Challenges faced in implementation of a telehealth enabled chronic wound care system.
In the rural Midwest region of Western Australia (WA), wound care is a major burden on the healthcare system. Optimal wound care was found to be impeded by issues that included the involvement of multiple healthcare providers, incomplete and inconsistent documentation, and limited access to expert review. A telehealth solution was trailed in 2007. To describe the systemic barriers encountered in implementing a telehealth program in rural WA and to provide recommendations for future telehealth initiatives. This study trialled the use of a shared electronic wound imaging and reporting system in combination with an expert remote wound consultation service for the management of patients with chronic wounds in the Midwest of WA. The trial sites included rural hospital out-patient clinics, a private domiciliary nursing service, residential aged care facilities, general practices and a podiatry clinic. The implementation conformed to accepted best practice in introducing telehealth initiatives. During the trial 12 sites had the relevant software installed and were able to access a central server. Although a total of 41 patients with chronic wounds were enrolled, four sites did not enroll any patients and only two sites successfully incorporated the system into regular practice. Major obstacles were workforce issues and significant delays in installing the software at some sites. Only 47% of the healthcare providers trained to use the software at the beginning of the trial were still employed when the trial ended. Prolonged periods of vacant positions at one remote clinic and an aged care facility made it impossible for the remaining providers to allocate time for using the wound care software. The disease burden of the patient group, funding models and workforce shortages frustrated the successful adoption of an evidence based strategy that was known to improve health outcomes.


Comparative FISH mapping of 32 loci reveals new homologous regions between donkey and horse karyotypes.
A total of 32 loci comprising specific genes, microsatellites and anonymous BAC clones from horse and cattle were mapped on donkey chromosomes. Of these, 13 markers were also mapped for the first time in the horse. This information, together with that previously available in donkey and horse updates the comparative status of the karyotypes of the two species. The findings of the present study for the first time show correlation between eleven equine acrocentric autosomes and the donkey chromosomes and in part enable detection of rearrangements between them. There are still 7-8 pairs of chromosomes/arms for which no correspondence is known. At least 20 chromosome rearrangements (inversions, fusions and fissions) are already identified that differentiate the two karyotypes. More will be known once complete correspondence is deduced between them. These observations match similar differences observed between human-gibbon and mouse-rat karyotypes that show considerable rearrangements in relation to each other. How donkey and horse karyotypes gathered these differences within a short period of 5-10 Myr since divergence from a common ancestor will be known only after an ancestral equid karyotype is deduced, and the direction of change leading to chromosome rearrangements is clearly understood.


Fullerene rotational dynamics generate disordered configurations that suppress thermal conductivity in superatomic crystals.
The thermal conductivity of fullerene-based superatomic crystals (SACs) is investigated using molecular dynamics simulations. The temperature-dependent predictions agree with the trends of previous measurements. The thermal conductivity behavior emerges as a result of the C60 molecule rotational dynamics and orientation, which are quantified using the root mean square displacements of the carbon atoms and the relative orientations of the C60s. At low temperatures, the C60s exhibit small rotations around equilibrium positions (i.e., librations). When the librating C60s are orientationally-ordered, as in the [C60] and [Co6Se8(PEt3)6][C60]2 SACs, thermal conductivity decreases with increasing temperature, as is typical for a crystal. When the librating C60s are orientationally-disordered, however, as in the [Co6Te8(PEt3)6][C60]2 SAC, thermal conductivity is lower and temperature independent, as is typical for an amorphous solid. At higher temperatures, where the C60s in all three SACs freely-rotate and are thus dynamically disordered, thermal conductivity is temperature independent. The abrupt changes driven by the C60 dynamics suggest that fullerene-based SACs can be designed to be thermal conductivity switches based on a variety of external stimuli.


Global detection of DNA repair outcomes induced by CRISPR-Cas9.
CRISPR-Cas9 generates double-stranded DNA breaks (DSBs) to activate cellular DNA repair pathways for genome editing. The repair of DSBs leads to small insertions or deletions (indels) and other complex byproducts, including large deletions and chromosomal translocations. Indels are well understood to disrupt target genes, while the other deleterious byproducts remain elusive. We developed a new in silico analysis pipeline for the previously described primer-extension-mediated sequencing assay to comprehensively characterize CRISPR-Cas9-induced DSB repair outcomes in human or mouse cells. We identified tremendous deleterious DSB repair byproducts of CRISPR-Cas9 editing, including large deletions, vector integrations, and chromosomal translocations. We further elucidated the important roles of microhomology, chromosomal interaction, recurrent DSBs, and DSB repair pathways in the generation of these byproducts. Our findings provide an extra dimension for genome editing safety besides off-targets. And caution should be exercised to avoid not only off-target damages but also deleterious DSB repair byproducts during genome editing.


Dielectrophoretic Microfluidic Chip Enables Single-Cell Measurements for Multidrug Resistance in Heterogeneous Acute Myeloid Leukemia Patient Samples.
The front-line treatment for adult acute myeloid leukemia (AML) is anthracycline-based combination chemotherapy. However, treatment outcomes remain suboptimal with relapses frequently observed. Among the mechanisms of treatment failure is multidrug resistance (MDR) mediated by the ABCB1, ABCC1, and ABCG2 drug-efflux transporters. Although genetic and phenotypic heterogeneity between leukemic blast cells is a well-recognized phenomenon, there remains minimal data on differences in MDR activity at the individual cell level. Specifically, functional assays that can distinguish the variability in MDR activity between individual leukemic blasts are lacking. Here, we outline a new dielectrophoretic (DEP) chip-based assay. This assay permits measurement of drug accumulation in single cells, termed same-single-cell analysis in the accumulation mode (SASCA-A). Initially, the assay was optimized in pretherapy samples from 20 adults with AML whose leukemic blasts had MDR activity against the anthracyline daunorubicin (DNR) tested using multiple MDR inhibitors. Parameters tested were initial drug accumulation, time to achieve signal saturation, fold-increase of DNR accumulation with MDR inhibition, ease of cell trapping, and ease of maintaining the trapped cells stationary. This enabled categorization into leukemic blast cells with MDR activity (MDR(+)) and leukemic blast cells without MDR activity (MDR(-ve)). Leukemic blasts could also be distinguished from benign white blood cells (notably these also lacked MDR activity). MDR(-ve) blasts were observed to be enriched in samples taken from patients who went on to enter complete remission (CR), whereas MDR(+) blasts were frequently observed in patients who failed to achieve CR following front-line chemotherapy. However, pronounced variability in functional MDR activity between leukemic blasts was observed, with MDR(+) cells not infrequently seen in some patients that went on to achieve CR. Next, we tested MDR activity in two paired AML patient samples. Pretherapy samples taken from patients that achieved CR to front-line chemotherapy were compared with samples taken at time of subsequent relapse. MDR(+) cells were frequently observed in leukemic blast cells in both pretherapy and relapsed samples, consistent with MDR as a mechanism of relapse in these patients. We demonstrate the ability of a new DEP microfluidic chip-based assay to identify heterogeneity in MDR activity in leukemic blasts. The test provides a platform for future studies to characterize the mechanistic basis for heterogeneity in MDR activity at the individual cell level.


[New aspects of differential vertigo diagnosis in childhood].
Eight cases of vertigo in childhood are reported. Four of them suffered from benign paroxysmal vertigo, and the other four from atypical migraine. History of a head injury and attacks of vertigo lasting only a few seconds are typical of benign paroxysmal vertigo. These children are older than 4 years and the vertigo is of peripheral origin. Vertigo due to atypical migraine is frequent in young children up to 4 years of age, shows a longer duration of vertigo (more than 1 min) and is of central origin.


Substrate specificity, membrane topology, and activity regulation of human alkaline ceramidase 2 (ACER2).
Human alkaline ceramidase 2 (ACER2) plays an important role in cellular responses by regulating the hydrolysis of ceramides in cells. Here we report its biochemical characterization, membrane topology, and activity regulation. Recombinant ACER2 was expressed in yeast mutant cells (Deltaypc1Deltaydc1) that lack endogenous ceramidase activity, and microsomes from ACER2-expressiong yeast cells were used to biochemically characterize ACER2. ACER2 catalyzed the hydrolysis of various ceramides and followed Michaelis-Menten kinetics. ACER2 required Ca(2+) for both its in vitro and cellular activities. ACER2 has 7 putative transmembrane domains, and its amino (N) and carboxyl (C) termini were found to be oriented in the lumen of the Golgi complex and cytosol, respectively. ACER2 mutant (ACER2DeltaN36) lacking the N-terminal tail (the first 36 amino acid residues) exhibited undetectable activity and was mislocalized to the endoplasmic reticulum, suggesting that the N-terminal tail is necessary for both ACER2 activity and Golgi localization. ACER2 mutant (ACER2DeltaN13) lacking the first 13 residues was also mislocalized to the endoplasmic reticulum although it retained ceramidase activity. Overexpression of ACER2, ACER2DeltaN13, but not ACER2DeltaN36 increased the release of sphingosine 1-phosphate from cells, suggesting that its mislocalization does not affect the ability of ACER2 to regulate sphingosine 1-phosphate secretion. However, overexpression of ACER2 but not ACER2DeltaN13 or ACER2DeltaN36 inhibited the glycosylation of integrin beta1 subunit and Lamp1, suggesting that its mistargeting abolishes the ability of ACER2 to regulation protein glycosylation. These data suggest that ACER2 has broad substrate specificity and requires Ca(2+) for its activity and that ACER2 has the cytosolic C terminus and luminal N terminus, which are essential for its activity, correct cellular localization, and regulation for protein glycosylation.


Molecular modeling of phytochrome using constitutive C-phycocyanin from Fremyella diplosiphon as a putative structural template.
Phytochrome, the ubiquitous photosensor in green plants, is similar to C-phycocyanin in a number of ways. We have produced a model of the phytochrome chromophore binding pocket based on the X-ray crystal structure of C-phycocyanin from Fremyella diplosiphon [Duerring et al. (1991) J. Mol. Biol. 217, 577-592]. Twenty residues around the chromophore binding site of C-phycocyanin were changed to the corresponding residues of Avena phytochrome A for the modeling. In the minimized model, Arg-318, Ala-319, the methylene of Ser-322, Leu-325, Gln-326, and Tyr-327 (using the numbering of the Avena sequence; Cys-323 is chromophore bound) form a pocket on one side of the chromophore. The other side of the chromophore lacks hydrogen-bond donors and is involved only in van der Waals contact with the chromophore. The overall structure of the model may be described as one peptide segment "anchoring" the chromophore hydrophobically, covalently, and electrostatically from several directions, while the other key peptide segment simply provides a hydrophobic surface for the chromophore to rest against. The red light absorbing (Pr) chromophore of the model is buried more deeply in the binding pocket than the far red light absorbing (Pfr) chromophore. This apparently reflects reduced compatibility of the chromophore with the pocket upon photoisomerization, which requires the insertion of hydrophilic parts of ring D into the hydrophobic core of the protein. This concept is consistent with the experimental evidence that photoisomerization of the Pr chromophore is followed by movement of the chromophore from its binding pocket. In the proposed model, increased exposure of hydrophobic portions of the Pfr chromophore compared to the Pr chromophore is consistent with the red shift observed in the first intermediate of the Pr to Pfr photoconversion. The proposed model may be tested by mutation experiments, thus providing a viable model to foster the current rapid progress of molecular biology in this field.


Quantitative analysis of the low molecular weight serum proteome using 18O stable isotope labeling in a lung tumor xenograft mouse model.
With advancements in the analytical technologies and methodologies in proteomics, there is great interest in biomarker discovery in biofluids such as serum and plasma. Current hypotheses suggest that the low molecular weight (LMW) serum proteome possesses an archive of clipped and cleaved protein fragments that may provide insight into disease development. Though these biofluids represent attractive samples from which new and more accurate disease biomarkers may be found, the intrinsic person-to-person variability in these samples complicates their discovery. Mice are one of the most extensively used animal models for studying human disease because they represent a highly controllable experimental model system. In this study, the LMW serum proteome was compared between xenografted tumor-bearing mice and control mice by differential labeling utilizing trypsin-mediated incorporation of the stable isotope of oxygen, 18O. The digestates were combined, fractionated by strong cation exchange chromatography, and analyzed by nanoflow reversed-phase liquid chromatography coupled online with tandem mass spectrometry, resulting in the identification of 6003 proteins identified by at least a single, fully tryptic peptide. Almost 1650 proteins were identified and quantitated by two or more fully tryptic peptides. The methodology adopted in this work provides the means for future quantitative measurements in comparative animal models of disease and in human disease cohorts.


First treatment and retreatment of medically refractive trigeminal neuralgia by stereotactic radiosurgery versus microvascular decompression: a systematic review and Meta-analysis.
Procedures to treat medically refractory trigeminal neuralgia (MRTN) include stereotactic radiosurgery (SRS) and microvascular decompression (MVD). The aim of this study was to compare outcomes of SRS versus MVD in the treatment of MRTN, with a subgroup focus on those being treated for the first time. Search strategy was performed using the PRISMA guidelines for article identification, screening, eligibility and inclusion. Relevant articles were identified from six electronic databases from their inception to June 2017. These articles were screened against established criteria for inclusion into this study. Meta-analysis was conducted by pooling results with odds ratios and subgroup analysis. From 13 relevant studies identified, 683 patients treated by SRS were compared with 670 patients treated by MVD for MRTN. Overall, the significant findings were that compared to MVD, SRS was associated with lower rates of short-term (OR = 0.16; 95%CI = 0.11-0.22; p < .001) and long-term pain freedom (OR = 0.31; 95%CI = 0.22-0.44; p < .001), fewer postoperative complications (OR = 0.06; 95%CI = 0.02-0.16; p < .001), more facial numbness and dysesthesia (OR = 1.64; 95%CI = 1.08-2.49; p = .02), and more pain recurrence (OR = 2.28; 95%CI = 1.32-3.93; p = .003). These trends were all reflected in MRTN patients being treated for the first time. Both SRS and MVD alleviate pain in MRTN patients. MVD results in superior rates of short- and long-term pain relief, facial numbness and dysesthesia control, and less recurrence amongst those in whom pain freedom was achieved, at the cost of greater postoperative complications when compared to SRS. Although no significant difference was found in terms of the need for retreatment surgery, there was a trend towards less procedures favoring MVD. First treatment by either technique represents the overall trends reported.


Spondylodiscitis (disc space infection) associated with negative microbiological tests: comparison of outcome of suspected disc space infections to documented non-tuberculous pyogenic discitis.
Discitis, an infection of the disc space, is an uncommon diagnosis that, if missed, can lead to spinal deformity and neurological deterioration, although as many as 30% of these patients will have negative microbiological cultures. It was unclear, however, whether the prognosis differed between patients who had positive or negative cultures. A retrospective case note review was carried out to assess the differences in presentation and outcome between these two groups. There were 26 and 43 patients in the negative and positive groups, respectively. Those with a positive culture were more likely to present with pyrexia, have a neurological deficit and not be independently mobile at presentation. The mean CRP recorded at the time of presentation was 96 and 157 in the negative and positive groups respectively (p = 0.004). Similarly, the mean ESR in the positive group was 88 compared with 69 in the negative group (p = 0.02). In conclusion, these patients may be at different ends of a clinical spectrum: those patients with a positive culture having a greater local and systemic inflammatory reaction to the disc space infection.


Measuring hospital adverse events: assessing inter-rater reliability and trigger performance of the Global Trigger Tool.
To determine the inter-rater reliability of the Institute for Healthcare Improvement's Global Trigger Tool (GTT) in a practice setting, and explore the value of individual triggers. Prospective assessment of application of the GTT to monthly random samples of hospitalized patients at four hospitals across three regions in the USA. Mayo Clinic campuses are in Minnesota, Arizona and Florida. A total of 1138 non-pediatric inpatients from all units across the hospital. GTT was applied to randomly selected medical records with independent assessments of two registered nurses with a physician review for confirmation. The Cohen Kappa coefficient was used as a measure of inter-rater agreement. The positive predictive value was assessed for individual triggers. Good levels of reliability were obtained between independent nurse reviewers at the case-level for both the occurrence of any trigger and the identification of an adverse event. Nurse reviewer agreement for individual triggers was much more varied. Higher agreement appears to occur among triggers that are objective and consistently recorded in selected portions of the medical record. Individual triggers also varied on their yield to detect adverse events. Cases with adverse events had significantly more triggers identified (mean 4.7) than cases with no adverse events (mean 1.8). The trigger methodology appears to be a promising approach to the measurement of patient safety. However, automated processes could make the process more efficient in identifying adverse events and has a greater potential of improving care delivery and patient 'outcomes'.


Uranium power and horizontal proliferation of nuclear weapons.
Only a few nonnuclear weapons states with uranium-fueled power plants have kept the weapons option open, and none has evidenced activities intended for diverting fissionable material from its civilian system. Analysis of alternative strategies shows that acquisition of nuclear weapons material would probably depend on military production facilities rather than diversion. Horizontal proliferation is primarily a political issue and is related only marginally to uranium power development. Restrictions of the Nuclear Non-Proliferation Act on the supply of equipment and fuel by the United States appear to have induced, in some nonnuclear weapons states, the building of small-scale facilities that can be modified for production of weapons material. More attention should be given to the international political, economic, and military factors that persuade such states to abjure nuclear weapons.


Maximum Tolerated Dose of Walking for Community-Dwelling People Recovering From Hip Fracture: A Dose-Response Trial.
To determine how much moderate-intensity physical activity, in the form of walking, could be prescribed for people living in the community after hip fracture in terms of safety, tolerability, and feasibility. Phase I dose-response design. Public community rehabilitation centers. Community-dwelling adults (N=21; 16 women; mean age, 75±9y) who were cognitively alert, attending community rehabilitation after hip fracture (mean days postfracture, 110±47d), able to walk with or without a gait aid, and for whom it was safe to participate in physical activity. Individually supervised doses of moderate-intensity walking completed in 1 week in addition to their usual levels of physical activity. Three participants were required to complete a dose of walking before dose escalation for the next cohort of 3 participants. Dose escalation ceased when >1 participant in a cohort had an adverse event or was unable to tolerate the dose or if the maximum dose of 150min/wk was achieved. Maximum tolerated dose of walking per week (in minutes), adverse events, mobility, and walking confidence. The maximum tolerated dose of walking for adults after hip fracture before significant discomfort was experienced (eg, breathlessness, pain, and fatigue) by any participant was 100min/wk. No adverse events occurred, but participants began to be unable to tolerate higher doses beyond 100min/wk. This provides preliminary evidence that community-dwelling older adults recovering from hip fracture can complete a sufficient amount of moderate-intensity physical activity to maintain and improve their health.


Are we teaching school health in residencies?
A school health survey was distributed to all family practice residency programs in the United States and Puerto Rico. The intent of the survey was to measure the degree to which residency programs include education on school health issues in their curriculum. The items on this survey measured four general areas: identification of family practice residency programs with school health components, curricular development, resident and faculty involvement, and the rationale for the lack of involvement of residency programs. The results of the survey indicate that approximately one-third of the residency programs presently have a school health component in their curriculum. Preschool physicals and sports medicine are the most popular activities within family practice residency programs. It was further determined that there is little question of the perceived importance of school health, but the question still remains as to what is the most efficient and effective manner of teaching residents these valuable skills.


A MMP derived versican neo-epitope is elevated in plasma from patients with atherosclerotic heart disease.
Extracellular matrix remodelling is a prerequisite for plaque rupture in atherosclerotic lesion. Versican, an extracellular matrix proteoglycan present in normal and atherosclerotic arteries is a substrate for matrix metalloproteinases (MMPs) present in macrophage rich areas. The aim of the current study was to develop an immunoassay to detect a specific MMP-12 derived versican degradation fragment (VCANM) and assess its potential as a biomarker for extracellular matrix remodelling in atherosclerosis. A mouse monoclonal antibody raised against VCANM was used for the development of a competitive ELISA for detection of the fragment in plasma. VCANM was measured in plasma of patients with different levels of heart diseases. Patients experiencing I) acute coronary syndrome, II) stable ischemic heart disease and III) demonstrating high levels of coronary calcium deposits had significantly higher plasma levels of VCANM compared to a control group of individuals with no detectable coronary calcium deposits. VCANM was also detected by immunohistochemistry in coronary artery sections of patients with different degrees of atherosclerosis. VCANM ability to separate patients with atherosclerotic diseases from healthy individuals suggested VCANM as a potential biomarker for the pathological arterial matrix remodelling associated with atherosclerosis.


A multifaceted quality improvement intervention for CVD risk management in Australian primary healthcare: a protocol for a process evaluation.
Cardiovascular disease (CVD) is the leading cause of death and disability worldwide. Despite the widespread availability of evidence-based clinical guidelines and validated risk predication equations for prevention and management of CVD, their translation into routine practice is limited. We developed a multifaceted quality improvement intervention for CVD risk management which incorporates electronic decision support, patient risk communication tools, computerised audit and feedback tools, and monthly, peer-ranked performance feedback via a web portal. The intervention was implemented in a cluster randomised controlled trial in 60 primary healthcare services in Australia. Overall, there were improvements in risk factor recording and in prescribing of recommended treatments among under-treated individuals, but it is unclear how this intervention was used in practice and what factors promoted or hindered its use. This information is necessary to optimise intervention impact and maximally implement it in a post-trial context. In this study protocol, we outline our methods to conduct a theory-based, process evaluation of the intervention. Our aims are to understand how, why, and for whom the intervention produced the observed outcomes and to develop effective strategies for translation and dissemination. We will conduct four discrete but inter-related studies taking a mixed methods approach. Our quantitative studies will examine (1) the longer term effectiveness of the intervention post-trial, (2) patient and health service level correlates with trial outcomes, and (3) the health economic impact of implementing the intervention at scale. The qualitative studies will (1) identify healthcare provider perspectives on implementation barriers and enablers and (2) use video ethnography and patient semi-structured interviews to understand how cardiovascular risk is communicated in the doctor/patient interaction both with and without the use of intervention. We will also assess the costs of implementing the intervention in Australian primary healthcare settings which will inform scale-up considerations. This mixed methods evaluation will provide a detailed understanding of the process of implementing a quality improvement intervention and identify the factors that might influence scalability and sustainability. 12611000478910.


Microeukaryotes in animal and plant microbiomes: Ecologies of disease?
Studies of animal and plant microbiomes are burgeoning, but the majority of these focus on bacteria and rarely include microeukaryotes other than fungi. However, there is growing evidence that microeukaryotes living on and in larger organisms (e.g. plants, animals, macroalgae) are diverse and in many cases abundant. We present here a new combination of 'anti-metazoan' primers: 574*f-UNonMet_DB that amplify a wide diversity of microeukaryotes including some groups that are difficult to amplify using other primer combinations. While many groups of microeukaryotic parasites are recognised, myriad other microeukaryotes are associated with hosts as previously unknown parasites (often genetically divergent so difficult to amplify using standard PCR primers), opportunistic parasites, commensals, and other ecto- and endo-symbionts, across the 'symbiotic continuum'. These fulfil a wide range of roles from pathogenesis to mutually beneficial symbioses, but mostly their roles are unknown and likely fall somewhere along this spectrum, with the potential to switch the nature of their interactions with the host under different conditions. The composition and dynamics of host-associated microbial communities are also increasingly recognised as important moderators of host health. This 'pathobiome' approach to understanding disease is beginning to supercede a one-pathogen-one-disease paradigm, which cannot sufficiently explain many disease scenarios.


Changes in self-reported sleep and cognitive failures: a randomized controlled trial of a stress management intervention.
This study evaluated the effectiveness of a stress management intervention combining individual cognitive behavioral therapy (CBT) with a brief workplace intervention on self-reported measures of sleep and cognitive functioning among patients on sick leave due to work-related stress complaints. Participants were patients referred to the regional Department of Occupational Medicine. Inclusion criteria were (i) sick leave due to work-related stress complaints and (ii) a diagnosis of adjustment disorder/reactions to stress or mild depression. Participants (N=137) were randomized to either an intervention (N=57) or control (N=80) group. The intervention comprised six sessions with a psychologist and the offer of a small workplace intervention. Questionnaires were answered at baseline and after 4, and 10 months. Symptoms were significantly reduced over time in both groups but there was no significant treatment effect on sleep or cognitive outcomes at any time point. From 0-4 months, there was a tendency for larger improvements in the intervention group with regards to sleep and cognitive failures in distraction. Although neither was significant, the results came close to significance depicting a small effect size (Cohen's d) on sleep complaints and distractions (but not memory). The specific intervention was not superior to the control condition in reducing symptoms of sleep problems and cognitive difficulties at any time point during the 10-month follow-up period. Substantial improvements in symptoms over time were seen in both groups.


Frequency and impact of individual symptoms on quality of life in dry eye disease in patients presenting to a tertiary care hospital.
To assess the frequency of symptoms and impact on quality of life in individuals with dry eye disease. The cross-sectional, study was conducted from December 2018 to June 2019 at the outpatient clinic of the Department of Ophthalmology, Pakistan Institute of Medical Sciences, Islamabad, Pakistan, and comprised dry eye disease patients aged at least 13 years with ocular burning sensation, lacrimation, grittiness, photophobia, redness or itching. Tear Film Breakup time <10 sec and / or positive ocular surface staining >5 corneal spots, >9 conjunctival spots, lid margin staining ≥ 2mm in length were used to diagnose the disease. Data was collected using the Dry Eye Quality of Life Score questionnaire. Data was analysed using SPSS 25. of the 191 patients, 135(71%) were females and 56(29%) were males. The overall mean age was 39.8±16.6 years. Itching 155(81.2%), watering 151(79.1%) and burning 124(64.9%) were the most frequent symptoms, with watering (p<0.001) and photophobia (p=0.012) significantly impacting quality of life. The most common symptoms among patients of dry eye disease affecting their quality of life were itching, watering, burning, photophobia and low mood.


Real-space observation of fluctuating antiferromagnetic domains.
Magnetic domains play a fundamental role in physics of magnetism and its technological applications. Dynamics of antiferromagnetic domains is poorly understood, although antiferromagnets are expected to be extensively used in future electronic devices wherein it determines the stability and operational speed. Dynamics of antiferromagnets also features prominently in the studies of topological quantum matter. Real-space imaging of fluctuating antiferromagnetic domains is therefore highly desired but has never been demonstrated. We use coherent x-ray diffraction to obtain videos of fluctuating micrometer-scale antiferromagnetic domains in Ni2MnTeO6 on time scales from 10-1 to 103 s. In the collinear phase, thermally activated domain wall motion is observed in the vicinity of the Néel temperature. Unexpectedly, the fluctuations persist through the full range of the higher-temperature helical phase. These observations illustrate the high potential significance of the dynamic domain imaging in phase transition studies and in magnetic device research.


Terminal deoxynucleotidyl transferase as a hematopoietic cell marker.
Terminal deoxynucleotidyl transferase (TdT) is an intracellular protein characteristic of certain primitive lymphocytes in normal thymus and bone marrow. It can be detected by enzyme assay or immunocytochemical staining. High levels of enzyme and increased numbers of TdT+ cells are found in some lymphoblastic leukemias and lymphomas. Analysis for TdT thus provides a useful adjunct in the differential diagnosis of leukemia.


Neuronal expression of P2X3 purinoceptors in the rat retina.
P2X3 purinoceptors are involved in fast, excitatory neurotransmission in the nervous system, and are expressed predominantly within sensory neurons. In this study, we examined the cellular and synaptic localization of the P2X3 receptor subunit in the retina of the rat using immunofluorescence immunohistochemistry and pre-embedding immunoelectron microscopy. In addition, we investigated the activity of ecto-ATPases in the inner retina using an enzyme cytochemical method. The P2X3 receptor subunit was expressed in the soma of a subset of GABA immunoreactive amacrine cells, some of which also expressed protein kinase C-alpha. In addition, punctate immunoreactivity was observed within both the inner and outer plexiform layers of the retina. Double labeling studies showed that P2X3 receptor puncta were associated with both rod and cone bipolar cell axon terminals in the inner plexiform layer. Ultrastructural studies indicated that P2X3 receptor subunits were expressed on putative A17 amacrine cells at sites of reciprocal synaptic input to the rod bipolar cell axon terminal. Moreover, we observed P2X3 immunolabeling on amacrine cell processes that were associated with cone bipolar cell axon terminals and other conventional synapses. In the outer retina, P2X3 immunoreactivity was observed on specialized junctions made by putative interplexiform cells. Ecto-ATPase activity was localized to the inner plexiform layer on the extracellular side of all plasma membranes, but was not apparent in the ganglion cell layer or the inner nuclear layer, suggesting that ATP dephosphorylation occurs exclusively in synaptic regions of the inner retina. These data provide further evidence that purines participate in retinal transmission, particularly within the rod pathway.


Association of intensive care unit delirium with sleep disturbance and functional disability after critical illness: an observational cohort study.
In medical intensive care unit (MICU) patients, the predictors of post-discharge sleep disturbance and functional disability are poorly understood. ICU delirium is a risk factor with a plausible link to sleep disturbance and disability. This study evaluated the prevalence of self-reported post-ICU sleep disturbance and increased functional disability, and their association with MICU delirium and other ICU factors. This was an observational cohort study of MICU patients enrolled in a biorepository and assessed upon MICU admission by demographics, comorbidities, and baseline characteristics. Delirium was assessed daily using the Confusion Assessment Method for the ICU. Telephone follow-up interview instruments occurred after hospital discharge and included the Pittsburgh Sleep Quality Index (PSQI), and basic and instrumental activities of daily living (BADLs, IADLs) for disability. We define sleep disturbance as a PSQI score > 5 and increased disability as an increase in composite BADL/IADL score at follow-up relative to baseline. Multivariable regression modeled the associations of delirium and other MICU factors on follow-up PSQI scores and change in disability scores. PSQI and BADL/IADL instruments were completed by 112 and 122 participants, respectively, at mean 147 days after hospital discharge. Of those surveyed, 63% had sleep disturbance by PSQI criteria, and 37% had increased disability by BADL/IADL scores compared to their pre-MICU baseline. Total days of MICU delirium (p = 0.013), younger age (p = 0.013), and preexisting depression (p = 0.025) were significantly associated with higher PSQI scores at follow-up. Lower baseline disability (p < 0.001), older age (p = 0.048), and less time to follow-up (p = 0.024) were significantly associated with worsening post-ICU disability, while the occurrence of MICU delirium showed a trend toward association (p = 0.077). After adjusting for important covariates, total days of MICU delirium were significantly associated with increased post-discharge sleep disturbance. Delirium incidence showed a trend toward association with increased functional disability in the year following discharge.


Variables affecting first-year student commitment during the transition to college in Ireland.
Third-level student attrition rates internationally are a cause for concern. Thus, student retention and academic success are top priorities for colleges, and students alike. In addition to this, student well-being is a growing public health concern. This study explores the need to incorporate well-being as a predictor in traditionally academically focused models of student persistence during the transition to college. A cross-sectional web-based survey was conducted with 574 first-year students aged 17-22 years attending a university in Ireland to examine the effect of student persistence and well-being variables on student commitment during the transition to college. This study highlights the connection between student academic, social and well-being variables during the transition to college within the fitted models that emerged from the data. Student well-being plays a key role ensuring student persistence during the transition to college. The results support our hypothesis that many student variables have the potential to substantially impact student commitment during the first-year transition to college. These variables include student depression, self-rated health, academic environment, peer connections and perceiving the academic programme to be an appropriate personal choice. It is therefore important for education institutions to acknowledge and address student persistence and student well-being in an integrative way and for traditionally academically focused student retention models to orientate to incorporate student well-being.


Glutamatergic calcium dynamics and deregulation of rat retinal ganglion cells.
A rise in intracellular calcium levels ([Ca(2+)](i)) is a key trigger for the lethal effects of the excitatory neurotransmitter glutamate in various central neurons, but a consensus has not been reached on the pathways that mediate glutamate-dependent increases of [Ca(2+)](i) in retinal ganglion cells (RGCs). Using Ca(2+) imaging techniques we demonstrated that, in the absence of external Mg(2+), the Ca(2+) signal evoked by glutamate was predominantly mediated by NMDA-type glutamate receptors (NMDA-Rs) in immunopanned RGCs isolated from neonatal or adult rats. Voltage-gated Ca(2+) channels and AMPA/kainate-Rs contributed a smaller portion of the Ca(2+) response at saturating concentrations of glutamate. Consistent with NMDA-R involvement, extracellular Mg(2+) inhibited RGC glutamate responses, while glycine had a potentiating effect. With Mg(2+) present externally, the effect of AMPA/kainate-R antagonists was enhanced and both NMDA- and AMPA/kainate-R antagonists greatly reduced the glutamate-induced increases of RGC [Ca(2+)](i). This finding indicates that the primary contribution of AMPA/kainate-Rs to RGC glutamatergic Ca(2+) dynamics is through the depolarization-dependent relief of the Mg(2+) block of NMDA-R channels. The effect of glutamate receptor antagonists on glutamatergic Ca(2+) signals from RGCs in adult rat retinal wholemounts yielded results similar to those obtained using immunopanned RGCs. Additional experiments on isolated RGCs revealed that during a 1 h glutamate (10-1000 microm) exposure, 18-28% of RGCs exhibited delayed Ca(2+) deregulation (DCD) and the RGCs that underwent DCD were positive for the death marker annexin V. RGCs with larger glutamate-evoked Ca(2+) signals were more likely to undergo DCD, and NMDA-R blockade significantly reduced the occurrence of DCD. Identifying the mechanisms underlying RGC excitotoxicity aids in our understanding of the pathophysiology of retinal ischaemia, and this work establishes a major role for NMDA-R-mediated increases in [Ca(2+)](i) in glutamate-related RGC death.


Characterization of saturable binding sites for rabies virus.
A specific, saturable receptor for rabies virus was analyzed on cultured cells of neural or non-neural origin. Viral attachment kinetics were enhanced by DEAE-dextran, an effect which in turn enhanced the apparent infectivity of the virus inoculum. Under optimized conditions, the attachment of metabolically labeled ERA strain rabies virus obeyed the laws of mass action, whereby the amount of virus bound to cells varied proportionally with the concentration of cells or virus. Attachment was sensitive to changes of temperature and pH, did not require divalent cations such as Mg2+ or Ca2+, and occurred despite prior treatment of cells with proteolytic or sialic acid-specific enzymes. Saturation of the cell surface with rabies virus could be accomplished with 3 X 10(3) to 15 X 10(3) attached virions per cell. Competition for the rabies receptor occurred with rabies nonpathogenic variant virus, RV194 -2, and vesicular stomatitis virus. Reovirus type 3, another neurotropic virus, failed to inhibit rabies virus binding, and West Nile virus only slightly inhibited rabies virus binding, suggesting independent cellular receptors were recognized by these viruses. Isolated rabies virus glycoprotein failed to compete in an equivalent manner. However, solubilization of BHK-21 cells with octylglucoside yielded a chloroform-methanol-soluble extract which blocked rabies virus attachment. The binding inhibition activity of this extract was resistant to proteases but could be destroyed by phospholipases and neuraminidase, suggesting a phospholipid or glycolipid component at the receptor site. These data provide evidence for a rhabdovirus-common mechanism for cellular attachment to cells in culture.


Genomics pipelines to investigate susceptibility in whole genome and exome sequenced data for variant discovery, annotation, prediction and genotyping.
Over the last few decades, genomics is leading toward audacious future, and has been changing our views about conducting biomedical research, studying diseases, and understanding diversity in our society across the human species. The whole genome and exome sequencing (WGS/WES) are two of the most popular next-generation sequencing (NGS) methodologies that are currently being used to detect genetic variations of clinical significance. Investigating WGS/WES data for the variant discovery and genotyping is based on the nexus of different data analytic applications. Although several bioinformatics applications have been developed, and many of those are freely available and published. Timely finding and interpreting genetic variants are still challenging tasks among diagnostic laboratories and clinicians. In this study, we are interested in understanding, evaluating, and reporting the current state of solutions available to process the NGS data of variable lengths and types for the identification of variants, alleles, and haplotypes. Residing within the scope, we consulted high quality peer reviewed literature published in last 10 years. We were focused on the standalone and networked bioinformatics applications proposed to efficiently process WGS and WES data, and support downstream analysis for gene-variant discovery, annotation, prediction, and interpretation. We have discussed our findings in this manuscript, which include but not are limited to the set of operations, workflow, data handling, involved tools, technologies and algorithms and limitations of the assessed applications.


Hispanic Baby Boomers: health inequities likely to persist in old age.
As the Baby-Boom generation enters the ranks of the elderly adults over the next 4 decades, the United States will witness an unprecedented growth in racial/ethnic diversity among the older adult population. Hispanics will comprise 20% of the next generation of older adults, representing the largest minority population aged 65 years and older, with those of Mexican-origin comprising the majority of Hispanics. Little is known about the health status of this population. DATA/METHODS: Data are for Baby Boomers born between 1946 and 1964 (ages 43-61) in the 2007 California Health Interview Survey. Logistic regression estimates the odds of diabetes, hypertension, obesity, fair/poor self-rated health (SRH), and functional difficulties among U.S.-born non-Hispanic Whites (NHW), U.S.-born Mexicans, naturalized Mexican immigrants, and noncitizen Mexican immigrants. The Mexican-origin populations are disadvantaged relative to NHW for all socioeconomic status (SES) and several health outcomes. The Mexican origin disadvantage in health attenuates when controlling for SES and demographics, but the disadvantage remains for diabetes, obesity, and fair/poor SRH. Baby Boomers of Mexican origin do not share the advantages of health, income, and educational attainment enjoyed by U.S.-born NHW. As this cohort moves into old age, the cumulative disadvantage of existing disparities are likely to result in continued or worse health disparities. Reductions in federal entitlement programs for the elderly adults that delay eligibility, scale back programs and services, or increase costs to consumers may exacerbate those inequities.


[The effect of hyper- and hypothermia on induced telomeric chromosome fusion].
After administration of colcemid and 5-BrdU in the cell culture, the cells pass through the first interphase to delay in mitosis. Then the cells overcome the colcemid blockade, and polykaryocytes with micronuclei are formed. The second interphase in followed by the second mitosis, during which dicentric chromosomes are observed. These dicentrics are the result of telomeric chromosome fusion. The action of hyperthermia (40 degrees C) during the whole period of colcemid and 5-BrdU treatment or that of the hyperthermia (40 degrees C) only during the first 17 hours (the first interphase and the first mitosis) lead to the increased frequency of dicentrics. Under condition of hypothermia (34 degrees C) the frequency of dicentric formation decreases. Changes in cultivation temperature during the latest 25 hr of colcemid and 5-BrdU action (the second interphase and the second mitosis) exert no influence on the dicentric formation frequency. Because there are no dicentrics in cells during the metaphase of the first mitosis, it is supposed that the temperature--sensitive period may be the latest steps of colcemid blockade, i.e. the period of formation of micronuclei.


Assessing disparities in medical students' knowledge and attitude about monkeypox: a cross-sectional study of 27 countries across three continents.
The recent monkeypox (Mpox) outbreak confirmed by the World Health Organization (WHO) underscores the importance of evaluating the knowledge and attitude of medical students toward emerging diseases, given their potential roles as healthcare professionals and sources of public information during outbreaks. This study aimed to assess medical students' knowledge and attitude about Mpox and to identify factors affecting their level of knowledge and attitude in low-income and high-income countries. A cross-sectional study was conducted on 11,919 medical students from 27 countries. A newly-developed validated questionnaire was used to collect data on knowledge (14 items), attitude (12 items), and baseline criteria. The relationship between a range of factors with knowledge and attitude was studied using univariate and multivariate analyses. 46% of the study participants were males; 10.7% were in their sixth year; 54.6% knew about smallpox; 84% received the coronavirus disease 2019 (COVID-19) vaccine; and 12.5% had training on Mpox. 55.3% had good knowledge of Mpox and 51.7% had a positive attitude towards it. Medical students in their third, fifth, or sixth year high- income countries who obtained information on Mpox from friends, research articles, social media and scientific websites were positive predictors for good knowledge. Conversely, being male or coming from high-income countries showed a negative relation with good knowledge about Mpox. Additionally, a positive attitude was directly influenced by residing in urban areas, being in the fifth year of medical education, having knowledge about smallpox and a history of receiving the coronavirus disease 2019 (COVID-19) vaccine. Receiving information about Mpox from social media or scientific websites and possessing good knowledge about Mpox were also predictors of a positive attitude. On the other hand, being male, employed, or receiving a training program about Mpox were inversely predicting positive attitude about Mpox. There were differences in knowledge and attitude towards Mpox between medical students in low and high-income countries, emphasizing the need for incorporating epidemiology of re-emerging diseases like Mpox into the medical curriculum to improve disease prevention and control.


A document classifier for medicinal chemistry publications trained on the ChEMBL corpus.
The large increase in the number of scientific publications has fuelled a need for semi- and fully automated text mining approaches in order to assist in the triage process, both for individual scientists and also for larger-scale data extraction and curation into public databases. Here, we introduce a document classifier, which is able to successfully distinguish between publications that are 'ChEMBL-like' (i.e. related to small molecule drug discovery and likely to contain quantitative bioactivity data) and those that are not. The unprecedented size of the medicinal chemistry literature collection, coupled with the advantage of manual curation and mapping to chemistry and biology make the ChEMBL corpus a unique resource for text mining. The method has been implemented as a data protocol/workflow for both Pipeline Pilot (version 8.5) and KNIME (version 2.9) respectively. Both workflows and models are freely available at: ftp://ftp.ebi.ac.uk/pub/databases/chembl/text-mining. These can be readily modified to include additional keyword constraints to further focus searches. Large-scale machine learning document classification was shown to be very robust and flexible for this particular application, as illustrated in four distinct text-mining-based use cases. The models are readily available on two data workflow platforms, which we believe will allow the majority of the scientific community to apply them to their own data. Graphical AbstractMultidimensional scaling analysis applied to document vectors derived from titles and abstracts in different corpora. Notably, there is large overlap between the documents in the different ChEMBL versions and BindingDB, while the background MEDLINE set is largely divergent.


Cascaded systems analysis of noise and detectability in dual-energy cone-beam CT.
Dual-energy computed tomography and dual-energy cone-beam computed tomography (DE-CBCT) are promising modalities for applications ranging from vascular to breast, renal, hepatic, and musculoskeletal imaging. Accordingly, the optimization of imaging techniques for such applications would benefit significantly from a general theoretical description of image quality that properly incorporates factors of acquisition, reconstruction, and tissue decomposition in DE tomography. This work reports a cascaded systems analysis model that includes the Poisson statistics of x rays (quantum noise), detector model (flat-panel detectors), anatomical background, image reconstruction (filtered backprojection), DE decomposition (weighted subtraction), and simple observer models to yield a task-based framework for DE technique optimization. The theoretical framework extends previous modeling of DE projection radiography and CBCT. Signal and noise transfer characteristics are propagated through physical and mathematical stages of image formation and reconstruction. Dual-energy decomposition was modeled according to weighted subtraction of low- and high-energy images to yield the 3D DE noise-power spectrum (NPS) and noise-equivalent quanta (NEQ), which, in combination with observer models and the imaging task, yields the dual-energy detectability index (d(')). Model calculations were validated with NPS and NEQ measurements from an experimental imaging bench simulating the geometry of a dedicated musculoskeletal extremities scanner. Imaging techniques, including kVp pair and dose allocation, were optimized using d(') as an objective function for three example imaging tasks: (1) kidney stone discrimination; (2) iodine vs bone in a uniform, soft-tissue background; and (3) soft tissue tumor detection on power-law anatomical background. Theoretical calculations of DE NPS and NEQ demonstrated good agreement with experimental measurements over a broad range of imaging conditions. Optimization results suggest a lower fraction of total dose imparted by the low-energy acquisition, a finding consistent with previous literature. The selection of optimal kVp pair reveals the combined effect of both quantum noise and contrast in the kidney stone discrimination and soft-tissue tumor detection tasks, whereas the K-edge effect of iodine was the dominant factor in determining kVp pairs in the iodine vs bone task. The soft-tissue tumor task illustrated the benefit of dual-energy imaging in eliminating anatomical background noise and improving detectability beyond that achievable by single-energy scans. This work established a task-based theoretical framework that is predictive of DE image quality. The model can be utilized in optimizing a broad range of parameters in image acquisition, reconstruction, and decomposition, providing a useful tool for maximizing DE-CBCT image quality and reducing dose.


Safety of vitrectomy for floaters.
To assess the risks of vitrectomy for the removal of primary and secondary vitreous opacities. Retrospective, nonrandomized, interventional case series. We reviewed the results of 116 consecutive cases of vitrectomy for vitreous floaters. Eighty-six cases were primary and 30 cases were secondary floaters. Main outcome measures were the incidence of iatrogenic retinal breaks and postoperative rhegmatogenous retinal detachments. We found iatrogenic retinal breaks in 16.4% of operations. There was no statistically significant difference in risk between cases of primary and secondary floaters. Intraoperative posterior vitreous detachment induction was found to increase significantly the risk of breaks. Retinal detachment occurred in 3 cases (2.5%), all after operations for primary floaters. One case of complicated retinal detachment ended with a low visual acuity of hand movements. Cataract occurred in 50% of phakic cases. Transient postoperative hypotony was found after 5.2% of our operations, and transient postoperative high intraocular pressure was encountered in 7.8%. An intraoperative choroidal hemorrhage occurred in 1 case, which resolved spontaneously. The mean visual acuity improved from 0.20 to 0.13 logarithm of the minimal angle of resolution units. The risk profile of vitrectomy for floaters is comparable with that of vitrectomy for other elective indications. Retinal breaks are a common finding during surgery and treatment of these breaks is crucial for the prevention of postoperative retinal detachment. Patients considering surgery for floaters should be informed specifically about the risks involved.


Dairy products, calcium, phosphorous, vitamin D, and risk of prostate cancer (Sweden)
Dairy products consistently have been associated with an increased risk of prostate cancer, yet the mechanism of this relationship remains unknown. Recent hypotheses propose that 1,25 dihydroxyvitamin D (1,25 D) is protective for prostate cancer. One study in the United States found that calcium consumption, which can lower circulating 1,25 D, was associated with higher risk of advanced prostate cancer, and we sought to address this hypothesis in a distinct population. We analyzed data from a population-based case-control study of prostate cancer conducted in Orebro, Sweden, with 526 cases and 536 controls. Using unconditional logistic regression models, we examined the relationship of dairy products, dietary calcium, phosphorous, and vitamin D with risk of total, extraprostatic, and metastatic prostate cancer. Calcium intake was an independent predictor of prostate cancer (relative risk (RR) = 1.91, 95 percent confidence interval (CI) 1.23-2.97 for intake > or = 1183 vs. < 825 mg/day), especially for metastatic tumors (RR = 2.64, 95 percent CI 1.24-5.61), controlling for age, family history of prostate cancer, smoking, and total energy and phosphorous intakes. High consumption of dairy products was associated with a 50 percent increased risk of prostate cancer. Our results support the hypothesis that high calcium intake may increase risk of prostate cancer, and this relation may underlie previously observed associations between dairy products and prostate cancer.


A transitional opioid program to engage hospitalized drug users.
Many opioid-dependent patients do not receive care for addiction issues when hospitalized for other medical problems. Based on 3 years of clinical practice, we report the Transitional Opioid Program (TOP) experience using hospitalization as a "reachable moment" to identify and link opioid-dependent persons to addiction treatment from medical care. A program nurse identified, assessed, and enrolled hospitalized, out-of-treatment opioid-dependent drug users based on their receipt of methadone during hospitalization. At discharge, patients transitioned to an outpatient interim opioid agonist program providing 30-day stabilization followed by 60-day taper. The nurse provided case management emphasizing HIV risk reduction, health education, counseling, and medical follow-up. Treatment outcomes included opioid agonist stabilization then taper or transfer to long-term opioid agonist treatment. From January 2002 to January 2005, 362 unique hospitalized, opioid-dependent drug users were screened; 56% (n = 203) met eligibility criteria and enrolled into the program. Subsequently, 82% (167/203) presented to the program clinic post-hospital discharge; for 59% (119/203) treatment was provided, for 26% (52/203) treatment was not provided, and for 16% (32/203) treatment was not possible (pursuit of TOP objectives precluded by medical problems, psychiatric issues, or incarceration). Program patients adhered to a spectrum of medical recommendations (e.g., obtaining prescription medications, medical follow-up). The Transitional Opioid Program (TOP) identified at-risk hospitalized, out-of-treatment opioid-dependent drug users and, by offering a range of treatment intensity options, engaged a majority into addiction treatment. Hospitalization can be a "reachable moment" to engage and link drug users into addiction treatment.


Blunt ocular trauma in patients over 70: Clinical characteristics and prognosis.
To describe the mechanisms and outcomes of open globe injuries (OGIs) in patients 70 years or older. A retrospective chart review of patients from two large academic centers in Israel who were underwent surgery for OGI. Data was collected from surgical and clinic charts. Anatomical and functional results were recorded as well any additional surgeries and complications. Thirty-two eyes of 32 patients were included. The mechanism of OGI consisted of blunt trauma with ocular rupture secondary to falls. All eyes underwent immediate primary closure of the corneal and scleral lacerations. Additional surgeries were performed in 34.3% of cases. In 31.2% of cases additional surgery was not required, and in 21.9% of cases the damage was so devastating that additional procedures were not recommended. Four patients (12.5%) chose not to undergo additional surgery. Mean visual acuity at presentation was 2.24 ± 0.72 logMAR and it improved to 1.92 ± 0.89 logMAR at final follow-up (p = 0.002). VA at presentation was significantly correlated to final VA (p < 0.001). OGI in the elderly represents a unique group. It is mostly related to falls, with a female predominance and a poor visual prognosis. In this patient population, general health concerns and increased risks of anesthesia require special attention. Care should be taken to educate elderly patients and their caretakers on how to avoid falls.


The effect of postoperative TACE on prognosis of HCC: an update.
Hepatocellular carcinoma (HCC) is a common malignancy in the world and is mainly treated by surgery resection. It is believed that even after radical resection, the recurrence and metastasis rates remain at a high level. This threatens the health and safety of patients. Postoperative adjuvant transcatheter arterial chemoembolization (TACE) is regarded as a common strategy for HCC patients at a high recurrence risk. However, there is a debate on the effects of postoperative TACE and range of applications in the medical world. Here we review the effects of postoperative TACE on the prognosis of HCC.


Maturation of the Autonomic Nervous System in Premature Infants: Estimating Development Based on Heart-Rate Variability Analysis.
This study aims at investigating the development of premature infants' autonomic nervous system (ANS) based on a quantitative analysis of the heart-rate variability (HRV) with a variety of novel features. Additionally, the role of heart-rate drops, known as bradycardias, has been studied in relation to both clinical and novel sympathovagal indices. ECG data were measured for at least 3 h in 25 preterm infants (gestational age ≤32 weeks) for a total number of 74 recordings. The post-menstrual age (PMA) of each patient was estimated from the RR interval time-series by means of multivariate linear-mixed effects regression. The tachograms were segmented based on bradycardias in periods after, between and during bradycardias. For each of those epochs, a set of temporal, spectral and fractal indices were included in the regression model. The best performing model has R 2 = 0.75 and mean absolute error MAE = 1.56 weeks. Three main novelties can be reported. First, the obtained maturation models based on HRV have comparable performance to other development models. Second, the selected features for age estimation show a predominance of power and fractal features in the very-low- and low-frequency bands in explaining the infants' sympathovagal development from 27 PMA weeks until 40 PMA weeks. Third, bradycardias might disrupt the relationship between common temporal indices of the tachogram and the age of the infant and the interpretation of sympathovagal indices. This approach might provide a novel overview of post-natal autonomic maturation and an alternative development index to other electrophysiological data analysis.


Methods to assess intended effects of drug treatment in observational studies are reviewed.
To review methods that seek to adjust for confounding in observational studies when assessing intended drug effects. We reviewed the statistical, economical and medical literature on the development, comparison and use of methods adjusting for confounding. In addition to standard statistical techniques of (logistic) regression and Cox proportional hazards regression, alternative methods have been proposed to adjust for confounding in observational studies. A first group of methods focus on the main problem of nonrandomization by balancing treatment groups on observed covariates: selection, matching, stratification, multivariate confounder score, and propensity score methods, of which the latter can be combined with stratification or various matching methods. Another group of methods look for variables to be used like randomization in order to adjust also for unobserved covariates: instrumental variable methods, two-stage least squares, and grouped-treatment approach. Identifying these variables is difficult, however, and assumptions are strong. Sensitivity analyses are useful tools in assessing the robustness and plausibility of the estimated treatment effects to variations in assumptions about unmeasured confounders. In most studies regression-like techniques are routinely used for adjustment for confounding, although alternative methods are available. More complete empirical evaluations comparing these methods in different situations are needed.


Invasive aspergillosis in pediatric patients.
This case-based review examines the growing literature on critical issues related to the epidemiology, diagnosis, and treatment of pediatric invasive aspergillosis. Immunocompromised children are at heightened risk for invasive aspergillosis. Children at highest risk include those with new-onset or relapsed hematologic malignancy and recipients of allogeneic stem cell transplants. Additional risk factors in stem cell transplant recipients include impaired lymphocyte engraftment and graft-versus-host disease. Pediatric invasive aspergillosis is associated with a high mortality rate (generally >50%) and requires prompt diagnosis and treatment to prevent dissemination and death. Tools available for diagnosis include radiologic examinations (primarily computed tomography), the galactomannan assay, bronchoalveolar lavage, and tissue biopsy. Age-related differences in computed tomography and galactomannan assay results have been suggested. Recommended primary therapy for pediatric invasive aspergillosis is voriconazole (7 mg/kg IV q12 hours). Currently approved alternative therapies include liposomal amphotericin B, amphotericin B lipid complex, and caspofungin. Posaconazole and itraconazole are also possibilities, but there is no established pediatric dose for posaconazole, and itraconazole dosing is difficult in children. In patients who do not benefit from initial antifungal therapy, options include switching to another agent with a different mechanism of action or combination therapy. Further research is required to better establish optimal approaches to the management of pediatric patients with invasive aspergillosis recalcitrant to initial primary therapy.


Infective endocarditis and infected aneurysm of splenic artery post colonoscopy.
Colonoscopy, a relatively non-invasive procedure, has been associated with several complications including perforation, hemorrhage and abdominal pain. Post-colonoscopy bacteremia can occur up to 4.4% of the time but is almost always transient without significant clinical sequelae. Post-colonoscopy infective endocarditis, on the other hand, is a rare occurrence associated with high rates of mortality and morbidity, and may be further complicated by aneurysm of splenic artery. Current definitive treatment of infected aneurysm is surgical ligation and excision with or without vascular anastomosis. If surgery is contraindicated, endovascular graft and transcatheter embolization may be the preferred treatment options. This is a case report of infective endocarditis and infected aneurysm of splenic artery presenting two weeks after elective colonoscopy.


ER Stress-Induced Secretion of Proteins and Their Extracellular Functions in the Heart.
Endoplasmic reticulum (ER) stress is a result of conditions that imbalance protein homeostasis or proteostasis at the ER, for example ischemia, and is a common event in various human pathologies, including the diseased heart. Cardiac integrity and function depend on the active secretion of mature proteins from a variety of cell types in the heart, a process that requires an intact ER environment for efficient protein folding and trafficking to the secretory pathway. As a consequence of ER stress, most protein secretion by the ER secretory pathway is decreased. Strikingly, there is a select group of proteins that are secreted in greater quantities during ER stress. ER stress resulting from the dysregulation of ER Ca2+ levels, for instance, stimulates the secretion of Ca2+-binding ER chaperones, especially GRP78, GRP94, calreticulin, and mesencephalic astrocyte-derived neurotrophic factor (MANF), which play a multitude of roles outside the cell, strongly depending on the cell type and tissue. Here we review current insights in ER stress-induced secretion of proteins, particularly from the heart, and highlight the extracellular functions of these proteins, ranging from the augmentation of cardiac cell viability to the modulation of pro- and anti-apoptotic, oncogenic, and immune-stimulatory cell signaling, cell invasion, extracellular proteostasis, and more. Many of the roles of ER stress-induced protein secretion remain to be explored in the heart. This article is part of a special issue entitled "The Role of Proteostasis Derailment in Cardiac Diseases."


Reliability of single sample experimental designs: comfortable effort level.
This study was designed to ascertain the intrasubject variability across multiple recording sessions-most often disregarded in reporting group mean data or unavailable because of single sample experimental designs. Intrasubject variability was assessed within and across several experimental sessions from measures of speaking fundamental frequency, vocal intensity, and reading rate. Three age groups of men and women--young, middle-aged, and elderly--repeated the vowel /a/, read a standard passage, and spoke extemporaneously during each experimental session. Statistical analyses were performed to assess each speaker's variability from his or her own mean, and that which consistently varied for any one speaking sample type, both within or across days. Results indicated that intrasubject variability was minimal, with approximately 4% of the data exhibiting significant variation across experimental sessions.


Geographical Disparities in Pooled Stroke Incidence and Case Fatality in Mainland China, Hong Kong, and Macao: Protocol for a Systematic Review and Meta-analysis.
Geographical variations in stroke incidence and case fatality in China have been reported. Nonetheless, pooled estimates in major Chinese regions are unknown. This systematic review and meta-analysis aims to investigate pooled estimates of incidence and short-term case fatality of stroke in Mainland China, Hong Kong, and Macao. Longitudinal studies published in English and indexed in PubMed/MEDLINE, Embase, CINAHL, and Web of Science, or in Chinese and indexed in SinoMed and CQVIP will be targeted. Articles reporting on adults living in China who experience first-ever stroke or die within 1 year from newly onset stroke will be included. The 95% confidence intervals of the event will be estimated using the exact method based on the Poisson distribution. The log incidence rates together with their corresponding log standard errors will be meta-analyzed using DerSimonian and Laird random effects models. Pooled case fatality rates will also be estimated using a random effect model. Time trends in pooled age-standardized stroke incidence and case fatality will be estimated. The heterogeneity of the included studies will be measured using the I2 statistic and meta-regressions will be run to analyze the effect of reported covariates on found heterogeneity. Risk of bias will be examined using the Newcastle-Ottawa Scale. Publication bias will be tested using funnel plots and Egger tests. Sensitivity analysis will be run by risk of bias. This study was funded and registered in 2020. The systematic searches, study selections, and quality assessments were completed in July 2021. Data extraction and analysis and manuscript writing are scheduled to be completed by December 2021. This will be the first study to provide regional differences in pooled estimates of stroke incidence with case fatality in Mainland China, Hong Kong, and Macao. This study will assist in addressing inequalities in stroke care across China. PROSPERO International Prospective Register of Systematic Reviews CRD42020170724; https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=170724. PRR1-10.2196/32566.


Devices for assessing microcirculation.
Devices for assessing microcirculation at the bedside have been a technological breakthrough in the critical care field. Thanks to this technology, a large body of scientific evidence has highlighted the relevance of microcirculatory disruptions during critical illness. The goal of this review is to analyze the current knowledge concerning microcirculation monitoring, mainly focused on clinically available devices. New evidence in oxygenation monitoring, advances in hand-held vital microscopes, and improvements in laser-based techniques ensure the possibility of detecting poor resuscitation, testing vascular reactivity, and assessing the effect of therapy during shock and resuscitation. Currently, there are several methods for microcirculatory monitoring. To properly apply and correctly interpret the information they provide, clinicians should know the fundamental principles and the strengths and weaknesses of the clinically available devices.


Varicella susceptibility and vaccine use among young adults enlisting in the United States Navy.
Primary varicella infection, or chicken pox, is a threat to all young adults who join the United States (U.S.) military if they fail to develop immunity prior to enlistment. Historically, outbreaks of chicken pox have caused marked morbidity and impaired military readiness. In December 1996, the U.S. Navy began performing serologic testing for varicella among all new recruits, and vaccinating those found to be sero-negative. We evaluated results of the screening program in its first 4 years, and used multivariable logistic regression modeling to describe factors associated with varicella susceptibility. Cases of chicken pox were tracked among all military services before and after program implementation. More than 190,000 young adults enlisted in the U.S. Navy between 1997 and 2000. Recruits originated from all 50 states and several foreign countries; 84% were male, and their average age was 19 years. Seven percent were found to be susceptible (sero-negative) to varicella. In multivariable modeling, race/ethnicity was associated with susceptibility, but age, gender, and home state were not. The overall incidence of chicken pox in the Navy was reduced by more than 80% after initiation of the screening-vaccination program. A successful varicella screening-vaccination program has been implemented in the U.S. Navy. Results of serologic screening undertaken on this large number of young adults may be useful in tracking the changing epidemiology of varicella in the general population in the post-vaccine era.


A Greater Improvement of Intrahepatic Fat Contents after 6 Months of Lifestyle Intervention Is Related to a Better Oxidative Stress and Inflammatory Status in Non-Alcoholic Fatty Liver Disease.
Non-alcoholic fatty liver disease (NAFLD) is a disorder characterized by the excessive accumulation of lipids in the liver parenchyma. To date, there is no effective pharmacological treatment against NAFLD. Objective: To assess the relationship between the improvement of the intrahepatic fat content (IFC) in patients with NAFLD and metabolic syndrome and biomarkers of oxidative stress and inflammation after 6 months of lifestyle intervention. Patients diagnosed with NAFLD (n = 60 adults; 40-60 years old) residing in the Balearic Islands, Spain, were distributed in tertiles attending the improvement of IFC calculated by magnetic resonance imaging (MRI). Anthropometrics, blood pressure, maximal oxygen uptake, and pro/antioxidant and inflammatory biomarkers were determined in plasma before and after the lifestyle intervention. The improvement in IFC levels was higher in tertile 3 with respect to tertiles 2 and 1. The greatest improvement in IFC is related to cardiorespiratory fitness and adherence to the Mediterranean diet (ADM). Higher reductions in weight, body mass index (BMI), and alanine aminotransferase (ALT) were observed in tertile 3 with respect to tertile 1 after 6 months of intervention. The improvement in catalase, irisin, and cytokeratin 18 plasma levels were higher in tertile 3, whereas no differences were observed in superoxide dismutase activity. Malondialdehyde and protein carbonyl levels, as biomarkers of oxidative damage, remained unchanged in all groups. The present data show that the reduction of IFC is associated with an improvement in pro/antioxidant and pro-inflammatory status and a better cardiorespiratory fitness in NAFLD patients.


Oral health knowledge, behaviour and practices among school children in Qatar.
The aim of this study was to assess the oral health knowledge behaviour and practices among school children in Qatar. A cross-sectional study was carried out in Qatar from October 2011 to April 2012. A total of 2200 school children aged 12-14 years were approached from 16 schools of different areas. The information about oral health knowledge and sources of information was obtained through a self-administrated questionnaire. Data analyses were performed. The overall response rate was (96%). Only (25.8%) of children reported a high level of oral health knowledge. After each meal, tooth brushing was observed by a very low percentage of children (3.7%). About 44.6% of children recognized dental floss as a cleaning device for between the teeth. A large number of children (32.5%) thought incorrectly that one must visit the dentist only in case of pain. A great majority was not aware of cariogenic potential of soft drinks (39%) and sweetened milk (97.8%). Less than half (38.9%) of children actually had heard about fluoride. Only (16.8%) correctly answered the question about sign of tooth decay. Slightly, less than half (48.4%) could not define the meaning of plaque. Parents were the most popular (69.1%), source of oral health information for the children. The oral health knowledge in Qatar is below the satisfactory level. Parents were the most popular source of oral health knowledge for the children followed by dentists, school teachers, and media.


[Hashimoto's encephalopathy: toxic or autoimmune mechanism?].
A 36-year-old woman presented with partial complex status epilepticus. Magnetic resonance imaging with T2-weighted sequences showed a high-intensity signal in the left posterior frontal area. Hashimoto's thyroiditis was then discovered. The disappearance of the high-intensity signal after corticosteroid therapy was suggestive of an autoimmune mechanism. However, improvement could be obtained only with a hormonal treatment, which supports the hypothesis of a pathogenetic role of the Tyrosine-Releasing Hormone (TRH).


A randomized trial alerting authors, with or without coauthors or editors, that research they cited in systematic reviews and guidelines has been retracted.
Retracted clinical trials may be influential in citing systematic reviews and clinical guidelines. We assessed the influence of 27 retracted trials on systematic reviews and clinical guidelines (citing publications), then alerted authors to these retractions. Citing publications were randomized to up to three e-mails to contact author with/without up to two coauthors, with/without the editor. After one year we assessed corrective action. We included 88 citing publications; 51% (45/88) had findings likely to change if retracted trials were removed, 87% (39/45) likely substantially. 51% (44/86) of contacted citing publications replied. Including three authors rather than the contact author alone was more likely to elicit a reply (P = 0.03). Including the editor did not increase replies (P = 0.66). Whether findings were judged likely to change, and size of the likely change, had no effect on response rate or action taken. One year after e-mails were sent only nine publications had published notifications. E-Mail alerts to authors and editors are inadequate to correct the impact of retracted publications in citing systematic reviews and guidelines. Changes to bibliographic and referencing systems, and submission processes are needed. Citing publications with retracted citations should be marked until authors resolve concerns.


Social networks, social support mechanisms, and quality of life after breast cancer diagnosis.
We examined mechanisms through which social relationships influence quality of life (QOL) in breast cancer survivors. This study included 3,139 women from the Pathways Study who were diagnosed with breast cancer from 2006 to 2011 and provided data on social networks (the presence of a spouse or intimate partner, religious/social ties, volunteering, and numbers of close friends and relatives), social support (tangible support, emotional/informational support, affection, positive social interaction), and QOL, measured by the FACT-B, approximately 2 months post diagnosis. We used logistic models to evaluate associations between social network size, social support, and lower versus higher than median QOL scores. We further stratified by stage at diagnosis and treatment. In multivariate-adjusted analyses, women who were characterized as socially isolated had significantly lower FACT-B (OR = 2.18, 95 % CI: 1.72-2.77), physical well-being (WB) (OR = 1.61, 95 % CI: 1.27-2.03), functional WB (OR = 2.08, 95 % CI: 1.65-2.63), social WB (OR = 3.46, 95 % CI: 2.73-4.39), and emotional WB (OR = 1.67, 95 % CI: 1.33-2.11) scores and higher breast cancer symptoms (OR = 1.48, 95 % CI: 1.18-1.87) compared with socially integrated women. Each social network member independently predicted higher QOL. Simultaneous adjustment for social networks and social support partially attenuated associations between social networks and QOL. The strongest mediator and type of social support that was most predictive of QOL outcomes was "positive social interaction." However, each type of support was important depending on outcome, stage, and treatment status. Larger social networks and greater social support were related to higher QOL after a diagnosis of breast cancer. Effective social support interventions need to evolve beyond social-emotional interventions and need to account for disease severity and treatment status.


The Efficacy of Probiotics, Prebiotic Inulin-Type Fructans, and Synbiotics in Human Ulcerative Colitis: A Systematic Review and Meta-Analysis.
Studies of probiotics, fructan-type prebiotics, and synbiotics in patients with ulcerative colitis (UC) show significant heterogeneity in methodology and results. Here, we study the efficacy of such interventions and the reasons for the heterogeneity of their results. Eligible random controlled trials were collected from the PUBMED and SCOPUS databases. A total of 18 placebo-controlled and active treatment-controlled (i.e., mesalazine) studies were selected with a Jadad score ≥ 3, including 1491 patients with UC. Data for prebiotics and synbiotics were sparse and consequently these studies were excluded from the meta-analysis. The UC remission efficacy of probiotics was measured in terms of relative risk (RR) and odds ratio (OR). Significant effects were observed in patients with active UC whenever probiotics containing bifidobacteria were used, or when adopting the US Food and Drug Administration (FDA)-recommended scales (UC Disease Activity Index and Disease Activity Index). By the FDA recommended scales, the RR was 1.55 (CI95%: 1.13⁻2.15, p-value = 0.007, I² = 29%); for bifidobacteria-containing probiotics, the RR was 1.73 (CI95%: 1.23⁻2.43, p-value = 0.002, I² = 35%). No significant effects were observed on the maintenance of remission for placebo-controlled or mesalazine-controlled studies. We conclude that a validated scale is necessary to determine the state of patients with UC. However, probiotics containing bifidobacteria are promising for the treatment of active UC.


Synthesis of citrulline from glutamine in pig enterocytes.
The synthesis of citrulline from glutamine was quantified in enterocytes from pre-weaning (14-21 days old) and post-weaning (29-58 days old) pigs. The cells were incubated at 37 degrees C for 30 min in Krebs-Henseleit bicarbonate buffer (pH 7.4) containing 0, 0.5, 2 and 5 mM glutamine. Oxygen consumption was linear during the 30 min incubation period. The rates of citrulline synthesis were low or negligible in enterocytes from 14-21-day-old pigs, but increased 10-20-fold in the cells from 29-58-day-old pigs. This marked elevation of citrulline synthesis coincided with an increase in the activity of pyrroline-5-carboxylate synthase with the animal's post-weaning growth. In contrast, decreases in the activities of phosphate-dependent glutaminase, ornithine aminotransferase, ornithine carbamoyltransferase and carbamoyl-phosphate synthase were observed as the age of the pigs increased. The concentrations of carbamoyl phosphate in enterocytes from pre-weaning pigs were higher than, or similar to, those in the cells from post-weaning pigs. It is possible that the low rate of citrulline synthesis from glutamine in enterocytes from pre-weaning pigs was due to a limited availability of ornithine, rather than that of carbamoyl phosphate. We suggest that this limited availability of ornithine in pre-weaning-pig enterocytes results from (i) the low rate of pyrroline-5-carboxylate synthesis from glutamate, due to the low activity of pyrroline-5-carboxylate synthase, and (ii) the competitive conversion of pyrroline-5-carboxylate into proline. Our present findings on the developmental aspect of citrulline synthesis in pig enterocytes may offer a biochemical mechanism for the previous observations that arginine is a nutritionally essential amino acid for suckling piglets, but not for adult pigs.


Notch activation enhances the microglia-mediated inflammatory response associated with focal cerebral ischemia.
Activation of Notch worsens ischemic brain damage as antisense knockdown or pharmacological inhibition of the Notch pathway reduces the infarct size and improves the functional outcome in a mouse model of stroke. We sought to determine whether Notch activation contributes to postischemic inflammation by directly modulating the microglial innate response. The microglial response and the attendant inflammatory reaction were evaluated in Notch1 antisense transgenic (Tg) and in nontransgenic (non-Tg) mice subjected to middle cerebral artery occlusion with or without treatment with a γ-secretase inhibitor (GSI). To investigate the impact of Notch on microglial effector functions, primary mouse microglia and murine BV-2 microglial cell line were exposed to oxygen glucose deprivation or lipopolysaccharide in the presence or absence of GSI. Immunofluorescence labeling, Western blotting, and reverse-transcription polymerase chain reaction were performed to measure microglial activation and production of inflammatory cytokines. The nuclear translocation of nuclear factor-κB in microglia was assessed by immunohistochemistry. The neurotoxic potential of microglia was determined in cocultures. Notch1 antisense mice exhibit significantly lower numbers of activated microglia and reduced proinflammatory cytokine expression in the ipsilateral ischemic cortices compared to non-Tg mice. Microglial activation also was attenuated in Notch1 antisense cultures and in non-Tg cultures treated with GSI. GSI significantly reduced nuclear factor-κB activation and expression of proinflammatory mediators and markedly attenuated the neurotoxic activity of microglia in cocultures. These findings establish a role for Notch signaling in modulating the microglia innate response and suggest that inhibition of Notch might represent a complementary therapeutic approach to prevent reactive gliosis in stroke and neuroinflammation-related degenerative disorders.


Brain-site-specific proteome changes induced by neuronal P60TRP expression.
p60 transcription regulator protein (p60TRP) facilitates the processing of the amyloid precursor protein towards the non-amyloidogenic pathway by inhibiting the β-secretase action. This protein was initially identified to be downregulated in the temporal lobe of brains from Alzheimer's disease patients. p60TRP is one of the G-protein-coupled receptor (GPCR)-associated proteins which directly influences the signalling capacity of GPCRs. In the present study, we investigated the brain-region-specific proteome profile of transgenic p60TRP mice to gain an insight into the molecular events mediated by the long-term effect of neuronal p60TRP overexpression on brain proteome changes and its potential implication for neuronal functions in the central nervous system. Using a proteomics research approach based on isobaric tags for relative and absolute quantitation, we identified 2,025 proteins, whereby 1,735 proteins were quantified, out of which 56 were found to be significantly altered in the cortex and/or hippocampus of neuronal transgenic neuronal p60TRP mice. Our data suggests that in vivo overexpression of neuronal p60TRP significantly affects cognitive and neuroprotective capacities.


Discovery and Mechanistic Characterization of Selective Inhibitors of H2S-producing Enzyme: 3-Mercaptopyruvate Sulfurtransferase (3MST) Targeting Active-site Cysteine Persulfide.
Very recent studies indicate that sulfur atoms with oxidation state 0 or -1, called sulfane sulfurs, are the actual mediators of some physiological processes previously considered to be regulated by hydrogen sulfide (H2S). 3-Mercaptopyruvate sulfurtransferase (3MST), one of three H2S-producing enzymes, was also recently shown to produce sulfane sulfur (H2Sn). Here, we report the discovery of several potent 3MST inhibitors by means of high-throughput screening (HTS) of a large chemical library (174,118 compounds) with our H2S-selective fluorescent probe, HSip-1. Most of the identified inhibitors had similar aromatic ring-carbonyl-S-pyrimidone structures. Among them, compound 3 showed very high selectivity for 3MST over other H2S/sulfane sulfur-producing enzymes and rhodanese. The X-ray crystal structures of 3MST complexes with two of the inhibitors revealed that their target is a persulfurated cysteine residue located in the active site of 3MST. Precise theoretical calculations indicated the presence of a strong long-range electrostatic interaction between the persulfur anion of the persulfurated cysteine residue and the positively charged carbonyl carbon of the pyrimidone moiety of the inhibitor. Our results also provide the experimental support for the idea that the 3MST-catalyzed reaction with 3-mercaptopyruvate proceeds via a ping-pong mechanism.


Cellular uptake and intracellular degradation of poly(alkyl cyanoacrylate) nanoparticles.
Poly(alkyl cyanoacrylate) (PACA) nanoparticles have shown promise as drug carriers both to solid tumors and across the blood-brain barrier. Efficient drug delivery requires both high cellular uptake of the nanoparticles and release of the drug from the nanoparticles. Release of hydrophobic drugs from PACA nanoparticles is primarily governed by nanoparticle degradation, and this process has been poorly studied at the cellular level. Here we use the hydrophobic model drug Nile Red 668 (NR668) to investigate intracellular degradation of PACA nanoparticles by measuring changes in NR668 fluorescence emission and lifetime, as the spectral properties of NR668 depend on the hydrophobicity of the dye environment. We also assess the potential of poly(butyl cyanoacrylate) (PBCA) and poly(octyl cyanoacrylate) (POCA) nanoparticles for intracellular drug delivery in the prostate cancer cell line PC3 and rat brain endothelial cell line RBE4 and the role of endocytosis pathways in PACA nanoparticle uptake in those cell lines. Fluorescence lifetime imaging, emission spectra analysis and Förster resonance energy transfer indicated that the intracellular degradation was in line with the degradation found by direct methods such as gas chromatography and scanning electron microscopy, showing that PBCA has a faster degradation rate compared to POCA. The combined P(BCA/OCA) nanoparticles had an intermediate degradation rate. The uptake of POCA and PBCA nanoparticles was much higher in RBE4 than in PC3 cells. Endocytosis inhibition studies showed that both clathrin- and caveolin-mediated endocytosis were involved in PACA nanoparticle uptake, and that the former played a predominant role, particularly in PC3 cells. In the present study, we used three different optical techniques to show that within a 24-hour period PBCA nanoparticles degraded significantly inside cells, releasing their payload into the cytosol, while POCA nanoparticles remained intact. This indicates that it is possible to tune the intracellular drug release rate by choosing appropriate monomers from the PACA family or by using hybrid PACA nanoparticles containing different monomers. In addition, we showed that the uptake of PACA nanoparticles depends not only on the monomer material, but also on the cell type, and that different cell lines can use different internalization pathways.


Soluble silica inhibits osteoclast formation and bone resorption in vitro.
Several studies have suggested that silicon (Si) may be essential for the normal development of connective tissue and the skeleton. Positive effects of Si from the diet as well as from Si-containing biomaterials, such as bioactive glass 45S5 (BG), have been demonstrated. Studies have reported that Si stimulates osteoblast proliferation and differentiation. However, the effects of Si on osteoclasts have not been directly addressed. The purpose of the present in vitro study was to clarify if Si has regulatory effects on osteoclast formation and bone resorption. The effects of BG, BG dissolution extracts and Si containing cell culture medium were investigated in a mouse calvarial bone resorption assay and osteoclast formation assays (mouse bone marrow cultures and RAW264.7 cell cultures). We conclude from our results that Si causes significant inhibition of osteoclast phenotypic gene expressions, osteoclast formation and bone resorption in vitro. In conclusion, the present study suggests that Si has a dual nature in bone metabolism with stimulatory effects on osteoblasts and inhibitory effects on osteoclasts. This suggested property of Si might be interesting to further explore in future biomaterials for treatments of bone defects in patients.


Small-Scale Sustainable Water Project Decreases Infections, Complements Short-Term Medical Missions.
Many communities in developing countries do not have access to clean water, which is essential to health. This project involved the installation of clean water filters in two rural Guatemalan villages, a small-scale sustainable community project proposed by a short-term medical mission team and accomplished in partnership with local missionaries. The percentage of infections in adult visitors to medical clinics was assessed pre- and post-installation of filters, and although not statistically significant, decreased.


Constitutive knockout of Surf1 is associated with high embryonic lethality, mitochondrial disease and cytochrome c oxidase deficiency in mice.
We report here the creation of a constitutive knockout mouse for SURF1, a gene encoding one of the assembly proteins involved in the formation of cytochrome c oxidase (COX). Loss-of-function mutations of SURF1 cause Leigh syndrome associated with an isolated and generalized COX deficiency in humans. The murine phenotype is characterized by the following hallmarks: (1) high post-implantation embryonic lethality, affecting approximately 90% of the Surf1(-/-) individuals; (2) early-onset mortality of post-natal individuals; (3) highly significant deficit in muscle strength and motor performance; (4) profound and isolated defect of COX activity in skeletal muscle and liver, and, to a lesser extent, heart and brain; (5) morphological abnormalities of skeletal muscle, characterized by reduced histochemical reaction to COX and mitochondrial proliferation; (6) no obvious abnormalities in brain morphology, reflecting the virtual absence of overt neurological symptoms. These results indicate a function for murine Surf1 protein (Surf1p) specifically related to COX and recapitulate, at least in part, the human phenotype. This is the first mammalian model for a nuclear disease gene of a human mitochondrial disorder. Our model constitutes a useful tool to investigate the function of Surf1p, help understand the pathogenesis of Surf1p deficiency in vivo, and evaluate the efficacy of treatment.