Real-world analysis of patients with haemophilia A and haemophilia A carriers in the United States: Demographics, clinical characteristics and costs.
Females with haemophilia A (HA [FHAs]) and HA carriers (HACs) have an increased risk of bleeding and complications compared to the general population. To examine the characteristics, billed annualised bleed rates (ABRb ), costs and healthcare resource utilisation for males with HA (MHAs), FHAs and HACs in the United States. Data were extracted from the IBM® MarketScan® Research Databases (Commercial and Medicaid) for claims during the index period (July 2016 to September 2018) and analysed across MHAs, FHAs and HACs. Dual diagnosis females (DDFs; both HA and HAC claims) were grouped as a separate cohort. MHAs were generally younger than females (all cohorts) by up to 19 years (Commercial) and 23 years (Medicaid). ABRb >0 was more frequent in females. Factor VIII claims were higher for MHAs versus female cohorts. Joint-related health issues were reported for 24.4 and 25.6% (Commercial) and 29.3 and 26.6% (Medicaid) of MHAs and FHAs, respectively; lower rates were reported in the other two cohorts. Heavy menstrual bleeding claims occurred for approximately a fifth (Commercial) to a quarter (Medicaid) of female cohorts. All-cause emergency department and inpatient visits in FHAs and DDFs were similar to, or more frequent than, those in MHAs; bleed-related inpatient visits were infrequent. In MHAs (Commercial), mean all-cause total costs ($214,083) were higher than in FHAs ($40,388), HACs ($15,647) and DDFs ($28,320) with similar trends for Medicaid patients. FHAs and HACs may be undermanaged and undertreated. Further research is needed to fully understand these cohorts' bleeding rates, long-term complications and costs.


Human guanylate-binding proteins in intracellular pathogen detection, destruction, and host cell death induction.
Cell-intrinsic defense is an essential part of the immune response against intracellular pathogens regulated by cytokine-induced proteins and pathways. One of the most upregulated families of proteins in this defense system are the guanylate-binding proteins (GBPs), large GTPases of the dynamin family, induced in response to interferon gamma. Human GBPs (hGBPs) exert their antimicrobial activity through detection of pathogen-associated molecular patterns and/or damage-associated molecular patterns to execute control mechanisms directed at the pathogen itself as well as the vacuolar compartments in which it resides. Consequently, hGBPs are also inducers of canonical and noncanonical inflammasome responses leading to host cell death. The mechanisms are both cell-type and pathogen-dependent with hGBP1 acting as a pioneer sensor for intracellular invaders. This review focuses on the most recent functional roles of hGBPs in pathways of pathogen detection, destruction, and host cell death induction.


Ethanol and leucine oxidation--III. Leucine oxidation by rat heart in vitro.
The activities of leucine aminotransferase (BCAT) and 2-oxoisocaproate dehydrogenase (OADH) were measured in rat heart in vitro. The effect upon these enzyme activities of both ethanol and acetaldehyde, administered either acutely or chronically, was determined. Enzyme activities were not significantly altered by either acetaldehyde or ethanol when given chronically. Ethanol administered acutely to rats decreased OADH activity but BCAT was unaffected. Acetaldehyde administered acutely did not alter significantly BCAT activity but significantly increased OADH activity.


Development and Technical Validation of an Immunoassay for the Detection of APP669-711 (Aβ-3-40) in Biological Samples.
The ratio of amyloid precursor protein (APP)669-711 (Aβ-3-40)/Aβ1-42 in blood plasma was reported to represent a novel Alzheimer's disease biomarker. Here, we describe the characterization of two antibodies against the N-terminus of Aβ-3-x and the development and "fit-for-purpose" technical validation of a sandwich immunoassay for the measurement of Aβ-3-40. Antibody selectivity was assessed by capillary isoelectric focusing immunoassay, Western blot analysis, and immunohistochemistry. The analytical validation addressed assay range, repeatability, specificity, between-run variability, impact of pre-analytical sample handling procedures, assay interference, and analytical spike recoveries. Blood plasma was analyzed after Aβ immunoprecipitation by a two-step immunoassay procedure. Both monoclonal antibodies detected Aβ-3-40 with no appreciable cross reactivity with Aβ1-40 or N-terminally truncated Aβ variants. However, the amyloid precursor protein was also recognized. The immunoassay showed high selectivity for Aβ-3-40 with a quantitative assay range of 22 pg/mL-7.5 ng/mL. Acceptable intermediate imprecision of the complete two-step immunoassay was reached after normalization. In a small clinical sample, the measured Aβ42/Aβ-3-40 and Aβ42/Aβ40 ratios were lower in patients with dementia of the Alzheimer's type than in other dementias. In summary, the methodological groundwork for further optimization and future studies addressing the Aβ42/Aβ-3-40 ratio as a novel biomarker candidate for Alzheimer's disease has been set.


Biochar-bacteria-plant partnerships: Eco-solutions for tackling heavy metal pollution.
Over the past 30 years, the ever-rising demands of the modern and growing population have led to the rapid development of agricultural and industrial sectors worldwide. However, this expansion has exposed the environment to various pollutants including heavy metal (HM)s. Almost all HMs are serious toxicants and can pose serious health risks to living organisms in addition to their bioaccumulative and non-biodegradable nature. Different techniques have been developed to restore the ecological functions of the HM-contaminated soil (HMCS). However, the major downfalls of the commonly used remediation technologies are the generation of secondary wastes, high operating costs, and high energy consumption. Phytoremediation is a prominent approach that is more innocuous than the existing remediation approaches. Some microbes-plant interactions enhance the bioremediation process, with heavy metal resistant-plant growth promoting bacteria (HMRPGPB) being widely used to assist phytoremediation of HMs. However, the most common of all major microbial assisted-phytoremediation disturbances is that the HM-contaminated soil is generally deficient in nutrients and cannot sustain the rapid growth of the applied HMRPGPB. In this case, biochar has recently been approved as a potential carrier of microbial agents. The biochar-HMRPGPB-plant association could provide a promising green approach to remediate HM-polluted sites. Therefore, this review addresses the mechanisms through which biochar and HMRPGPB can enhance phytoremediation. This knowledge of biochar-HMRPGPB-plant interactions is significant with respect to sustainable management of the HM-polluted environment in terms of both ecology and economy, and it offers the possibility of further development of new green technologies.


Assessment of protein domain fusions in human protein interaction networks prediction: application to the human kinetochore model.
In order to understand how biological systems function it is necessary to determine the interactions and associations between proteins. Some proteins, involved in a common biological process and encoded by separate genes in one organism, can be found fused within a single protein chain in other organisms. By detecting these triplets, a functional relationship can be established between the unfused proteins. Here we use a domain fusion prediction method to predict these protein interactions for the human interactome. We observed that gene fusion events are more related to physical interaction between proteins than to other weaker functional relationships such as participation in a common biological pathway. These results suggest that domain fusion is an appropriate method for predicting protein complexes. The most reliable fused domain predictions were used to build protein-protein interaction (PPI) networks. These predicted PPI network models showed the same topological features as real biological networks and different features from random behaviour. We built the PPI domain fusion sub-network model of the human kinetochore and observed that the majority of the predicted interactions have not yet been experimentally characterised in the publicly available PPI repositories. The study of the human kinetochore domain fusion sub-network reveals undiscovered kinetochore proteins with presumably relevant functions, such as hubs with many connections in the kinetochore sub-network. These results suggest that experimentally hidden regions in the predicted PPI networks contain key functional elements, associated with important functional areas, still undiscovered in the human interactome. Until novel experiments shed light on these hidden regions; domain fusion predictions provide a valuable approach for exploring them.


Serum alkaline phosphatase isoenzymes in epileptic children receving anticonvulsant drugs.
In 40 epileptic children on long-term anticonvulsant treatment, serum alkaline phosphatase (AP) isoenzymes were separated semiquantitatively using a combination of L-phenylalanine inhibition and heat inactivation. Though mean total serum AP activity was significantly increased compared to age matched controls, only 4 individual values exceeded the upper limit (mean + 2SD) of the reference sample. In epileptics the mean activity of the heat-sensitive non L-phenylalanine sensitive AP fraction (non-LPSAP) was significantly increased (P less than 0.01) and the mean Q-value (i.e. percentage ratio of heat-stable non-LPSAP/non-LPSAP) was significantly decreased (P less than 0.05), thus indicating an enhancement of the bone isoenzymes during anticonvulsant treatment. In 4 patients the isoenzyme pattern was abnormal although total serum AP activity was normal and in 3 of them the deviation indicated enhanced bone isoenzyme. The data provide evidence that in anticonvulsant treated children the bone isoenzyme, rather than hepatobiliary isoenzyme fraction, may be increased even when total serum AP activity is normal. Thus, semiquantitative separation of serum AP isoenzymes may be a helpful guide as to whether or not an eplieptic child should be given vitamin D.


Clinical practice: vocal nodules in dysphonic children.
Common among children, vocal symptoms are a cause of concern for parents who seek elucidation of their diagnosis and treatment. Vocal nodules are the major cause of dysphonias in children and are related to vocal abuse. We conducted a literature review considering clinical, physiopathological, epidemiological, and histological aspects of vocal nodules, as well as diagnostic methods, highlighting the main studies addressing this issue. The controversial points of treatments were also discussed.


Novel cytogenic and neurovascular niches due to blood-brain barrier compromise in the chronic pain brain.
The mechanisms by which painful injuries are linked to the multitude of pain-related comorbidities and neuroplastic changes in the brain remain poorly understood. Here we propose a model that relies on epi-neuronal communication through the vascular system to effect various brain structures. Specifically, we hypothesize that the differential vulnerability of the blood-brain barrier (BBB) in different brain regions is associated with region-specific neuroplastic and neurovascular changes that are in turn associated with particular pain-related comorbidities. We will present our hypothesis by focusing on two main points: (A) chronic pain (CP) is associated with differential BBB compromise. (B) Circulating mediators leaking through the BBB create cytogenic and neovascular niches associated with pain-related co-morbidities. Pre-clinically, our hypothesis can be tested by observing, in parallel, BBB compromise, (neo)vascularization, neurogenesis, and their co-localization in animal pain models using imaging, microscopy, biochemical and other tools. Furthermore, the BBB can be experimentally damaged in specific brain regions, and the consequences of those lesions studied on nociception and associated comorbidities. Recently developed imaging techniques allow the analysis of blood brain barrier integrity in patients providing a route for translation of the laboratory findings. Though perhaps more limited, post-mortem examination of brains with available pain histories constitutes a second approach to addressing this hypothesis. Understanding changes in BBB permeability in chronic pain conditions has clear implications both for understanding the pathogenesis of chronic pain and for the design of novel treatments to prevent chronic pain and its consequences. More broadly, this hypothesis may help us to understand how peripheral injuries impact the brain via mechanisms other than commonly studied efferent sensory pathways.


Primary internal fixation and bone grafting for open fractures of the hand.
Primary bone grafting and internal fixation is a reliable option for the treatment of segmental hand defects resulting from high-energy crush or avulsion injuries. Immediate reconstruction can be performed safely if radical debridement and adequate soft tissue coverage of bone can be achieved. When these two criteria cannot be met, staged reconstruction should be performed. The low infection rate (0%), the high union rate (92%), and the return to full, unrestricted, previous employment for all patients in the authors' series supports the use of primary bone grafting in an acute setting. This single-stage approach to fracture management also permits earlier and more aggressive hand therapy that, in turn, can lower the high functional morbidity so often associated with these injuries.


Intra-abdominal desmoid tumor difficult to distinguish from a gastrointestinal stromal tumor: report of two cases.
Desmoid tumors are benign fibroblastic neoplasms with no metastatic potential, but a propensity for local recurrence even after complete surgical resection. These lesions can develop at any site in the body, and commonly occur in the intra-abdominal area. Intra-abdominal desmoid tumors usually occur at the mesentery or retroperitoneum, and may morphologically mimic gastrointestinal stromal tumors (GISTs). Distinguishing between these tumors is important, because the therapies differ substantially, but is often difficult even with the use of CD117 staining. We herein report the cases of two patients with sporadic intra-abdominal desmoid tumors that were differentiated from GIST by immunohistological examination using beta-catenin and CD34. Desmoid tumors specifically express nuclear beta-catenin, and show no expression of CD34. We recommend staining for beta-catenin and CD34 when an intra-abdominal desmoid tumor is suspected.


Oral contraceptives and risk of breast cancer and ovarian cancer in women with a BRCA1 or BRCA2 mutation: a meta-analysis of observational studies.
It remains inconclusive whether the use of oral contraceptives (OCs) alters the risks of breast or ovarian cancer in women with a breast cancer susceptibility gene 1 (BRCA1) or breast cancer susceptibility gene 2 (BRCA2) mutation. We investigated the association between OC use and the risks of breast or ovarian cancer in this group by using a meta-analysis. PubMed and EMBASE were searched using keywords until February 2021 to identify relevant studies that evaluated the association between OC ever use and the risks of breast or ovarian cancer in women with a BRCA1 or BRCA2 mutation. Twelve studies for breast cancer and eight studies for ovarian cancer were identified. In the random-effects meta-analysis, the ever use of OCs was significantly associated with an increased risk of breast cancer [odds ratio (OR), relative risk (RR) or hazard ratio (HR) = 1.24; 95% confidence interval (CI) 1.08-1.41] and a decreased risk of ovarian cancer (OR/RR/HR = 0.53, 95% CI 0.41-0.67). Consistent findings were observed when BRCA1 and BRCA2 mutation carriers were analyzed separately. The increased risk of breast cancer was observed only in the long-term (>5 years) users of OCs, while the decreased risk of ovarian cancer was observed regardless of the duration of OC use. The current study suggests that the ever use of OCs in BRCA mutation carriers is significantly associated with an increased risk of breast cancer and a decreased risk of ovarian cancer. Therefore, the use of OCs as chemoprevention of ovarian cancer should be cautious in BRCA mutation carriers.


Salvage radiation therapy for residual superficial esophageal cancer after endoscopic mucosal resection.
To analyze the outcomes of radiation therapy for patients with residual superficial esophageal cancer (rSEC) after endoscopic mucosal resection (EMR). From May 1996 to October 2002, a total of 30 rSEC patients without lymph node metastasis received radiation therapy at Tohoku University Hospital and associated hospitals. The time interval from EMR to start of radiation therapy ranged from 9 to 73 days (median interval, 40 days). Radiation doses ranged from 60 Gy to 70 Gy (mean dose, 66 Gy). Chemotherapy was used in 9 of 30 patients (30%). The 2-year, 3-year, and 5-year overall survival rates and cause-specific survival rates were 91%, 82%, and 51%, respectively, and 95%, 85%, and 73%, respectively. The 2-year, 3-year, and 5-year local control rates for mucosal cancer were 91%, 91%, and 91%, respectively, and those for submucosal cancer were 89%, 89%, and 47%, respectively. These differences in survival rates for patients with two types of cancer were not statistically significant. Local recurrence and lymph node recurrence were more frequent in patients with submucosal cancer than in patients with mucosal cancer (p = 0.38 and p = 0.08, respectively). Esophageal stenosis that required balloon dilatation developed in 3 of the 30 patients, and radiation pneumonitis that required steroid therapy developed in 1 patient. Radiation therapy is useful for preventing local recurrence after incomplete EMR.


The effect of recipient BMI on waitlist and post-transplant outcomes after the 2018 heart transplant allocation policy change.
The effects of recipient body mass index (BMI) on waitlist strategies, waitlist outcomes, and post-transplant outcomes among adult patients listed for heart transplantation under the updated 2018 allocation system have not been well characterized. The United Network of Organ Sharing data set between October 2015 and March 2021 was analyzed, and patients were grouped based on recipient BMI and whether listing occurred in the old (pre-October 2018) or new allocation system. Listing strategies differed by BMI group, but trends of increased use of temporary mechanical support and decreased use of durable support remained among all BMI groups, except those with BMI > 35 kg/m2 . Waitlist outcomes improved among all BMI cohorts in the new allocation system, including among patients with BMI 30-34.9 and >35 kg/m2 , although patients with higher BMIs continued to have longer waitlist times. Post-transplant outcomes in the new allocation system are worse for patients with BMI > 30 kg/m2 (hazard ratio: 1.47; confidence interval: 1.19-1.82; p < .001). The 2018 change to the heart transplant allocation system was associated with similar changes in the use of mechanical support for listing strategy across BMI ranges, except in the most obese, and improved waitlist outcomes across all BMI ranges. Post-transplant outcomes in the new allocation system are worse for patients with BMI > 30 kg/m2 compared to patients with BMI < 30 kg/m2 . These findings have important clinical implications for our understanding of the ongoing influence of BMI on waitlist courses and post-transplant outcomes among patients listed for heart transplantation.


Clinical and Diagnostic Significance of Sialic Acids Determination in Biological Material.
Sialic acids (SA) are neuraminic acid derivatives, located at the terminal position in the chains of monosaccharide residues of various glycoconjugates. SA play a dual role: they either mask recognition sites, or, on the contrary, represent biological targets that can be recognized by receptor proteins and serve as ligands. The desialylation/sialylation processes can be considered as a dynamic modification regulated by sialyltransferases and sialidases in response to external or internal stimuli. This review describes the structural and functional diversity and the potential use of SA fractions as biomarkers for various pathological conditions. Almost any extreme impact on the body and inflammatory processes are accompanied by an increase in the level of both total and free SA in the blood and tissues. Possible reasons for the increase of sialoglycoconjugate metabolism indicators in biological material include: (i) activation of the hepatocyte synthesis and secretion of various acute-phase proteins, many of which are sialoglycoproteins, (ii) impaired membrane integrity and destruction of body cells, (iii) high activity of sialidases (neurominidases) and sialyltransferases. Most acute and chronic liver diseases are characterized by the decrease in the total level of SA in the blood serum (because many plasma proteins are synthesized and glycosylated in hepatocytes). Aberrant sialylation results in changes of sialoglycoconjugate structure, its ability to perform biological functions and sialoglycoconjugate half-life. Glycosylation is the most common post-translational modification of proteins in the virus, which not only promotes the formation of specific conformation of viral proteins, but also modulates their interaction with receptors and affects host cell recognition, viral replication and infectivity. Serum total SA concentration increases in some benign and inflammatory conditions, which indicates a lack of specificity and limits their use for early detection and screening of neoplastic diseases. Clinical and diagnostic value of determining the sialoglycoconjugate metabolic indicators, including changes in the content of both SA fractions and specific proteins in various biological fluids and tissues, consists in establishing the causes and mechanisms of biochemical changes in the body in certain diseases. In combination with the measurement of existing markers, they can be used to improve diagnosis, staging and monitoring of therapeutic response in some pathological conditions where the need for specificity is less than for specific diagnostics.


Nonlinear finite element analyses: advances and challenges in dental applications.
To discuss the development and current status of application of nonlinear finite element method (FEM) in dentistry. The literature was searched for original research articles with keywords such as nonlinear, finite element analysis, and tooth/dental/implant. References were selected manually or searched from the PUBMED and MEDLINE databases through November 2007. The nonlinear problems analyzed in FEM studies were reviewed and categorized into: (A) nonlinear simulations of the periodontal ligament (PDL), (B) plastic and viscoelastic behaviors of dental materials, (C) contact phenomena in tooth-to-tooth contact, (D) contact phenomena within prosthodontic structures, and (E) interfacial mechanics between the tooth and the restoration. The FEM in dentistry recently focused on simulation of realistic intra-oral conditions such as the nonlinear stress-strain relationship in the periodontal tissues and the contact phenomena in teeth, which could hardly be solved by the linear static model. The definition of contact area critically affects the reliability of the contact analyses, especially for implant-abutment complexes. To predict the failure risk of a bonded tooth-restoration interface, it is essential to assess the normal and shear stresses relative to the interface. The inclusion of viscoelasticity and plastic deformation to the program to account for the time-dependent, thermal sensitive, and largely deformable nature of dental materials would enhance its application. Further improvement of the nonlinear FEM solutions should be encouraged to widen the range of applications in dental and oral health science.


Radical Transurethral Resection of Bladder Tumor in Organ-confined Muscle-invasive Bladder Cancer: Yes!
Muscle-invasive bladder cancer is a potentially lethal disease often impacting elder and comorbid patients. Neoadjuvant chemotherapy followed by radical cystectomy is associated with morbidity and is an option that many patients refuse. Maximal transurethral resection of bladder tumor (TURBT) as part of a bladder preservation strategy can achieve surgical cure and may improve long-term recurrence-free survival. We encourage bladder preservation after maximal TURBT for appropriate patients.


Cryo-electron microscopy structures of pyrene-labeled ADP-Pi- and ADP-actin filaments.
Since the fluorescent reagent N-(1-pyrene)iodoacetamide was first used to label skeletal muscle actin in 1981, the pyrene-labeled actin has become the most widely employed tool to measure the kinetics of actin polymerization and the interaction between actin and actin-binding proteins. Here we report high-resolution cryo-electron microscopy structures of actin filaments with N-1-pyrene conjugated to cysteine 374 and either ADP (3.2 Å) or ADP-phosphate (3.0 Å) in the active site. Polymerization buries pyrene in a hydrophobic cavity between subunits along the long-pitch helix with only minor differences in conformation compared with native actin filaments. These structures explain how polymerization increases the fluorescence 20-fold, how myosin and cofilin binding to filaments reduces the fluorescence, and how profilin binding to actin monomers increases the fluorescence.


Prevalence and predictors of unsupervised walking and physical activity in a community population of women with fibromyalgia.
Physical exercise is recognized as a component of the evidence-based guidelines for treatment of fibromyalgia. Walking is a low-moderate intensity exercise easily adaptable to a fibromyalgia patient's situation. The present study aims to estimate the prevalence of unsupervised walking for exercise in women with fibromyalgia, to describe their level of physical activity and to identify their predictors among socio-demographic, symptom perception and medical advice to walk. A cross-sectional survey with 920 women (all members of fibromyalgia associations) completed the International Physical Activity Questionnaire-Short Form and self-reported scales to assess symptom perception, walking, medical advice to walk and physical comorbidity. The prevalence of reported walking regularly as physical exercise was 30.8 % and it was predicted by medical advice (odds ratio, OR 1.876), age (OR 1.021) and fatigue intensity (OR 0.912). The prevalence of physical activity was 16 % for high-intensity activity, 40 % for moderate activity and 44 % for low activity. Predictors of low versus moderate and high physical activity were pain intensity (OR 1.171) and fatigue impact perception (OR 1.076). Evidence shows a low percentage of women with fibromyalgia walking regularly for physical exercise. Most reported low or moderate physical activity. The results indicate the importance of doctors' advice in promoting walking. Symptom perception and socio-demographic characteristics were weak predictors. Further work is required to examine other determinants of these low levels.


Real-time driver drowsiness feedback improves driver alertness and self-reported driving performance.
Driver drowsiness has been implicated as a major causal factor in road accidents. Tools that allow remote monitoring and management of driver fatigue are used in the mining and road transport industries. Increasing drivers' own awareness of their drowsiness levels using such tools may also reduce risk of accidents. The study examined the effects of real-time blink-velocity-derived drowsiness feedback on driver performance and levels of alertness in a military setting. A sample of 15 Army Reserve personnel (1 female) aged 21-59 (M=41.3, SD=11.1) volunteered to being monitored by an infra-red oculography-based Optalert Alertness Monitoring System (OAMS) while they performed their regular driving tasks, including on-duty tasks and commuting to and from duty, for a continuous period of 4-8 weeks. For approximately half that period, blink-velocity-derived Johns Drowsiness Scale (JDS) scores were fed back to the driver in a counterbalanced repeated-measures design, resulting in a total of 419 driving periods under "feedback" and 385 periods under "no-feedback" condition. Overall, the provision of real-time feedback resulted in reduced drowsiness (lower JDS scores) and improved alertness and driving performance ratings. The effect was small and varied across the 24-h circadian cycle but it remained robust after controlling for time of day and driving task duration. Both the number of JDS peaks counted for each trip and their duration declined in the presence of drowsiness feedback, indicating a dynamic pattern that is consistent with a genuine, entropy-reducing feedback mechanism (as distinct from random re-alerting) behind the observed effect. Its mechanisms and practical utility have yet to be fully explored. Direct examination of the alternative, random re-alerting explanation of this feedback effect is an important step for future research.


RNF213 p.R4810K Variant Carriers with Intracranial Arterial Stenosis Have a Low Atherosclerotic Burden.
The ring finger protein 213 gene (RNF213) p.R4810K variant is a major susceptibility gene for intracranial arterial stenosis in East Asia. We hypothesized that if intracranial arterial stenosis is induced by a non-atherosclerotic mechanism similar to moyamoya disease, the patients with RNF213 p.R4810K variant may have a lower cumulative atherosclerotic burden than the non-carriers. A total of 112 participants with intracranial arterial stenosis were enrolled in this multicenter cross-sectional study. We compared the prevalence of atherosclerotic risk factors and three different cardiovascular risk scores (Essen Stroke Risk Score, Framingham Risk Score, and Suita Risk Score) between the RNF213 p.R4810K variant carriers and non-carriers. Patients with moyamoya disease were excluded from the study. The RNF213 p.R4810K variant carriers were younger than the non-carriers (P<0.001). The prevalence of each atherosclerotic risk factor was not significant, but it tended to be lower in the variant carriers. The Essen Stroke Risk Score (carriers: 2.3±1.5 vs. non-carriers: 2.9±1.5, P=0.047), Framingham Risk Score (10.7±6.4 vs. 15.3±6.2, P=0.001), and Suita Risk Score (35.4±15.8 vs. 48.7±15.2, P<0.001) were significantly lower in the variant carriers. Among the three risk scores, the Suita score showed the highest predictive accuracy for the variant carriers. RNF213 p.R4810K variant carriers have a lower cumulative atherosclerotic burden than non-carriers among patients with intracranial arterial stenosis. New therapeutic approaches beyond the standard management of atherosclerotic risk factors are required to prevent the development of intracranial arterial stenosis.


Identification of potential regulatory long non-coding RNA-associated competing endogenous RNA axes in periplaque regions in multiple sclerosis.
Slow-burning inflammation at the lesion rim is connected to the expansion of chronic multiple sclerosis (MS) lesions. However, the underlying processes causing expansion are not clearly realized. In this context, the current study used a bioinformatics approach to identify the expression profiles and related lncRNA-associated ceRNA regulatory axes in the periplaque region in MS patients. Expression data (GSE52139) from periplaque regions in the secondary progressive MS spinal cord and controls were downloaded from the Gene Expression Omnibus database (GEO), which has details on mRNAs and lncRNAs. Using the R software's limma package, the differentially expressed lncRNAs (DElncRNAs) and mRNAs (DEmRNAs) were found. The RNA interactions were also found using the DIANA-LncBase, miRTarBase, and HMDD databases. The Pearson correlation coefficient was used to determine whether there were any positive correlations between DEmRNAs and DElncRNAs in the ceRNA network. Finally, lncRNA-associated ceRNA axes were created based on co-expression and connections between DElncRNA, miRNA, and DEmRNA. We used the Enrichr tool to enrich the biological process, molecular function, and pathways for DEmRNAs and DElncRNAs. A network of DEmRNAs' protein-protein interactions was developed, and the top five hub genes were found using Cytoscape and STRING. The current study indicates that 15 DEmRNAs, including FOS, GJA1, NTRK2, CTNND1, and SP3, are connected to the MS ceRNA network. Additionally, four DElncRNAs (such as TUG1, ASB16-AS1, and LINC01094) that regulated the aforementioned mRNAs by sponging 14 MS-related miRNAs (e.g., hsa-miR-145-5p, hsa-miR-200a-3p, hsa-miR-20a-5p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-27a-3p, hsa-miR-29b-3p, hsa-miR-29c-3p, hsa-miR-34a-5p) were found. In addition, the analysis of pathway enrichment revealed that DEmRNAs were enriched in the pathways for the "MAPK signaling pathway", "Kaposi sarcoma-associated herpesvirus infection", "Human immunodeficiency virus one infection", "Lipid and atherosclerosis", and "Amphetamine addiction". Even though the function of these ceRNA axes needs to be investigated further, this study provides research targets for studying ceRNA-mediated molecular mechanisms related to periplaque demyelination in MS.


Lack of Association of Initial Vasopressor Dosing with Survival and Cardiac Re-Arrest Likelihood After Return of Spontaneous Circulation.
Cardiac arrest occurs in approximately 350,000 patients outside the hospital and approximately 30,000 patients in the emergency department (ED) annually in the United States. When return of spontaneous circulation (ROSC) is achieved, hypotension is a common complication. However, optimal dosing of vasopressors is not clear. The objective of this study was to determine if initial vasopressor dosing was associated with cardiac re-arrest in patients after ROSC. This was a retrospective, single-center analysis of adult patients experiencing cardiac arrest prior to arrival or within the ED. Patients were assigned to one of four groups based on starting dose of vasopressor: low dose (LD; < 0.25 µg/kg/min), medium dose (MD; 0.25-0.49 µg/kg/min), high dose (HD; 0.5-0.99 µg/kg/min), and very high dose (VHD; ≥ 1 µg/kg/min). Data collection was performed primarily via manual chart review of medical records. The primary outcome was incidence of cardiac re-arrest within 1 h of vasopressor initiation. Multivariate logistic regression analysis was conducted to identify any covariates strongly associated with the primary outcome. No difference in cardiac re-arrest incidence was noted between groups. The VHD group was significantly more likely to require a second vasopressor (p = 0.003). The HD group had lower survival rates to hospital discharge compared with the LD and MD groups (p = 0.0033 and p = 0.0147). In the multivariate regression, longer duration of pre-vasopressor re-arrests and hyperkalemic cardiac arrest etiology were significant predictors of cardiac re-arrest after vasopressor initiation. Initial vasopressor dosing was not found to be associated with risk of cardiac re-arrest or, conversely, risk of adverse events.


Acceptor Engineering for Optimized ROS Generation Facilitates Reprogramming Macrophages to M1 Phenotype in Photodynamic Immunotherapy.
Reprogramming tumor-associated macrophages to an antitumor M1 phenotype by photodynamic therapy is a promising strategy to overcome the immunosuppression of tumor microenvironment for boosted immunotherapy. However, it remains unclear how the reactive oxygen species (ROS) generated from type I and II mechanisms, relate to the macrophage polarization efficacy. Herein, we design and synthesize three donor-acceptor structured photosensitizers with varied ROS-generating efficiencies. Surprisingly, we discovered that the extracellular ROS generated from type I mechanism are mainly responsible for reprogramming the macrophages from a pro-tumor type (M2) to an anti-tumor state (M1). In vivo experiments prove that the photosensitizer can trigger photodynamic immunotherapy for effective suppression of the tumor growth, while the therapeutic outcome is abolished with depleted macrophages. Overall, our strategy highlights the designing guideline of macrophage-activatable photosensitizers.


The number of retrieved oocytes does not decrease during consecutive gonadotrophin-stimulated IVF cycles.
Our aim was to study whether there is a decreasing trend in the number of retrieved oocytes in women who had all undergone at least seven consecutive IVF cycles. A nationwide retrospective cohort study was conducted among women whose first IVF cycle was stimulated with gonadotrophins in The Netherlands between 1983 and 1995. Among these eligible women, we identified all women who had received at least seven consecutive IVF cycles (n = 330). Poisson regression analysis was used to assess the trend in the number of retrieved oocytes over the first six IVF cycles. The unadjusted results showed a slight but non-significant decrease in the number of retrieved oocytes over six IVF cycles. The change in cycle 6 compared with cycle 1 was -0.06 oocytes (5.8% decrease) (P = 0.21). When adjusting for the number of ampoules and the stimulation protocol (fertility drug used combined with use of GnRH agonists), there was a considerable and highly significant decrease from cycle 1 to cycle 6 [change in cycle 6 compared with cycle 1: -0.19 oocytes (17.4% decrease), (P < 0.0001)]. However, when adjusting for age of the women, this decrease almost completely disappeared [change in cycle 6 compared with cycle 1: -0.05 oocytes (5% decrease), (P = 0.50)]. The results suggest that there is no decrease in the number of retrieved oocytes over subsequent cycles when simultaneously accounting for the increasing age of the women, differences in the number of ampoules of gonadotrophins used, type of stimulation protocol and year of IVF treatment.


The induction of organ-specific antibodies during the graft-vs.-host reaction.
During the parent (P) into F1 hybrid graft-vs.-host reaction (GVHR), nuclear, leukocyte and erythrocyte autoantibodies are commonly seen. The specificity of these autoantibodies is reminiscent of those found in systemic lupus erythematosus (SLE) patients and SLE-prone mice. Organ-specific antibodies, however, including thyro-globulin (Tg) antibodies do not arise spontaneously. There have been conflicting reports about the ability of exogenous Tg to induce an anti-Tg response during the GVHR. We have re-examined this question in greater detail. Using the murine P----F1 GVHR system, the results of this work demonstrate that mouse thyroglobulin (MTg)-specific antibodies can be induced during a GVHR. However, mice must both be undergoing a GVHR, and have received exogenous MTg. The highest autoantibody response occurs if mice are injected with mouse thyroid extract or purified MTg at the time of P----F1 cell transfer. The anti-MTg response is MTg dose dependent. The ability to induce anti-MTg antibody was not major histocompatibility complex restricted, for both the DBA/2----B6D2F1 (low responder H-2 haplotypes to MTg), and AKR or DBA/2----AKD2F1 (high/low responder----high responder haplotype) GVHR gave similar responses. The anti-MTg titers peaked between days 7-10 and declined thereafter. In contrast, antibodies to dsDNA were not present at this early time, but developed after several weeks. We conclude that organ-specific autoantibodies can be induced during a GVHR if the appropriate antigen(s) are presented near the time of GVHR induction.


Alterations of the fetal capillary bed in the guinea pig placenta following long-term hypoxia.
Oxygen supply is an important regulator of the fetal placental capillarization. To determine the effects of long-term hypoxia on the fetal placental vessel arrangement pregnant guinea pigs were kept under hypoxic conditions (12% O2 for 45 days). Vessel casts showed a significant difference in branching and orientation of the vessels between the controls and the hypoxic animals. The hypoxic group had a less orientated capillary bed with increased branching and coiling. By light- and transmission electron microscopical studies, there was a decreased diffusion distance, a decreased diameter of the fetal capillaries, and an increased number of capillary cross-sections. These results are consistent with the hypothesis that chronic hypoxia is responsible for increased branching and coiling of the capillaries resulting in a dense network of short and narrow capillaries in the placenta.


Bulky DNA-adduct formation induced by Ni(II) in vitro and in vivo as assayed by 32P-postlabeling.
Various small oxidation products (e.g. 8-hydroxydeoxyguanosine) can be induced in DNA by nickel compounds. In this study, the 32P-postlabeling assay was applied to determine whether Ni(II) compounds are able to induce bulky DNA-adduct formation in vitro and in vivo. In vitro studies detected two major and several minor adducts in DNA incubated with NiCl2 and H2O2 at 37 degrees C for 1 h. Formation of the two major adducts increased with incubation time (0-24 h) and NiCl2 concentration (0-800 microM). Adduct levels were greatly reduced by hydroxyl free-radical scavengers, i.e. 0.4 M sodium formate or 0.05 M p-nitrosodimethylaniline, and by a singlet oxygen scavenger, 0.05 M sodium azide. The in vitro effects of NiCl2 on DNA were significantly enhanced by (1) addition of 3 mM ascorbic acid, (2) replacement of H2O with D2O in the reaction, and (3) prior denaturation of DNA. Adduct formation presumably involved a Fenton-type reaction, in which DNA crosslinks may arise by reaction with hydroxyl free radicals and singlet oxygen. For in vivo studies, male 6-8 wk old B6C3F1 mice were used. In untreated mice, several I-compounds (putative indigenous DNA modifications that increase with age) were detected in liver, kidney, and lung. Two of these (spots 1 and 2) were chromatographically identical to the two major spots induced by Ni(II) in vitro. The intensities of spots 1 and 2 in kidney and of some other spots in liver and lung were increased 1 and 2 h after i.p. injection with a single dose of 170 mumols/kg NiAc2. The effects of NiAc2 were reduced or undetectable in the three tissues 24 h after treatment. These observations indicate the capacity of Ni(II) to induce and modulate bulky DNA modifications both in vitro and in vivo.


Corrigendum to "Effects of Number of Repetitions and Number of Hours of Shaping Practice during Constraint-Induced Movement Therapy: A Randomized Controlled Trial".
[This corrects the article DOI: 10.1155/2018/5496408.].


Octanoic acid-rich enteral nutrition prevented lipopolysaccharide-induced acute liver injury through c-Jun N-terminal kinase-dependent autophagy.
Acute liver injury (ALI) is an essential component of sepsis associated with poor outcomes. Octanoic acid (OA), a medium-chain fatty acid, has a protective effect on sepsis-induced organ damage, and autophagy is an adaptive response to sepsis. However, the underlying mechanism by which OA prevents ALI remains unknown. Therefore, we investigated whether OA-rich enteral nutrition (EN) prevented lipopolysaccharide (LPS)-induced ALI through the c-Jun N-terminal kinase (JNK)-dependent autophagy. Firstly, Sprague Dawley rats were randomly divided into four groups (sham, LPS, LPS + EN, and LPS + EN + OA) to detect the effect of OA-rich EN on LPS-induced ALI. Then, rats were randomly divided into five groups (sham, LPS, LPS + EN + OA, LPS + EN + OA + anisomycin (AN), and LPS + SP600125) to explore the mechanism by which OA-rich EN prevented ALI. EN and OA-rich EN were conducted through gastric tubes for 3 days. The liver protective effects were measured by liver histopathological changes, enzymes, inflammatory cytokines of serum and liver, the levels of autophagy, and JNK activity. OA-rich EN inhibited JNK activity, up-regulated autophagy and prevented LPS-induced ALI. Inhibition of JNK activity conferred by SP promoted autophagy and prevented LPS-induced ALI. Moreover, the protective effect of autophagy and inhibition of JNK activity conferred by OA-rich EN on ALI was counteracted by AN. OA-rich EN prevented LPS-induced ALI through JNK-dependent autophagy. This result suggested that OA-rich EN may be a therapeutic potential for ALI in patients with sepsis.


Impaired coronary flow reserve in young patients affected by severe psoriasis.
Our study aimed to evaluate the effects of psoriasis (Pso) on coronary microvascular function and whether there is a relationship between disease activity scores and coronary blood flow abnormalities. 56 young patients (pts) with Pso (42 M, aged 37±3 years) without clinical evidence of cardiovascular diseases, and 56 controls matched for age and gender were studied. Coronary flow velocity in the left anterior descending coronary artery was detected by transthoracic echocardiography at rest and during adenosine infusion. Coronary flow reserve (CFR) was the ratio of hyperaemic diastolic flow velocity (DFV) to resting DFV. A CFR≤2.5 was considered abnormal. In pts with Pso, CFR was lower than in controls (3.2±0.9 vs. 3.7±0.7, p=0.02). CFR was abnormal (≤2.5) in 12 pts (22% vs. 0% controls, p<0.0001). Moreover, in pts with CFR≤2.5, Psoriasis Area Severity Index (PASI), a clinical score for Pso severity, was higher (11±6 vs. 7±3, p=0.006) compared to pts with CFR>2.5. At multivariable analysis PASI remained the only determinant of CFR≤2.5 (p=0.02). CFR in young pts with severe Pso without coronary disease is reduced suggesting a coronary microvascular dysfunction, independently related to the severity and extension of Pso. This early microvascular impairment might be hypothesized as the consequence of prolonged and sustained systemic inflammation and might explain the increased cardiovascular risk conferred by Pso.


Purification and characterization of glutathione transferases with an activity toward nitroglycerin from human aorta and heart. Multiplicity of the human class Mu forms.
Although recent studies suggest involvement of glutathione transferase (GST) of blood vessels in vasodilation by nitroglycerin, GST forms in blood vessels remain to be studied. In this study, three GST forms (pI values 8.3, 6.6, and 4.8) were purified from human aorta and four (pI values 6.0, 5.6, 5.3, and 4.6) from the heart by affinity chromatography followed by chromatofocusing. The major form of both aorta (pI 4.8) and heart (pI 4.6) was identified as GST-pi, and the other five forms were immunologically related to GST-mu, suggesting that the five belong to the Mu class. Among nine human GST forms, including three in the Alpha class purified from the liver, GST-mu, aorta pI 8.3 form, and GST-I (a form of the Alpha class, corresponding to GST-epsilon (B1B1)) showed high activities toward nitroglycerin, 1.08, 0.85, and 0.78 units/mg protein, respectively. GST-pi did not exhibit the activity. The Km values of the aorta form (pI 8.3) for glutathione (GSH) and nitroglycerin were calculated as 0.12 and 1.1 mM, respectively. The Km values of GST-mu and GST-I for GSH were 0.29 and 0.09 mM, and those for nitroglycerin were 2.5 and 0.3 mM, respectively. The activity of the pI 8.3 form as well as GST-mu toward nitroglycerin was inhibited by bromosulfophthalein, which is known to inhibit the relaxation of rabbit aorta induced by nitroglycerin, at the lower concentration (IC50, 2 microM) than was GST-I (IC50, 32 microM). Two-dimensional gel electrophoresis and N-terminal amino acid sequence analysis revealed that five forms in the Mu class are homo- or heterodimers of five different subunits named M1 (pI 7.0/Mr 27,000), M2 (6.6/27,000), M3 (6.0/27,000), N1 (6.5/26,500), and N2 (5.9/26,500). The subunit structures of the five forms are as follows: pI 8.3 form, M1M2; 6.6 form, M2N1; 6.0 form, M3M3; 5.6 form, M3N2; and 5.3 form, N2N2. M3 and N2 seem to correspond to the subunits of GST-mu, and -4 (Board, P. G., Suzuki, T., and Shaw, D. C. (1988) Biochim. Biophys. Acta 953, 214-217), respectively. These subunits except N1 are different from each other at two or three positions in the first 20 residues of N-terminal amino acid sequence. These results indicate the presence of five different subunits in the human Mu class and also suggest that GST-M1M2 and -M2N1 found in the aorta are involved in the expression of the pharmacologic effect of nitroglycerin.


Genetic variation of 20 autosomal STR loci in the Han nationality in Central Yunnan, Southwest China.
A total of 20 autosomal short tandem repeat (STR) loci from 2220 unrelated healthy individuals of Han population living in the central area of Yunnan province, Southwest China were amplified with the PowerPlex® 21 System. After Hardy-Weinberg equilibrium examination, the allele frequencies and forensic statistical parameters of 20 STR loci were evaluated. A total of 298 alleles and 1225 genotypes were observed for all the 20 loci. The allele frequencies varied from 0.0002 to 0.5130. The combined power of discrimination and the combined probability of exclusion of all 20 STR loci were 0.99999999521565 and 0.999999999999999999999999637, respectively. Meanwhile, genetic distances between Central Yunnan Han and 17 previously published populations were compared and a neighbor-joining (NJ) phylogenetic tree was developed and visualized by using MEGA 7 based on Nei's standard genetic distance. The results demonstrated that these loci were highly polymorphic in the Han nationality in Central Yunnan, Southwest China and could be applied in forensic medicine and population genetics.


Q & A. Philip Ingham.
Philip Ingham grew up in Liverpool and graduated from Cambridge University in 1977. He did his D. Phil in Developmental Genetics at Sussex University and postdoctoral work in Strasbourg, France before joining the laboratory of David Ish-Horowicz at the ICRF Mill Hill Laboratories. Here he applied the emerging technique of tissue in situ hybridisation to the analysis of the Drosophila segmentation genes. After a short spell at the MRC Laboratory of Molecular Biology in Cambridge, he rejoined the ICRF as a Research Scientist at the Developmental Biology Unit in Oxford. His group pioneered the analysis of the Hedgehog signalling pathway in Drosophila and in collaboration with the labs of Andy McMahon and Cliff Tabin at Harvard University, discovered the Hedgehog gene family in vertebrates. In 1996 he was appointed Professor of Developmental Genetics at the University of Sheffield where he has established the Centre for Developmental Genetics.


The impact of the COVID-19 pandemic on rabies reemergence in Latin America: the case of Arequipa, Peru.
Over the past decades, there has been tremendous progress towards eliminating canine rabies in Latin America. Major components of rabies prevention programs in Latin America leading to these successes have been constant and intense surveillance for rabid dogs and uninterrupted yearly mass dog vaccination campaigns. However, vital measures to control COVID-19 in Latin America have had the negative trade-off of jeopardizing these rabies elimination and prevention activities. In this paper, we aimed to assess the effect of interrupting canine rabies surveillance and mass dog vaccination campaigns on rabies trends. We built a deterministic compartment model of dog rabies dynamics parameterized for conditions found in Arequipa, Peru, where there is an ongoing dog rabies epidemic. Our model suggests that a decrease in canine vaccination coverage as well as decreased surveillance leading to an increased length of survival of infected dogs could lead to a sharp rise in canine rabies and, subsequently, human rabies risk. We examined our results over the best estimate of the basic reproductive number in Arequipa (R0 = 1.44) and a range of plausible values for R0 (1.36 - 2). The rising trend was consistent. It is very possible that COVID-19 will continue to challenge our public health departments in the short- and medium-term. Innovative strategies to conduct dog vaccination and rabies surveillance during these trying times should be considered to safeguard the achievements made in Latin America towards the elimination of dog-mediated human rabies.


Interaction of alpha1-adrenoceptor subtypes with different G proteins induces opposite effects on cardiac L-type Ca2+ channel.
We examined the effect of alpha(1)-adrenoceptor subtype-specific stimulation on L-type Ca2+ current (I(Ca)) and elucidated the subtype-specific intracellular mechanisms for the regulation of L-type Ca2+ channels in isolated rat ventricular myocytes. We confirmed the protein expression of alpha(1A)- and alpha(1B)-adrenoceptor subtypes at the transverse tubules (T-tubules) and found that simultaneous stimulation of these 2 receptor subtypes by nonsubtype selective agonist, phenylephrine, showed 2 opposite effects on I(Ca) (transient decrease followed by sustained increase). However, selective alpha(1A)-adrenoceptor stimulation (> or =0.1 micromol/L A61603) only potentiated I(Ca), and selective alpha(1B)-adrenoceptor stimulation (10 mumol/L phenylephrine with 2 micromol/L WB4101) only decreased I(Ca). The positive effect by alpha(1A)-adrenoceptor stimulation was blocked by the inhibition of phospholipase C (PLC), protein kinase C (PKC), or Ca2+/calmodulin-dependent protein kinase II (CaMKII). The negative effect by alpha(1B)-adrenoceptor stimulation disappeared after the treatment of pertussis toxin or by the prepulse depolarization, but was not attributable to the inhibition of cAMP-dependent pathway. The translocation of PKCdelta and epsilon to the T-tubules was observed only after alpha(1A)-adrenoceptor stimulation, but not after alpha(1B)-adrenoceptor stimulation. Immunoprecipitation analysis revealed that alpha(1A)-adrenoceptor was associated with G(q/11), but alpha(1B)-adrenoceptor interacted with one of the pertussis toxin-sensitive G proteins, G(o). These findings demonstrated that the interactions of alpha(1)-adrenoceptor subtypes with different G proteins elicit the formation of separate signaling cascades, which produce the opposite effects on I(Ca). The coupling of alpha(1A)-adrenoceptor with G(q/11)-PLC-PKC-CaMKII pathway potentiates I(Ca). In contrast, alpha(1B)-adrenoceptor interacts with G(o), of which the betagamma-complex might directly inhibit the channel activity at T-tubules.


Lytic and latent EBV gene expression in transplant recipients with and without post-transplant lymphoproliferative disorder.
Epstein-Barr virus (EBV) is associated with post-transplant lymphoproliferative disorder (PTLD), which has significant morbidity and mortality in transplant recipients. To devise prophylactic measures, we need predictors of PTLD and a better understanding of the physiopathogenesis of the disease. To identify a molecular pattern of EBV gene products in blood that is specific to PTLD and can be used for the diagnosis of this disease. We evaluated the ratio between latent and replicating EBV nucleic acids in individuals with PTLD by comparison with transplant recipients without PTLD and immunocompetent hosts with EBV DNA-emia. Subjects were prospectively identified between July 2009 and October 2010 at the University of Colorado Hospital. EBV DNA, LMP-2A Latency III and BZLF1 Lytic genes mRNA were quantified using real-time PCR. We found that PTLD subjects (N = 7) had significantly higher EBV DNA-emia compared with non-transplant immunocompetent subjects (N = 69; p<0.0001), and transplant recipients without PTLD (N = 105; p<0.0001). The ratios between LMP-2A and BZLF1 mRNA in transplant recipients were significantly lower than in non-transplant subjects (p = 0.04). However, PTLD and non-PTLD transplant recipients displayed similar ratios. These results suggest that EBV replication makes a larger contribution to the circulating EBV DNA in transplant recipients compared with immunocompetent hosts. Transplant recipients seem to lose control over EBV replication, which may contribute to the development of PTLD.


Characterization of ML-005, a Novel Metaproteomics-Derived Esterase.
A novel gene encoding for a lipolytic enzyme, designated ML-005, was recently identified using a functional metaproteomics approach. We heterologously expressed this protein in Escherichia coli and biochemically characterized it. ML-005 exhibited lipolytic activity toward short-chained substrates with the preferred substrate being p-nitrophenyl-butyrate, suggesting that ML-005 is an esterase. According to homology analysis and site-directed mutagenesis, the catalytic triad of the enzyme was identified as Ser-99, Asp-164, and His-191. Its optimal pH was determined to be at pH 8. Optimal activity was observed at 45°C. It also exhibited temperature, pH and salt tolerance. Residual relative activity after incubating at 50-60°C for 360 min was above 80% of its initial activity. It showed tolerance over a broad range of pH (5-12) and retained most of its initial activity. Furthermore, incubating ML-005 in 1 - 5M NaCl solution had negligible effect on its activity. DTT, EDTA, and ß-mercaptoethanol had no significant effect on ML-005's activity. However, addition of PMSF led to almost complete inactivation consistent with ML-005 being a serine hydrolase. ML-005 remains stable in the presence of a range of metal ions, but addition of Cu2+ significantly reduces its relative activity. Organic solvents have an inhibitory effect on ML-005, but it retained 21% of activity in 10% methanol. SDS had the most pronounced inhibitory effect on ML-005 among all detergents tested and completely inactivated it. Furthermore, the Vmax of ML-005 was determined to be 59.8 μM/min along with a Km of 137.9 μM. The kcat of ML-005 is 26 s-1 and kcat/Km is 1.88 × 105 M-1 s-1.


Crassulacean acid metabolism in the shade. Studies on an epiphytic fern, Pyrrosia longifolia, and other rainforest species from Australia.
Crassulacean acid metabolism (CAM) was studied in a tropical epiphytic fern, Pyrrosia longifolia, from a fully sun-exposed and from a very shaded site in Northern Queensland, Australia. Measurements of instantaneous net CO2 exchange showed carbon gain via CO2 dark fixation with some net CO2 uptake also occuring during late afternoon, in both sun and shade fronds. Maximum rates of net CO2 uptake and the nocturnal increase in titratable acidity were lower in shade than in sun fronds. δ13C values of sun and shade fronds were not significantly different, and ranged between-14 and-15‰ suggesting that, in the long term, carbon gain was mainly via CO2 dark fixation. Sun fronds had a higher light compensation point of photosynthesis than shade fronds but the same quantum yield. Yet there was no acclimation of photosynthetic O2 evolution, (measured at 5% CO2) in sun and shade fronds and photosynthesis saturated at between 200 and 400 μmol quanta m-2 s-1. Use of higher light intensities for photosynthesis of sun fronds was probably precluded by low nutrient availability. Total nitrogen was less than 1% of dry weight in fully expanded sun and shade fronds. Exposure of shade fronds to full sunlight for 6 h led to a 60% decline in the quantum yield of photosynthesis and to a decline in variable fluorescence measured at room temperature. Photoinhibition by high light was also observed in Hoya nicholsoniae, a rainforest climber growing in deep shade. This species also exhibited CAM as demonstrated by nocturnal net CO2 uptake, nocturnal acidification and a δ13C value of-14‰. Photosynthetic O2 evolution in this species was saturated at 2.5% of full sunlight. Two species of Dendrobium (Orchidaceae) from sun-exposed sites, one species exhibiting CAM and the other one exhibiting net CO2 uptake exclusively during daytime via conventional C3 photosynthesis, showed similar light response curves and the same quantum yield for photosynthetic O2 evolution.


Cost effectiveness of entecavir versus lamivudine with adefovir salvage in HBeAg-positive chronic hepatitis B.
To evaluate the cost effectiveness of treatment of hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) with entecavir compared with lamivudine with adefovir salvage, based primarily on the results of a recent 2-year, randomised, multicentre, clinical trial (n = 709). Previous economic analyses have been limited by the lack of comparative clinical data for entecavir and lamivudine beyond 1-year duration and for salvage therapy. We conducted a cost-utility analysis using a Markov model from a US-payer perspective over a lifetime time horizon. The hypothetical cohort was 35-year-old patients with HBeAg-positive CHB. We evaluated 2 years of treatment with entecavir 0.5mg/day versus lamivudine 100mg/day, plus addition of adefovir 10mg/day for patients who developed virologic breakthrough due to resistance to either drug. In a scenario analysis, we considered adefovir plus lamivudine combination therapy for treatment-naive patients. Clinical and economic inputs ($US, year 2006 values) were derived from publicly available data, and probabilistic sensitivity analyses were conducted to evaluate uncertainty in the results. The estimated 10-year cumulative incidence of cirrhosis for patients initiated on entecavir was 2.3% lower than for those on lamivudine (20.5% vs 22.8%). The discounted incremental cost per QALY gained was $US7600 in the base-case analysis, and the 95% central range from probabilistic sensitivity analysis was $US2500-$US19 100. Combination therapy for treatment-naive patients led to an increase in costs without improvement in patient outcomes compared with entecavir monotherapy. Our analysis suggests entecavir improves health outcomes in a cost-effective manner compared with lamivudine with adefovir salvage or combination therapy, and highlights the importance of using evidence-based effectiveness estimates in economic studies of CHB therapies.


Distribution of two splice variants of the glutamate transporter GLT-1 in rat brain and pituitary.
We have performed immunocytochemistry on rat brains using a highly specific antiserum directed against the originally described form of the glutamate transporter GLT-1 (referred to hereafter as GLT-1alpha), and another against a C-terminal splice variant of this protein, GLT-1B. Both forms of GLT-1 were abundant in rat brain, especially in regions such as the hippocampus and cerebral cortex, and macroscopic examination of sections suggested that both forms were generally regionally coexistent. However, disparities were evident; GLT-1alpha was present in the intermediate lobe of the pituitary gland, whereas GLT-1B was absent. Similar marked disparities were also noted in the external capsule, where GLT1A labeling was abundant but GLT-1B was only occasionally encountered. Conversely, GLT-1B was more extensively distributed, relative to GLT-1alpha, in areas such as the deep cerebellar nuclei. In most regions, such as the olfactory bulbs, both splice variants were present but differences were evident in their distribution. In cerebral cortex, patches were evident where GLT-1B was absent, whereas no such patches were evident for GLT-1alpha. At high resolution, other discrepancies were evident; double-labeling of areas such as hippocampus indicated that the two splice variants may either be differentially expressed by closely apposed glial elements or that the two splice variants may be differentially targeted to distinct membrane domains of individual glial cells.


Immunocytochemical demonstration of cytokeratin in gastrointestinal carcinoids and their probable precursor cells.
Occurrence and distribution of cytokeratin, neuron specific enolase (NSE) and actin were studied by the immunoperoxidase-antiperoxidase (PAP)-technique using specific antibodies in formalin-fixed, paraffin-embedded material from 6 cases of neurogenic appendicopathy with numerous endocrine cells in the mucosal stroma (SEC), 5 cases of microcarcinoidosis of the stomach, 12 gastrointestinal carcinoids and 4 bronchial carcinoids. Cytokeratin was detectable in all tumor cells. In addition, the epithelial endocrine cells (EEC) and the SEC of intestinal origin were cytokeratin positive. EEC, SEC and cells of microcarcinoids and carcinoids showed a positive immunoreactivity with antibodies to NSE, whereas actin antibodies did not reveal significant staining of these cells. These results strongly suggest that carcinoids of the gastrointestinal tract originate from SEC that have migrated downwards into the stroma from the epithelial layer (" Endophytie " according to Feyrter ).


9-Methyl-7-bromoeudistomin D, a potent inducer of calcium release from sarcoplasmic reticulum of skeletal muscle.
The Ca2+-releasing action of several derivatives of eudistomin D isolated from a marine tunicate was compared with that of caffeine. It was found that 9-methyl-7-bromoeudistomin D was approximately 1000 times more potent than caffeine in causing Ca2+ release from the sarcoplasmic reticulum.


[Statistical findings on the variability of chemo-antibiotic resistance in minor Salmonella strains particularly frequent in the Piedmont area].
462 Strains of Salmonella spp. were tested; they had been arranged in species according to the frequency of appearance in Regione Piemonte: 358 of them were isolated in the period 1975-'76 and 104 in 1977. The susceptibility of the two groups considered to 40 antibiotics was determined. The results obtained evidenced significant variations of this parameter.


Removal of superficial inhibition releases hyperexcitability in middle and deep horizontal slices from rat somatosensory neocortex.
Synaptic physiology was studied in horizontal slices of rat somatosensory neocortex. Intrinsic properties of pyramidal neurons from horizontal slices resembled those recorded in cells from coronal slices, but cells in superficial horizontal slices displayed more prominent fast and slow inhibition, while cells from deeper slices showed disinhibition. This disinhibition in deeper horizontal slices resulted in epileptogenesis in 81% of middle and 35% of deep layer horizontal slices. Brief exposure to glutamate antagonists and dantrolene was ineffective in preventing epileptic activity, but limited pentobarbital exposure reduced the proportion of deep slices manifesting epileptiform activity by 75%. Thus, within cortex inhibition dominates superficially and excitation predominates in deeper layers. While the cortex is vulnerable to hyperexcitability when superficial cortex is compromised, enhancing fast inhibition can reset the excitation-inhibition balance, and prevent epileptogenesis.


The effect of hydroxyapatite-hPRP, and coral-hPRP on bone healing in rabbits: radiological, biomechanical, macroscopic and histopathologic evaluation.
There is a continuing search for bone substitutes to avoid or minimize the need for autogenous bone grafts. Human platelet-rich plasma (hPRP) is used to stimulate bone formation in vivo. Hydroxyapatite, a crystalline phase of calcium phosphate found naturally in bone minerals, has shown tremendous promise as a graft material. Coral is an osteoconductive material used as a bone graft extender. This study examined the effect of human platelet-rich plasma in combination with hydroxyapatite and coral on osteogenesis in vivo using rabbit model bone healing. A critical size defect of 10 mm elongation was created in the radial diaphysis of 36 rabbit and either supplied with human platelet-rich plasma (12 rabbits), and in combination with hydroxyapatite (12 rabbits), or coral (12 rabbits). Radiographs of each forelimb were taken postoperatively on 1st day and then at the 2nd, 4th, 6th and 8th weeks post injury to evaluate bone defect healing. The operated radiuses were removed on the 56th postoperative day and were grossly and histopathologically evaluated. In addition, biomechanical test was conducted on the operated and normal forearms of another half of the rabbits in each group. This study demonstrated that high concentrations of xenogenic platelets lead to superior and faster bone formation in comparison with hydroxyapatite-hPRP and coral-hPRP. Hydroxyapatite-hPRP and coral-hPRP resulted to almost similar results in bone healing process at this stage.


Effects of category diversity on learning, memory, and generalization.
In this study, we examined the effect of within-category diversity on people's ability to learn perceptual categories, their inclination to generalize categories to novel items, and their ability to distinguish new items from old. After learning to distinguish a control category from an experimental category that was either clustered or diverse, participants performed a test of category generalization or old-new recognition. Diversity made learning more difficult, increased generalization to novel items outside the range of training items, and made it difficult to distinguish such novel items from familiar ones. Regression analyses using the generalized context model suggested that the results could be explained in terms of similarities between old and new items combined with a rescaling of the similarity space that varied according to the diversity of the training items. Participants who learned the diverse category were less sensitive to psychological distance than were the participants who learned a more clustered category.


Proteasome inhibitors induce apoptosis of prostate cancer cells by inducing nuclear translocation of IkappaBalpha.
Proteasome inhibitors are known to suppress the proteasome-mediated degradation of IkappaBalpha in stimulated cells. This results in the cytoplasmic retention of NFkappaB and its reduced nuclear transcriptional activity. In this study, we show that in the metastatic prostate cancer cells, the proteasome inhibitors exhibit a novel, previously unrecognized effect: they increase the cellular levels of IkappaBalpha, which then translocates to the nucleus, associates with the nuclear p65 NFkappaB, thus inhibiting the constitutive NFkappaB DNA binding activity and inducing apoptosis. The proteasome inhibition-induced nuclear translocation of IkappaBalpha is dependent on de novo protein synthesis, occurs also in other cell types, and does not require IkappaBalpha phosphorylation on Ser-32. Since NFkappaB activity is constitutively increased in many human cancers as well as in inflammatory disorders, the proteasome inhibition-induced nuclear translocation of IkappaBalpha could thus provide a new therapeutic strategy aimed at the specific inhibition of NFkappaB activity by the nuclear IkappaBalpha.


Left ventricular dysfunction following ligation of the ductus arteriosus in the preterm baboon.
We evaluated the effect of early closure of the patent ductus arteriosus (PDA) on cardiac performance in a preterm baboon model of hyaline membrane disease. Eleven baboon fetuses were delivered at 75% gestation, resuscitated, and treated with conventional ventilation. The animals were divided into two groups, and underwent left thoracotomy at 2 hr of age. Group 1 (N = 5) had ductal infiltration with formalin to maintain ductal patency. Group 2 (N = 6) had ductal ligation. Hemodynamic parameters were compared between groups before and after surgical intervention. Examination of cardiovascular function was performed at intervals over a 24-hr period using two-dimensional, M-mode, and pulsed Doppler echocardiography. Ventricular performance was significantly worse in the ligation animals as demonstrated by reduced left and right ventricular outputs and reduced left ventricular shortening. However, pulmonary disease and arterial blood gases were not different. Early ligation was also associated with significantly increased systemic vascular resistance. We conclude that early ductal ligation leads to impaired ventricular performance in the premature primate. We hypothesize that the PDA may protect the immature ventricle by preventing pressure overload during adaptation to postnatal life.


RAS and RAF mutations in banal melanocytic aggregates contiguous with primary cutaneous melanoma: clues to melanomagenesis.
Distinguishing banal melanocytic aggregates contiguous with malignant melanoma can be a histological challenge but is essential because of the potential for a spurious Breslow measurement. Our aim was to ascertain whether the histological distinction between the two relates to differences in the prevalence of mutations in genes significant in melanomagenesis. Mutations in BRAF codon 600, NRAS1 codons 12/13, NRAS2 codons 60/61 and KRAS codons 12/13 were ascertained in 18 cases of primary cutaneous malignant melanoma contiguous with banal melanocytic aggregates using laser capture microdissection. Overall, 12 of 18 cases (67%) exhibited a mutation in at least one gene. BRAF V600E appeared to be the most commonly mutated gene in both the melanocytic aggregate (seven of 18, 39%) and the melanoma (four of 18, 22%). Both populations demonstrated a similar BRAF genomic profile in 11 of 18 cases (61%) (two BRAF V600E, nine BRAF-WT), a similar KRAS genomic profile in 14 of 18 cases (78%) (one KRAS G12V, 13 KRAS-WT) and a similar NRAS2 genomic profile in 14 of 18 cases (all WT). Of interest, we noted a relatively high prevalence of KRAS mutations (five of 18, 28%). The frequency of KRAS mutations in the melanocytic aggregate (five of 18, 28%) was second to BRAF V600E, while in melanoma, the frequency was also second to BRAF V600E but equalled that of NRAS2 (1 of 18, 6%). No NRAS1 mutations were observed. BRAF and RAS mutations appeared to be mutually exclusive with only three of 18 cases (17%) demonstrating a mutation in both genes (melanocytic aggregate only). Our findings hint towards the interpretation of banal melanocytic aggregates serving as precursor lesions.


Magnet hospitals: insights and issues.
This article explores some of the major lessons that have been learned over more than 2 decades of experience with the Magnet Hospital recognition program. Both the background for the work and some of the issues related to the effort are discussed.


Hematopoietic cell transplantation ameliorates clinical phenotype and progression of the CNS pathology in the mouse model of late onset Krabbe disease.
Krabbe disease is a genetic demyelinating disease caused by a deficiency of galactosylceramidase. The majority of cases are of infantile onset with rapid clinical course. A rare late onset form with milder clinical symptoms also exists. The latter form has been reported to respond well to the bone marrow transplantation (BMT) therapy. We tested whether the BMT could be an effective therapy for the mouse model of the late onset form, saposin-A-/- (SAP-A-/-) mice. We used green fluorescent protein transgenic mice as the donors. Chimeric SAP-A-/- mice that received BMT showed very little evidence of neurologic symptoms. At postnatal day 190 when severe demyelination was evident in naive SAP-A-/- mice, demyelination was virtually absent in the brain of chimeric SAP-A-/- mice. Presence of residual enzyme activity, at the time of rapid myelination in SAP-A-/- mice, appears to limit initial inflammatory responses and macrophage infiltration, thereby preventing progression of demyelination in the CNS in SAP-A-/- mice. In contrast, the peripheral nerves showed features of hypertrophic neuropathy with hypomyelination and onion bulb formation, suggesting that there are different cellular responses to the BMT in the CNS and PNS.


The stress sensitization hypothesis: understanding the course of bipolar disorder.
The influence of psychosocial stress on the course of bipolar disorder has been increasingly recognized. The authors tested hypotheses about both stress and early adversity "sensitization" on the course of bipolar disorder over a one-year period. The participants were 58 adults (29 male and 29 female) with a diagnosis of bipolar I disorder. They were evaluated every three months for one year. Stressful life events and the presence of early adversity were assessed by structured interview. There was no significant interaction between stress and episode number in the prediction of bipolar recurrence. The interaction of early adversity severity and stressful life events significantly predicted recurrence in a manner consistent with the sensitization hypothesis. Participants with early adversity reported lower levels of stress prior to recurrence than those without early adversity. Individuals with early adversity also had a significantly younger age of bipolar onset. The sample size was small and the number of past episodes was determined retrospectively, mainly through self-report. Severe early adversity may result in a greater effect of stress on bipolar recurrence and earlier onset of bipolar disorder, suggesting the need for further studies of stress mechanisms in bipolar disorder and of treatments designed to intervene early among those at risk.


Benefit-to-Risk Balance of Weight Loss Interventions in Older Adults with Obesity.
Obesity in the older adult is a burgeoning health epidemic that leads to increased morbidity, disability, and institutionalization. This review presents a brief overview of geriatric-specific consequences of obesity by highlighting the risks and benefits of intentional weight loss. Intentional weight loss reduces the extent of adiposity-related illnesses, yet the approach in older adults is fraught with challenges. Interventions combining caloric restriction and physical exercise (aerobic and resistance) maximize fat loss and minimize loss of muscle and bone. Interventions are also effective at improving physical function, reducing medication burden, and improving symptomatic osteoarthritis in this population. Approaches can mitigate the risks of isolated caloric restriction on muscle and bone in a safe and effective manner. Effective weight loss strategies should be considered in older adults. While there are potential risks, practical clinical approaches can minimize the potential harms while maximizing their benefits.


Multi-dimensional nanoscale liquid chromatography and nano-electrospray ion-trap mass spectrometry for detection of Clostridium botulinum type C and the produced botulinum neurotoxin type C complex.
Botulinum neurotoxin types C, D and their mosaic forms C/D and D/C produced mainly by Clostridium botulinum types C and D cause botulism in animals and belong to the most toxic substances for poultry and fish. In addition to intoxications, also toxoinfections with C. botulinum types C and D play a role that should not be underestimated, especially in veterinary medicine. Contrary to other botulinum neurotoxin complexes (BT x), the biosynthesis of these types is phage-encoded. Currently, the gold standard for neurotoxin detection in cases of clinical botulism is the mouse bioassay. In the last few years, alternatives for replacing this mouse bioassay have become increasingly interesting for the detection and characterisation of botulinum neurotoxins. Therefore, immunological techniques based mainly on antibodies, PCR or mass spectral methods have been developed. In this context, the most promising development is that of different endopeptidase assays. In our study, we were able to show that the 2D-nano-LC-MS/MS method presented by Klaubert et al. 2009 especially for detecting BT x A, B, E and F in complex culture media can also be used for detecting BT x C. The focus was therefore on transferring this method to detecting BT x C and pointing out necessary modifications of this current method. For method development, we used different culture preparations and sample conditions. To find out whether BT x C is just as stable against acetic peptic pretreatment as other BT x, we used sample preparations with and without peptic pretreatment. The decisive difference to previous publications is the detection of produced BT x C directly from culture supernatant of different strains of C. botulinum type C. In addition, we present a new approach of detecting protein fragments from C3 and C2 toxin and some specific host cell proteins of the bacterium Clostridium spp. in order to specify the carrier bacterium, therefore verifying the presence of an intact neurotoxin-encoding phage also without directly detecting BT x C and thus the possibility to produce neurotoxin. Herein, we describe a new method to examine environmental samples or suspected feed samples in cases of toxoinfections as well as finding out the causes of clinical botulism. This new approach is particularly interesting for veterinary medicine, especially for diseases like chronic botulism in cows or equine grass sickness.


Chlorates and perchlorates as potential high-energy materials: chlorate- and perchlorate-substituted methanes.
The structures and properties of thirty-three of the thirty-four possible chlorato-Cl-, chlorato-O- and perchlorato-derivatives of methane have been computed using the range-separated hybrid generalized gradient approximation density functional method ωB97X-D with the 6-311+G(2d,p) basis set. These results indicate that the chlorato-O-substituent confers more stability to a molecule than does the chlorato-Cl-substituent; the perchlorato-substituent is approximately intermediate in this regard when decomposition energies are calculated. The C-Cl bond lengths in the chlorato-Cl-substituents generally elongate and the C-O distances in the chlorato-O- and perchlorato-substituents tend to shorten as the number of chlorate/perchlorate substituents increases. In addition, as the C-O bond shortens, the CO-Cl bond lengthens. The calculated Mulliken and Löwdin bond orders for these bonds exhibit the opposite of the trends exhibited by the bond lengths, as expected: As the bond lengthens, the bond order decreases, and vice versa. The single molecule that could not be optimized as a stable methane derivative, (chlorato-Cl-)tris(chlorato-O-)methane, rearranged during all optimization attempts to an isomer of the neutral Cl2O5 molecule and a hitherto unknown molecule, bis(chlorato-O-)carbonyl, (O2ClO)2C=O.


Development of Gluten-Free Rice Flour Noodles That Suit the Tastes of Japanese People.
Gluten-free rice flour noodles with a flavor and texture profile preferred by the Japanese people were developed. The rice noodles contained potato starch (PS) as a binder. "Koshihikari" was selected from several candidate varieties based on its pasting properties. Since the Japanese people prefer the chewy texture of wheat flour "Udon" noodles, first, the stress-strain characteristics of "Udon" noodles in Japan were quantified, using a mechanical test. Next, different formulations of rice noodles were prepared by changing the amount of PS blended into the noodles. The mechanical tests on wheat and rice noodles show that rice noodles made from 85% rice flour and 15% PS have a texture similar to that of "Udon" noodles. Brown rice noodles containing roasted brown rice flour were also developed. Since brown rice flour hinders the binding of the dough, it was necessary to increase the amount of PS to increase the binding of roasted brown rice flour. Finally, noodles with 70% white rice flour, 10% brown rice flour, and 20% PS were produced. The gas chromatography-mass spectrometry analysis of the volatile compounds contained in white rice noodles and brown rice noodles identified the volatile compounds characteristic each of type.


Sulindac sulfide, but not sulindac sulfone, inhibits colorectal cancer growth.
Sulindac sulfide, a metabolite of the nonsteroidal antiinflammatory drug (NSAID) sulindac sulfoxide, is effective at reducing tumor burden in both familial adenomatous polyposis patients and in animals with colorectal cancer. Another sulindac sulfoxide metabolite, sulindac sulfone, has been reported to have antitumor properties without inhibiting cyclooxygenase activity. Here we report the effect of sulindac sulfone treatment on the growth of colorectal carcinoma cells. We observed that sulindac sulfide or sulfone treatment of HCA-7 cells led to inhibition of prostaglandin E2 production. Both sulindac sulfide and sulfone inhibited HCA-7 and HCT-116 cell growth in vitro. Sulindac sulfone had no effect on the growth of either HCA-7 or HCT-116 xenografts, whereas the sulfide derivative inhibited HCA-7 growth in vivo. Both sulindac sulfide and sulfone inhibited colon carcinoma cell growth and prostaglandin production in vitro, but sulindac sulfone had no effect on the growth of colon cancer cell xenografts in nude mice.


A Preoperative Risk Model for Postoperative Pneumonia After Coronary Artery Bypass Grafting.
Postoperative pneumonia is the most prevalent of all hospital-acquired infections after isolated coronary artery bypass graft surgery (CABG). Accurate prediction of a patient's risk of this morbid complication is hindered by its low relative incidence. In an effort to support clinical decision making and quality improvement, we developed a preoperative prediction model for postoperative pneumonia after CABG. We undertook an observational study of 16,084 patients undergoing CABG between the third quarter of 2011 and the second quarter of 2014 across 33 institutions participating in the Michigan Society of Thoracic and Cardiovascular Surgeons Quality Collaborative. Variables related to patient demographics, medical history, admission status, comorbid disease, cardiac anatomy, and the institution performing the procedure were investigated. Logistic regression through forward stepwise selection (p < 0.05 threshold) was utilized to develop a risk prediction model for estimating the occurrence of pneumonia. Traditional methods were used to assess the model's performance. Postoperative pneumonia occurred in 3.30% of patients. Multivariable analysis identified 17 preoperative factors, including demographics, laboratory values, comorbid disease, pulmonary and cardiac function, and operative status. The final model significantly predicted the occurrence of pneumonia, and performed well (C-statistic: 0.74). These findings were confirmed through sensitivity analyses by center and clinically important subgroups. We identified 17 readily obtainable preoperative variables associated with postoperative pneumonia. This model may be used to provide individualized risk estimation and to identify opportunities to reduce a patient's preoperative risk of pneumonia through prehabilitation.


Syntheses, characterization and electrochemical and spectroscopic properties of ruthenium-iron complexes of 2,3,5,6-tetrakis(2-pyridyl)pyrazine and ferrocene-acetylide ligands.
Heterodimetallic Ru-Fe complexes [(tppz)(PPh3)2RuL](ClO4) (L = C[triple bond, length as m-dash]CFc, [](ClO4); C[triple bond, length as m-dash]C-C6H4-C[triple bond, length as m-dash]CFc), [](ClO4); C[triple bond, length as m-dash]C-C6H4-C6H4-C[triple bond, length as m-dash]CFc, [](ClO4)) were synthesized by the reactions of [(tppz)(PPh3)2RuCl](ClO4) (tppz = 2,3,5,6-tetrakis(2-pyridyl)pyrazine) with ferrocence-acetylide ligands and characterized by ESI-MS, and (1)H and (31)P NMR spectroscopies. The structure of [](PF6) was determined by X-ray crystallography. The electrochemical studies show that compounds [](ClO4)-[](ClO4) possess two widely separated anodic peaks, ascribable to one-electron oxidation of Fc and Ru(II), respectively. This assignment is further corroborated by the results of UV-vis-NIR, XPS, and theoretical calculation studies. Compound [](ClO4) exhibits significant RuFe metal-metal interactions across the Ru-C[triple bond, length as m-dash]C-Fc backbone. As revealed by electrochemical, spectroscopic and theoretical computational studies, one-electron oxidized species [](ClO4)2 is between the electronically delocalized and valence-trapped state and shows a typical Robin-Day class II mixed-valence behavior.


Trophic exploitation in grassland food chains: simple models and a field experiment.
This study provides insight into the importance of top carnivores (top-down control) and nutrient inputs (bottom-up control) in structuring food chains in a terrestrial grassland system. Qualitative predictions about food chain structure are generated using 4 simple models, each differing in assumptions about some key component in the population dynamics of the herbivore trophic level. The four model systems can be classified broadly into two groups (1) those that assume plant resource intake by herbivores is limited by search rate and handling time as described by classic Lotka-Volterra models; and (2) those that assume plant resource intake by herbivores is limited externally by the supply rate of resources as described by alternatives to Lotka-Volterra formulations. The first class of models tends to ascribe greater importance to top-down control of food chain structure whereas the second class places greater weight on bottom-up control. I evaluated the model predictions using experimentally assembled grassland food chains in which I manipulated nutrient inputs and carnivore (wolf spider) abundance to determine the degree of top-down and bottom-up control of grassland plants and herbivores (grasshoppers). The experimental results were most consistent with predictions of the second class of models implying a predominance of bottom-up control of food chain structure.


Phase I/II study of Fluosol-DA and 100% oxygen as an adjuvant to radiation in the treatment of advanced squamous cell tumors of the head and neck.
Fluosol-DA 20% (Fluosol) is an emulsion of perfluorodecalin and perfluorotripropylamine, which has the ability to carry oxygen and has been shown to enhance the ability of radiation to control tumors in animal studies. Since November 1984, patients with unresectable squamous cell carcinomas of the head and neck have been enrolled in a study to evaluate the safety and potential efficacy of this adjuvant therapy. Forty-six patients were entered of which 37 completed radiation and are evaluable. Patients were infused weekly with Fluosol and then breathed 100% oxygen for a minimum of 30 minutes prior to and during radiation. Eleven patients received 5 infusions of 8 mL/Kg, four patients 6 infusions of 8 mL/Kg, five patients 5 infusions of 9 mL/Kg, seven patients 7 infusions of 7 mL/Kg and eight patients 8 infusions of 7 mL/Kg. Nine patients had Stage III disease, 20 patients Stage IV disease and 8 patients had failed previous therapy with chemotherapy and/or surgery. The radiation doses delivered ranged from 6600 cGy to 7500 cGy. The overall complete response rate for this group was 76%. All 9 Stage III patients were complete responders, 13 of 20 Stage IV responded and 6 of 8 with previous therapy were complete responders. The survival rate at 1 year was 67% for absolute and 78% as determinant. Of those patients achieving a complete response, 75% continued free of disease 1 year after therapy. Out of 254 total test doses, 11 patients experienced a reaction to the test dose of Fluosol. Of 235 total infusions 6 patients experienced a reaction during the Fluosol infusion with 7 patients experiencing post infusion reactions. These were readily controlled with diphenhydramine or acetominophen. Elevated liver enzymes were observed in some patients with a mean time to normalization of 102 days for alkaline phosphatase, 39 days for SGOT, and 46 days for SGPT.


Patent foramen ovale and perioperative stroke in noncardiac surgery: a systematic review and meta-analysis.
Patent foramen ovale (PFO) is associated with perioperative stroke in noncardiac surgery. The magnitude of this association was assessed in a systematic review and meta-analysis. Electronic databases were searched up to June 2022 for studies assessing the association between patent foramen ovale and perioperative stroke in adult patients undergoing noncardiac surgery. The primary analysis was limited to studies reporting effect estimates adjusted for significant clinical confounders. We calculated the adjusted odds ratio (aOR) and 95% confidence interval (CI). We included nine retrospective and two prospective observational studies, including 21 257 082 patients. The presence of a patent foramen ovale was independently associated with stroke at 30 days after surgery (aOR=6.68 [95% CI: 3.51-12.73]; P<0.001) and at longest follow-up available (aOR=7.36 [95% CI: 3.56-15.21]; P<0.001). The odds of stroke at 30 days varied according to surgical specialty: neurosurgery (aOR=4.52 [95% CI: 3.17-6.43]), vascular surgery (aOR=7.15 [95% CI: 2.52-20.22]), thoracic surgery (aOR=10.64 [95% CI: 5.97-18.98]), orthopaedic surgery (aOR=11.85 [95% CI: 5.38-26.08]), general surgery (aOR=14.40 [95% CI: 10.88-19.06]), and genitourinary surgery (aOR=17.28 [95% CI: 10.36-28.84]). The presence of a patent foramen ovale is associated with a large and consistent increase in odds of stroke across all explored surgical settings. Prospective trials should further explore this association by systematically assessing patent foramen ovale and stroke prevalence and identifying a specific population at risk. This is crucial for the elaboration of prevention plans and may improve perioperative outcomes.


The Adh gene promoters of Drosophila melanogaster and Drosophila orena are functionally conserved and share features of sequence structure and nuclease-protected sites.
The sibling species Drosophila melanogaster and D. orena show similar patterns of alcohol dehydrogenase expression, both spatially and temporally. These two species diverged from a common ancestor 6 million to 15 million years ago, and the DNA sequences of the promoter regions of their Adh genes show a mosaic pattern of conservation and change. By interspecific transformation of D. orena sequences into D. melanogaster, we demonstrate a functional equivalence between these sequences. Using both D. melanogaster embryo extracts and purified transcription factor Adf-1, we compare the protection of these promoter sequences from nuclease, demonstrating considerable conservation.


Initiation, promotion and complete carcinogenesis by N-methyl-N'-nitro-N-nitrosoguanidine or ethylnitrosourea in the Sencar mouse skin tumorigenesis model.
Five doses of either N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or ethylnitrosourea (ENU) were tested as complete carcinogens, tumor initiators and tumor promoters in the SENCAR skin tumorigenesis model. As tumor initiators, MNNG-induced papillomas at all doses tested, while ENU was active from 10-40 mumol. As complete carcinogens, MNNG from doses of 0.5-5.0 mumol and ENU from doses of 10 mumol-40 mumol were potent inducers of both papillomas and carcinomas indicating that these agents are active as both tumor initiators and tumor promoters.


Retraction: A highly concise and practical route to clavaminols, sphinganine and (+)-spisulosine via indium mediated allylation of α-hydrazino aldehyde and a theoretical insight into the stereochemical aspects of the reaction.
[This retracts the article DOI: 10.1039/C3RA43048K.].


The evolution of the adenylate-forming protein family in beetles: multiple luciferase gene paralogues in fireflies and glow-worms.
Bioluminescence in beetles is dependent upon the enzyme luciferase. It has been hypothesised luciferase evolved from a fatty acyl-CoA synthetase gene deriving a novel bioluminescent function (neofunctionalization) after a gene duplication event. We evaluated this hypothesis within a phylogenetic framework using independent evidence obtained from the genome of Tribolium castaneum, published luciferase genes and novel luciferase and luciferase-like sequences. This phylogenetic study provides evidence for a large gene family of luciferase and luciferase-like paralogues in bioluminescent and non-bioluminescent beetles. All luciferase sequences formed a clade supporting a protoluciferase existing prior to the divergence of the Lampyridae, Elateridae and Phengodidae (Elateroidea). Multiple luciferase genes were identified from members of the Photurinae and the Luciolinae indicating complex gene duplication events within lampyrid genomes. The majority of luciferase residues were identified to be under purifying selection as opposed to positive selection. We conclude that beetle luciferase may have arisen from a process of subfunctionalization as opposed to neofunctionalization early on in the evolution of the Elateroidea.


The PINK1-PARKIN Mitochondrial Ubiquitylation Pathway Drives a Program of OPTN/NDP52 Recruitment and TBK1 Activation to Promote Mitophagy.
Damaged mitochondria are detrimental to cellular homeostasis. One mechanism for removal of damaged mitochondria involves the PINK1-PARKIN pathway, which poly-ubiquitylates damaged mitochondria to promote mitophagy. We report that assembly of ubiquitin chains on mitochondria triggers autophagy adaptor recruitment concomitantly with activation of the TBK1 kinase, which physically associates with OPTN, NDP52, and SQSTM1. TBK1 activation in HeLa cells requires OPTN and NDP52 and OPTN ubiquitin chain binding. In addition to the known role of S177 phosphorylation in OPTN on ATG8 recruitment, TBK1-dependent phosphorylation on S473 and S513 promotes ubiquitin chain binding in vitro as well as TBK1 activation, OPTN mitochondrial retention, and efficient mitophagy in vivo. These data reveal a self-reinforcing positive feedback mechanism that coordinates TBK1-dependent autophagy adaptor phosphorylation with the assembly of ubiquitin chains on mitochondria to facilitate efficient mitophagy, and mechanistically links genes mutated in Parkinson's disease and amyotrophic lateral sclerosis in a common selective autophagy pathway.


Rhodotorula mucilaginosa Fungemia and Pleural Tuberculosis in an Immunocompetent Patient: An Uncommon Association.
We report an atypical case of Rhodotorula mucilaginosa fungemia coexisting with pleural tuberculosis, in an immunocompetent host. The patient was an inhaled drug abuser and worked in a fruit market. The diagnosis of Rhodotorula mucilaginosa infection was established by the isolation of the yeast in two blood cultures followed by a good response to amphotericin B treatment. Persistent evening fever and pleural effusion led to the second diagnosis-pleural tuberculosis. In the last 5 years, this was the only case of Rhodotorula mucilaginosa fungemia in our hospital and the first case in the literature that documents Rhodotorula mucilaginosa fungemia associated with pleural tuberculosis.