Aptamer-based regulation of transcription circuits.
We propose synthetic DNA/RNA transcription circuits based on specific aptamer recognition. By mimicking transcription factor regulation, combined with specific enzyme/DNA aptamer binding, multiple biomolecules including DNA, RNA, polymerase, restriction enzymes and methylase were used as regulators. In addition, multi-level cascading networks and methylation-switch circuits were also established. This regulation strategy has the potential to expand the toolkit of in vitro synthetic biology.


Idiopathic Condylar Resorption: A Survey and Review of the Literature.
Idiopathic condylar resorption (ICR) is a relatively uncommon condition. Its diagnosis, etiology, and management options are controversial. Furthermore, it is difficult for 1 provider to collect a large cohort of ICR patients to develop a statistically significant study of these concerns. Therefore, the purpose of this study was to survey experienced temporomandibular joint (TMJ) surgeons who have managed ICR cases relative to these concerns, as well as review the management outcome literature. SurveyMonkey (Palo Alto, CA) was used to canvas the 88 international TMJ surgeons on the TMJ Concepts (Ventura, CA) InterNetwork. This network connects a group of surgeons across the world who consistently perform TMJ surgery. The intent was to provide a global snapshot of the demographic, epidemiologic, diagnostic workup, and outcome data related to the management of ICR cases. The surveys from surgeons who did not respond to all 12 questions were not included in the results. After application of the exclusion criteria, complete data on a cohort of 100 patients were obtained and used for the study. A history of hormonal imbalance was reported in only 10% of patients; however, 42 of 94 women were reportedly taking birth control pills presumably affecting their menstrual cycles. The most common reason for consultation was Class II malocclusion (98% of patients). Treatment modalities varied and included orthodontics, orthotics, TMJ total joint replacement, orthognathic surgery, and disc repositioning. ICR management proved to be controversial among the surveyed surgeons. Multiple treatment options have been described in the literature, including medical management, orthognathic surgery only, TMJ and orthognathic surgery, and total joint prosthesis reconstruction. Long-term, controlled, multicenter clinical studies should be developed to evaluate outcomes of all nonsurgical and surgical management options for the ICR patient.


Mechanical analysis of an axially symmetric cylindrical phantom with a spherical heterogeneity for MR elastography.
Cylindrical homogenous phantoms for magnetic resonance (MR) elastography in biomedical research provide one way to validate an imaging systems performance, but the simplified geometry and boundary conditions can cloak complexity arising at tissue interfaces. In an effort to develop a more realistic gel tissue phantom for MRE, we have constructed a heterogenous gel phantom (a sphere centrally embedded in a cylinder). The actuation comes from the phantom container, with the mechanical waves propagating toward the center, focusing the energy and thus allowing for the visualization of high-frequency waves that would otherwise be damped. The phantom was imaged and its stiffness determined using a 9.4 T horizontal MRI with a custom build piezo-elastic MRE actuator. The phantom was vibrated at three frequencies, 250, 500, and 750 Hz. The resulting shear wave images were first used to reconstruct material stiffness maps for thin (1 mm) axial slices at each frequency, from which the complex shear moduli μ were estimated, and then compared with forward modeling using a recently developed theoretical model which took μ as inputs. The overall accuracy of the measurement process was assessed by comparing theory with experiment for selected values of the shear modulus (real and imaginary parts). Close agreement is shown between the experimentally obtained and theoretically predicted wave fields.


Medico-legal reasoning in disability assessment: a focus group and validation study.
Decisions on disability pensions are based, among others, on medical reports. The way these medical assessments are performed is largely unclear. The aim of the study was to determine which grounds are used by social insurance physicians (SIPs) in these assessments and to determine if the identification of these grounds can help improve the quality of assessments in social insurance practice. The article describes a focus group study and a questionnaire study with SIPs in four different countries. Using focus group discussions of SIPs discussing the same case in Belgium, the Netherlands, Norway and Slovenia (N = 29) we determined the arguments and underlying grounds as used by the SIP's. We used a questionnaire study among other SIPs (N = 60) in the same countries to establish a first validation of these grounds. Grounds in the focus groups were comparable between the countries studied. The grounds were also recognized by SIPs who had not participated in the focus groups. SIPs agreed most on grounds with regard to the claimant's health condition, and about the claimant's duty to explore rehabilitation and work resumption, but less on accepting permanent incapacity when all options for treatment were exhausted. Grounds that SIPs use refer to a limited group of key elements of disability evaluation. SIPs interpret disability in social insurance according to the handicapped role and strive at making their evaluation fair trials. ICF is relevant with regard to the health condition and to the process of evaluation. Identification of grounds is a valuable instrument for controlling the quality of disability evaluation. The grounds also appear to be internationally comparable which may enhance scientific study in this area.


Insight into the bioabsorption of Fe-based materials and their current developments in bone applications.
Iron (Fe) and Fe-based materials have been vigorously explored in orthopedic applications in the past decade mainly owing to their promising mechanical properties including high yield strength, elastic modulus and ductility. Nevertheless, their corrosion products and low corrosion kinetics are the major concerns that need to be improved further despite their appealing mechanical strengths. The current studies on porous Fe-based scaffolds show an improved corrosion rate but the in vitro biocompatibility is still problematic in general. Unlike the Mg implants, the biodegradation and bioabsorption of Fe-based implants are still not well described. This vague issue could implicate the development of Fe-based materials as potential medical implants as they have not reached the clinical trial stage yet. Thus, there is a need to understand in-depth the Fe corrosion behavior and its bioabsorption mechanism to facilitate the material design of Fe-based scaffolds and further improve its biocompatibility. This manuscript provides an important insight into the basic bioabsorption of the multi-ranged Fe-based corrosion products with a review of the latest progress on the corrosion & in vitro biocompatibility of porous Fe-based scaffolds together with the remaining challenges and the perspective on the future direction.


Immediate effects of noxious and innocuous thermal stimulation on brain activation in patients with stroke.
Case-control studies have shown that noxious thermal stimulation (TS) can improve arm function in patients with stroke. However, the neural mechanisms underlying this improvement are largely unknown. We explored functional neural activation due to noxious and innocuous TS intervention applied to the paretic arm of patients with stroke. Sixteen participants with unilateral cortical infarctions were allocated to one of two groups: noxious TS (8 patients; temperature combination: hot pain 46°C to 47°C, cold pain 7°C-8°C) or innocuous TS (n = 8; temperature combination: hot 40°C-41°C, cold 20°C-21°C). All subjects underwent fMRI scanning before and after 30 min TS intervention and performed a finger tapping task with the affected hand. Immediate brain activation effects were assessed according to thermal type (noxious vs. innocuous TS) and time (pre-TS vs post-TS). Regions activated by noxious TS relative to innocuous TS (P < .05, adjusted for multiple comparisons) were related to motor performance and sensory function in the bilateral primary somatosensory cortices, anterior cingulate cortex, insula, thalamus, hippocampus and unilateral primary motor cortex, secondary somatosensory cortex at the contralateral side of lesion, and unilateral supplementary motor area at the ipsilateral side of lesion. Greater activation responses were observed in the side contralateral to the lesion, suggesting a significant intervention effect. Our preliminary findings suggest that noxious TS may induce neuroplastic changes unconstrained to the local area.Trial registration: NCT01418404.


Targeting DNA repair in gliomas.
Gliomas represent a disparate group of malignancies with varying clinical outcomes despite a tremendous amount of time, effort, and resources dedicated to their management and understanding. The most aggressive entity, glioblastoma, has a dismal prognosis with poor local control despite intense local and systemic treatment, including radiation therapy. Given the heterogeneity in genotype, phenotype, and patient outcomes, researchers and clinicians have turned their attention toward attacking DNA damage response and repair mechanisms in gliomas in an effort to develop novel chemo and radiosensitizers. However, despite extensive work in both the laboratory and the clinic, no sensitizers have yet to emerge as clear options in the treatment of glioma, often because of meager preclinical data or an inability to penetrate the blood-brain barrier. This review will examine current understanding of molecular DNA repair targets in glioma and their potential exploitation to improve local control and, ultimately, overall survival of patients afflicted with these diseases.


An accurate and powerful method for copy number variation detection.
Integration of multiple genetic sources for copy number variation detection (CNV) is a powerful approach to improve the identification of variants associated with complex traits. Although it has been shown that the widely used change point based methods can increase statistical power to identify variants, it remains challenging to effectively detect CNVs with weak signals due to the noisy nature of genotyping intensity data. We previously developed modSaRa, a normal mean-based model on a screening and ranking algorithm for copy number variation identification which presented desirable sensitivity with high computational efficiency. To boost statistical power for the identification of variants, here we present a novel improvement that integrates the relative allelic intensity with external information from empirical statistics with modeling, which we called modSaRa2. Simulation studies illustrated that modSaRa2 markedly improved both sensitivity and specificity over existing methods for analyzing array-based data. The improvement in weak CNV signal detection is the most substantial, while it also simultaneously improves stability when CNV size varies. The application of the new method to a whole genome melanoma dataset identified novel candidate melanoma risk associated deletions on chromosome bands 1p22.2 and duplications on 6p22, 6q25 and 19p13 regions, which may facilitate the understanding of the possible roles of germline copy number variants in the etiology of melanoma. http://c2s2.yale.edu/software/modSaRa2 or https://github.com/FeifeiXiaoUSC/modSaRa2. Supplementary data are available at Bioinformatics online.


Genomic analysis of Hox clusters in the sea lamprey Petromyzon marinus.
The sea lamprey Petromyzon marinus is among the most primitive of extant vertebrates. We are interested in the organization of its Hox gene clusters, because, as a close relative of the gnathostomes, this information would help to infer Hox cluster organization at the base of the gnathostome radiation. We have partially mapped the P. marinus Hox clusters using phage, cosmid, and P1 artificial chromosome libraries. Complete homeobox sequences were obtained for the 22 Hox genes recovered in the genomic library screens and analyzed for cognate group identity. We estimate that the clusters are somewhat larger than those of mammals (roughly 140 kbp vs. 105 kbp) but much smaller than the single Hox cluster of the cephalochordate amphioxus (at more than 260 kb). We never obtained more than three genes from any single cognate group from the genomic library screens, although it is unlikely that our screen was exhaustive, and therefore conclude that P. marinus has a total of either three or four Hox clusters. We also identify four highly conserved non-coding sequence motifs shared with higher vertebrates in a genomic comparison of Hox 10 genes.


Comparative and functional genomics of lipases in holometabolous insects.
Lipases have key roles in insect lipid acquisition, storage and mobilisation and are also fundamental to many physiological processes underpinning insect reproduction, development, defence from pathogens and oxidative stress, and pheromone signalling. We have screened the recently sequenced genomes of five species from four orders of holometabolous insects, the dipterans Drosophila melanogaster and Anopheles gambiae, the hymenopteran Apis mellifera, the moth Bombyx mori and the beetle Tribolium castaneum, for the six major lipase families that are also found in other organisms. The two most numerous families in the insects, the neutral and acid lipases, are also the main families in mammals, albeit not in Caenorhabditis elegans, plants or microbes. Total numbers of the lipases vary two-fold across the five insect species, from numbers similar to those in mammals up to numbers comparable to those seen in C. elegans. Whilst there is a high degree of orthology with mammalian lipases in the other four families, the great majority of the insect neutral and acid lipases have arisen since the insect orders themselves diverged. Intriguingly, about 10% of the insect neutral and acid lipases have lost motifs critical for catalytic function. Examination of the length of lid and loop regions of the neutral lipase sequences suggest that most of the insect lipases lack triacylglycerol (TAG) hydrolysis activity, although the acid lipases all have intact cap domains required for TAG hydrolysis. We have also reviewed the sequence databases and scientific literature for insights into the expression profiles and functions of the insect neutral and acid lipases and the orthologues of the mammalian adipose triglyceride lipase which has a pivotal role in lipid mobilisation. These data suggest that some of the acid and neutral lipase diversity may be due to a requirement for rapid accumulation of dietary lipids. The different roles required of lipases at the four discrete life stages of holometabolous insects may also contribute to the diversity of lipases required by insects. In addition, insects use lipases to perform roles for which there are no correlates in mammals, including as yolk and male accessory gland proteins.


Soil carbon stocks in temperate grasslands differ strongly across sites but are insensitive to decade-long fertilization.
Enhancing soil carbon (C) storage has the potential to offset human-caused increases in atmospheric CO2 . Rising CO2 has occurred concurrently with increasing supply rates of biologically limiting nutrients such as nitrogen (N) and phosphorus (P). However, it is unclear how increased supplies of N and P will alter soil C sequestration, particularly in grasslands, which make up nearly a third of non-agricultural land worldwide. Here, we leverage a globally distributed nutrient addition experiment (the Nutrient Network) to examine how a decade of N and P fertilization (alone and in combination) influenced soil C and N stocks at nine grassland sites spanning the continental United States. We measured changes in bulk soil C and N stocks and in three soil C fractions (light and heavy particulate organic matter, and mineral-associated organic matter fractions). Nutrient amendment had variable effects on soil C and N pools that ranged from strongly positive to strongly negative, while soil C and N pool sizes varied by more than an order of magnitude across sites. Piecewise SEM clarified that small increases in plant C inputs with fertilization did not translate to greater soil C storage. Nevertheless, peak season aboveground plant biomass (but not root biomass or production) was strongly positively related to soil C storage at seven of the nine sites, and across all nine sites, soil C covaried with moisture index and soil mineralogy, regardless of fertilization. Overall, we show that site factors such as moisture index, plant productivity, soil texture, and mineralogy were key predictors of cross-site soil C, while nutrient amendment had weaker and site-specific effects on C sequestration. This suggests that prioritizing the protection of highly productive temperate grasslands is critical for reducing future greenhouse gas losses arising from land use change.


A reproducible laser-wounded skin equivalent model to study the effects of aging in vitro.
Skin aging involves both chronological and photoaging processes. The effects of these processes are often overlapping and include changes in both the stratified epithelium and the fibroblast-rich dermis. Wound healing is frequently delayed with aging and can result in scarring. A skin equivalent model can be used to study the role of cells and the extracellular matrix in the process of wound healing. Current studies using this model employ a full-thickness wound placed atop a nonwounded dermis to mimic a partial-thickness wound. However, a true reproducible partial-thickness wound model has yet to be described. In this study, we investigated whether a laser-wounded skin equivalent would be a useful partial-thickness wound healing model. Three lasers were compared for the ability to generate a reproducible wound: an erbium-YAG, a high-powered excimer, and a low-powered excimer laser. Reepithelialization ability was tested using newborn and adult skin keratinocytes, adult esophageal keratinocytes, and cdk4-overexpressing newborn keratinocytes. Keratinocyte compartmentalization and basement membrane formation were assessed by immunofluorescence. The erbium-YAG and high-powered excimer laser cut reproducible wounds but left the remaining surface either discolored due to thermal damage and/or ragged; keratinocytes were unable to migrate into the wound area. The low-powered excimer laser cut reproducible wounds, leaving the cut surface intact and visibly unaltered; keratinocytes reepithelialized the wound in a collagenase-dependent manner within 3 days; and return of compartmentalization and basement membrane occurred within 14 days. The laser-wounded skin equivalent is an adjustable, reproducible partial-thickness wound model where keratinocyte biology akin to in vivo can be studied, and will be useful to study the effects of aging on wound healing.


Assessment of benefit in tele-ophthalmology using a consensus panel.
A videoconferencing link was established from a hospital in South Africa to Moorfields Eye Hospital in London. A clinician in South Africa used a video slit-lamp and videoconferencing equipment to capture images and communicate with a specialist in London. Over 12 months, 113 cases were discussed in teleconsultations. Case-notes were subsequently obtained for 90 cases (80%). Three consultant ophthalmologists took part in the consensus process, one from the UK and two from South Africa. The consensus panel achieved on average 78-96% agreement on the items rated. In approximately half the cases, the panel judged that teleconsultation had had an effect on diagnosis. In nine cases (10%) there was potential for definite improvement in visual health as a result of tele-ophthalmology and in a further 48 cases (53%) there was potential for possible improvement in visual health. The consensus process identified 35 cases (39%) where there was potential improvement in general health as a result of tele-ophthalmology. Despite the methodological limitations of our approach, it was possible to learn a great deal about the effectiveness of the telemedicine intervention.


Critical shunt-induced subdural hematoma treated with combined pressure-programmable valve implantation and endoscopic third ventriculostomy.
The authors present 2 patients with VP shunt-induced subdural hematomas (SDH) treated with pressure-programmable valve implantation and endoscopic third ventriculostomies (TV). The first patient is an 11-year-old girl who developed a shunt-induced SDH. Revision of the shunt valve with a higher-pressure valve resulted in a prolonged deterioration of her consciousness. External ventricular drainage at low pressure led to clinical improvement. A pressure-programmable valve set at 50 mm H(2)O was implanted, and the pressure gradually increased. At a pressure of 120 mm H(2)O symptoms recurred, even though the subdural collection was beginning to decrease in size. An endoscopic TV was performed, and the valve pressure was then increased to 200 mm H(2)O without any neurological symptoms. The second patient, a 7-year-old boy with shunt-induced SDH, had recurrent SDH, even after shunt revision with placement of a higher-pressure valve, which resulted in prolonged lethargy. A pressure-programmable valve was implanted with concurrent endoscopic TV. Gradual valve pressure increases up to 200 mm H(2)O could be performed without recurrent symptoms. Eventually, the shunt system was ligated to resolve residual positional headache, and the TV has been patent for more than 3 years. In both patients, the pressure-programmable valve was useful, since the optimal CSF drainage pressure changed during the period of recovery from symptomatic subdural collections. Concurrent TV appeared to enable increasing the valve pressure gradually without any neurological symptoms. The advantages of this combined approach are discussed.


Gut microbiome is associated with multiple sclerosis activity in children.
To identify features of the gut microbiome associated with multiple sclerosis activity over time. We used 16S ribosomal RNA sequencing from stool of 55 recently diagnosed pediatric-onset multiple sclerosis patients. Microbiome features included the abundance of individual microbes and networks identified from weighted genetic correlation network analyses. Prentice-Williams-Peterson Cox proportional hazards models estimated the associations between features and three disease activity outcomes: clinical relapses and both new/enlarging T2 lesions and new gadolinium-enhancing lesions on brain MRI. Analyses were adjusted for age, sex, and disease-modifying therapies. Participants were followed, on average, 2.1 years. Five microbes were nominally associated with all three disease activity outcomes after multiple testing correction. These included butyrate producers Odoribacter (relapse hazard ratio = 0.46, 95% confidence interval: 0.24, 0.88) and Butyricicoccus (relapse hazard ratio = 0.49, 95% confidence interval: 0.28, 0.88). Two networks of co-occurring gut microbes were significantly associated with a higher hazard of both MRI outcomes (gadolinium-enhancing lesion hazard ratios (95% confidence intervals) for Modules 32 and 33 were 1.29 (1.08, 1.54) and 1.42 (1.18, 1.71), respectively; T2 lesion hazard ratios (95% confidence intervals) for Modules 32 and 33 were 1.34 (1.15, 1.56) and 1.41 (1.21, 1.64), respectively). Metagenomic predictions of these networks demonstrated enrichment for amino acid biosynthesis pathways. Both individual and networks of gut microbes were associated with longitudinal multiple sclerosis activity. Known functions and metagenomic predictions of these microbes suggest the important role of butyrate and amino acid biosynthesis pathways. This provides strong support for future development of personalized microbiome interventions to modify multiple sclerosis disease activity.


[Efferent innervation of pulmonary blood vessels and bronchi in rat (an immunohistochemical study)].
In this investigation the peculiarities of innervation of bronchi and blood vessels of the lung were studied in 20 rats using immunohistochemical demonstration of synaptophysin and alpha-actin. The results obtained have showen that the densest innervation is typical for bronchial walls, particularly, for the muscular lamina. Synaptophysin-immunoreactive terminals (SFIT) were detected in the bronchi in close association with both circular bundles of smooth muscle cells and microganglia. Dense network of SFIT was found in the pulmonary vein--in its middle tunic formed by cardiomyocytes. In contrast to the bronchi and pulmonary vein, large branches of the pulmonary artery contained no SFIT. We briefly discuss the problem of the origin of the nerve fibers described and their functions and suggest that SFIT are formed by efferent fibers (axons) of neurons arising from either the intrapulmonary parasympathetic ganglia.


Recombinant Desmodus rotundus salivary plasminogen activator crosses the blood-brain barrier through a low-density lipoprotein receptor-related protein-dependent mechanism without exerting neurotoxic effects.
Desmoteplase, a recombinant form of the plasminogen activator DSPAalpha1 from Desmodus rotundus, may offer improved clinical benefits for acute ischemic stroke treatment over the current therapy, recombinant tissue plasminogen activator (rtPA). Accumulating evidence suggests that clinical use of rtPA could be limited by unfavorable properties, including its ability to cross the blood-brain barrier (BBB), thus potentially adding to the pro-excitotoxic effect of endogenous tPA in cerebral parenchyma. Here, to investigate whether desmoteplase may display a safer profile than the structurally-related tPA, both agents were compared for their ability to cross the BBB and promote neurotoxicity. First, the passage of vascular DSPA and rtPA was investigated in vitro in a model of BBB, subjected or not to oxygen and glucose deprivation. Second, we studied DSPA- and rtPA-mediated effects in an in vivo paradigm of excitotoxic necrosis. The rtPA and desmoteplase cross the intact BBB by LRP-mediated transcytosis. Under conditions of oxygen and glucose deprivation, translocation rates of both compounds increased; however, unlike rtPA, desmoteplase transport remained LRP-dependent. Additionally, neither intracerebral nor intravenous desmoteplase administration enhanced NMDA-induced excitotoxic striatal damage in vivo. Interestingly, intravenous but not intrastriatal coadministration of desmoteplase and rtPA reduced the pro-excitotoxic effect of rtPA. We show that desmoteplase crosses the BBB but does not promote neuronal death. Moreover, intravenous administration of desmoteplase antagonizes the neurotoxicity induced by vascular rtPA. This action may be caused by competition of desmoteplase with rtPA for LRP binding at the BBB, thus effectively blocking rtPA access to the brain parenchyma.


A qualitative study to understand the Duchenne muscular dystrophy experience from the parent/patient perspective.
Duchenne muscular dystrophy (DMD) is a rare, severe, fatal neuromuscular disease characterized by progressive atrophy and muscle weakness, resulting in loss of ambulation, decreased upper body function, and impaired cardiorespiratory function. This study aimed to generate qualitative evidence to describe the primary symptoms and impacts of DMD in ambulatory and non-ambulatory patients as reported by patient/caregiver dyads. Information was also gathered on expectations for future DMD treatments. Forty-six dyads (caregiver and patients with DMD aged 4 to 22 years) participated in 60-min semi-structured video interviews. Interview transcripts were analyzed using thematic analysis. Differences in experiences with DMD by ambulation status were examined. Mean ages of ambulatory (n = 28) and non-ambulatory participants (n = 18) were 8.7 and 11.3 years, respectively, with an average age of diagnosis of 3.7 years (SD = 2.3). The primary symptoms reported by both groups were lack of strength (ambulatory: n = 28, 100.0%; non-ambulatory: n = 17, 94.4%) and fatigue (ambulatory: n = 24, 85.7%; non-ambulatory: n = 14, 77.8%). Physical function was the domain that was most impacted by DMD, with participants describing progressive decline of physical function due to loss of physical strength as the primary defining feature of the disease across all stages of ambulatory ability. For those who maintained ambulatory ability at the time of the interview, physical function impacts described impaired mobility (e.g., climbing stairs: n = 16, 57.1%; running: n = 13, 46.4%), impaired upper body function, in particular fine motor skills like holding a pen/pencil or buttoning clothes (n = 17, 60.7%), problem with transfers (e.g., getting off the floor: n = 10, 35.7%), and activities of daily living (ADLs; n = 15, 53.6%). For non-ambulatory participants, the functional impacts most frequently described were problems with transfers (e.g., getting in/out of bed: n = 13, 72.2%; getting in/out of chair or position in bed: both n = 10, 55.6%), impaired upper body function (reaching: n = 14, 77.8%), and ADLs (n = 15, 83.3%). Meaningful treatment goals differed by ambulatory status; for ambulatory participants, goals included maintaining current functioning (n = 20, 71.4%), improving muscle strength (n = 7, 25.9%), and reducing fatigue (n = 6, 22.2%). For non-ambulatory participants, these included increased upper body strength (n = 8, 42.1%) and greater independence in ADLs (n = 6, 31.6%). A preliminary conceptual model was developed to illustrate the primary symptoms and physical function impacts of DMD and capture their relationship to disease progression. This study contributes to the limited qualitative literature by characterizing impacts of physical limitations and symptoms of DMD on disease progression and thus providing insights into the lived experience with DMD. Differences in treatment goals were also identified based on ambulatory status. Taken together, these findings can help inform patient-centered measurement strategies for evaluating outcomes in DMD clinical research.


Coevolutionary networks: a novel approach to understanding the relationships of humans with the infectious agents.
Human organism is interpenetrated by the world of microorganisms, from the conception until the death. This interpenetration involves different levels of interactions between the partners including trophic exchanges, bi-directional cell signaling and gene activation, besides genetic and epigenetic phenomena, and tends towards mutual adaptation and coevolution. Since these processes are critical for the survival of individuals and species, they rely on the existence of a complex organization of adaptive systems aiming at two apparently conflicting purposes: the maintenance of the internal coherence of each partner, and a mutually advantageous coexistence and progressive adaptation between them. Humans possess three adaptive systems: the nervous, the endocrine and the immune system, each internally organized into subsystems functionally connected by intraconnections, to maintain the internal coherence of the system. The three adaptive systems aim at the maintenance of the internal coherence of the organism and are functionally linked by interconnections, in such way that what happens to one is immediately sensed by the others. The different communities of infectious agents that live within the organism are also organized into functional networks. The members of each community are linked by intraconnections, represented by the mutual trophic, metabolic and other influences, while the different infectious communities affect each other through interconnections. Furthermore, by means of its adaptive systems, the organism influences and is influenced by the microbial communities through the existence of transconnections. It is proposed that these highly complex and dynamic networks, involving gene exchange and epigenetic phenomena, represent major coevolutionary forces for humans and microorganisms.


Bone bruises in anterior cruciate ligament injured knee and long-term outcomes. A review of the evidence.
Bone bruises are frequently associated with anterior cruciate ligament (ACL) tears as a result of trauma or direct shear stress of the bone. To review the evidence regarding the characteristics of the bone bruise associated with ACL tears, its relevance on clinical outcomes, and its progression over time. In particular, the long-term effects of the bone bruise on the knee osteochondral architecture and joint function were evaluated. Review; level of evidence: 4. An electronic search was performed on PubMed. Combinations of keywords included: "bone bruise AND knee"; "bone bruise AND anterior cruciate ligament"; "bone bruise AND osteo-chondral defects". Any level of evidence studies concerning bone bruises in patients with partial or complete ACL tears were retrieved. A total of 25 studies were included; three of them investigated biomechanical parameters, seven were concerned with clinical outcomes, and 15 were radiological studies. Evaluation of the bone bruise is best performed using a fat-saturated T2-weighted fast spin echo exam or a short tau inversion recovery sequence where fat saturation is challenging. The location of the injury has been demonstrated to be more frequent in the lateral compartment of the joint (lateral femoral condyle and lateral tibial plateau). It is associated with ACL tears in approximately 70% of cases, often with collateral ligament or meniscal tears. Mid- and long-term outcomes demonstrated a complete healing of the marrow lesions at magnetic resonance imaging, but chondral defects detected with T1ρ sequences are still present 1 year after the ACL injury. Functional examination of the knee, through clinical International Knee Documentation Committee scores, did not show any correlation with the bone bruise. Although bone bruise presence yields to higher pain levels, no correlation with functional outcomes was reported. Most studies have a short-term follow-up (<2 years) compared to the length of time it takes to develop post-traumatic osteoarthritis, so it still remains unclear whether the initial joint injury and bone bruise have a direct relationship to long-term function.


Extranodal extension of lymph node metastasis influences recurrence in prostate cancer: a systematic review and meta-analysis.
The extranodal extension (ENE) of nodal metastasis involves the extension of neoplastic cells through the lymph node capsule into the perinodal adipose tissue. This morphological feature has recently been indicated as an important prognostic factor in various cancer types, but its role in prostate cancer is still unclear. We aimed to clarify it, performing the first meta-analysis on this issue, comparing prognostic parameters in surgically treated, node-positive prostate cancer patients with (ENE+) vs. without (ENE-) ENE. Data were summarized using risk ratios (RRs) for number of deaths/recurrences and hazard ratios (HRs), with 95% confidence intervals (CI), for the time-dependent risk related to ENE positivity. Six studies followed-up 1,113 patients with N1 prostate cancer (658 ENE+ vs. 455 ENE-) for a median of 83 months. The presence of ENE was associated with a significantly higher risk of biochemical recurrence (RR = 1.15; 95%CI: 1.03-1.28; I2 = 0%; HR = 1.40, 95%CI: 1.12-1.74; I2 = 0%) and "global" (biochemical recurrence and distant metastasis) recurrence (RR = 1.15; 95%CI: 1.04-1.28; I2 = 0%; HR = 1.41, 95%CI: 1.14-1.74; I2 = 0%). ENE emerged as a potential prognostic moderator, earmarking a subgroup of patients at higher risk of recurrence. It may be considered for the prognostic stratification of metastatic patients. New possible therapeutic approaches may explore more in depth this prognostic parameter.


[The aetiology of rheumatoid arthritis (author's transl)].
An ineffective aetiology for rheumatoid arthritis and other connective tissue diseases has been frequently postulated but never proven. Despite the failure to obtain firm evidence of viral infection in most patients with these disorders for several reasons this theory should not be discarded. Firstly several mechanisms have been discovered by which virus infections can persist in lymphocytes and other tissue thereby provoking inflammation without the production of complete readily detectable virus particles. Secondly there are numerous ways in which host resistance to virus can be subverted with the perpetuation of an ineffective or inappropriate immune response. Thirdly the immunopathological features of these diseases are entirely compatible with an infective aetiology. The main problem is likely to prove the difficulty in attributing a primary pathogenetic role to any isolated virus rather than regarding it as a passenger virus which has been non-specifically activated by the disease process. However preoccupation with a viral aetiology should not blind one to other possibilities since many environmental allergens can produce immunopathological disease of highly protean nature.


Pyrogenic toxin superantigen site specificity in toxic shock syndrome and food poisoning in animals.
Staphylococcus aureus and Streptococcus pyogenes express pyrogenic toxin superantigens (PTSAgs) that are associated with toxic shock syndrome (TSS) and staphylococcal food poisoning (SFP). Most PTSAgs cause TSS in deep-tissue infections, whereas only TSS toxin 1 (TSST-1) is associated with menstrual, vaginal TSS. In contrast, SFP has been linked only with staphylococcal enterotoxins (SEs). Because of the differential abilities of PTSAgs to cause systemic or localized symptoms in a site-dependent manner, the present study was undertaken to assess the toxins' abilities to cross mucosal barriers. The activity of three PTSAgs when delivered orally, vaginally, or intravenously to rabbits and orally to monkeys was investigated. TSST-1 induced shock via all three routes in rabbits. Although active when administered intravenously, SEC1 and streptococcal pyrogenic exotoxin A (SPEA) did not cause symptoms when administered orally or vaginally. Only SEC1 induced emesis in the monkey feeding assay. TSST-1, albeit less stable than SEC1 and SPEA to pepsin, induced diarrhea in monkeys. Our results may explain the unique association of TSST-1 with menstrual TSS and why SPEA is only rarely associated with TSS after pharyngitis, despite being highly associated with TSS after subcutaneous infections. Finally, our studies indicate that enterotoxicity in SFP is not the result of superantigenicity.


Prenatal diagnosis of imperforate hymen with hydrometrocolpos.
Imperforate hymen is one of the most simple and most common anomalies in the female genital organs, occurring in 0.1% of girls born at term. Some are recognized because of mucocolpos at birth, but the diagnosis is usually not detected before puberty. Rarely, obstetric sonography may prenatally detect imperforate hymen due to presentation of hydrocolpos or hydrometrocolpos in the fetus if fetal cervical and vaginal secretions accumulate in response to circulating maternal estrogens. Nonetheless, reports of prenatal ultrasound diagnosis of hydrometrocolpos are still very rare. In this article we report a prenatally diagnosed of imperforate hymen with hydrometrocolpos.


Quality of life in survivors of adult haematological malignancy.
Survivors of haematological malignancies endure long-term effects of both treatment and disease. This paper examines factors that influence their quality of life through reporting on the results of a survey. Survey using previously validated quality of life questionnaires for use in cancer management. Participants were adults aged 18 and over who had completed treatment for a haematological malignancy and were between 1 and 5 years post-treatment. A total of 131 participants, median age of 66, completed questionnaires (66% response rate). Significant associations were found between age, global quality of life, physical and role functioning. Men reported better physical functioning and lower symptom scores than women. Employed participants reported better quality of life. Increasing age was associated with lowest quality of life. Best role functioning was also noted in participants who lived beyond 2.5 years following treatment completion. The survey suggested a gender difference with men reporting better physical functioning, fewer symptoms of pain and less loss of sleep compared with women. This study contributes to the underdeveloped area of care for and research into adult haematological cancer survivors. Knowledge and understanding of the factors that affect the quality of life of such adults may provide an insight into implementation measures.


[The single-layer uterine suture in cesarean section. A comparative study].
Caesarean section had to be carried out in 297 cases (4.9 per cent) out of 6064 births (infant mortality 0.44 per cent). A comparison between 96 Caesarean sections with a one-layer suture of the myometrium (32 per cent) and 201 Caesarean sections with a two-layer suture (68 per cent) clearly shows less morbidity (19 per cent vs. 37 per cent) in case of a one-layer uterus suture. In those cases the postoperative loss of blood, the involution of the uterus and the healing of the scar are also more favourable. Afebrile courses clearly predominate in one-layer sutures (92 per cent to 76 per cent in two-layer sutures). Most important of all are the ability and experience of the surgeon, not the use of antibiotica.


Low paraoxonase in Persian Gulf War Veterans self-reporting Gulf War Syndrome.
Exposure to organophosphate (OP's) insecticides and nerve gases during the Persian Gulf War has been implicated in the development of Gulf War Syndrome. Paraoxonase (PON1) present in human serum detoxifies OP's. We determined the levels of PON1 in the serum of Gulf War Veterans and compared these to those found in a control population. One hundred fifty-two Gulf War Veterans from the UK who self-reported the presence of Gulf War Syndrome via a questionnaire and 152 age and gender matched controls were studied. PON1 activity, concentration, and genotype were determined. In the Gulf War Veterans, paraoxon hydrolysis was less than 50% of that found in the controls (100.3 (14.8-233.8) vs 214.6 (50.3-516.2) nmol/min/ml, P < 0.001). This low activity was independent of the effect of PON1 genotype. The serum PON1 concentration was also lower in the Gulf War Veterans (75.7 (18.1-351.3) vs 88.2 (34.5-527.4) microg/ml, P < 0.00025), which was again independent of PON1 genotype. There was no difference in the rate of diazoxon hydrolysis between the groups (10. 2 +/- 4.1 micromol/min/ml vs 9.86 +/- 4.4, P = NS). A decreased capacity to detoxify OP insecticides resulting from low serum PON1 activity may have contributed to the development of Gulf War Syndrome.


Sinorhizobium medicae WSM419 Genes That Improve Symbiosis between Sinorhizobium meliloti Rm1021 and Medicago truncatula Jemalong A17 and in Other Symbiosis Systems.
Some soil bacteria, called rhizobia, can interact symbiotically with legumes, in which they form nodules on the plant roots, where they can reduce atmospheric dinitrogen to ammonia, a form of nitrogen that can be used by growing plants. Rhizobium-plant combinations can differ in how successful this symbiosis is: for example, Sinorhizobium meliloti Rm1021 forms a relatively ineffective symbiosis with Medicago truncatula Jemalong A17, but Sinorhizobium medicae WSM419 is able to support more vigorous plant growth. Using proteomic data from free-living and symbiotic S. medicae WSM419, we previously identified a subset of proteins that were not closely related to any S. meliloti Rm1021 proteins and speculated that adding one or more of these proteins to S. meliloti Rm1021 would increase its effectiveness on M. truncatula A17. Three genes, Smed_3503, Smed_5985, and Smed_6456, were cloned into S. meliloti Rm1021 downstream of the E. coli lacZ promoter. Strains with these genes increased nodulation and improved plant growth, individually and in combination with one another. Smed_3503, renamed iseA (increased symbiotic effectiveness), had the largest impact, increasing M. truncatula biomass by 61%. iseA homologs were present in all currently sequenced S. medicae strains but were infrequent in other Sinorhizobium isolates. Rhizobium leguminosarum bv. viciae 3841 containing iseA led to more nodules on pea and lentil. Split-root experiments with M. truncatula A17 indicated that S. meliloti Rm1021 carrying the S. medicae iseA is less sensitive to plant-induced resistance to rhizobial infection, suggesting an interaction with the plant's regulation of nodule formation. IMPORTANCE Legume symbiosis with rhizobia is highly specific. Rhizobia that can nodulate and fix nitrogen on one legume species are often unable to associate with a different species. The interaction can be more subtle. Symbiotically enhanced growth of the host plant can differ substantially when nodules are formed by different rhizobial isolates of a species, much like disease severity can differ when conspecific isolates of pathogenic bacteria infect different cultivars. Much is known about bacterial genes essential for a productive symbiosis, but less is understood about genes that marginally improve performance. We used a proteomic strategy to identify Sinorhizobium genes that contribute to plant growth differences that are seen when two different strains nodulate M. truncatula A17. These genes could also alter the symbiosis between R. leguminosarum bv. viciae 3841 and pea or lentil, suggesting that this approach identifies new genes that may more generally contribute to symbiotic productivity.


Estrogen receptor beta (ERβ) produces autophagy and necroptosis in human seminoma cell line through the binding of the Sp1 on the phosphatase and tensin homolog deleted from chromosome 10 (PTEN) promoter gene.
Testicular germ cell tumors are the most common tumor in male and the least studied. We focused on human seminoma using the TCAM2 cell line. Through ERβ, 10 nM estradiol (E2) was able to induce PTEN gene expression and promoter transactivation. Transient transfections, ChIP and EMSA assays evidenced the 5'-flanking region of PTEN gene promoter E2-responsive. The ERβ binding to the Sp1 on PTEN promoter decreased cell survival. The presence of ERβ or PTEN is necessary to induce the loss of cell survival upon E2, addressing their cooperation in this action. pAKT and AKT expression decreased under E2 and DPN, while known apoptotic markers appeared to be unchanged. The PI3K/AKT pathway inhibition also leads to autophagy: E2 and DPN enhanced the expression of autophagy-related markers such as PI3III, Beclin 1, AMBRA and UVRAG. MDC and TEM assays confirmed E2-induced autophagy. The absence of DNA fragmentation, caspase 9 and PARP1 cleavages suggested that necroptosis and/or parthanatos may occur. FACS analysis, LDH assay and RIP1 expression attested this hypothesis. Our study reveals a unique mechanism through which ERβ/PTEN signaling induces cell death in TCAM2 by autophagy and necroptosis. These data, supporting estrogen-dependency of human seminoma, propose ERβ ligands for therapeutic use in the treatment of this pathological condition.


Automated cover testing for binocular misalignment in awake monkeys using spectacle- mounted liquid crystal shutters.
In humans, pathologic misalignment of the eyes (strabismus, or hetero-tropia) is readily detected using a clinical technique in which the visual axis of each eye is blocked briefly by a hand held occluder (i.e., the cover test, AKA the single cover test, or unilateral cover test). This manual technique is impractical for use in non-human primates. This paper reports the electronic hardware and computer software we have devised and adapted to perform an automated cover test in awake macaque monkeys (Macaca mulatta or rhesus, and Macaca nemistrina or pigtail) to facilitate the study of strabismus in this species. Liquid crystal shutters (Displaytech LV2500OP-OEM) are mounted in aluminum spectacle frames which are positioned in front of the monkey's eyes by attaching the frame holder to an acrylic skull cap. The shutters are electrically actuated alternately to an "open" (transparent) or "closed" (opaque) state, occluding each eye in turn. The actuation signal (+/-5 volts) is generated by a single logic chip. The chip is driven by commercial data control and acquisition software (Spike 2, Cambridge Electronic Design) running on a Macintosh computer. Eye position and movement during this cover tests is monitored and recorded by the computer using surgically implanted scleral coils. Monkeys are rewarded for fixating small, moveable targets so that binocular misalignment can be measured with high precision at different or cardinal positions of horizontal and vertical gaze. The shutters produced occlusion of each eye as effective as that of an opaque plastic occluder used in previous experiments that required monocular viewing. Heterotropias were detected and recorded in monkeys and closely resembled those observed in human patients. It was also possible to detect heterophorias by actuating the shutters alternately. The shutters operated reliably, with negligible delay and transition times, providing automated, repeatable cover tests that are, in fact, in many respects superior to the standard clinical manual technique. In addition, their application to humans, using infrared electro-oculography instead of scleral coils to monitor eye position and movement, holds promise for future applications.


COVID-19 mortality in the United States: It's been two Americas from the start.
During the summer of 2021, a narrative of "two Americas" emerged: one with high demand for the COVID-19 vaccine and the second with widespread vaccine hesitancy and opposition to masks and vaccines. We analyzed "excess mortality" rates (the difference between total deaths and what would have been expected based on earlier time periods) prepared by the CDC for the United States from January 3, 2020 to September 26, 2021. Between Jan. 3, 2020 and Sept. 26, 2021, there were 895,693 excess deaths associated with COVID-19, 26% more than reported as such. The proportion of deaths estimated by the excess mortality method that was reported as COVID-19 was highest in the Northeast (92%) and lowest in the West (72%) and South (76%). Of the estimated deaths, 43% occurred between Oct. 4, 2020 and Feb. 27, 2021. Before May 31, 2020, approximately 56% of deaths were in the Northeast, where 17% of the population resides. Subsequently, 48% of deaths were in the South, which makes up 38% of the population. Since May 31, 2020, the South experienced COVID-19 mortality 26% higher than the national rate, whereas the Northeast's rate was 42% lower. If each region had the same mortality rate as the Northeast, more than 316,234 COVID-19 deaths between May 31, 2020 and Sept. 26, 2021 were "avoidable." More than half (63%) of the avoidable deaths occurred between May 31, 2020 and February, 2021, and more than half (60%) were in the South. Regional differences in COVID-19 mortality have been strong throughout the pandemic. The South has had higher mortality rates than the rest of the U.S. since May 31, 2020, and experienced 62% of the avoidable deaths. A comprehensive COVID-19 policy, including population-based restrictions as well as vaccines, is needed to control the pandemic.


Role of Oxidative Stress in Peyronie's Disease: Biochemical Evidence and Experiences of Treatment with Antioxidants.
Peyronie's disease (PD) is a chronic inflammatory condition affecting adult males, involving the tunica albuginea of the corpora cavernosa of the penis. PD is frequently associated with penile pain, erectile dysfunction, and a secondary anxious-depressive state. The etiology of PD has not yet been completely elucidated, but local injury is generally recognized to be a triggering factor. It has also been widely proven that oxidative stress is an essential, decisive component in all inflammatory processes, whether acute or chronic. Current conservative medical treatment comprises oral substances, penile injections, and physical therapy. This article intends to show how antioxidant therapy is able to interfere with the pathogenetic mechanisms of the disease. This article consists of a synthetic narrative review of the current scientific literature on antioxidant therapy for this disease. The good results of the antioxidant treatment described above also prove that the doses used were adequate and the concentrations of the substances employed did not exceed the threshold at which they might have interacted negatively with the mechanisms of the redox regulation of tissue. We believe new, randomized, controlled studies are needed to confirm the efficacy of treatment with antioxidants. However, we consider the experiences of antioxidant treatment which can already be found in the literature useful for the clinical practice of urologists in the treatment of this chronic inflammatory disease.


Cancer risk after renal transplantation in Japan.
Excess of cancer in patients receiving renal transplantation is well-known in Western countries, but information in Japan remains limited. Our study examined whether excess risk is found in patients receiving renal transplantation in Japan. Between 1970 and 1995, 1155 males and 589 females underwent renal transplantation in 6 hospitals, and a total of 12,982 person-years of observation was accumulated. Malignancies developed in 2.6% of patients; O/E ratio was 2.78. Median interval from renal transplantation to tumor development was 58 months. The interval in the patients receiving medication with cyclosporine-A (CyA) (median, 42.5 months) was significantly shorter than that with non-CyA (median, 95.5 months). Median age at the diagnosis of malignancy was 40 years, which is much younger than that in the general population. Relative risk was highest in renal cancer, followed by thyroid cancer, malignant lymphoma and uterine cancer. A distribution of malignancies was different from that reported from Western countries. These findings showed the excess risk of malignancies in Japan with renal transplants, especially in male patients, similar to that observed in Western countries, though the types of malignancy were different.


Measurement of absolute K X-ray emission intensities in the decay of 103mRh.
A new experiment was designed to measure the photon emission intensities in the decay of 103mRh. The rhodium samples were activated in the ISIS experimental nuclear reactor at CEA Saclay. The procedure includes an absolute activity measurement by liquid scintillation counting using the Triple-to-Double Coincidence Ratio method, followed by X-ray spectrometry using a high-purity germanium detector to determine the photon emission intensities. The new result (IX = 0.0825 (17)) is derived with a significant reduction of the uncertainty.


Development of an agar-diffusion method for the assay of quaternary ammonium germicides.
Five quaternary ammonium germicides (QAGs) were tested for their adsorption by agar. This was found to be in the following ascending order: alkylbenzylmethyl-ammonium chloride, alkyltrimethylammonium bromide, cetyltrimethylammonium bromide, cetylpyrimidinium chloride and cetylbenzyldimethylammonium chloride. An inverse relationship was established between the extent of agar binding of the QAGs and their inhibition zones. In an attempt to develop a sensitive cup-plate assaying technique suitable for QAGs, important factors affecting the agar-diffusion of QAGs were investigated. These included the influence of various polysorbates, buffer ions, agars and test organisms. Furthermore, the effect of the pH and/or the concentration of the selected polysorbate and the buffer were studied. The best medium developed for the sensitive agar-diffusion assay of QAGs was nutrient agar-Tris (0.05 M, pH 8) provided that distilled water and polysorbate 20 (0.5%) were used as diluents for the mixed alkyl and the pure cetyl QAGs, respectively.


[Suppression of hepatitis C virus (HCV) replication with serine palmitoyltransferase inhibitor].
Hepatitis C virus (HCV) persists chronically in most infected patients, eventually causing chronic hepatitis, liver cirrhosis, and in some cases hepatocellular carcinoma. The combination therapy of PEG-IFN and ribavirin improves efficacy in many patients, although it does not lead to sufficient achievements in genotype 1b patients. To aid in invention of new anti-HCV reagents, we focused on host factors that contributed to HCV lifecycle. We identified serine palmitoyltransferase inhibitor as an anti-HCV reagent through high-throughput screening using HCV replicon cells. We investigated the mechanism of anti-HCV effect of SPT inhibitor. It has been reported that sphingolipids and cholesterol compose the lipid raft where replication of HCV occurs. We investigated the influence of SPT inhibitor to lipid rafts by analyzing the detergent-resistant membrane (DRM). The analysis showed that SPT inhibitor moved HCV RNA-dependent RNA polymerase (NS5B) to detergent-soluble fraction from DRM, and Biacore analysis indicated binding of sphingomyelin to NS5B. These results suggest that SPT inhibitor disrupts the interaction between NS5B and sphingomyelin. Moreover, we evaluated the anti-HCV effect of SPT inhibitor in vivo with humanized chimeric mice. SPT inhibitor led to rapid decline in serum HCV-RNA of about 1-2 log within 8 days. Furthermore, combination therapy of SPT inhibitor and PEG-IFN achieved about 3 log reduction in serum HCV-RNA.


Impact of facility type and volume in locally advanced esophageal cancer.
We hypothesized that academic facilities and high-volume facilities would be independently associated with improved survival and a greater propensity for performing surgery in locally advanced esophageal cancer. We identified patients diagnosed with stage IB-III esophageal cancer during 2004-2016 from the National Cancer Database. Facility type was categorized as academic or community, and facility volume was based on the number of times a facility's unique identification code appeared in the dataset. Each facility type was dichotomized into high- and low-volume subgroups using the cutoff of 20 esophageal cancers treated/year. We fitted multivariable regression models in order to assess differences in surgery selection and survival between facilities according to type and volume. Compared to patients treated at high-volume community hospitals, those at high-volume academic facilities were more likely to undergo surgery (odds ratio: 1.865, p < 0.001) and were associated with lower odds of death (odds ratio: 0.784, p = 0.004). For both academic and community hospitals, patients at high-volume facilities were more likely to undergo surgery compared to those at low-volume facilities, p < 0.05. For patients treated at academic facilities, high-volume facilities were associated with lower odds of death (odds ratio: 0.858, p = 0.02) compared to low-volume facilities, while there was no significant difference in the odds of death between high- and low-volume community hospitals (odds ratio: 1.018, p = 0.87). Both facility type and case volume impact surgery selection and survival in locally advanced esophageal cancer. Compared to community hospitals, academic facilities were more likely to perform surgery and were associated with improved survival.


Tubulogenesis in a simple cell cord requires the formation of bi-apical cells through two discrete Par domains.
Apico-basal polarization is a crucial step in the de novo formation of biological tubes. In Ciona notochord, tubulogenesis occurs in a single file of cells in the absence of cell proliferation. This configuration presents a unique challenge for the formation of a continuous lumen. Here, we show that this geometric configuration is associated with a novel polarization strategy: the generation of bipolar epithelial cells possessing two apical/luminal domains instead of one, as in the conventional epithelium. At the molecular level, cells establish two discrete Par3/Par6/aPKC patches, and form two sets of tight junctions, in opposite points of the cells. The key molecule controlling the formation of both domains is Par3. Changing the position of the cells within the organ fundamentally changes their polarity and the number of apical domains they develop. These results reveal a new mechanism for tubulogenesis from the simplest cell arrangement, which occurs in other developmental contexts, including vertebrate vascular anastomosis.


A comparative investigation of modern feature selection and classification approaches for the analysis of mass spectrometry data.
Many analytical approaches such as mass spectrometry generate large amounts of data (input variables) per sample analysed, and not all of these variables are important or related to the target output of interest. The selection of a smaller number of variables prior to sample classification is a widespread task in many research studies, where attempts are made to seek the lowest possible set of variables that are still able to achieve a high level of prediction accuracy; in other words, there is a need to generate the most parsimonious solution when the number of input variables is huge but the number of samples/objects are smaller. Here, we compare several different variable selection approaches in order to ascertain which of these are ideally suited to achieve this goal. All variable selection approaches were applied to the analysis of a common set of metabolomics data generated by Curie-point pyrolysis mass spectrometry (Py-MS), where the goal of the study was to classify the Gram-positive bacteria Bacillus. These approaches include stepwise forward variable selection, used for linear discriminant analysis (LDA); variable importance for projection (VIP) coefficient, employed in partial least squares-discriminant analysis (PLS-DA); support vector machines-recursive feature elimination (SVM-RFE); as well as the mean decrease in accuracy and mean decrease in Gini, provided by random forests (RF). Finally, a double cross-validation procedure was applied to minimize the consequence of overfitting. The results revealed that RF with its variable selection techniques and SVM combined with SVM-RFE as a variable selection method, displayed the best results in comparison to other approaches.


Salt preference and the risk of gastrointestinal cancers.
The relationship between the consumption of salt and the risk of gastrointestinal cancer was examined using data from a case-control study conducted in northern Italy. The study was conducted on 526 incident cases of gastric cancer, 621 of colon cancers, 382 of rectal cancers, and 1,223 controls admitted to hospital for acute, nonneoplastic, nondigestive tract disorders. Compared with those for people with low salt intakes, the relative risks (and 95% confidence intervals) for those with intermediate and high salt intakes were 1.3 (1.0-1.8) and 1.2 (0.8-1.7) for stomach, 1.0 (0.8-1.3) and 1.1 (0.8-1.5) for colon, and 1.2 (0.9-1.6) and 0.9 (0.6-1.3) for rectum. None of the trends in risk were statistically significant, and the risk estimates were not materially modified by allowance for major identified potential confounding factors. Thus, this study gives little support to the existence of any strong association between salt intake and gastrointestinal cancers. However, in view of the results obtained and of the uncertainties of salt intake measurement, the possibility exists of a real, although moderate, association of salt intake with gastric cancer.


Fundus autofluorescence findings in a mouse model of retinal detachment.
Fundus autofluorescence (fundus AF) changes were monitored in a mouse model of retinal detachment (RD). RD was induced by transscleral injection of hyaluronic acid (Healon) or sterile balanced salt solution (BSS) into the subretinal space of 4-5-day-old albino Abca4 null mutant and Abca4 wild-type mice. Images acquired by confocal scanning laser ophthalmoscopy (Spectralis HRA) were correlated with spectral domain optical coherence tomography (SD-OCT), infrared reflectance (IR), fluorescence spectroscopy, and histologic analysis. Results. In the area of detached retina, multiple hyperreflective spots in IR images corresponded to punctate areas of intense autofluorescence visible in fundus AF mode. The puncta exhibited changes in fluorescence intensity with time. SD-OCT disclosed undulations of the neural retina and hyperreflectivity of the photoreceptor layer that likely corresponded to histologically visible photoreceptor cell rosettes. Fluorescence emission spectra generated using flat-mounted retina, and 488 and 561 nm excitation, were similar to that of RPE lipofuscin. With increased excitation wavelength, the emission maximum shifted towards longer wavelengths, a characteristic typical of fundus autofluorescence. In detached retinas, hyper-autofluorescent spots appeared to originate from photoreceptor outer segments that were arranged within retinal folds and rosettes. Consistent with this interpretation is the finding that the autofluorescence was spectroscopically similar to the bisretinoids that constitute RPE lipofuscin. Under the conditions of a RD, abnormal autofluorescence may arise from excessive production of bisretinoid by impaired photoreceptor cells.


[Dehiscence of the superior semicircular canal: approach and CT scan classifications].
The syndrome of dehiscence of the superior semicircular canal (DCSS) is primarily associated with vertigo and/or hearing loss. The dehiscence may be completely asymptomatic and represent an incidental finding on radiological investigation. To demonstrate the advantages of a volume rendered CT study of the petrous temporal bone of patients with hearing loss, and to demonstrate the effectiveness of its systematic application in the protocols of examination. To propose a radiological classification of DCSS with a therapeutic application. The examination technique which was performed in incremental mode (axial and frontal sections) and in "volume rendered" mode, on a high resolution apparatus is described. The authors studied 154 scans of the petrous temporal bone obtained by this technique. They correlated the cases of DCSS with the indications for the radiological examination. Each 3d CT scan was studied and the type of fistula described. The authors propose a classification of fistulae into three types, depending on 3d CT scan appearance. Out of 154 CT scans of the petrous temporal bone (77 patients), 13 cases of DCSS were discovered. DCSS was bilateral in 4 cases. The primary indication for investigation was the assessment of conductive or mixed hearing loss. The "volumetric" technique was compared with standard imaging techniques and/or reconstructed images in the superior canal plane. The correlation was perfect in all the cases. The description of the fistulae allowed a classification into 3 types: Type I (symmetrical fistula, 8 cases); Type II (asymmetrical fistula, 3 cases) corresponding to the canal dome; Type III (2 cases) involving the foot of the canal. The increased frequency of DCSS in this series (prevalence of 17% against 0.5% in post mortem studies) is probably explained by the selection bias of the patients and also by the systematic application of this novel radiological technique. We propose to include this protocol in all CT scans of the temporal bone, particularly when investigating symptoms consistent with a syndrome of Minor or the Tullio phenomenom. This system of classification makes it possible to describe the fistula and to specify its location. This should prove to be a valuable aid for pre-operative planning and intra-operative localisation of the fistula.


Silylation of C-H bonds in aromatic heterocycles by an Earth-abundant metal catalyst.
Heteroaromatic compounds containing carbon-silicon (C-Si) bonds are of great interest in the fields of organic electronics and photonics, drug discovery, nuclear medicine and complex molecule synthesis, because these compounds have very useful physicochemical properties. Many of the methods now used to construct heteroaromatic C-Si bonds involve stoichiometric reactions between heteroaryl organometallic species and silicon electrophiles or direct, transition-metal-catalysed intermolecular carbon-hydrogen (C-H) silylation using rhodium or iridium complexes in the presence of excess hydrogen acceptors. Both approaches are useful, but their limitations include functional group incompatibility, narrow scope of application, high cost and low availability of the catalysts, and unproven scalability. For this reason, a new and general catalytic approach to heteroaromatic C-Si bond construction that avoids such limitations is highly desirable. Here we report an example of cross-dehydrogenative heteroaromatic C-H functionalization catalysed by an Earth-abundant alkali metal species. We found that readily available and inexpensive potassium tert-butoxide catalyses the direct silylation of aromatic heterocycles with hydrosilanes, furnishing heteroarylsilanes in a single step. The silylation proceeds under mild conditions, in the absence of hydrogen acceptors, ligands or additives, and is scalable to greater than 100 grams under optionally solvent-free conditions. Substrate classes that are difficult to activate with precious metal catalysts are silylated in good yield and with excellent regioselectivity. The derived heteroarylsilane products readily engage in versatile transformations enabling new synthetic strategies for heteroaromatic elaboration, and are useful in their own right in pharmaceutical and materials science applications.


Treatment engagement of depressed patients with and without psychosis in a partial hospital program: Dropout, symptom reduction, and satisfaction.
The ways patients with psychosis and depression engage in therapeutic treatment is not well understood. To determine if an intensive outpatient psychotherapy program could benefit patients experiencing psychotic symptoms, it is important to know how these individuals engage with psychotherapeutic treatment. The present study from the Rhode Island Hospital Methods to Improve Diagnostic Assessment and Services (MIDAS) project compared dropout rates, treatment response, and satisfaction among 219 individuals with psychosis and major depressive disorder (MDD) to 2,545 individuals with MDD at a general, Acceptance and Commitment Therapy-based partial hospital program (PHP). Those with psychosis were significantly less likely to complete treatment. Approximately one-fifth of all patients experienced at least a 50% reduction in depressive and anxiety symptoms. The vast majority of patients with psychosis were highly satisfied with treatment. Findings suggest patients with psychosis have a higher risk of premature dropout. Patients with psychosis demonstrated a reduction in symptoms during PHP treatment and self-reported high satisfaction with treatment. This study calls for the implementation of practices to reduce premature dropout for patients with psychosis, and for future research on the effectiveness of general psychiatric treatment for those with psychotic symptoms.


Knowledge and Attitude Regarding National Tobacco Control Law and Practice of Tobacco Smoking among Bangladesh Police.
Tobacco is an important public health problem of Bangladesh having both high production and consumption. Police are engaged in enforcement of the tobacco control law. For effective enforcement of national tobacco control law (NTCL), they should have sound knowledge of tobacco control law and their attitude to the tobacco control law must be a positive one. The objective of the study was to explore the status of knowledge and attitude regarding national tobacco control law and practice of tobacco smoking among Bangladesh Police. The descriptive cross sectional study was conducted among 440 members of Bangladesh Police during the period of March 2015 to November 2015. Police personnel of different ranks were interviewed randomly using a semi-structured questionnaire from the selected SP offices, police lines, police stations and police camps. Among the respondents 414(94.1%) were male and 26(5.9%) were female. About half (214, 48.6%) of the respondents never smoked in life, while 226(51.4%) smoked in life even one or two puffs. There was no female among the ever smokers. One quarter (114, 25.9%) of the respondents ever smoked but quitted and 112(25.4%) respondents were currently smoker. Among the current smokers 31(7.0%) were occasional, while 81 (18.4%) were daily smoker. It was observed that there was lack of knowledge among police personnel regarding various rules and clauses of national tobacco control law. Most of the respondents (434, 98.6%) showed positive attitude regarding rationality of prohibition of smoking in public places and public transports. The most of the respondents (418, 95.0%) agreed that there should not be any advertisement of tobacco products in cinema hall, radio and television channel. A highest majority (377, 85.7%) of the respondents gave their opinion that Government should increase taxes on tobacco products. Most of the respondents (433, 94.4%) opined against the selling of tobacco products to the minor. Almost every respondent (436, 99.1%) wanted proper implementation of tobacco control law. Nearly half of the respondents (213, 48.4%) thought that smoking among police was one of the obstacles in effective implementation of tobacco control law. An overwhelming majority of the respondents (435, 98.9%) agreed that police personnel should not smoke tobacco for keeping their health and fitness and thereby for better performance. Almost all respondents (436, 99.1%) gave a positive opinion that police personnel may be a role model for the public by not smoking tobacco. Most of the respondents (427, 97.1%) think that orientation or training program on tobacco control law should be arranged for police personnel. More than one fourth (25.5%) of the police personnel in Bangladesh are currently smokers and this should be intervened. Smoke-free police station initiative is started but not being 100% implemented. Lack of knowledge regarding various rules and clauses of NTCL was sufficient enough to draw intervention in this regard. Positive attitude of police personnel towards NTCL and smoke free police station initiative will help to implement tobacco control law properly.


Preparation and characterization of Ni-nitrilotriacetic acid bearing poly(methacrylic acid) coated superparamagnetic magnetite nanoparticles.
Stable superparamagnetic magnetite (Fe3O4) nanoparticles were synthesized via co-precipitation in the presence of poly(methacrylic acid) (PMAA) in aqueous solution. The polymer coated Fe3O4 nanoparticles were characterized using transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction, thermal analysis, and vibrating sample magnetometry (VSM) techniques. These measurements reveal the presence of magnetite nanoparticles with a size of approximately 8 nm inside the PMAA matrix. The magnetization value of these superparamagnetic nanoparticles at room temperarure and 7 T was measured as about 40 emu/g. PMAA-coated Fe3O4 nanoparticles were further assembled with Ni-chelate through a reaction between a primary amine-bearing NTA (nitrilotriacetic acid) ligand and carboxy-functional groups of PMAA. NTA-PMAA-coated magnetite nanoparticles were then loaded with nickel ions and characterized using FTIR. The average amount of binded Ni on the surface of the NTA-modified PMAA coated Fe3O4 was calculated as 1.65 +/- 0.3 x 10(-6) mol nickel(II) ions per g of the magnetic particles from the inductively coupled plasma optical emission spectroscopy (ICP-OES) measurements.


The factor structure of social cognition in schizophrenia: Weak evidence for separable domains.
This study examined the factor structure of social cognition in a Norwegian sample of individuals diagnosed with schizophrenia (n = 83). Eight variables from three social cognitive tests from three theoretical domains were included: emotion processing, social perception and theory of mind. Factor analysis with maximum likelihood extraction and oblique rotation resulted in two factors using Kaiser's criterion. Although the two-factor model had better fit than a unifactorial model, it did not represent the data well. Two social cognitive variables did not load on either factor. The two extracted factors did not correspond to an expected distinction between low and high level of processing or between affective and cognitive processes. A non-negligible number of nonredundant residuals between observed and computed correlations suggested poor model fit. In conclusion, this study failed to identify separable dimensions of social cognition in spite of including measures from different theoretical domains.


A new pro-prodrug aminoacid-based for trans-ferulic Acid and silybin intestinal release.
The aim of this work was the preparation and characterization of a pro-prodrug able to simultaneously transport silybin, a drug possessing various pharmacological effects, and trans-ferulic acid, a known antioxidant. More specifically, l-phenylalanine-N-(4-hydroxy-3-methoxy-phenyl) prop-2-en-O-(2R,3R)-3,5,7-trihydroxy-2-((2R,3R)-3-(4-hydroxy-3-methoxyphenyl)-2-(hydroxymethyl)-2,3-dihydro-benzo-(1,4)-dioxin-6-yl)croman-4-one was synthesized by using the aminoacid l-phenylalanine (l-Phe) as carrier. Indeed, l-Phe is characterized by an intrinsic chemical reactivity due to the presence of an amino group, placed on the chiral center, and of a carboxylic group. The synthesis has been characterized first by adding silybin by means of carboxylic group and then, with the aim to confer antioxidant properties to this new carrier, by linking trans-ferulic acid to l-Phe via amino group. The so obtained derivative was then characterized by FT-IR, and 1H-NMR spectroscopies. Furthermore, its ability to inhibit lipid peroxidation induced by tert-butyl hydroperoxide in rat liver microsomes, was evaluated. The 1,1-diphenyl-2-picrylhydrazyl radical-scavenging effect, was also assessed. The release of silybin and trans-ferulic acid was determined in simulated gastric and intestinal fluids over the time. The results showed that the covalent bond between both (i) silybin; or (ii) trans-ferulic acid and the amino acid was degraded by enzymatic reactions. In addition, the pro-prodrug, showed strong antioxidant and scavenger activities. Due to these properties, this new pro-prodrug could be applied for the treatment of intestinal pathologies and it might improve the therapeutic potential of silybin which is strongly limited by its low solubility.


Expression of endothelial nitric oxide synthase in developing rat kidney.
Endothelium-derived nitric oxide (NO) is synthesized within the developing kidney and may play a crucial role in the regulation of renal hemodynamics. The purpose of this study was to establish the expression and intrarenal localization of the NO-synthesizing enzyme endothelial NO synthase (eNOS) during kidney development. Rat kidneys from 14 (E14)-, 16 (E16)-, 18 (E18)-, and 20-day-old (E20) fetuses and 1 (P1)-, 3 (P3)-, 7 (P7)-, 14 (P14)-, and 21-day-old (P21) pups were processed for immunocytochemical and immunoblot analysis. In fetal kidneys, expression of eNOS was first observed in the endothelial cells of the undifferentiated intrarenal capillary network at E14. At E16, strong eNOS immunoreactivity was observed in the endothelial cells of renal vesicles, S-shaped bodies (stage II glomeruli), and stage III glomeruli at the corticomedullary junction. At E18-20, early-stage developing glomeruli located in the subcapsular region showed less strong eNOS immunoreactivity than those of E16. The eNOS-positive immature glomeruli were observed in the nephrogenic zone until 7 days after birth. In fetal kidneys, eNOS was also expressed in the medulla in the endothelial cells of the capillaries surrounding medullary collecting ducts. After birth, eNOS immunostaining gradually increased in the developing vascular bundles and peritubular capillaries in the medulla and was highest at P21. Surprisingly, eNOS was also expressed in proximal tubules, in the endocytic vacuolar apparatus, only at P1. The strong expression of eNOS in the early stages of developing glomeruli and vasculature suggests that eNOS may play a role in regulating renal hemodynamics of the immature kidney.


Risks of infection, hospital and ICU admission, and death from COVID-19 in people with asthma: systematic review and meta-analyses.
To determine if and to what degree asthma may predispose to worse COVID-19 outcomes in order to inform treatment and prevention decisions, including shielding and vaccine prioritisation. Systematic review and meta-analysis. Electronic databases were searched (October 2020) for clinical studies reporting at least one of the following stratified by asthma status: risk of infection with SARS-CoV-2; hospitalisation, intensive care unit (ICU) admission or mortality with COVID-19. Adults and children who tested positive for or were suspected to have COVID-19. Main outcome measures were the following stratified by asthma status: risk of infection with SARS-CoV-2; hospitalisation, ICU admission or mortality with COVID-19. We pooled odds ratios (ORs) and presented these with 95% confidence intervals (CI). Certainty was assessed using GRADE (Grading of Recommendations, Assessment, Development and Evaluations). 30 (n=112 420) studies were included (12 judged high quality, 15 medium, 3 low). Few provided indication of asthma severity. Point estimates indicated reduced risks in people with asthma for all outcomes, but in all cases the evidence was judged to be of very low certainty and 95% CIs all included no difference and the possibility of increased risk (death: OR 0.90, 95% CI 0.72 to 1.13, I2=58%; hospitalisation: OR 0.95, 95% CI 0.71 to 1.26; ICU admission: OR 0.96, 95% CI 0.75 to 1.24). Findings on hospitalisation are also limited by substantial unexplained statistical heterogeneity. Within people with asthma, allergic asthma was associated with less COVID-19 risk and concurrent chronic obstructive pulmonary disease was associated with increased risk. In some studies, corticosteroids were associated with increased risk, but this may reflect increased risk in people with more severe asthma. Though absence of evidence of a clear association between asthma and worse outcomes from COVID-19 should not be interpreted as evidence of absence, the data reviewed indicate that risks from COVID-19 in people with asthma, as a whole, may be less than originally anticipated.


[Structural-functional research on polymyxins. 1H-NMR spectra of polymyxin B and its shortened analog].
Polymyxin B and its shortened analog were studied comparatively by 1H-NMR spectroscopy. Analysis of the signal chemical shifts, constants of spin-spin interaction of 3J HN-C alpha H and temperature coefficients of the NH signal chemical shifts revealed absolute structural identity of both molecules cyclic parts. This proved that there was no conformative interaction between the cyclic and linear parts of the polymyxin B molecule. Comparison of the results with the data on the biological activity showed that the hydrophobic N-end moiety of the polymyxin B molecule played a specific role in its antibacterial effect and toxicity.


Effects of hematoma on the short-term fate of experimental microvenous autografts.
The early effects of hematoma on the healing pattern of interpositional vein grafts were studied in 60 rats. In all animals, vein grafts approximately 10-mm in length were used to bridge defects created in the femoral artery and vein on the same side, with a 1:1 graft-to-host diameter match. Grafted vessels were completely covered with hematoma and separated from the surrounding tissue in 30 animals, while the other 30 rats served as controls. Both groups were divided into six subgroups of five animals each, according to the age of the grafts: 1, 3, 5, 7, 14, and 21 days. Graft patency and healing were assessed both clinically and histologically 20/30 (66.6 percent) of the interarterial and 27/30 (90 percent) of the intervenous vein grafts in Group 1 (hematoma group), and 28/30 (93.3 percent) of the interarterial and 29/30 196.6 percent) of the intervenous vein grafts in Group 2 (controls) were found patent. The patency rate of the interarterial vein grafts in the hematoma group was statistically significantly lower than in the control group and than in the intervenous vein grafts of the same group (p<0.005). The majority of interarterial vein-graft failures occurred after postoperative day 7 in Group 1. Patent specimens were examined under scanning electron and light microscopy. The authors concluded that perivascular hematoma caused spasm and flow disturbance, prolonged vessel-wall ischemia, and severe vessel-wall injury in the arterial circulation of the microvenous grafts. It also delayed the healing process and subjected the grafts to the development of occlusive mural thrombus. In contrast, the healing pattern of the intervenous vein grafts was not markedly influenced by perivascular hematoma, and equilibrium between thrombogenic and antithrombogenic factors on the graft surface was restored over a short period of time.


Investigating the numerical effects of ascertainment bias in linkage analysis: development of methods and preliminary results.
It is general practice to have nonsingle ascertainment of pedigrees for linkage studies, along with intrafamilial sampling that is dependent on who among the related individuals was initially ascertained (Proband dependent or PD sampling). Vieland and Hodge [1995; 1996] have shown that under these conditions, the likelihood used in calculating the lod score is not strictly correct and can produce asymptotically biased estimates of the recombination fraction, theta. However they speculated that this bias would be small in most applications. This paper presents preliminary work aimed at quantifying the numerical magnitude of the bias introduced by PD sampling and nonsingle ascertainment in linkage analysis. We considered five generating models where we varied the ascertainment procedure, intrafamilial sampling scheme, and the sample size for each model. In this limited initial set of simulations, asymptotic bias in theta appears to be trivial, while PD sampling procedures can increase the efficiency of theta. These preliminary results support the view that the advantages of unsystematic ascertainment may offset any small estimation bias that may arise.


Recombination, statistical power, and genetic studies of sexual isolation in Drosophila.
Genetic studies of sexual isolation in Drosophila have generally failed to fully evaluate the effects of their sample size and recombination between markers on their conclusions. In this study we evaluate recombinational distances between markers in Drosophila pseudoobscura and D. persimilis, a species pair in which numerous genetic mapping studies have been performed. We conclude that, contrary to assertions, the inversions that distinguish these two species still allow for much recombination within most of their chromosome arms in F1 hybrid females. Such recombination may have caused previous mapping studies in these species to miss (or grossly underestimate) the effects of several genomic regions. We also evaluate the effects of sample size and recombination on genetic studies of sexual isolation in other Drosophila species groups. We conclude that some of these studies may have been heavily biased toward detecting only genes of large effect. Future studies of sexual isolation should be preceded by detailed statistical power analyses that determine the effects of recombination and sample size in the species pair being studied to avoid these complications.


Efficacy of Pulsatile Flow Perfusion in Adult Cardiac Surgery: Hemodynamic Energy and Vascular Reactivity.
Background: The role of pulsatile (PP) versus non-pulsatile (NP) flow during a cardiopulmonary bypass (CPB) is still debated. This study's aim was to analyze hemodynamic effects, endothelial reactivity and erythrocytes response during a CPB with PP or NP. Methods: Fifty-two patients undergoing an aortic valve replacement were prospectively randomized for surgery with either PP or NP flow. Pulsatility was evaluated in terms of energy equivalent pressure (EEP) and surplus hemodynamic energy (SHE). Systemic (SVRi) and pulmonary (PVRi) vascular resistances, endothelial markers levels and erythrocyte nitric-oxide synthase (eNOS) activity were collected at different perioperative time-points. Results: In the PP group, the resultant EEP was 7.3% higher than the mean arterial pressure (MAP), which corresponded to 5150 ± 2291 ergs/cm3 of SHE. In the NP group, the EEP and MAP were equal; no SHE was produced. The PP group showed lower SVRi during clamp-time (p = 0.06) and lower PVRi after protamine administration and during first postoperative hours (p = 0.02). Lower SVRi required a higher dosage of norepinephrine in the PP group (p = 0.02). Erythrocyte eNOS activity results were higher in the PP patients (p = 0.04). Renal function was better preserved in the PP group (p = 0.001), whereas other perioperative variables were comparable between the groups. Conclusions: A PP flow during a CPB results in significantly lower SVRi, PVRi and increased eNOS production. The clinical impact of increased perioperative vasopressor requirements in the PP group deserves further evaluation.


Using a Complex Network Methodology to Track, Evaluate, and Transform Fundamental Care.
In this study, we drew on methods originating in complex adaptive systems and social network analysis to develop a novel way to quantify fundamental care. Data were obtained from a public statement from the Australian Royal Commission into Aged Care Quality and Safety. Results support the importance of using a systemic approach to assess the multiple dimensions of the fundamentals of care. Our method allows measurement of the problem within its system, providing a detailed quantification of care events and identifying excellence and improvement opportunities. We illustrate the strengths of this approach using principal component analysis and heat mapping. The application of the proposed methodology in healthcare decision-making, planning, and quality improvement is discussed.


Biochemical and genetic requirements for function of the immune response regulator BOTRYTIS-INDUCED KINASE1 in plant growth, ethylene signaling, and PAMP-triggered immunity in Arabidopsis.
Arabidopsis thaliana BOTRYTIS-INDUCED KINASE1 (BIK1) regulates immune responses to a distinct class of pathogens. Here, mechanisms underlying BIK1 function and its interactions with other immune response regulators were determined. We describe BIK1 function as a component of ethylene (ET) signaling and PAMP-triggered immunity (PTI) to fungal pathogens. BIK1 in vivo kinase activity increases in response to flagellin peptide (flg22) and the ET precursor 1-aminocyclopropane-1-carboxylic acid (ACC) but is blocked by inhibition of ET perception. BIK1 induction by flg22, ACC, and pathogens is strictly dependent on EIN3, and the bik1 mutation results in altered expression of ET-regulated genes. BIK1 site-directed mutants were used to determine residues essential for phosphorylation and biological functions in planta, including PTI, ET signaling, and plant growth. Genetic analysis revealed flg22-induced PTI to Botrytis cinerea requires BIK1, EIN2, and HUB1 but not genes involved in salicylate (SA) functions. BIK1-mediated PTI to Pseudomonas syringae is modulated by SA, ET, and jasmonate signaling. The coi1 mutation suppressed several bik1 phenotypes, suggesting that COI1 may act as a repressor of BIK1 function. Thus, common and distinct mechanisms underlying BIK1 function in mediating responses to distinct pathogens are uncovered. In sum, the critical role of BIK1 in plant immune responses hinges upon phosphorylation, its function in ET signaling, and complex interactions with other immune response regulators.


Chemotherapy for gynecological malignancies in organ transplantation patients: report of two cases.
Long-term risk of gynecological malignancies in organ transplantation patients has increased compared with that of the general population owing to the use of immunosuppressive agents. Treatment, especially chemotherapy, in these patients should take into consideration their renal function and the effects of immunosuppressive agents. We here present two case reports of patients with chemotherapy-treated gynecological malignancies who had previously received organ transplantation. The first case, a rare occurrence of simultaneous carcinomas of the uterine corpus and ovary, is the first such report in the English literature describing chemotherapy for concurrent serous papillary ovarian carcinoma and endometrioid endometrial carcinoma in a renal transplant patient. The second case report, describing chemotherapy for cervical cancer following two organ transplantation, also rare, is the first such report in the English literature and the first report of cervical cancer after heart-kidney transplantation.


"Stand Up and Do Something": Exploring Students' Perspectives on Bystander Intervention.
Bystander education is an increasingly popular primary prevention strategy for preventing sexual and relationship violence on college campuses. Although the evidence base for this primary prevention approach has grown substantially in recent years, more research is needed that accounts for changes in students' readiness to help and helping behaviors over time. In addition, because there is still much to learn about the factors that contribute to these changes in behavior, it is important to hear from students in their own words about their experiences with bystander education. This article presents qualitative data from a group of students who participated in a bystander intervention education program over an 18-month period and explores what conclusions can be drawn about the impact of this program on student attitudes, self-perception, and behaviors related to preventing sexual violence on a college campus. Themes that emerged from the data include students' increased knowledge about sexual violence, perceived changes in attitude about sexual assault on campus, students' increased desire to help in situations that could escalate to sexual assault, actual helping, and students' developing sense of themselves as persons of integrity and action.


A dosing regimen for immediate N-acetylcysteine treatment for acute paracetamol overdose.
Current treatment of paracetamol (acetaminophen) poisoning involves initiating a 3-phase N-acetylcysteine (NAC) infusion after comparing a plasma concentration, taken ≥ 4 h post-overdose, to a nomogram. This may result in dosing errors, a delay in treatment, or possibly more adverse effects - due to the use of a high dose rate for the first infusion when treatment is initiated. Our aim was to investigate a novel dosing regimen for the immediate administration of NAC on admission at a lower infusion rate. We used a published population pharmacokinetic model of NAC to simulate a scenario where a patient presents to the hospital 2 h post-overdose. The conventional regimen is commenced 6 h post-overdose when the 4-h plasma paracetamol concentration is available. We investigated an NAC infusion using a lower dosing rate initiated immediately on presentation. We determined a dosing rate that gave an area under the curve (AUC) of the concentration-time curve that was the same or greater than that from the conventional regimen on 90% of occasions. Lower dosing rates of NAC initiated immediately resulted in a similar exposure to NAC. An infusion of 110 mg/kg over the first 5 h (22 mg/kg/h) followed by the last two phases of the conventional regimen, or 200 mg/kg over 9 h (22.6 mg/kg/h) followed by the last phase of the conventional regimen could be used. The novel dosing regimen allowed immediate treatment of a patient using a lower dosing rate. This greatly simplifies the current dosing regimen and may reduce NAC adverse effects while ensuring the same amount of NAC is delivered.


Multiple Parameters Beyond Lipid Binding Affinity Drive Cytotoxicity of Cholesterol-Dependent Cytolysins.
The largest superfamily of bacterial virulence factors is pore-forming toxins (PFTs). PFTs are secreted by both pathogenic and non-pathogenic bacteria. PFTs sometimes kill or induce pro-pathogen signaling in mammalian cells, all primarily through plasma membrane perforation, though the parameters that determine these outcomes are unclear. Membrane binding, calcium influx, pore size, and membrane repair are factors that influence PFT cytotoxicity. To test the contribution of membrane binding to cytotoxicity and repair, we compared the closely related, similarly-sized PFTs Perfringolysin O (PFO) from Clostridium perfringens and Streptolysin O (SLO) from Streptococcus pyogenes. Cell death kinetics for PFO and SLO were different because PFO increased in cytotoxicity over time. We introduced known L3 loop mutations that swap binding affinity between toxins and measured hemolytic activity, nucleated cell death kinetics and membrane repair using viability assays, and live cell imaging. Altered hemolytic activity was directly proportional to toxin binding affinity. In contrast, L3 loop alterations reduced nucleated cell death, and they had limited effects on cytotoxicity kinetics and membrane repair. This suggests other toxin structural features, like oligomerization, drives these parameters. Overall, these findings suggest that repair mechanisms and toxin oligomerization add constraints beyond membrane binding on toxin evolution and activity against nucleated cells.