Mitochondrial release of AIF and EndoG requires caspase activation downstream of Bax/Bak-mediated permeabilization.
Mitochondrial outer-membrane permeabilization by pro-apoptotic Bcl-2 family members plays a crucial role in apoptosis induction. However, whether this directly causes the release of the different mitochondrial apoptogenic factors simultaneously is currently unknown. Here we report that in cells or with isolated mitochondria, pro-apoptotic Bcl-2 proteins cause the release of cytochrome c, Smac/Diablo and HtrA2/Omi but not endonuclease G (EndoG) and apoptosis-inducing factor (AIF). In cells treated with Bax/Bak-dependent pro-apoptotic drugs, neither the caspase inhibitor zVAD-fmk nor loss of Apaf-1 affected the efflux of cytochrome c, Smac/Diablo and HtrA2/Omi, but both prevented the release of EndoG and AIF. Our findings identify the mitochondrial response to pro-apoptotic stimuli as a selective process leading to a hierarchical ordering of the effectors involved in cell death induction. Moreover, as in Caenorhabditis elegans, EndoG and AIF act downstream of caspase activation. Thus EndoG and AIF seem to define a 'caspase-dependent' mitochondria-initiated apoptotic DNA degradation pathway that is conserved between mammals and nematodes.


Effects of the contact isolation application on anxiety and depression levels of the patients.
This study was aimed to assess the effects of contact isolation application on anxiety and depression levels of the patients, the effects of certain sociodemographics and patient characteristics on anxiety and depression levels, and the thoughts of the isolated patients about contact isolation. This non-randomized quasi-experimental study was carried out with 60 isolated and 57 non-isolated patients with hospital infection. The data were acquired from Hospital Anxiety and Depression Scale (HADS-A (anxiety) and HADS-D (depression)) and patient information form. There was no statistically significant difference between the anxiety and depression levels of the isolated and non-isolated patients. In the isolated patients, the depression points were higher in patients who were, women, received primary education and had lower income levels. Of the patients, 86.4% of them told that they were happy to be in the isolation room. Contact isolation application did not affect anxiety and depression levels of the patients. However, personal attributes increased the development of depression. In contact isolated patients, personal attributes should be taken into consideration in nursing care planning to prevent development of depression.


Multiplex and Label-Free Relative Quantification Approach for Studying Protein Abundance of Drug Metabolizing Enzymes in Human Liver Microsomes Using SWATH-MS.
We describe a sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS) based method for label-free, simultaneous, relative quantification of drug metabolism enzymes in human liver microsomes (HLM; n = 78). In-solution tryptic digestion was aided by a pressure cycling method, which allowed a 90 min incubation time, a significant reduction over classical protocols (12-18 h). Digested peptides were separated on an Acquity UHPLC Peptide BEH C18 column using a 60 min gradient method at a flow rate of 0.100 mL/min. The quadrupole-time-of-flight mass spectrometer (ESI-QTOFMS) was operated in positive electrospray ionization mode, and data were acquired by data-dependent acquisition (DDA) and SWATH-MSALL mode. A pooled HLM sample was used as a quality control to evaluate variability in digestion and quantification among different batches, and inter-batch %CV for various proteins was between 3.1 and 7.8%. Spectral library generated from the DDA data identified 1855 distinct proteins and 25 681 distinct peptides at a 1% global false discovery rate (FDR). SWATH data were queried and analyzed for 10 major cytochrome P450 (CYP) enzymes using Skyline, a targeted data extraction software. Further, correlation analysis was performed between functional activity, protein, and mRNA expression for ten CYP enzymes. Pearson correlation coefficient (r) between protein and activity for CYPs ranged from 0.314 (CYP2C19) to 0.767 (CYP2A6). A strong correlation was found between CYP3A4 and CYP3A5 abundance and activity determined using midazolam and testosterone (r > 0.600, p < 0.001). Protein-to-activity correlation was moderate (r > 0.400-0.600, p < 0.001) for CYP1A2, CYP2A6, CYP2B6, CYP2C9, and CYP2E1 and significant but poor (r < 0.400, p < 0.05) for CYP2C8, CYP2C19, and CYP2D6. The findings suggest the suitability of SWATH-MS based method as a valuable and relatively fast analytical technique for relative quantification of proteins in complex biological samples. We also show that protein abundance is a better surrogate than mRNA to predict the activity of CYP activity.


Women with endometriosis are more likely to suffer from migraines: a population-based study.
Previous research suggests that a co-morbid relationship exists between migraine and endometriosis; however, results have been inconsistent. In addition, female hormones, which are important in the pathogenesis and management of endometriosis, have been reported to precipitate migraine attacks and may confound the results. The aim of this population-based cohort study was to explore the relationship between migraine and endometriosis in women of reproductive age (18-51 years). Data were derived from the National Health Insurance Research Database of Taiwan, which contains outpatient and inpatient records from 2000 to 2007. Our study cohort included 20,220 endometriosis patients and 263,767 controls without endometriosis. We analyzed the prevalence of migraine in these women as recorded during the eight years of the database. Our results found that patients with endometriosis were more likely to suffer migraine headaches compared to controls (odds ratio [OR], 1.70; 95% confidence interval [CI] [1.59, 1.82]; p<0.001). In addition, the co-morbid association between migraine and endometriosis remained significant after the data were controlled for age and frequently utilized hormone therapies (OR, 1.37; 95% CI, [1.27, 1.47]; p<0.001). The results of this cohort study support the existence of a co-morbid relationship between migraine and endometriosis, even after adjusting for the possible effects of female hormone therapies on migraine attacks.


Dielectric Response of Cytoplasmic Water and Its Connection to the Vitality of Human Red Blood Cells. II. The Influence of Storage.
Maintaining an appropriate inventory of packaged blood products is a critical part of modern medicine. Consequently, the assessment of red blood cell (RBC) functionality is instrumental for the monitoring of the quality of stored RBC (sRBC) in the blood bank. We present a comprehensive study of sRBC lesion kinetics in SAGM (saline, adenine, glucose, mannitol) solution, using microwave dielectric spectroscopy (0.5-50 GHz) and cell deformability. As part of the research, we have isolated the microwave dielectric response of cytoplasmic water in sRBC. The extracted dielectric parameters are sensitive to the age of the cells and, in particular, to the critical moment of transition from discocyte to echinocyte. From the analysis of the dielectric relaxation as a function of storage-duration, we postulate that the behavior is rooted in the delicate interplay between bound and bulk water in the cellular interior. In particular, the microwave dielectric response reflects the moment when the continuous diffusion of oxygen to the cell and the oxygenation of hemoglobin affects the role played by water in the maintenance of cell integrity. These results open a possible new avenue for the noninvasive inspection of stored red blood cells, permitting a true inventory system for the modern blood bank.


Morphological study of the fetal parotid duct and buccinator muscle and the relationship to salivary secretion.
The parotid glands secrete about 25% of all saliva produced. In the presence of a stimulus, the amount of saliva secreted from the parotid gland increases to 50%. A decrease in the amount of produced saliva due to aging and parotiditis results in a dry mouth. Therefore, the parotid duct is important to maintaining a healthy oral cavity. In human adults, the parotid duct, approximately 6-8-cm long, travels over the masseter muscle and penetrates the buccinator muscle to enter the oral cavity. Although there have been various studies regarding the parotid gland, only few suggest a functional role of the parotid duct, especially its area of penetration of the buccinator muscle. In this study, 34 fetal specimens ranging from 4 to 10 months of age at death were dissected for anatomical and histological examinations. The area of the parotid duct penetrating the buccinator muscle was fully formed in 5-month-old fetuses. This study found buccinator muscle fibers invading the parotid duct wall near its opening in 6-month-old fetuses and older. Our results support the claim that the buccinator muscle may act as a sphincter, playing a role in regulating and possibly preventing the reflux of salivary secretions into the parotid duct.


Enzymatic transition states: thermodynamics, dynamics and analogue design.
Kinetic isotope effects and computational chemistry have defined the transition state structures for several members of the N-ribosyltransferase family. Transition state analogues designed to mimic their cognate transition state structures are among the most powerful enzyme inhibitors. In complexes of N-ribosyltransferases with their transition state analogues, the dynamic nature of the transition state is converted to an ordered, thermodynamic structure closely related to the transition state. This phenomenon is documented by peptide bond H/D exchange, crystallography and computational chemistry. Complexes with substrate, transition state and product analogues reveal reaction coordinate motion and catalytic interactions. Isotope-edited spectroscopic analysis and binding specificity of these complexes provides information about specific enzyme-transition state contacts. In combination with protein dynamic QM/MM models, it is proposed that the transition state is reached by stochastic dynamic excursions of the protein groups near the substrates in the closed conformation. Examples from fully dissociated (D(N) *A(N)), hybrid (D(N)A(N)) and symmetric nucleophilic displacement (A(N)D(N)) transition states are found in the N-ribosyltransferases. The success of transition state analogue inhibitor design based on kinetic isotope effects validates this approach to understanding enzymatic transition states.


Chemokines and other GPCR ligands synergize in receptor-mediated migration of monocyte-derived immature and mature dendritic cells.
Dendritic cells (DCs) are potent antigen presenting cells, described as the initiators of adaptive immune responses. Immature monocyte-derived DCs (MDDC) showed decreased CD14 expression, increased cell surface markers DC-SIGN and CD1a and enhanced levels of receptors for the chemokines CCL3 (CCR1/CCR5) and CXCL8 (CXCR1/CXCR2) compared with human CD14⁺ monocytes. After further MDDC maturation by LPS, the markers CD80 and CD83 and the chemokine receptors CXCR4 and CCR7 were upregulated, whereas CCR1, CCR2 and CCR5 expression was reduced. CCL3 dose-dependently synergized with CXCL8 or CXCL12 in chemotaxis of immature MDDC. CXCL12 augmented the CCL3-induced ERK1/2 and Akt phosphorylation in immature MDDC, although the synergy between CCL3 and CXCL12 in chemotaxis of immature MDDC was dependent on the Akt signaling pathway but not on ERK1/2 phosphorylation. CCL2 also synergized with CXCL12 in immature MDDC migration. Moreover, two CXC chemokines not sharing receptors (CXCL12 and CXCL8) cooperated in immature MDDC chemotaxis, whereas two CC chemokines (CCL3 and CCL7) sharing CCR1 did not. Further, the non-chemokine G protein-coupled receptor ligands chemerin and fMLP synergized with respectively CCL7 and CCL3 in immature MDDC signaling and migration. Finally, CXCL12 and CCL3 did not cooperate, but CXCL12 synergized with CCL21 in mature MDDC chemotaxis. Thus, chemokine synergy in immature and mature MDDC migration is dose-dependently regulated by chemokines via alterations in their chemokine receptor expression pattern according to their role in immune responses.


Prevention of anxiety disorders.
Research into the prevention of anxiety has increased dramatically in the past few years. Prevention programs have been directed at broad, nonspecific anxiety and at more specific anxiety types, such as panic disorder and post-traumatic stress disorder. Prevention of anxiety is still a relatively new field, but there has been a recent surge of literature reporting on different prevention programs. Universal prevention trials have shown modest but promising results, and school-based programs offered to all students also help to reduce stigmatization and common barriers to accessing treatment (eg, time, location, and cost). In contrast, targeted programs tend to show somewhat larger effects but rely on identification of relevant populations. Specific programs for the prevention of panic disorder and post-traumatic stress disorder have also demonstrated some preliminary success. This paper reviews the recent studies of prevention of anxiety and discusses several key issues, specifically (1) identification of at-risk participants for prevention programs, (2) motivation for participation, (3) optimal age for intervention, and (4) who should deliver the program.


Heterogeneity in tuberculosis pathology, microenvironments and therapeutic responses.
Tuberculosis (TB) lesions are extremely complex and dynamic. Here, we review the multiple types and fates of pulmonary lesions that form following infection by Mycobacterium tuberculosis and the impact of this spatial and temporal heterogeneity on the bacteria they harbor. The diverse immunopathology of granulomas and cavities generates a plethora of microenvironments to which M. tuberculosis bacilli must adapt. This in turn affects the replication, metabolism, and relative density of bacterial subpopulations, and consequently their respective susceptibility to chemotherapy. We outline recent developments that support a paradigm shift in our understanding of lesion progression. The simple model according to which lesions within a single individual react similarly to the systemic immune response no longer prevails. Host-pathogen interactions within lesions are a dynamic process, driven by subtle and local differences in signaling pathways, resulting in diverging trajectories of lesions within a single host. The spectrum of TB lesions is a continuum with a large overlap in the lesion types found in latently infected and active TB patients. We hope this overview will guide TB researchers in the design, choice of read-outs, and interpretation of future studies in the search for predictive biomarkers and novel therapies.


Comparative transcriptomics of stem rust resistance in wheat NILs mediated by Sr24 rust resistance gene.
Stem rust of wheat is a deleterious fungal disease across the globe causing severe yield losses. Although, many stem rust resistance genes (Sr) are being used in wheat breeding programs, new emerging stem rust pathotypes are a challenge to important Sr genes. In recent years, multiple studies on leaf and yellow rust molecular mechanism have been done, however, for stem rust such studies are lacking. Current study investigated stem rust induced response in the susceptible wheat genotype C306 and its Near Isogenic Line (NIL) for Sr24 gene, HW2004, using microarray analysis to understand the transcriptomic differences at different stages of infection. Results showed that HW2004 has higher basal levels of several important genes involved in pathogen detection, defence, and display early activation of multiple defence mechanisms. Further Gene Ontology (GO) and pathway analysis identified important genes responsible for pathogen detection, downstream signalling cascades and transcription factors (TFs) involved in activation and mediation of defence responses. Results suggest that generation of Reactive Oxygen Species (ROS), cytoskeletal rearrangement, activation of multiple hydrolases, and lipid metabolism mediated biosynthesis of certain secondary metabolites are collectively involved in Sr24-mediated defence in HW2004, in response to stem rust infection. Novel and unannotated, but highly responsive genes were also identified, which may also contribute towards resistance phenotype. Furthermore, certain DEGs also mapped close to the Sr24-linked marker on Thinopyrum elongatum translocated fragment on wheat 3E chromosome, which advocate further investigations for better insights of the Sr24-mediated stem rust resistance.


Low-dose Computed Tomography in a Pregnant Woman with a Ruptured Pseudoaneurysm of the Abdominal Aorta.
Imaging the pregnant patient presents a unique challenge to radiologist due to the risk of radiation to the conceptus (embryo/fetus). A rare case of a successfully recognized and treated pseudoaneurysm (PA) of the abdominal aorta is to be presented. The pseudoaneurysm occurred in the third trimester and had a favorable outcome for the mother and the baby. Emergent abdominal ultrasound (US) is the first modality in diagnostic algorithm for the rupture of aortic aneurysm in a pregnant woman. It provides the most rapid diagnostic information, although intestinal gas and abdominal tenderness may limit its accuracy. To confirm the findings, magnetic resonance angiography (MRA) or CT angiography (CTA) can be used. In our case, the diagnosis was established using a color Doppler ultrasonography of the abdomen and was later confirmed by a low dose CT scan of the abdominal aorta. MRA in such cases have some disadvantages. At many health centers, the monitoring of patients with acute ruptures is more difficult in the MR suite than at the CT scanner. MRA angiographic images are also subject to degradation by multiple artifacts and the visualization of the distal vasculature is suboptimal and inferior to the one done by CTA. Due to fetal movements, a small quantity of fresh blood can be overlooked by MR. MRA is often not available on a 24-hours basis, and the time required for making a diagnosis can preclude the use of MRA in an unstable patient. For this reason, we used a low dose CTA protocol to confirm the diagnosis. Low dose scanning protocols in CT can obtain sufficient diagnostic information while reducing the risk of radiation. A particular focus is put on the outline of new concepts for dose management and optimization. We used new approaches based on tube current modulation. The birth was induced by an urgent Caesarean section followed by a resection of a pseudoaneurysm and a reconstruction of the aorta with an end-to-end vascular prosthesis.


Superconductivity--the state that came in from the cold.
The exploration of high transition temperature copper-oxide-based superconductors has proceeded vigorously and internationally during the first year following the initial publication of the work of Bednorz and Müller. Progress in understanding the physics that underlies the phenomena has been slowed by difficulties resulting from the delicate and complex crystal chemistry of the material. Reports of superconducting behavior well above 100 kelvin have not been confirmed to date, although there is some suggestive evidence. A survey of the present state of the science and the possibilities for electronic and electrical power technologies is given.


Fundamental approach in the management of Drava Bahula Amlapitta with Bhringaraja (Eclipta alba).
The disease Amlapitta has been selected for the clinical trials because it presents two type of manifestations depending upon the involvement of Agni (Ushnagunadhikya) and Jala (Dravagunadhikya) Mahabhuta. The present research work was focused at Drava Guna, with an aim to assess the efficacy of a drug with quality of Ruksha and Ushna predominance like Bhringaraja in treating Amlapitta with Pitta Drava Guna Vriddhi. Randomized open clinical trials were conducted on 22 patients of Amlapitta who were screened on the basis of clinical findings and allocated in to two groups. The criteria for selection were the signs and symptoms of Dravagunadhikya Amlapitta, irrespective of sex, religion, etc. Group A consisting of 15 cases received the trial drug Bhringaraja tablet (4 Tab. two times, 1 tablet=500 mg) and 7 cases in Group B received rice powder tablet as a placebo (4 Tab. two times, 1 tablet=500 mg) for 4 weeks. Special scoring pattern was adopted for the assessment of Amlapitta. Routine pathological tests such as blood, urine, stool, etc. were also carried out. In Group A, 55.33% patients showed marked improvement, whereas moderate improvement was observed in 26.67% patients. Complete cure was found in 06.67% of the patients and mild improvement in the chief complaints was observed in 13.33% patients. All the selected symptoms showed statistically significant results (P<0.01) except the Vidbheda in treated Group A, while in Group B, all symptoms showed statistically insignificant results except the Utklesha and Amlodgara. Total effect of the therapy showed statistically significant effect of the test drug. These results support the hypothesis.


Appetite suppression and weight reduction by a centrally active aminosterol.
The rise in obesity and its complications has generated enormous interest in the regulation of feeding and body weight. We show that a spermine metabolite of cholesterol (MSI-1436) decreases body weight, specifically fat, by suppressing feeding and preventing the reduction in energy expenditure, hormonal changes, and patterns of neuropeptide expression normally associated with weight loss. MSI-1436 enters the brain after peripheral injection and is more potent when injected into the cerebral ventricle (intracerebroventricular [ICV]). Systemic or ICV MSI-1436 administration induced similar patterns of Fos immunoreactivity in the brain, especially the paraventricular hypothalamic nucleus (PVN). This brain region integrates neural signals from hypothalamic and brain stem nuclei and regulates feeding behavior, autonomic function, and neuroendocrine function. Microinjection of MSI-1436 into the PVN potently suppressed feeding and reduced body weight for several days. Unlike caloric restriction, MSI-1436 decreased mRNA levels of agouti-related peptide and neuropeptide Y in the hypothalamus. These findings indicate that MSI-1436 acts in the brain to regulate food intake and energy expenditure, likely through suppression of orexigenic hypothalamic pathways.


Roles of Communication Problems and Communication Strategies on Resident-Related Role Demand and Role Satisfaction.
This study investigated the impact of dementia-related communication difficulties and communication strategies used by staff on resident-related indicators of role demand and role satisfaction. Formal/paid long-term care staff caregivers (N = 109) of residents with dementia completed questionnaires on dementia-related communication difficulties, communication strategies, role demand (ie, residents make unreasonable demands), and role satisfaction (measured by relationship closeness and influence over residents). Three types of communication strategies were included: (a) effective repair strategies, (b) completing actions by oneself, and (c) tuning out or ignoring the resident. Analyses using structural equation modeling revealed that communication problems were positively linked with role demand. Repair strategies were positively linked with relationship closeness and influence over residents. Completing actions by oneself was positively linked to role demand and influence over residents, whereas tuning out was negatively linked with influence over residents. The findings underscore that effective caregiver communication skills are essential in enhancing staff-resident relationships.


Prognostic value of carcinoembryonic antigen and vascular endothelial growth factor tumor tissue content in colorectal cancer.
This study was designed to assess the prognostic significance of the combined measurement of vascular endothelial growth factor (VEGF) and carcinoembryonic antigen (CEA) tissue content with respect to relapse-free and overall survival of patients with colorectal cancer (CRC). Quantitative evaluation of VEGF and CEA content was performed on protein extracts obtained from tissue biopsies from 69 CRC patients and 15 healthy donors. VEGF significantly correlated with CEA content of either tumor tissues (rho = 0.55, p < 0.0001) or corresponding normal mucosa (rho = 0.34, p < 0.005). General regression analyses demonstrated that CEA was an independent predictor of VEGF tissue content either in CRC biopsies (regression coefficient = 0.57, p < 0.0001) or normal mucosa (regression coefficient = 0.25, p < 0.05). Cox proportional hazards survival analysis showed that tumor tissue content of both VEGF and CEA had an independent prognostic value in predicting both relapse-free (hazards ratio = 5.98, p = 0.002) and overall (hazards ratio = 4.73, p = 0.007) survival, irrespective of Dukes' stage. Kaplan-Meier analysis demonstrated that an elevated tumor content of both CEA and VEGF had a negative prognostic value in respect to either relapse-free (log-rank test: 10.4, p = 0.001) or overall survival (log-rank test: 7.33, p = 0.007). Tumor tissue VEGF and CEA content determination might add useful prognostic information in the management of patients with CRC.


Remote Grading of the Anterior Chamber Angle Using Goniophotographs and Optical Coherence Tomography: Implications for Telemedicine or Virtual Clinics.
To evaluate the agreement and accuracy of grading goniophotographs and anterior segment optical coherence tomography (AS-OCT) results for assessment of the anterior chamber angle, and elicit factors driving concordance between perceived grade and ground truth. Three clinicians evaluated the goniophotographs and AS-OCT results of 75 patients. Graders' impressions of the angle grade, trabecular pigmentation, and iris contour were compared with the ground truth gonioscopic examination result when physically performed by a senior optometrist. Percentage agreement and kappa statistics were calculated. Binary logistic regression was used to elicit factors for accurate grading. Exact angle matches and binary (open or closed) evaluations were above guessing rate for all graders. There was a systematic bias toward underestimating the angle structure across all graders, especially at the superior angle, by approximately 1 ordinal unit. Kappa statistics showed fair-moderate agreement for exact (0.387-0.520) and binary (0.347-0.520) angle evaluations. Agreement was unchanged when using a multimodal approach (0.373-0.523). Factors driving concordance were primarily related to the extremes of the anterior chamber angle configuration (shallow or deep structures, and iris contour). However, prediction models did not fully explain the levels of concordance with the ground truth (maximum R 2 amongst models 0.177). Although moderate agreement between graders and ground truth could be obtained under binary evaluations, angle grades were generally underestimated. Factors affecting concordance were primarily the extremes of the ground truth angle and iris contour. We highlight factors affecting accuracy of grading goniophotography and AS-OCT images of the anterior chamber angle.


Atrial natriuretic factor (auriculin): structure and biological effects.
Atrial natriuretic factor (ANF) is a recently discovered peptide present in secretory granules specifically found in atrial muscle cells. Multiple structurally related peptides have been isolated from atrial tissues, all of which are derived from a common 152-amino-acid precursor. ANF induces profound natriuresis and diuresis in experimental animals and also causes relaxation of precontracted vascular smooth muscle. ANF has striking renal hemodynamic actions (most consistently an increased glomerular filtration rate), which probably explain its natriuretic effects. ANF also can inhibit renin secretion in vivo and causes direct inhibition of basal and stimulated aldosterone production. It lowers arterial blood pressure, probably reflecting in part its vasorelaxant actions, and this effect is particularly marked in renin-dependent (and possibly other vasoconstricted) models of hypertension. Although the exact structure and regulation of the presumed circulating form(s) of ANF remain to be clarified, available information suggests that it may be a new, previously unrecognized factor in the regulation of fluid volume and renal and cardiovascular function.


Bone turnover in preterm infants.
Total parenteral nutrition is associated with osteopenia in preterm infants. Insufficient calcium and phosphate are likely causes: aluminum contamination is another possible contributing factor as this adversely affects bone formation and mineralization. The study was designed to evaluate changes in biochemical markers of bone turnover in 22 preterm infants receiving total parenteral nutrition in comparison with 19 term infants. We collected urine and serum samples from 22 preterm infants, mean gestational age 29 wk, within 48 h and again at 3 wk of life. We also collected urine samples from 19 term infants, mean gestational age 39 wk, during the first day of life. Bone resorption was assessed by the measurement of urinary pyridinium cross-links by HPLC and ELISA and the N-telopeptide of type I collagen by ELISA. Bone formation was assessed in premature infants by the measurement of serum osteocalcin. The N-telopeptide of type I collagen was higher in the preterm infants compared with term at baseline (p < 0.01). There was no difference between the pyridinium cross-links in the preterm and term infants. All the biochemical markers of bone turnover increased significantly in the preterm infants during the first 3 wk of life, e.g. N-telopeptide was a 153% change from baseline (p < 0.001). Aluminum in the total parenteral nutrition solutions did not cause a decrease in bone formation at the level administered (3-6 microg, 0.1-0.2 micromol x kg(-1) x d(-1)).


Root and xylem anatomy varies with root length, root order, soil depth and environment in intermediate wheatgrass (Kernza®) and alfalfa.
Deep roots (i.e. >1 m depth) are important for crops to access water when the topsoil is dry. Root anatomy and hydraulic conductance play important roles in the uptake of soil water, particularly water located deep in the soil. We investigated whether root and xylem anatomy vary as a function of root type, order and length, or with soil depth in roots of two deep-rooted perennial crops: intermediate wheatgrass [Thinopyrum intermedium (Kernza®)] and alfalfa (Medicago sativa). We linked the expression of these anatomical traits to the plants' capacity to take up water from deep soil layers. Using laser ablation tomography, we compared the roots of the two crops for cortical area, number and size of metaxylem vessels, and their estimated root axial hydraulic conductance (ERAHCe). The deepest roots investigated were located at soil depths of 2.25 and at 3.5 m in the field and in rhizoboxes, respectively. Anatomical differences were characterized along 1-m-long individual roots, among root types and orders, as well as between environmental conditions. For both crops, a decrease in the number and diameter, or both, of metaxylem vessels along individual root segments and with soil depth in the field resulted in a decrease in ERAHCe. Alfalfa, with a greater number of metaxylem vessels per root throughout the soil profile and, on average, a 4-fold greater ERAHCe, took up more water from the deep soil layers than intermediate wheatgrass. Root anatomical traits were significantly different across root types, classes and growth conditions. Root anatomical traits are important tools for the selection of crops with enhanced exploitation of deep soil water. The development and breeding of perennial crops for improved subsoil exploitation will be aided by greater understanding of root phenotypes linked to deep root growth and activity.


"There are still broken or fragmented systems": Qualitative assessment of needs to strengthen adolescent mental health services in American Samoa.
Mental health challenges appear common among adolescents in American Samoa. There is a community-identified need to better understand the health burden and identify opportunities to strengthen the mental health system to better meet adolescent mental health needs. The goal of this qualitative study was to gather community stakeholders' perceptions of common mental health problems among adolescents in American Samoa, identify existing services for adolescent mental health, and highlight strength-based opportunities to enhance care. Using the Fa'afaletui research framework, semi-structured in-depth interviews (n = 28) were conducted between October 2020 and February 2021 using Zoom. Key informants were adults living in American Samoa, sampled for diversity in profession, age, gender, education, and region of residence. Transcripts were coded in duplicate and analysed using deductive thematic analysis. Participants reported that suicide, depression, anxiety, and post-traumatic stress disorder are the conditions of highest burden. Participants identified an impressive amount of infrastructure and community mobilization, but also limitations in these services in addressing adolescents' health needs. Four themes arose related to the need to: (1) strengthen existing infrastructure; (2) reduce barriers to treatment access; (3) improve sustainability; and (4) encourage collaboration among leaders. Policymakers should prioritize the allocation of resources to strengthen infrastructure, public education, and stakeholder collaboration to improve adolescent mental health in American Samoa.


Incidence and Clinical Presentation of Pheochromocytoma and Sympathetic Paraganglioma: A Population-based Study.
Pheochromocytoma and sympathetic paraganglioma (PPGL) are rare catecholamine-secreting tumors but recent studies suggest increasing incidence. Traditionally, PPGL are described to present with paroxysmal symptoms and hypertension, but existing data on clinical presentation of PPGL come from referral centers. We aimed to describe time trends in clinical presentation and incidence of PPGL in a population-based study. We conducted a nationwide retrospective cohort study of a previously validated cohort of 567 patients diagnosed with PPGL in Denmark 1977-2015. We collected clinical data from medical records of a geographic subcohort of 192 patients. We calculated age-standardized incidence rates (SIRs) and prevalence for the nationwide cohort and descriptive statistics on presentation for the subset with clinical data. SIRs increased from 1.4 (95% CI 0.2-2.5) per million person-years in 1977 to 6.6 (95% CI 4.4-8.7) per million person-years in 2015, corresponding to a 4.8-fold increase. The increase was mainly due to incidentally found tumors that were less than 4 cm and diagnosed in patients older than 50 years with no or limited paroxysmal symptoms of catecholamine excess. On December 31, 2015, prevalence of PPGL was 64.4 (CI 95% 57.7-71.2) per million inhabitants. Of 192 patients with clinical data, 171 (89.1%) had unilateral pheochromocytoma, while unilateral paraganglioma (n = 13, 6.8%) and multifocal PPGL (n = 8, 4.2%) were rare. Incidence of PPGL has increased 4.8-fold from 1977 to 2015 due to a "new" group of older patients presenting with smaller incidentally found PPGL tumors and few or no paroxysmal symptoms.


Tumor angiogenesis in gastric cancer.
Gastric cancer (GC) is still a major health problem, being one of the leading causes of cancer-related death in the world. Although the overall incidence of GC is decreasing in the United States and Western Europe, it is still high in many countries from Asia, South America, and Eastern Europe. The process of angiogenesis or the formation of new blood vessels plays an important role in cancer progression, as it allows oxygen supply, nutrients, and factors to grow tumor cells. In our study, we found that gastric neoplasms have high vascularity, with anarchic distribution, uneven in tumor stroma, sometimes with congestion vessels and microhemorrhages. Most vessels were capillaries, with a discontinuous endothelium, poorly structured basement membrane, without junctions between endothelial cells, hyperpermeable, creating the possibility of local edema in the tumor microenvironment (TME), and also extravasation of the plasma, leukocytes and even red blood cells. Angiogenesis vessels showed a low number of pericytes, which shows that they are young vessels, both morphologically and functionally immature. Tumor cells can synthesize biochemical factors [vascular endothelial growth factor-A (VEGF-A)] that stimulate the formation of new vessels (angiogenesis) to ensure their growth and metastasis. Some connective cells (tumor-associated mast cells, tumor-associated fibroblasts) are also involved in the angiogenesis process, which stimulate the progression of tumor cells and remodel the TME.


The use of chimpanzee-modified faunal assemblages to investigate early hominin carnivory.
Chimpanzees regularly hunt and consume prey smaller than themselves. It seems therefore likely that early hominins also consumed small vertebrate meat before they started using and producing stone tools. Research has focused on cut marks and large ungulates, but there is a small body of work that has investigated the range of bone modifications produced on small prey by chimpanzee mastication that, by analogy, can be used to identify carnivory in pre-stone tool hominins. Here, we review these works along with behavioral observations and other neo-taphonomic research. Despite some equifinality with bone modifications produced by baboons and the fact that prey species used in experiments seldom are similar to the natural prey of chimpanzees, we suggest that traces of chimpanzee mastication are sufficiently distinct from those of other predators that they can be used to investigate mastication of vertebrate prey by early hominins.


A decision model and cost-effectiveness analysis of colorectal cancer screening and surveillance guidelines for average-risk adults.
Guidelines for colorectal cancer screening and surveillance in people at average risk and at increased risk have recently been published by the American Gastroenterological Association. The guidelines for the population at average risk were evaluated using cost-effectiveness analyses. Since colorectal cancers primarily arise from precancerous adenomas, a state transition model of disease progression from adenomatous polyps was developed. Rather than assuming that polyps turn to cancer after a fixed interval (dwell time), such transitions were modeled to occur as an exponential function of the age of the polyps. Screening strategies included periodic fecal occult blood test, flexible sigmoidoscopy, double-contrast barium enema, and colonoscopy. Screening costs in 1994 dollars were estimated using Medicare and private claims data, and clinical parameters were based upon published studies. Cost per life-year saved was $12,636 for flexible sigmoidoscopy every 5 years and $14,394 for annual fecal occult blood testing. The assumption made for polyp dwell time critically affected the attractiveness of alternative screening strategies. Sigmoidoscopy every 5 years and annual fecal blood testing were the two most cost-effective strategies, but with low compliance, occult blood testing was less cost-effective. Lowering colonoscopy costs greatly improved the cost-effectiveness of colonoscopy every 10 years.


Life in the slow drain: Landscape structure affects farm ditch water quality.
Agrichemical contamination is a major threat to aquatic ecosystems in farmland. There is a need to better understand the influence of the surrounding landscape on farm wetlands to recommend land management options that minimize water quality impacts from agricultural practices. We tested hypothesized relationships between landscape structure and multiple water quality measures in farm drainage ditches in a multi-landscape study in Eastern Ontario, Canada. We measured physicochemical water quality (levels of atrazine, glyphosate, neonicotinoid insecticides, inorganic nitrogen, and dissolved oxygen), and biological water quality indicators (aquatic macroinvertebrate richness, leaf litter decomposition, and Ceriodaphnia dubia population responses) in 27 farm ditches, and measured the amounts of forest cover and high-intensity crop cover (landscape composition), and field edge cover (landscape configuration) in 1-km radius landscapes surrounding each ditch sampling site. We used confirmatory path analysis to simultaneously model the direct and indirect relationships between the landscape predictors and water quality variables. Landscape composition measures were the strongest predictors of water quality: pesticides decreased as surrounding forest cover increased, and nitrogen increased with increasing amounts of high-intensity crop cover. Crop cover was also indirectly negatively related to macroinvertebrate richness via its effects on nitrogen and dissolved oxygen. We found no effects of landscape configuration on agrichemical levels, but there was some support for a positive relationship between macroinvertebrate richness and field edge cover. Our results indicate that aquatic macroinvertebrate richness is strongly impacted by fertilizer use in our region, and that macroinvertebrate richness is a more sensitive biotic indicator of farmland water quality than leaf litter decomposition or C. dubia responses. We conclude that, in our region, landscape management to improve farmland water quality should focus primarily on landscape composition. Such management should aim to increase amounts of non-crop cover such as forest, and reduce amounts of crop cover with high agrichemical inputs.


Cutaneous Borreliosis With a T-Cell-Rich Infiltrate and Simultaneous Involvement by B-Cell Chronic Lymphocytic Leukemia With t(14;18)(q32;q21).
Pseudolymphomatous infiltrates in Borrelia infection of the skin most commonly manifest with dense B-cell infiltrates and plasma cells. Cutaneous infiltrates of B-cell chronic lymphocytic leukemia (B-CLL) may accumulate at sites of infection, including Borrelia infection. We report an unusual constellation in a patient with synchronously diagnosed B-CLL and Borrelia infection of skin presenting with a dense dermal T-cell-rich infiltrate masking specific leukemic infiltrates of neoplastic B cells in the context of B-CLL harboring t(14;18)(q32;q21). Specific cutaneous involvement by B-CLL was confirmed by the detection of t(14;18)(q32;q21) (BCL2-IGH) using FISH in neoplastic B cells within the skin infiltrates. Borrelia burgdorferi (sensu lato) DNA detected by nested polymerase chain reaction in the skin biopsy and serological findings proved Borrelia infection. Complete resolution of the cutaneous infiltrates was observed after antibiotic treatment. This case demonstrates that Borrelia infection of the skin may present with dense T-cell-rich infiltrates mimicking cutaneous T-cell lymphoma and masking the synchronous presence of neoplastic B cells in the context of B-CLL.


Prolonged survival without neurological improvement in patients with AIDS-related progressive multifocal leukoencephalopathy on potent combined antiretroviral therapy.
To evaluate the benefit of combined antiretroviral therapy including protease inhibitors (CART) on survival time and neurological progression in patients with AIDS-related progressive multifocal leukoencephalopathy (PML), 81 consecutive PML cases, collected between January 1990 and June 1998, were reviewed. Fifteen patients were neuropathologically proven. JC virus detection in CSF was positive in 59 patients. At PML diagnosis, median CD4 cell count was low (median, 35 cells/microL) and plasma HIV load, determined in 41 patients, was high (median, 4.8 log10 copies/ml). Following PML diagnosis, there was a significant difference (P<10(-4)) in survival between patients who were untreated or treated with nucleoside analogs (n=50, median: 80 days) and patients who were started early on CART (n=23, median: 246 days). A third group of eight patients who received CART late during the course of PML was considered separately. At the study endpoint, 18 of all the CART-treated patients (n=31) were still alive. Plasma HIV load was undetectable in 67% of them. The median increase in CD4 cell count was 112 cells/microL from CART onset. In contrast, no significant improvement in neurological status was observed. Our results demonstrate a benefit of CART on survival of AIDS-related PML patients and suggest the need for an early, specific anti-JC virus treatment to limit the neurological deterioration.


The need for integrated risk management in capitated provider groups.
Financial risk is not just ensuring that there is enough capitation revenue to cover claims for services. The changing world of health care is devolving risk and exposures to provider groups that were previously covered by insurance companies, HMOs, and other payers. Capitated providers must first understand the risk they face and then manage it. There is more to be gained in terms of independence than to be feared in additional administrative work.


Maternal physiologic parameters in relationship to systemic inflammatory response syndrome criteria: a systematic review and meta-analysis.
To establish the normal maternal range in healthy pregnant women for each component of the systemic inflammatory response syndrome (SIRS) criteria and compare these ranges with existing SIRS criteria. PubMed, Embase, and ClinicalTrials.gov databases were searched to identify studies of healthy parturients from the first trimester through 12 weeks postpartum that reported maternal temperature, respiratory rate, PaCO2, heart rate, white blood cell count data, or a combination of these. Data were extracted from studies providing maternal values for components of SIRS criteria. The mean, standard deviation, and two standard deviations from the mean for all criteria parameters published in the literature were reported. Eighty-seven studies met inclusion criteria and included 8,834 patients and 15,237 data points: temperature (10 studies and 2,367 patients), respiratory rate (nine studies and 312 patients), PaCO2 (12 studies and 441 patients), heart rate (39 studies and 1,374 patients), and white blood cell count (23 studies and 4,553 patients). Overlap with SIRS criteria occurred in healthy pregnant women during the second trimester, third trimester, and labor for each of the SIRS criteria except temperature. Every mean value for PaCO2 during pregnancy (and up to 48 hours postpartum) was below 32 mm Hg. Two standard deviations above the mean for temperature, respiratory rate, and heart rate were 38.1°C, 25 breaths per minute, and 107 beats per minute, respectively. Current SIRS criteria often overlap with normal physiologic parameters during pregnancy and the immediate postpartum period; thus, alternative criteria must be developed to diagnose maternal sepsis.


Preparation and enzymatic application of flower-like hybrid microcapsules through a biomimetic mineralization approach.
Fast and quantitative adsorption of molecules may confer hybrid microcapsules high performance in specific applications, such as biocatalysis, biosensing, drug delivery, and so on. Engineering the chemical/topological structures of the capsule wall renders great potential to achieve this goal. In the present study, flower-like microcapsules are prepared through a biomimetic mineralization approach. Briefly, protein-hybrid microflowers are firstly synthesized through enzyme-induced precipitation of Cu3(PO4)2, which are subsequently utilized as templates for alternative deposition of the protamine layer and the silica layer through biomimetic silicification assisted by layer-by-layer (LbL) assembly. After treatment with EDTA, the flower-like protamine/silica hybrid (FPSH) microcapsules are obtained. Additionally, the hybrid capsule wall also endows the FPSH microcapsules with desirable pH, temperature and storage stability. Hopefully, our approach may provide a promising alternative to increase the efficiency of catalysis, drug delivery, etc.


Measuring socioeconomic inequalities in the use of dental care services among older adults in Brazil.
The primary objectives are to assess socioeconomic inequality in the use of dental care among older Brazilian adults and to analyse the extent to which certain determinants contribute to that inequality. A cross-sectional study using data from the National Oral Health Survey conducted in 2010. All individuals answered a structured questionnaire containing questions on their use of dental care and socioeconomic conditions and underwent a clinical oral examination by a dentist. Concentration indices were decomposed to determine the contribution of socioeconomic factors to inequalities. Being in the fifth wealth, quintile was associated with higher odds of having recently visited a dentist (reference: 1st quintile, odds-ratio (OR) 2.26, 95% confidence interval (CI) 1.51-3.38). In addition, being in the top two quintiles of wealth was negatively associated with the use of public dental services. Having eight or more years of schooling was associated with higher odds of both having a recent dental visit and receiving preventive care (relative to having 0-3 years of education), and negatively associated with using public dental services. Results indicate pro-rich inequalities in recent dental visits and preventive dental care. Further, there was a pro-poor inequality in the use of public dental care services. The recent use of dental care and the use of preventive care are disproportionately concentrated among wealthier older adults, whereas the use of public services is more common among poorer individuals. Wealth inequalities in dental care use were mainly explained by socioeconomic factors, such as wealth and education, rather than oral health factors, such as needing treatment or a dental prosthesis.


Physiological and immunohistochemical characterization of cisplatin-induced neuropathy in mice.
We investigated the neuropathic effects of cisplatin in two groups of mice treated with 5 or 10 mg/kg/week of cisplatin for 7 or 8 weeks. Peripheral nerve functions were evaluated by sweat imprints, and electrophysiological, rotarod, and nociceptive tests. Protein gene product 9.5 (PGP), calcitonin gene-related peptide (CGRP), and vasoactive intestinal peptide (VIP) were immunohistochemically localized in footpads. Tibial nerves were analyzed morphometrically. Functional deficits developed progressively with higher cumulative doses, more markedly in mice treated with high than in those with low doses. From cumulative doses of 10 mg/kg, significant declines in sensory nerve conduction velocity and sudomotor responses were found, whereas motor and nociceptive functions were involved later. There were no morphometrical changes in tibial nerves. A marked decrease of CGRP- and VIP-immunoreactive nerves occurred in samples from treated mice, whereas PGP-labeled profiles decreased mildly at late stages. Impairment of the content of neuropeptides with neurosecretor role was detectable earlier than functional abnormalities. Immunohistochemical analysis of skin biopsies offers a useful diagnostic tool for peripheral neuropathies.


Differential gene expression in multiple neurological, inflammatory and connective tissue pathways in a spontaneous model of human small vessel stroke.
Cerebral small vessel disease (SVD) causes a fifth of all strokes plus diffuse brain damage leading to cognitive decline, physical disabilities and dementia. The aetiology and pathogenesis of SVD are unknown, but largely attributed to hypertension or microatheroma. We used the spontaneously hypertensive stroke-prone rat (SHRSP), the closest spontaneous experimental model of human SVD, and age-matched control rats kept under identical, non-salt-loaded conditions, to perform a blinded analysis of mRNA microarray, qRT-PCR and pathway analysis in two brain regions (frontal and mid-coronal) commonly affected by SVD in the SHRSP at age five, 16 and 21 weeks. We found gene expression abnormalities, with fold changes ranging from 2.5 to 59 for the 10 most differentially expressed genes, related to endothelial tight junctions (reduced), nitric oxide bioavailability (reduced), myelination (impaired), glial and microglial activity (increased), matrix proteins (impaired), vascular reactivity (impaired) and albumin (reduced), consistent with protein expression defects in the same rats. All were present at age 5 weeks thus predating blood pressure elevation. 'Neurological' and 'inflammatory' pathways were more affected than 'vascular' functional pathways. This set of defects, although individually modest, when acting in combination could explain the SHRSP's susceptibility to microvascular and brain injury, compared with control rats. Similar combined, individually modest, but multiple neurovascular unit defects, could explain susceptibility to spontaneous human SVD.


Mathematical modeling of the brain: principles and challenges.
The use of mathematics in the study of phenomena and systems of interest to medicine has become quite popular in recent years, but not much progress has been made as a result of these efforts. The aim of this article is to identify the reasons for this failure and to suggest procedures for more successful outcomes. We review and assess a variety of mathematical modeling procedures, from microscopic (at the level of molecular behavior) to macroscopic standpoints, from lumped-parameters to distributed-parameters approaches. Using examples that are as simple as possible, we elucidate the difference between the predictive and the explanatory powers of mathematical models, as well as the uses (and abuses) of analogy in their construction. Mathematical medicine is a truly interdisciplinary area that brings together medical researchers, engineers, and applied mathematicians whose vast differences in expertise and background make collaboration difficult. The lack of a common language and a common way of understanding what a mathematical model is, and what it can do, is identified as the main source of the slow progress to date, and constructive suggestions are made to improve the situation.


Impact of COVID-19 on occupational injuries and illnesses among nursing care facility workers: Analysis of California workers' compensation data, 2019-2021.
The coronavirus disease 2019 (COVID-19) pandemic greatly affected healthcare workers, both physically and psychologically, by increasing their workload and stress. This may also have increased their risk of occupational injuries. This study analyzed workers' compensation (WC) claims among California nursing care facility workers during 2019-2021, to assess the impact of the pandemic on occupational injury risk. This study used data from the California Workers' Compensation Information System. WC claims in prepandemic and pandemic periods were described and compared between nursing care facilities and other settings. Nursing facility cases were described by demographic, job, and injury characteristics. In 2019-2021, we identified 41,134 claims as nursing facility cases in California. Annual claims increased by 64.6% from 2019 to 2020 and then decreased in 2021, returning to levels similar to pre-COVID. In contrast, non-healthcare settings had decreases in claims during the pandemic. COVID-related claims accounted for 50.5% of claims in nursing facilities in 2020-2021; this proportion was much higher compared with other healthcare (35.7%) or nonhealthcare settings (14.3%). Non-COVID claims decreased during the pandemic but mental disorder or stress claims increased in nursing facilities, particularly in 2020 (+42.5%). Our findings show that nursing care facilities were more severely affected by the pandemic than other settings. We did not find evidence of an increased risk of occupational injuries during the pandemic, except for increased claims for stress or mental disorders. Our findings indicate a clear need to address psychological stress and mental health among nursing facility workers during a pandemic.


The Significance of Touch in Pediatric Physiotherapy.
Change in theoretical framework over the last decades and recent research in pediatric physiotherapy, has created a debate surrounding therapeutic touch. What is the role of or is there a need for handling and hands-on facilitated guidance (facilitation)? Does it limit and/or interfere with children's learning and development? It is frequently argued that therapeutic touch represents a passive and/or static approach that restricts disabled children's participation during interaction and activity in clinical encounters leading to decreased home, school and community participation. Touch may even appear as coercive and controlling. In this context, therapeutic touch is largely associated with physical hands-on activities. However, therapeutic touch can also be understood as an intersubjective phenomenon that arises from a deep connection between movement, perception, and action. We believe the significance of therapeutic touch and its impact on physiotherapy for children has not been considered from this broader, holistic perspective. In this theoretical paper, we will apply enactive concepts of embodiment, sensory-motor agency, coordination, and emergence to explore the concept and importance of touch in physiotherapists' clinical face-to face encounters with children. We will frame the discussion within the context of the typical sensorimotor development of children from the fetal stage to birth on and into adulthood. Moreover, we will rely on biological, physiological, and phenomenological insights to provide an extended understanding of the importance of touch and the significance of touch in clinical practice.


Mitochondria, autophagy and age-associated neurodegenerative diseases: New insights into a complex interplay.
Mitochondria represent the major bioenergetic hub coordinating cellular and organismal homeostasis. The underlying causes of many pathologies tormenting humans converge on impaired mitochondrial maintenance. Mitochondria-specific autophagy (mitophagy), a cellular catabolic process targeting mitochondria, holds a prominent role in mitochondrial quality control. In addition to core autophagic machinery components, mitophagy exploits a variety of molecules that identify damaged or superfluous mitochondria and mediate their elimination. Signaling pathways integrating environmental and genetic stimuli interact with key mitophagy effectors to activate cellular stress response mechanisms, ultimately modulating health and lifespan. Here, we review the signaling cascades and molecular mechanisms that govern the process of mitophagy and discuss their involvement in ageing and neurodegeneration. This article is part of a Special Issue entitled: Mitochondrial Dysfunction in Aging.


Delayed roentgenographic signs associated with acute pneumonia in children.
Three cases are reported in order to emphasize occult presentations of pneumonia in children. In each hospitalized child, the initial chest roentgenogram was negative, only to become positive up to five years after presentation. Possible explanations for delayed roentgenographic signs of pneumonia are discussed. Less common signs and symptoms of pneumonia in children of various ages are outlined in order to increase the awareness of primary care physicians to atypical presentation of pneumonia in childhood.


Synthesis of sialoglycoconjugates during dimethylsulfoxide-induced erythrodifferentiation of friend leukemia cells.
We have previously shown that erythroid differentiation of Friend murine leukemia cells by dimethylsulfoxide results in a decrease in sialic acid content and net negative surface charge. The mechanism responsible for the decrease in sialic acid content was examined by measuring the synthesis of sialic acid from N-acetylmannosamine and its catabolic removal from sialoconjugates during the maturation process. A decrease in the incorporation of N-[3H]acetylmannosamine into sialoglycoconjugates occurred as early as 12 h after exposure to dimethylsulfoxide. Radioactivity incorporated into sialoglycoconjugates was relatively stable in untreated and dimethyl-sulfoxide-treated cells, implying that catabolic removal of sialic acid residues was not a factor in the decreased surface sialic acid content of differentiated erythroleukemia cells. In addition, no difference existed between control and treated cells in sialyltransferase activity. Significant decreases occurred, however, in the incorporation of radioactivity from N-[3H]acetylmannosamine into N-acetylneuraminic acid, CMP-N-acetylneuraminic acid and a material tentatively identified as N-acetylmannosamine-6-phosphate, 48 h after the addition of dimethylsulfoxide. The decrease in sialic acid biosynthesis in differentiated erythroleukemia cells was reflected by an 83% decrease in the amount of radioactively-labeled sialic acid released by neuraminidase treatment of cells exposed to dimethylsulfoxide. These findings are consistent with a cellular aging phenomenon triggered by the polar solvent-induced differentiation of the leukemic cells into more mature forms.


In vivo proliferation of differentiated pancreatic islet beta cells in transgenic mice expressing mutated cyclin-dependent kinase 4.
It has previously been hypothesised that highly differentiated endocrine cells do not proliferate or regenerate. However, recent studies have revealed that cyclin-dependent kinase 4 (CDK4) is necessary for the proliferation of pancreatic islet beta cells. The aim of this study was to determine whether activation of CDK4 can potentially be used as a radical treatment for diabetes without malignant transformation. We generated transgenic mice expressing mutant CDK4 under the control of the insulin promoter to examine the effect of activated CDK4 overexpression in the postnatal development of pancreatic islets. In the transgenic mice, total CDK4 protein expression was increased by up to 5-fold, with a concomitant increase in CDK4 activity indicated by the detection of phosphorylated Rb protein in pancreatic islets. Histopathologically, many cells tested positive for proliferating cell nuclear antigen, and pancreatic islets displayed hyperplasia due to the extreme proliferation of beta cells containing a large number of insulin granules. Pancreatic islet alpha, delta and PP cells did not increase. Over an 18-month observation period, the transgenic mice did not develop insulinoma. Levels of expression of GLUT1 and c-myc were comparable to those in the littermates of the transgenic mice. GLUT2 expression was identified in the pancreatic islets of transgenic mice. No significant differences in telomerase activities were detected between transgenic mice and their littermates. Transgenic mice were superior to their littermates in terms of glucose tolerance and insulin secretion in response to the intraperitoneal injection of glucose, and hypoglycaemia was not observed. Activated CDK4 stimulates postnatal pancreatic beta cell proliferation, during which the highly differentiated phenotypes of pancreatic islet beta cells are preserved without malignant transformation.


A Pharmacological Chaperone Molecule Induces Cancer Cell Death by Restoring Tertiary DNA Structures in Mutant hTERT Promoters.
Activation of human telomerase reverse transcriptase (hTERT) is necessary for limitless replication in tumorigenesis. Whereas hTERT is transcriptionally silenced in normal cells, most tumor cells reactivate hTERT expression by alleviating transcriptional repression through diverse genetic and epigenetic mechanisms. Transcription-activating hTERT promoter mutations have been found to occur at high frequencies in multiple cancer types. These mutations have been shown to form new transcription factor binding sites that drive hTERT expression, but this model cannot fully account for differences in wild-type (WT) and mutant promoter activation and has not yet enabled a selective therapeutic strategy. Here, we demonstrate a novel mechanism by which promoter mutations activate hTERT transcription, which also sheds light on a unique therapeutic opportunity. Promoter mutations occur in a core promoter region that forms tertiary structures consisting of a pair of G-quadruplexes involved in transcriptional silencing. We show that promoter mutations exert a detrimental effect on the folding of one of these G-quadruplexes, resulting in a nonfunctional silencer element that alleviates transcriptional repression. We have also identified a small drug-like pharmacological chaperone (pharmacoperone) molecule, GTC365, that acts at an early step in the G-quadruplex folding pathway to redirect mutant promoter G-quadruplex misfolding, partially reinstate the correct folding pathway, and reduce hTERT activity through transcriptional repression. This transcription-mediated repression produces cancer cell death through multiple routes including both induction of apoptosis through inhibition of hTERT's role in regulating apoptosis-related proteins and induction of senescence by decreasing telomerase activity and telomere length. We demonstrate the selective therapeutic potential of this strategy in melanoma cells that overexpress hTERT.


[The correlation between procalcitonin, C-reactive protein and severity scores in patients with sepsis and their value in assessment of prognosis].
To investigate the correlation between procalcitonin (PCT), C-reactive protein (CRP) and acute physiology and chronic health evaluation II (APACHE II) score and sequential organ failure assessment (SOFA) score, and to investigate the value in assessment of PCT and CRP in prognosis in patients with sepsis. Clinical data of patients admitted to intensive care unit (ICU) of Changzheng Hospital Affiliated to the Second Military Medical University from January 2011 to June 2014 were retrospectively analyzed. 201 sepsis patients who received PCT and CRP tests, and evaluation of APACHE II score and SOFA score were enrolled. The values of PCT, CRP, APACHE II score and SOFA score between survivals (n = 136) and non-survivals (n = 65) were compared. The values of PCT and CRP among groups with different APACHE II scores and SOFA scores were compared. The relationships between PCT, CRP and APACHE II score and SOFA score were analyzed by Spearman correlation analysis. Receiver operating characteristic (ROC) curve was plotted to assess the prognostic value of PCT and CRP for prognosis of patients with sepsis. Compared with survival group, the values of PCT [μg/L: 11.03 (19.17) vs. 1.39 (2.61), Z = -4.572, P < 0.001], APACHE II score (19.16±5.32 vs. 10.01±3.88, t = -13.807, P < 0.001) and SOFA score (9.66±4.28 vs. 4.27±3.19, t = -9.993, P < 0.001) in non-survival group were significantly increased, but the value of CRP was not significantly different between non-survival group and survival group [mg/L: 75.22 (110.94) vs. 56.93 (100.75), Z = -0.731, P = 0.665]. The values of PCT were significantly correlated with APACHE II score and SOFA score (r1 = 0.373, r2 = 0.392, both P < 0.001), but the values of CRP were not significantly correlated with APACHE II score and SOFA score (r1 = -0.073, P1 = 0.411; r2 = -0.106, P2 = 0.282). The values of PCT rose significantly as the APACHE II score and SOFA score became higher, but the value of CRP was not significantly increased. When APACHE II score was 0-10, 11-20, and > 20, the value of PCT was 1.45 (2.62), 1.96 (9.04), and 7.41 (28.9) μg/L, respectively, and the value of CRP was 57.50 (83.40), 59.00 (119.70), and 77.60 (120.00) mg/L, respectively. When SOFA score was 0-5, 6-10, and > 10, the value of PCT was respectively 1.43 (3.09), 3.41 (9.75), and 5.43 (29.60) μg/L, and the value of CRP was 49.30 (86.20), 76.00 (108.70), and 75.60 (118.10) mg/L, respectively. There was significant difference in PCT between any two groups with different APACHE II and SOFA scores (P < 0.05 or P < 0.01), but no significant differences in CRP were found. The area under the ROC curve (AUC) of PCT for prognosis was significantly greater than that of CRP [0.872 (95% confidence interval 0.811-0.943) vs. 0.512 (95% confidence interval 0.427-0.612), P < 0.001]. When the cut-off value of PCT was 3.36 μg/L, the sensitivity was 66.8%, and the specificity was 45.4%. When the cut-off value of CRP was 44.50 mg/L, the sensitivity was 82.2%, and the specificity was 80.3%. Compared with CRP, PCT was more significantly correlated with APACHE II score and SOFA score. PCT can be a better indicator for evaluation of degree of severity, and also prognosis in sepsis patients.


Is a nicotinic influence involved in denervation-induced depolarization of muscle?
The neurotrophic role of acetylcholine (ACh) in the denervation-dependent decline of muscle resting membrane potential (RMP) was evaluated. Freshly dissected rat hemidiaphragms with short or long (2 cm) nerve stumps attached ("-N" and "+N" preparations, respectively) were incubated in organ culture in the presence or absence of the nicotinic blockers, alpha-bungarotoxin (alpha-BTX) or d-tubocurarine (curare). Subsequently, RMPs and miniature endplate potentials (MEPPs) of the junctional region were measured. Spontaneous MEPPs disappeared with a half-life of 12 and 20 hr in -N and +N preparations, respectively. A 10- to 15-mV depolarization of RMP was observed between 15 and 20 hr in -N muscles and between 24 and 28 hr in +N muscles. This time course of disappearance of spontaneous potentials and of membrane depolarization agrees well with that observed in vivo. Although nicotinic transmission was blocked from the initiation of the incubation period in alpha-BTX- or curare-treated muscles, no acceleration of RMP decline in -N muscles in vitro was observed. Moreover, in +N preparations the effect of the nerve stump in delaying RMP depolarization persisted despite the continuous presence of alpha-BTX or curare. Since excess ACh triggers a lysosomal proteolytic response at the nerve-muscle junction and since this may occur early in denervation, the possible role of a nicotinic-induced proteolytic mechanism was tested in vitro with the potent protease inhibitor leupeptin. This inhibitor did not delay or prevent the denervation-dependent alterations.(ABSTRACT TRUNCATED AT 250 WORDS)


[Role of spot gene product in the degradation of pppGpp in bacteria].
In Baccillus subtilis cells, in contrast to Escherichia coli cells, chelating agent 1,10-phenanthroline induces the expansion of guanosine-5'-diphosphate,3'-diphosphate( (ppGpp) pool, as well as the pool of its precursor guanosine-5'-triphosphate,3'-diphosphate (pppGpp). Under these conditions the degradation rate of both nucleotides decreases greatly, which is the main cause of their accumulation in the cells. In E. coli phenanthroline inactivates the product of spoT gene, which is responsible for ppGpp degradation, as a result of combining Mn2+ ions necessary for the activity of this enzyme. The addition of Mn2+ ions to B. subtilis cells, treated with phenanthroline, leads to the decline in (p)ppGpp pools. Antibiotic tetracycline, which has the chelating properties at the concentration of 1 mg/ml, also inactivates spoT gene product in E. coli and slows down the decay of ppGpp, but not of pppGpp. The addition of high concentrations of tetracycline to B. subtilis cells leads to severe inhibition of the degradation of both nucleotides. Therefore in B. subtilis spoT gene product is involved in the degradation of pppGpp, as well as ppGpp. In E. coli cells with defective gpp gene product, taking part in the conversion of pppGpp to ppGpp, phenanthroline and tetracycline also inhibit the breakdown of both nucleotides. The similarity of B. subtilis and E. coli gpp cells in respect of spoT gene product functions and of enhanced pppGpp fraction in the total amount of guanosine polyphosphates during aminoacyl-tRNA limitation makes it plausible that in B. subtilis cells the product of gpp gene is missing or has low activity.


Self-discrepancy theory as a transdiagnostic framework: A meta-analysis of self-discrepancy and psychopathology.
Self-discrepancy theory (SDT) is a model of the relations between the self and affect which has been applied to the study of different types of psychopathology including depression, anxiety, and eating disorders. Although the theory itself is compatible with a transdiagnostic perspective on psychopathology, to date no systematic review of the literature has examined that possibility. We conducted a meta-analysis that synthesized the literature on self-discrepancy and psychopathology across a heterogeneous range of 70 studies. Results showed a small-to-medium association between self-discrepancy and psychopathology that was highly robust and similar in magnitude across domains. Furthermore, self-discrepancy was related to higher levels of a range of negative emotions and lower levels of a range of positive emotions. Meta-regression models showed that the effects were greater for actual:ideal discrepancy compared with actual:ought discrepancy for both depression and anxiety, which was contrary to the tenets of SDT which suggests specific associations between actual:ideal discrepancy and depression and actual:ought discrepancy and anxiety. Measurement type (i.e., idiographic vs. nomothetic) was a significant predictor of the effects for depression and anxiety, such that nomothetic measures evidenced greater associations compared with idiographic measures. Our findings could suggest that self-discrepancy represents a contributory factor related to a number of psychiatric disorders. However, the tenet of SDT suggesting unique associations between actual:ideal and actual:ought discrepancy and anxiety and depression respectively was not supported. Implications are discussed for future research on self-discrepancy and psychopathology including the study of mechanistic frameworks. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Impact of sex differences in co-morbidities and medication adherence on outcome in 25 776 heart failure patients.
Health insurance claims (HIC) databases in the Netherlands capture unselected patient populations, which makes them suitable for epidemiological research on sex differences. Based on a HIC database, we aimed to reveal sex differences in heart failure (HF) outcomes, with particular focus on co-morbidities and medication. The Achmea HIC database included 14 517 men and 11 259 (45%) women with a diagnosis treatment code for chronic HF by January 2015. We related their sex, co-morbidities, and medication adherence (medication possession rate >0.8) with the primary endpoint (PE) of all-cause mortality or HF admission during a median follow-up of 3.3 years, using Cox regression. Median age of men and women was 72 and 76 years, respectively. Prevalence of co-morbidities and use of disease-modifying drugs was higher in men; however, medication adherence was similar. At the end of follow-up, 35.1% men and 31.8% women had reached the PE. The adjusted hazard ratio for men was 1.25 (95% confidence interval: 1.19-1.30). A broad range of co-morbidities was associated with the PE. Overall, these associations were stronger in women than in men, particularly for renal insufficiency, chronic obstructive pulmonary disease/asthma, and diabetes. Non-adherence to disease-modifying drugs was related with a higher incidence of the PE, with similar effects between sexes. In a representative sample of the Dutch population, as captured in a HIC database, men with chronic HF had a 25% higher incidence of death or HF admission than women. The impact of co-morbidities on the outcome was sex dependent, while medication adherence was not.


Identification of Streptococcus pneumoniae by the Phadebact coagglutination test.
The Phadebact Pneumococcus Test is a coagglutination slide test for the serological identification of Streptococcus pneumoniae. Of 200 alpha-hemolytic streptococcal isolates, coagglutination test results agreed with those of optochin susceptibility and bile solubility in 189 cases, 105 of which were identified as S. pneumoniae by all three methods. The Phadebact test was 100% (113 of 113) sensitive and 98% (85 of 87) specific and was more sensitive than counterimmunoelectrophoresis in detecting the presence of pneumococcal antigen in cerebrospinal fluid. In fluids seeded with known amounts of pneumococcal antigen, it consistently detected lower levels than did counterimmunoelectrophoresis. The test provides a rapid and simple method for the definitive identification of S. pneumoniae.


Bidirectional microglia-neuron communication in the healthy brain.
Unlike other resident neural cells that are of neuroectodermal origin, microglia are resident neural cells of mesodermal origin. Traditionally recognized for their immune functions during disease, new roles are being attributed to these cells in the development and maintenance of the central nervous system (CNS) including specific communication with neurons. In this review, we highlight some of the recent findings on the bidirectional interaction between neurons and microglia. We discuss these interactions along two lines. First, we review data that suggest that microglial activity is modulated by neuronal signals, focusing on evidence that (i) neurons are capable of regulating microglial activation state and influence basal microglial activities; (ii) classic neurotransmitters affect microglial behavior; (iii) chemotactic signals attract microglia during acute neuronal injury. Next, we discuss some of the recent data on how microglia signal to neurons. Signaling mechanisms include (i) direct physical contact of microglial processes with neuronal elements; (ii) microglial regulation of neuronal synapse and circuit by fractalkine, complement, and DAP12 signaling. In addition, we discuss the use of microglial depletion strategies in studying the role of microglia in neuronal development and synaptic physiology. Deciphering the mechanisms of bidirectional microglial-neuronal communication provides novel insights in understanding microglial function in both the healthy and diseased brain.


Computer programs for handling nucleic acid sequences.
Programs have been developed that will accurately display restriction enzyme maps, open translational reading frames and the results of specific searches on a high resolution graphics terminal (Retrographics VT640). Many of our earlier programs have been upgraded to allow online access to GENBANK, the NIH data bank of DNA sequences and also to deal with sequences of any length. Progress has also been made in the automation of the DNA assembly programs.


Breakdown of random matrix universality in Markov models.
Biological systems need to react to stimuli over a broad spectrum of timescales. If and how this ability can emerge without external fine-tuning is a puzzle. We consider here this problem in discrete Markovian systems, where we can leverage results from random matrix theory. Indeed, generic large transition matrices are governed by universal results, which predict the absence of long timescales unless fine-tuned. We consider an ensemble of transition matrices and motivate a temperature-like variable that controls the dynamic range of matrix elements, which we show plays a crucial role in the applicability of the large matrix limit: as the dynamic range increases, a phase transition occurs whereby the random matrix theory result is avoided, and long relaxation times ensue, in the entire "ordered" phase. We furthermore show that this phase transition is accompanied by a drop in the entropy rate and a peak in complexity, as measured by predictive information [Bialek, Nemenman, and Tishby Neural Comput. 13, 2409 (2001)NEUCEB0899-766710.1162/089976601753195969]. Extending the Markov model to a Hidden Markov model (HMM), we show that observable sequences inherit properties of the hidden sequences, allowing HMMs to be understood in terms of more accessible Markov models. We then apply our findings to fMRI data from 820 human subjects scanned at wakeful rest. We show that the data can be quantitatively understood in terms of the random model, and that brain activity lies close to the phase transition when engaged in unconstrained, task-free cognition-supporting the brain criticality hypothesis in this context.


Interactions between Mediterranean Diet Supplemented with Dairy Foods and the Gut Microbiota Influence Cardiovascular Health in an Australian Population.
The impact of a Mediterranean diet on the intestinal microbiome has been linked to its health benefits. We aim to evaluate the effects of a Mediterranean diet supplemented with dairy foods on the gut microbiome in Australians at risk of cardiovascular disease. In a randomised controlled cross-over study, 34 adults with a systolic blood pressure ≥120 mmHg and with risk factors for cardiovascular disease were randomly allocated to a Mediterranean diet with 3-4 daily serves of dairy foods (Australian recommended daily intake (RDI) of 1000-1300 mg per day (MedDairy)) or a low-fat (LFD) control diet. Between each 8-week diet, participants underwent an 8-week washout period. Microbiota characteristics of stool samples collected at the start and end of each diet period were determined by 16S rRNA amplicon sequencing. MedDairy-associated effects on bacterial relative abundance were correlated with clinical, anthropometric, and cognitive outcomes. No change in the overall faecal microbial structure or composition was observed with either diet (p > 0.05). The MedDairy diet was associated with changes in the relative abundance of several bacterial taxa, including an increase in Butyricicoccus and a decrease in Colinsella and Veillonella (p < 0.05). Increases in Butyricicoccus relative abundance over 8 weeks were inversely correlated with lower systolic blood pressure (r = -0.38, p = 0.026) and positively correlated with changes in fasting glucose levels (r = 0.39, p = 0.019), specifically for the MedDairy group. No significant associations were observed between the altered taxa and anthropometric or cognitive measures (p > 0.05). Compared to a low-fat control diet, the MedDairy diet resulted in changes in the abundance of specific gut bacteria, which were associated with clinical outcomes in adults at risk of CVD.


In vivo dosimetry and shielding disk alignment verification by EBT3 GAFCHROMIC film in breast IOERT treatment.
Intraoperative electron radiation therapy (IOERT) cannot usually benefit, as conventional external radiotherapy, from software systems of treatment planning based on computed tomography and from common dose verify procedures. For this reason, in vivo film dosimetry (IVFD) proves to be an effective methodology to evaluate the actual radiation dose delivered to the target. A practical method for IVFD during breast IOERT was carried out to improve information on the dose actually delivered to the tumor target and on the alignment of the shielding disk with respect to the electron beam. Two EBT3 GAFCHROMIC films have been positioned on the two sides of the shielding disk in order to obtain the dose maps at the target and beyond the disk. Moreover the postprocessing analysis of the dose distribution measured on the films provides a quantitative estimate of the misalignment between the collimator and the disk. EBT3 radiochromic films have been demonstrated to be suitable dosimeters for IVD due to their linear dose-optical density response in a narrow range around the prescribed dose, as well as their capability to be fixed to the shielding disk without giving any distortion in the dose distribution. Off-line analysis of the radiochromic film allowed absolute dose measurements and this is indeed a very important verification of the correct exposure to the target organ, as well as an estimate of the dose to the healthy tissue underlying the shielding. These dose maps allow surgeons and radiation oncologists to take advantage of qualitative and quantitative feedback for setting more accurate treatment strategies and further optimized procedures. The proper alignment using elastic bands has improved the absolute dose accuracy and the collimator disk alignment by more than 50%.


Oscillatory reactivity to effortful cognitive processing in the subthalamic nucleus and internal pallidum: a depth electrode EEG study.
This study investigates how complex motor-cognitive activities are processed in the subthalamic nucleus (STN) and internal globus pallidum (GPi), as adverse neuropsychiatric effects may accompany deep brain stimulation (DBS), mainly in Parkinson's disease (PD) and STN-DBS. Dystonia patients with GPi-DBS electrodes (n = 5) and PD subjects (n = 5) with STN-DBS electrodes performed two tasks: (1) copying letters; and (2) writing any letter other than that appearing on the monitor. The cognitive load of the second task was greater than that of the first. Intracranial local field potentials (LFPs) were analysed. A beta power decrease was the main correlate of the enhanced cognitive load during the second task in both structures, with a lateralization to the left side, mainly in the GPi. A gamma power increase linked with the increased cognitive activity was observed only in the STN. Differences were also observed in the theta and alpha bandpasses. Beta ERD reactivity seems to be essential during the processing of complex motor-cognitive tasks, increases with enhanced cognitive effort, and was observed in both the STN and GPi. Oscillatory reactivity to effortful cognitive processing in other frequency bands was less consistent, with differences between the studied nuclei. Lateralization of activity related to cognitive factors was observed mainly in the GPi.


The modulation of topoisomerase I-mediated DNA cleavage and the induction of DNA-topoisomerase I crosslinks by crotonaldehyde-derived DNA adducts.
Crotonaldehyde is a representative alpha,beta-unsaturated aldehyde endowed of mutagenic and carcinogenic properties related to its propensity to react with DNA. Cyclic crotonaldehyde-derived deoxyguanosine (CrA-PdG) adducts can undergo ring opening in duplex DNA to yield a highly reactive aldehydic moiety. Here, we demonstrate that site-specifically modified DNA oligonucleotides containing a single CrA-PdG adduct can form crosslinks with topoisomerase I (Top1), both directly and indirectly. Direct covalent complex formation between the CrA-PdG adduct and Top1 is detectable after reduction with sodium cyanoborohydride, which is consistent with the formation of a Schiff base between Top1 and the ring open aldehyde form of the adduct. In addition, we show that the CrA-PdG adduct alters the cleavage and religation activities of Top1. It suppresses Top1 cleavage complexes at the adduct site and induces both reversible and irreversible cleavage complexes adjacent to the CrA-PdG adduct. The formation of stable DNA-Top1 crosslinks and the induction of Top1 cleavage complexes by CrA-PdG are mutually exclusive. Lastly, we found that crotonaldehyde induces the formation of DNA-Top1 complexes in mammalian cells, which suggests a potential relationship between formation of DNA-Top1 crosslinks and the mutagenic and carcinogenic properties of crotonaldehyde.


Surviving rectal cancer at the cost of a colostomy: global survey of long-term health-related quality of life in 10 countries.
Colorectal cancer management may require an ostomy formation; however, a stoma may negatively impact health-related quality of life (HRQoL). This study aimed to compare generic and stoma-specific HRQoL in patients with a permanent colostomy after rectal cancer across different countries. A cross-sectional cohorts of patients with a colostomy after rectal cancer in Denmark, Sweden, Spain, the Netherlands, China, Portugal, Australia, Lithuania, Egypt, and Israel were invited to complete questionnaires regarding demographic and socioeconomic factors along with the Colostomy Impact (CI) score, European Organisation for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire (QLQ-C30) and five anchor questions assessing colostomy impact on HRQoL. The background characteristics of the cohorts from each country were compared and generic HRQoL was measured with the EORTC QLQ-C30 presented for the total cohort. Results were compared with normative data of reference European populations. The predictors of reduced HRQoL were investigated by multivariable logistic regression, including demographic and socioeconomic factors and stoma-related problems. A total of 2557 patients were included. Response rates varied between 51-93 per cent. Mean time from stoma creation was 2.5-6.2 (range 1.1-39.2) years. A total of 25.8 per cent of patients reported that their colostomy impairs their HRQoL 'some'/'a lot'. This group had significantly unfavourable scores across all EORTC subscales compared with patients reporting 'no'/'a little' impaired HRQoL. Generic HRQoL differed significantly between countries, but resembled the HRQoL of reference populations. Multivariable logistic regression showed that stoma dysfunction, including high CI score (OR 3.32), financial burden from the stoma (OR 1.98), unemployment (OR 2.74), being single/widowed (OR 1.35) and young age (OR 1.01 per year) predicted reduced stoma-related HRQoL. Overall HRQoL is preserved in patients with a colostomy after rectal cancer, but a quarter of the patients interviewed reported impaired HRQoL. Differences among several countries were reported and socioeconomic factors correlated with reduced quality of life.


Live subgroup B respiratory syncytial virus vaccines that are attenuated, genetically stable, and immunogenic in rodents and nonhuman primates.
Optimal immunization of neonates against disease caused by respiratory syncytial virus (RSV) probably will require multiple doses of a vaccine containing viruses of both subgroups A and B. Live subgroup B RSV mutants were generated containing multiple attenuating mutations, ts (temperature-sensitive) and non-ts (host range), that were introduced by prolonged passage in cell culture or by chemical mutagenesis. The cold-passaged (cp)-52 mutant was restricted in replication compared to wild type virus in rodents and nonhuman primates. In addition, the attenuation phenotype of cp-52 was stable after prolonged replication in immunosuppressed rodents. One or two ts mutations were then introduced into the cp-52 mutant to generate additional candidate vaccine strains that were more attenuated in vivo than the cp-52 parental virus. Tests in humans are being done to determine if one or more of the RSV B-1 mutants exhibit a satisfactory balance between attenuation and immunogenicity.


Screening of the Pandemic Response Box Reveals an Association between Antifungal Effects of MMV1593537 and the Cell Wall of Cryptococcus neoformans, Cryptococcus deuterogattii, and Candida auris.
There is an urgent unmet need for novel antifungals. In this study, we searched for novel antifungal activities in the Pandemic Response Box, a collection of 400 structurally diverse compounds in various phases of drug discovery. We identified five molecules which could control the growth of Cryptococcus neoformans, Cryptococcus deuterogattii, and the emerging global threat Candida auris. After eliminating compounds which demonstrated paradoxical antifungal effects or toxicity to mammalian macrophages, we selected compound MMV1593537 as a nontoxic, fungicidal molecule for further characterization of antifungal activity. Scanning electron microscopy revealed that MMV1593537 affected cellular division in all three pathogens. In Cryptococcus, MMV1593537 caused a reduction in capsular dimensions. Treatment with MMV1593537 resulted in increased detection of cell wall chitooligomers in these three species. Since chitooligomers are products of the enzymatic hydrolysis of chitin, we investigated whether surface chitinase activity was altered in response to MMV1593537 exposure. We observed peaks of enzyme activity in C. neoformans and C. deuterogattii in response to MMV1593537. We did not detect any surface chitinase activity in C. auris. Our results suggest that MMV1593537 is a promising, nontoxic fungicide whose mechanism of action, at least in Cryptococcus spp, requires chitinase-mediated hydrolysis of chitin. IMPORTANCE The development of novel antifungals is a matter of urgency. In this study, we evaluated antifungal activities in a collection of 400 molecules, using highly lethal fungal pathogens as targets. One of these molecules, namely, MMV1593537, was not toxic to host cells and controlled the growth of isolates of Cryptococcus neoformans, C. deuterogattii, C. gattii, Candida auris, C. albicans, C. parapsilosis, and C. krusei. We tested the mechanisms of antifungal action of MMV1593537 in the Cryptococcus and C. auris models and concluded that the compound affects the cell wall, a structure which is essential for fungal life. At least in Cryptococcus, this effect involved chitinase, an enzyme which is required for remodeling the cell wall. Our results suggest that MMV1593537 is a candidate for future antifungal development.


Cost-Effectiveness of Mobile App-Guided Training in Extended Focused Assessment with Sonography for Trauma (eFAST): A Randomized Trial.
Ultrasound training is associated with a long learning curve and use of substantial faculty resources. Self-directed ultrasound training may decrease the need for faculty-led teaching. Mobile apps seem promising for use in self-directed ultrasound training, but no studies have examined the cost-effectiveness of mobile app-guided training versus traditional formats such as textbook-guided training. This study evaluated the cost-effectiveness of mobile app-guided versus textbook-guided ultrasound training. First-year residents (n = 38) with no previous ultrasound experience were randomized into mobile app-guided versus textbook-guided self-directed ultrasound training groups. Participants completed a transfer test involving four patient cases and a theoretical test on diagnostic accuracy. Two ultrasound experts assessed the residents' performance using the Objective Structured Assessment of Ultrasound Skills (OSAUS) scale. The costs of developing mobile app and textbook material were calculated and used for the analysis of cost-effectiveness. 34 participants completed the transfer test. There was no statistically significant difference in test performance or diagnostic accuracy between the mobile app-guided (mean-OSAUS 42.3 % [95 %CI38.5 - 46.0 %]) and textbook-guided groups (mean-OSAUS 45.3 % [95 %CI39.3 - 51.3 %]) (d.f. [1.33] = 0.45, p = 0.41). However, development costs differed greatly for each instructional format. Textbook-guided training was significantly more cost-effective than mobile app-guided training (Incremental Cost Effectiveness Ratio -861 967 [95 %CI-1071.7 to-3.2] USD/pct. point change in OSAUS score). Mobile app-guided ultrasound training is less cost-effective than textbook-guided self-directed training. This study underlines the need for careful evaluation of cost-effectiveness when introducing technological innovations for clinical skills training.


Bias and prevalence effects on kappa viewed in terms of sensitivity and specificity.
Paradoxical effects of bias and prevalence on the kappa coefficient are examined using the concepts of sensitivity and specificity. Results that appear paradoxical when viewed as a 2 x 2 table of frequencies do not appear paradoxical when viewed as a pair of sensitivity and specificity measures where each observer is treated as a predictor of the other observer. An adjusted kappa value can be obtained from these sensitivity/specificity measures but simulation studies indicate that it would result in substantial overestimation of reliability when bias or prevalence effects are observed. It is suggested that investigators concentrate on obtaining populations with trait prevalence near 50% rather than searching for statistical indices to rescue or excuse inefficient experiments.


The Effect of Vocabulary Intervention on Text Comprehension: Who Benefits?
Purpose Vocabulary intervention can improve comprehension of texts containing taught words, but it is unclear if all middle school readers get this benefit. This study tests 2 hypotheses about variables that predict response to vocabulary treatment on text comprehension: gains in vocabulary knowledge due to treatment and pretreatment reading comprehension scores. Method Students in Grade 6 (N = 23) completed a 5-session intervention based on robust vocabulary instruction (RVI). Knowledge of the semantics of taught words was measured pre- and posttreatment. Participants then read 2 matched texts, 1 containing taught words (treated) and 1 not (untreated). Treated texts and taught word lists were counterbalanced across participants. The difference between text comprehension scores in treated and untreated conditions was taken as a measure of the effect of RVI on text comprehension. Results RVI resulted in significant gains in knowledge of taught words (dRM = 2.26) and text comprehension (dRM = 0.31). The extent of gains in vocabulary knowledge after vocabulary treatment did not predict the effect of RVI on comprehension of texts. However, untreated reading comprehension scores moderated the effect of the vocabulary treatment on text comprehension: Lower reading comprehension was associated with greater gains in text comprehension. Readers with comprehension scores below the mean experienced large gains in comprehension, but those with average/above average reading comprehension scores did not. Conclusion Vocabulary instruction had a larger effect on text comprehension for readers in Grade 6 who had lower untreated reading comprehension scores. In contrast, the amount that children learned about taught vocabulary did not predict the effect of vocabulary instruction on text comprehension. This has implications for the identification of 6th-grade students who would benefit from classroom instruction or clinical intervention targeting vocabulary knowledge.