D-glucose, a non-metabolizable D-glucose analogue, with an efficiency similar to Sz. pombe wild-type cells. The ability of the S. cerevisiae mutant RE700A to accumulate 2DG in an delta mu H+ dependent manner after transformation with GHT1 provides evidence that the Sz. pombe transporter catalyses an energy-dependent uptake of glucose.


Sustaining primary care teams in the midst of a pandemic.
The challenges wrought by the COVID-19 pandemic on health systems have tested primary care clinicians, who are on the front lines of care delivery. To ensure the longevity of the primary care workforce, strong interprofessional teams are one important solution to alleviating burnout and increasing clinician and patient satisfaction, but the pandemic has demonstrated that the operating manual needs to be adapted for virtual work. Essential principles of primary care, including preventative care, communication and collaboration, and building strong relationships, can be applied to strengthen virtual primary care teams.


Does application of abduction brace after arthroscopic rotator cuff repair improve blood flow around posterosuperior rotator cuff and repair site, affecting pain levels and clinical and structural outcomes? A pilot randomized controlled trial.
As per some cadaveric studies, blood flow in posterosuperior rotator cuff tendons improves in the abducted shoulder position compared with the neutral position. In a clinical post-rotator cuff repair scenario, the impact of abduction on altered blood flow in and around the posterosuperior rotator cuff tendons is unknown in terms of clinical outcomes and structural healing. This study included 42 eligible patients aged between 40 and 70 years with clinically diagnosed and radiologically confirmed rotator cuff tears undergoing arthroscopic rotator cuff repair. Patients were randomly allocated to undergo application of either an abduction brace (group 1) or an arm pouch (group 2). On postoperative day 1, power Doppler scanning was performed on the index shoulder in adduction and 30° of abduction in each patient; the allocated treatment (abduction brace or arm pouch) was then applied. Power Doppler scanning was repeated at 6 weeks in the immobilization position assigned to the patient (abduction or adduction). The vascular flow in 6 regions was noted as per the criteria of Fealy et al. A visual analog scale score was assessed preoperatively and at 1, 3, 6, 12, and 56 weeks postoperatively. Clinical assessment was performed with the Constant-Murley score at 1 year, and structural healing of the cuff was assessed using ultrasonography at 3 and 12 months. On the first postoperative day, blood flow was significantly higher in all 6 areas of the shoulder in group 1 than in group 2. The mean total vascular score was significantly higher in group 1 than in group 2 on postoperative day 1 (P = .0001) and remained so at 6 weeks (P = .0001). However, significantly higher vascular flow was noted only in the peribursal region at 6 weeks in group 1 (P = .04). No significant difference in the visual analog scale score was noted between the 2 groups at any given point of follow-up. Furthermore, no clinical and structural healing differences were noted between the 2 groups at final follow-up. Higher blood flow in and around the posterosuperior rotator cuff owing to an abducted shoulder position with an abduction brace in the first 6 weeks postoperatively fails to offer any advantage in terms of lower pain levels, better clinical scores, or superior cuff healing.


Management of glioblastoma at safety-net hospitals.
Safety-net hospitals (SNHs) provide disproportionate care for underserved patients. Prior studies have identified poor outcomes, increased costs, and reduced access to certain complex, elective surgeries at SNHs. However, it is unknown whether similar patterns exist for the management of glioblastoma (GBM). We sought to determine if patients treated at HBHs receive equitable care for GBM, and if safety-net burden status impacts post-treatment survival. The National Cancer Database was queried for GBM patients diagnosed between 2010 and 2015. Safety-net burden was defined as the proportion of Medicaid and uninsured patients treated at each hospital, and stratified as low (LBH), medium (MBH), and high-burden (HBH) hospitals. The impact of safety-net burden on the receipt of any treatment, trimodality therapy, gross total resection (GTR), radiation, or chemotherapy was investigated. Secondary outcomes included post-treatment 30-day mortality, 90-day mortality, and overall survival. Univariate and multivariate analyses were utilized. Overall, 40,082 GBM patients at 1202 hospitals (352 LBHs, 553 MBHs, and 297 HBHs) were identified. Patients treated at HBHs were significantly less likely to receive trimodality therapy (OR = 0.75, p < 0.001), GTR (OR = 0.84, p < 0.001), radiation (OR = 0.73, p < 0.001), and chemotherapy (OR = 0.78, p < 0.001) than those treated at LBHs. Patients treated at HBHs had significantly increased 30-day (OR = 1.25, p = 0.031) and 90-day mortality (OR = 1.24, p = 0.001), and reduced overall survival (HR = 1.05, p = 0.039). GBM patients treated at SNHs are less likely to receive standard-of-care therapies and have increased short- and long-term mortality. Additional research is needed to evaluate barriers to providing equitable care for GBM patients at SNHs.


[Hepatocystis of Malaysia.--IV. Description of Hepatocystis rodhaini n. sp. parasite of Microchiroptera (author's transl)].
A new Haemoproteid of Malaysian Microchiroptera (Hepatochstis rodhaini n. sp.) is described; it is classified in the genus Hepatocystis because of the morphology of its gametocytes and tissue schizonts.


Clinical-pathological correlation in massive periretinal proliferation.
In the owl monkey, proliferation of pigment epithelial and glial cells occurred in varying degrees in nearly all eyes with retinal detachments. These cells grew and formed membranes on all available intraocular structures in the posterior part of the eye, such as vitreous structures and inner and outer retinal surfaces. Clinically, this process covered a wide variety of lesions, including fixed retinal folds. Depending on the location, size, and extent of the pigment epithelial or glial membrane, different types of fixed folds occurred. In an exaggerated end stage of this proliferative process, a clinical picture may develop identical to what has been called massive vitreous retraction or massive preretinal retraction in humans. To emphasize the importance and ubiquity of cellular proliferations, we called this entity massive periretinal proliferation.


LINC00624/TEX10/NF-κB axis promotes proliferation and migration of human prostate cancer cells.
Prostate cancer (PCa) is a malignant epithelial tumor with a high rate of biochemical or local recurrence. Studies have suggested that LINC00624 plays an important oncogenic role in liver cancer. However, whether it exerts similar effects in PCa progression remains unknown. In this study, we explored the effects of LINC00624 on the malignant progression of PCa and sought to identify the relevant signaling pathways. The results showed that LINC00624 was highly expressed in PCa tissues and cells and was associated with poor prognosis in PCa patients. In vitro and in vivo assays further showed that LINC00624 knockdown could decrease the proliferative and migratory ability of PCa cells. Mechanistically, we found that LINC00624 and TEX10 formed a co-regulatory axis that stimulated NF-κB activity. Our data suggest that LINC00624 acts as an oncogene in PCa progression and has potential as a novel biomarker for PCa.


[Value of pelvic osteotomy in the management of the hip in children with cerebral palsy].
The authors analyzed the effectiveness of pelvic osteotomy for the treatment of hip dislocation or subluxation in CP children. Fifteen pelvic osteotomies were performed on 11 children aged 30 months to 12 years (mean age 6 years). Seven children suffered from Little's disease. Follow-up averaged 10 years (from 2 to 16). Six children were reviewed at the end of growth (9 cases). A Salter or triple osteotomy with adductor release was performed. Four cases had also a proximal femoral osteotomy. Following surgery, the mean Reimers' percentage decreased from 55 to 0 per cent. Correction was perfect in 11 cases, incomplete in 4 but excentration remained under 20 per cent. At the end of growth, 6 out of 9 hips remained stable without any further surgery, 2 hips developed slight excentration (less than 20 per cent) and one recurrent subluxation was treated by proximal femoral osteotomy. Six hips remained stable but their growth was not ended at last review. This surgical procedure is compared to others described in literature. Pelvic osteotomy with tenotomy improves acetabular cover and corrects muscle imbalance. Its risks seem overestimated in the literature. Both pelvic and femoral osteotomies are useful in specific cases. Triple or Salter osteotomy with adducter release seems to be a useful procedure to restore good hip morphology at the end of growth. No posterior dislocations following this osteotomy were seen in spite of the important correction. Proximal femoral osteotomy is not always necessary.


2,3,7,8-Tetrachlorodibenzo-p-dioxin affects retinol esterification in rat hepatic stellate cells and kidney.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), a highly toxic environmental contaminant, interferes with retinoid homeostasis. To elucidate the underlying mechanism, the activities of lecithin : retinol and acyl-CoA : retinol acyltransferase (LRAT and ARAT) were determined in liver, kidney, and hepatic parenchymal and nonparenchymal cell fractions from rats 7 days after a single oral dose of 10 μg TCDD/kg body weight (b.w). Severely depressed LRAT activity in hepatic stellate cells, and greatly increased LRAT activity in kidneys, as well as decreased ARAT activity in stellate cells, were seen in TCDD-treated rats. Although the relevance of decreased ARAT activity under physiological conditions is not clear, the changed LRAT activities most likely contributes significantly to the TCDD-induced effects on tissue retinyl ester levels. It is intriguing that TCDD affects LRAT activity in hepatic stellate cells and kidney in opposite directions. The results suggest that effects of TCDD on retinyl ester tissue levels could be due to a specific interaction with retinoid metabolism.


Turnover of cellular carbohydrates in normal and Rous sarcoma virus-transformed cells.
We have analyzed the distribution of glucosamine-labeled polymers on the cell surface, in the growth medium, and inside the cell and the net turnover of these polymers during the process of malignant transformation of chick embryo fibroblasts by Rous sarcoma virus. The distribution of label and the turnover kinetics for hyaluronic acid, total glycoproteins, and chondroitins were found to be identical in both normal and transforming cultures. We conclude that nonspecific degradative processes are probably not involved in causing transformation-related alterations in cell surface carbohydrates, althougn degradation of specific macromolecules is not excluded.


Nocturnal circulating tumor cells: The ultimate selective filter in cancer progression?
The survival duration of circulating tumor cells (CTCs) in the vasculature is a critical parameter in the establishment of the metastatic cascade. Diamantopoulou and colleagues demonstrate that the metastatic capacity of CTCs is strongly influenced by circadian rhythms, suggesting a rationale for the time-controlled interrogation and treatment of metastatic cancers.


Study of defence injuries in homicidal deaths - An autopsy study.
In order to determine specific patterns and distribution of defence injuries, this study was conducted on 121 homicidal deaths which showed defence injuries in 40 (33%) cases. Of these 40 victims, 72.5% were males and 27.5% were females. Maximum numbers of victims were in the age group of 20-29 years. In 70% of cases, more than one assailant was involved. It was found that in 77.5% cases, sharp weapons were alone used, whereas, in 10% and 12.5% of victims, blunt weapons and multiple (sharp and blunt) weapons, respectively, were used. Fatal wounds were seen most commonly on the head and neck region. In 42.5% of cases, defence injuries were seen on the right side only, whereas in 27.5% of cases both sides were involved. Victim's right forearm and hand were more commonly involved because these are nearest to the perpetrator and consistent with the preponderance of right-handed individuals in the population.


Mitral Regurgitation Severity and Left Ventricular Systolic Dimension Predict Survival in Young Cavalier King Charles Spaniels.
Development and progression of myxomatous mitral valve disease (MMVD) in dogs are difficult to predict. Identification at a young age of dogs at high risk of adverse outcome in the future is desirable. To study the predictive value of selected clinical and echocardiographic characteristics associated with MMVD obtained at a young age for prediction of long-term cardiac and all-cause mortality in Cavalier King Charles Spaniels (CKCS). 1125 privately owned CKCS. A retrospective study including CKCS examined at the age of 1-3 years. Long-term outcome was assessed by telephone interview with owners. The value of variables for predicting mortality was investigated by Cox proportional hazard and Kaplan-Meier analyses. Presence of moderate to severe mitral regurgitation (MR) (hazard ratio (HR) = 3.03, 95% confidence interval (95% CI) = 1.48-6.23, P = 0.0025) even intermittent moderate to severe MR (HR = 2.23, 95% CI = 1.48-6.23, P = 0.039) on color flow Doppler echocardiography was significantly associated with increased hazard of cardiac death. An interaction between MR and sex was significant for all-cause mortality (P = 0.035), showing that males with moderate to severe MR had a higher all-cause mortality compared to males with no MR (HR = 2.38, 95% CI = 1.27-4.49, P = 0.0071), whereas no difference was found between female MR groups. The risk of cardiac (HR = 1.37, 95% CI = 1.14-1.63, P < 0.001) and all-cause (HR = 1.13, 95% CI = 1.02-1.24, P = 0.016) mortality increased with increasing left ventricular end-systolic internal dimension normalized for body weight (LVIDSN ). Moderate to severe MR, even if intermittent, and increased LVIDSN in dogs <3 years of age were associated with cardiac death later in life in CKCS.


Development of a pentylenetetrazole-induced seizure model to evaluate kinase inhibitor efficacy in the central nervous system.
c-Jun N-terminal kinases (JNKs) are implicated in cell death in neurodegenerative disorders. Therefore, JNK inhibitors could act as neuroprotective agents. To evaluate potential candidates, reproducible and quantitative CNS in vivo models are required. To that end, a pentylenetetrazole-induced seizure model was explored. c-Jun phosphorylation was detected in hippocampal extracts by blotting c-Jun immunoprecipitates with phosphorylation-specific antibodies. Pentylenetetrazole administration induced rapid and reproducible increases in c-Jun phosphorylation. However, special attention had to be paid to the composition of the extraction buffer to ensure stabilization of protein phosphorylation, as demonstrated using internal standards of phosphorylated recombinant c-Jun. As JNK and its upstream activator MKK4 are activated by phosphorylation, these events were also evaluated. In principle, kinase inhibitors could act at the level of JNK or upstream kinases to inhibit c-Jun phosphorylation. MKK4 phosphorylation was dramatically increased in response to pentylenetetrazole but, again, only when appropriate phosphatase inhibitors were in the extraction buffer. In contrast, JNK was found to be constitutively phosphorylated and unaltered upon pentylenetetrazole treatment. The JNK inhibitor SP600125 was shown to inhibit c-Jun phosphorylation without affecting MKK4 phosphorylation. Our procedures enable analysis of JNK pathway signalling in a CNS model and, also, should be applicable to that of other protein phosphorylation events in vivo.


Effective PFAS degradation by electrochemical oxidation methods-recent progress and requirement.
The presence of per- and poly-fluoroalkyl substances (PFASs) in water is of global concern due to their high stability and toxicity even at very low concentrations. There are several technologies for the remediation of PFASs, but most of them are inadequate either due to limited effectiveness, high cost, or production of a large amount of sludge. Electrochemical oxidation (EO) technology shows great potential for large-scale application in the degradation of PFASs due to its simple procedure, low loading of chemicals, and least amount of waste. Here, we have reviewed the recent progress in EO methods for PFAS degradation, focusing on the last 10 years, to explore an efficient, cost-effective, and environmentally benign remediation technology. The effects of important parameters (e.g., anode material, current density, solution pH, electrolyte, plate distance, and electrical connector type) are summarized and evaluated. Also, the energy consumption, the consequence of different PFASs functional groups, and water matrices are discussed to provide an insight that is pivotal for developing new EO materials and technologies. The proposed degradation pathways of shorter-chain PFAS by-products during EO of PFAS are also discussed.


Arylamidases of rat liver and chemically induced hepatomas. II. Preparation and characterization of monospecific antisera against two distinct arylamidase-active antigens.
Monospecific antisera were prepared against the most prominent arylamidase (alpha-aminoacyl-peptide hydrolase (microsomal), EC 220.127.116.11) active antigen in plasma membranes (the plasma membrane arylamidase) and lysomal content (the lysosomal content arylamidase), respectively. Plasma membrane extract and lysosomal content were allowed to react in crossed immunoelectrophoresis against their homologous antisera. The electrophoretic plates were washed extensively, dried and subsequently stained for arylamidase activity. The particular immunoprecipitates were thus identified and could be excised to be used for immunizations. The two resulting antisera precipitated the arylamidase used for immunization, but failed to be monospecific as they precipitated additional antigens. These antisera with restricted specificity against some plasma membrane and lysosomal content antigens, respectively, were used to produce immunoprecipitates intended for new attempts to prepare monospecific antisera by a second cycle of immunizations. A monospecific antiserum against the plasma membrane arylamidase was thus obtained, while a third cycle of immunizations was needed to get a monospecific anti-lysosomal content antiserum. The plasma membrane arylamidase showed ATPase activity also after precipitation with the monospecific antiserum, thus still retaining its characteristics as a multienzyme complex.


Rare complication of coupled VASER liposuction and Renuvion technologies: a case report.
This study presents the case of a women, who underwent VASER-assisted liposuction of the abdomen coupled with Renuvion skin tightening by J-Plasma for skin retraction. She developed pain and moderate surgical emphysema. Radiological findings showed moderate subcutaneous emphysema. There were no signs of viscus perforation, or pneumothorax.


Three-dimensional display of calculated velocity profiles for physiological flow waveforms.
To improve the understanding of the nature of pulsatile flow, three-dimensional idealized velocity profiles corresponding to measured physiological mean flow velocity waveforms were displayed at selected instants throughout the flow cycle. The Fourier harmonics for each waveform were determined, and their corresponding velocity profiles at each instant of time were calculated with the Womersley equations. Velocity profiles were calculated by summing the contributions from each harmonic. Calculated profiles were displayed in a three-dimensional perspective for both normal carotid and femoral arteries and for simple sinusoidal flow with a superimposed steady component. The potential value of such displays is discussed in terms of gaining an improved understanding of the nature of pulsatile flow and clarifying the interpretation of Doppler ultrasound recordings.


Transient Receptor Potential channels, TRPV1 and TRPA1 in melanocytes synergize UV-dependent and UV-independent melanogenesis.
Melanogenesis is essential for pigmentation and deregulated melanogenesis causes pigmentary diseases. Psoralen and ultraviolet A (PUVA) therapy strongly stimulates pigmentation, but the underlying molecular mechanisms are elusive. Melanin content of cultured human melanocytes was spectrophotometrically measured. Patch-clamp recordings were made in human melanocytes or HEK 293 cells transiently expressing wild type or mutant human TRPV1 and TRPA1 channels. Endogenous expression of TRPV1 and TRPA1 in melanocytes was analysed by western blotting and was knocked down with siRNA. In vivo pigmentary responses were measured by a colorimeter in mouse ear skin. The expression of TRPV1 and TRPA1 in human pigmented lesions was examined by immunohistochemical staining. PUVA strongly stimulated melanogenesis and PUVA-induced TRPV1 and TRPA1 channel activation in melanocytes and the resulting Ca2+ influx were required for the stimulated melanogenesis both in vitro and in vivo. Agonists-induced TRPV1 and TRPA1 activation alone did not stimulate melanogenesis, but it synergized UVA or intrinsic cAMP and NO signalling pathways to stimulate UV-dependent or UV-independent melanogenesis. Moreover, the expressions of TRPV1 and TRPA1 were increased in human melanocytic lesions and inhibition of both channels decreased melanin content in melanoma cells. TRPV1 and TRPA1 are key molecular sensors and enhancers of extrinsic and intrinsic melanogenic signals in both physiological and pathological conditions, and activation of both channels in melanocytes contributes to PUVA therapy-induced pigmentation. Our work provides a common mechanism of melanogenic regulation and highlights TRPV1 and TRPA1 as potential therapeutic targets for pigmentary disorders.


Reductions in acute medication use and healthcare resource utilization in patients with chronic migraine: a secondary analysis of a phase 3, randomized, double-blind, placebo-controlled study of galcanezumab with open-label extension (REGAIN).
To analyze secondary objectives of the REGAIN study related to acute headache medication use and healthcare resource utilization (HCRU) in patients with chronic migraine treated with galcanezumab, a monoclonal antibody to calcitonin gene-related peptide. Adults with chronic migraine (N = 1,113) were randomized (2:1:1) and treated with double-blind monthly injections of placebo, galcanezumab-120 mg, or galcanenzumab-240 mg for 3 months, followed by a 9-month open-label extension with 120 or 240 mg/month galcanezumab. Headache and medication information was collected by daily eDiary. HCRU was reported for the 6 months before randomization, monthly thereafter, and converted to rate per 100-patient-years. At baseline, 63-64% of patients met criteria for acute headache medication overuse. At Month 3, incidence of headache medication overuse in the galcanezumab groups (33% and 33%) was significantly lower than in the placebo group (46%, both p < .001) and was 16% and 23% in the previous-galcanezumab groups at Month 12. From a baseline of 14.5 to 15.5, reduction in mean number of monthly migraine headache days with acute headache medication use was also significantly greater in the galcanezumab groups at Month 3 (-4.2 and -4.9) than in placebo (-2.6, both p < .001), with reductions of -6.8 and -7.6 in the previous-galcanezumab groups at Month 12. Migraine-specific HCRU rates decreased for all groups, with no significant between-group differences at Month 3. At Month 12, in the two previous-galcanezumab groups, emergency room visits decreased by 58% and 75%, hospital admissions by 100%, and healthcare professional visits by 54% and 67%. Only 3 months of double-blind, placebo-controlled data, a longer HCRU recall period for baseline than postbaseline, and patients receiving care in the clinical trial itself, may limit generalizability. Treatment with galcanezumab resulted in significant reductions in headache medication overuse and migraine headache days requiring acute medication use, with notable reductions in migraine-specific HCRU.


The theory of planned behaviour explains intentions to use antiresorptive medication after a fragility fracture.
Our objective was to ascertain whether the Theory of Planned Behaviour (TPB) explains patient intentions to use antiresorptive medication after a fracture. A qualitative study was conducted with English-speaking members of the Canadian Osteoporosis Patient Network (COPN) who had sustained a fragility fracture at 50+ years of age and were not taking antiresorptive medication at the time of that fracture. Questions during a 1-h telephone interview were guided by the domains of the TPB: they addressed the antecedent constructs regarding antiresorptive medication (attitudes, subjective norms, and perceived behavioural control) as well as intentions regarding antiresorptive medication use. We created a coding template a priori based on the TPB domains and applied this template to the interview data. Twenty-six eligible participants (24 females, 2 males) aged 51-89 completed an interview. The TPB appeared to be predictive of intentions in 19 (73%) participants. In the majority of participants where the TPB did not appear to be predictive (57%), a positive attitude toward antiresorptive medication was the most important antecedent variable in determining intentions. The TPB appeared to be predictive of intentions to use antiresorptive medication among individuals who had experienced a fragility fracture. Attitudes towards medication were especially important.


Use of comprehensive geriatric assessment in older cancer patients: recommendations from the task force on CGA of the International Society of Geriatric Oncology (SIOG).
As more and more cancers occur in elderly people, oncologists are increasingly confronted with the necessity of integrating geriatric parameters in the treatment of their patients. The International Society of Geriatric Oncology (SIOG) created a task force to review the evidence on the use of a comprehensive geriatric assessment (CGA) in cancer patients. A systematic review of the evidence was conducted. Several biological and clinical correlates of aging have been identified. Their relative weight and clinical usefulness is still poorly defined. There is strong evidence that a CGA detects many problems missed by a regular assessment in general geriatric and in cancer patients. There is also strong evidence that a CGA improves function and reduces hospitalization in the elderly. There is heterogeneous evidence that it improves survival and that it is cost-effective. There is corroborative evidence from a few studies conducted in cancer patients. Screening tools exist and were successfully used in settings such as the emergency room, but globally were poorly tested. The article contains recommendations for the use of CGA in research and clinical care for older cancer patients. A CGA, with or without screening, and with follow-up, should be used in older cancer patients, in order to detect unaddressed problems, improve their functional status, and possibly their survival. The task force cannot recommend any specific tool or approach above others at this point and general geriatric experience should be used.


Virulence characteristics of translocating Escherichia coli and the interleukin-8 response to infection.
Four efficiently translocating Escherichia coli (TEC) strains isolated from the blood of humans (HMLN-1), pigs (PC-1) and rats (KIC-1 and KIC-2) were tested for their ability to adhere and translocate across human gut epithelial Caco-2 and HT-29 cells, to elicit a proinflammatory response and for the presence of 47 pathogenic E. coli virulence genes. HMLN-1 and PC-1 were more efficient in adhesion and translocation than rat strains, had identical biochemical phenotype (BPT) and serotype (O77:H18) and phylogenetic group (D). KIC-2 adhered more than KIC-1, belonged to different BPT and serotype but the same phylogenetic group as KIC-1. TEC strains elicited significantly higher IL-8 response in both cell lines (P < 0.05) and monocytic THP-1 (P < 0.0001) cells than non-TEC strains. KIC-2 induced the highest IL-8 response which may be associated with its immunostimulatory flagellin. Apart from adhesin genes fimH and bmaE that were carried by all strains, HMLN-1 and PC-1 carried capsule synthesis gene kpsMT III and KIC-2 carried the EAST1 toxin gene. The lack of known virulence genes and the ability of TEC to efficiently adhere and translocate whilst causing proinflammatory response suggests that these strains may carry as yet unidentified genes that enable their translocating ability.


Cost-effectiveness of high-sensitivity faecal immunochemical test and colonoscopy screening for colorectal cancer.
Colorectal cancer screening can decrease morbidity and mortality. However, there are widespread differences in the implementation of programmes and choice of strategy. The primary objective of this study was to estimate lifelong costs and health outcomes of two of the currently most preferred methods of screening for colorectal cancer: colonoscopy and sensitive faecal immunochemical test (FIT). A cost-effectiveness analysis of colorectal cancer screening in a Swedish population was performed using a decision analysis model, based on the design of the Screening of Swedish Colons (SCREESCO) study, and data from the published literature and registries. Lifelong cost and effects of colonoscopy once, colonoscopy every 10 years, FIT twice, FIT biennially and no screening were estimated using simulations. For 1000 individuals invited to screening, it was estimated that screening once with colonoscopy yielded 49 more quality-adjusted life-years (QALYs) and a cost saving of €64 800 compared with no screening. Similarly, screening twice with FIT gave 26 more QALYs and a cost saving of €17 600. When the colonoscopic screening was repeated every tenth year, 7 additional QALYs were gained at a cost of €189 400 compared with a single colonoscopy. The additional gain with biennial FIT screening was 25 QALYs at a cost of €154 300 compared with two FITs. All screening strategies were cost-effective compared with no screening. Repeated and single screening strategies with colonoscopy were more cost-effective than FIT when lifelong effects and costs were considered. However, other factors such as patient acceptability of the test and availability of human resources also have to be taken into account.


Programmed assembly of long DNA synthons: design, mechanism, and online monitoring.
Synthesis of custom de novo DNA sequences is highly demanded by fast-growing field of synthetic biology. Usually DNA sequences with length more than 1 kb are assembled from smaller synthetic DNA fragments (synthons) obtained by PCR assembly. The ability to synthesize longer synthons sufficiently reduces efforts and time for DNA synthesis. We developed a novel rational oligonucleotide design and programmed approach for the assembly of synthetic DNA synthons up to 1550 bp. The developed procedure was thoroughly investigated by synthesis of cholesterol oxidase gene from Streptomyces lavendulae (1544 bp). Our approach is based on combined design, oligonucleotide concentration gradient, and specialized assembly program that directs assembly reaction to full-length gene in a stepwise manner. The process includes conventional thermodynamically balanced assembly, thermodynamically balanced inside-out elongation, and further amplification. The ability of DNA polymerase to perform programmed assembly is highly influenced by the presence of 5' → 3'-exonuclease activity. Oligonucleotide probing of PCR assembly products allowed us to shed light on the nature of high molecular weight spurious by-products and to understand the mechanism of their formation. For the first time, we applied light scattering techniques for tracking of oligonucleotide annealing, analysis of gene assembly products, and even for real-time monitoring of gene assembly process.


[Value of positive resection margins in patients with pT2 prostate cancer. Implications for adjuvant treatment].
To analyse the impact on the recurrence-free biochemical survival of tumour involvement of surgical resection margins in patients with localized prostate cancer (pT2) in the prostatectomy specimen and its implications for adjuvant treatment. Retrospective study of 536 patients with stage pT2 prostate cancer, treated with radical prostatectomy between 1996 and 2007. Subsequent to the prostatectomy, the following variables were collected: Gleason score, pathological stage, capsular invasion, surgical margins and perineural invasion. We performed a univariate analysis and subsequently adjusted it by means of a Cox proportional hazard model (enter method). 21.7% presented positive surgical margins and 20.9% developed biochemical recurrence after a mean follow-up of 57 months. 37.9% of the patients with pathological involvement of the resection surgical margins presented biochemical recurrence against 16% that did not have it (p<0.001). In the multivariate analysis, only the surgical margin (p<0.001) and the Gleason score greater or equal to 8 (p<0.001) behaved as independent biochemical recurrence factors. On stratifying the series according to these two variables, we found that the patients with positive surgical margins and a Gleason score of ≤ 7 have a recurrence probability at 5 and 10 years of 35% and 50% against 74% and 87% in the group with positive surgical margins and a Gleason score of ≥ 8 (p=0.002). Patients with pT2 prostate cancer, positive surgical margins and a Gleason score of ≥ 8 will benefit from adjuvant radiotherapy. 50% of the patients with positive margins and a Gleason score of ≤ 7 will not recur, which means that the indication of adjuvant radiotherapy continues to be controversial.