Etiology of posterior subcapsular cataracts based on a review of risk factors including aging, diabetes, and ionizing radiation.
Since the exact development of posterior subcapsular cataracts (PSCs) is poorly understood, we review various risk factors and propose a two-stage etiology for PSCs. The biological mechanisms associated with age-related cataracts (primarily nuclear cataracts, cortical cataracts and PSCs) were reviewed in relation to selected risk factors that induce PSCs (including atopy, diabetes, hypoparathyroidism, myopia, retinitis, solar radiation, steroid use, uveitis, vitrectomy and ionizing radiation). We particularly focused on ionizing radiation, as this is known to be a risk factor specific to PSCs. Based on an analysis of the reviewed material, we propose a detailed explanation of the etiology of PSCs. Lens epithelial cells (LECs) and lens fiber cells are normally hypoxic and therefore very sensitive to changes in oxidative stress, as quantified by the radiation oxygen effect. We hypothesize that the development of PSC opacities is a two-stage process. Stage I, early in life, is driven by risk factors that promote oxidative stress and ion-pump disruption, harming lens fibers and causing aberrant LECs to proliferate and ectopically migrate as Wedl cells (perhaps by processes associated with an epithelial to mesenchymal transition) to the posterior pole region. After a latent period, in Stage II, the development of PSCs advances mainly due to chronic inflammation and other premature aging-related mechanisms that promote mature vacuolar or plaque PSC. This two-stage hypothesis of PSC etiology accounts for risk factors, such as aging, diabetes and ionizing radiation, which directly affects LECs and the lens. In addition, these risk factors can damage other ocular regions, such as the retina and vitreous, that also indirectly contribute to the development of PSCs. It is possible that the incidence of PSCs may be reduced by reversing the effects of Stage I through various means, including ocular antioxidants.


A bright FIT-PNA hybridization probe for the hybridization state specific analysis of a C → U RNA edit via FRET in a binary system.
Oligonucleotide probes that show enhanced fluorescence upon nucleic acid hybridization enable the detection and visualization of specific mRNA molecules, in vitro and in cellulo. A challenging problem is the analysis of single nucleotide alterations that occur, for example, when cellular mRNA is subject to C → U editing. Given the length required for uniqueness of the targeted segment, the commonly used probes do not provide the level of sequence specificity needed to discriminate single base mismatched hybridization. Herein we introduce a binary probe system based on fluorescence resonance energy transfer (FRET) that distinguishes three possible states i.e. (i) absence of target, (ii) presence of edited (matched) and (iii) unedited (single base mismatched) target. To address the shortcomings of read-out via FRET, we designed donor probes that avoid bleed through into the acceptor channel and nevertheless provide a high intensity of FRET signaling. We show the combined use of thiazole orange (TO) and an oxazolopyridine analogue (JO), linked as base surrogates in modified PNA FIT-probes that serve as FRET donor for a second, near-infrared (NIR)-labeled strand. In absence of target, donor emission is low and FRET cannot occur in lieu of the lacking co-alignment of probes. Hybridization of the TO/JO-PNA FIT-probe with the (unedited RNA) target leads to high brightness of emission at 540 nm. Co-alignment of the NIR-acceptor strand ensues from recognition of edited RNA inducing emission at 690 nm. We show imaging of mRNA in fixed and live cells and discuss the homogeneous detection and intracellular imaging of a single nucleotide mRNA edit used by nature to post-transcriptionally modify the function of the Glycine Receptor (GlyR).


Projected impact of a reduction in sugar-sweetened beverage consumption on diabetes and cardiovascular disease in Argentina: A modeling study.
Sugar-sweetened beverage (SSB) consumption is associated with obesity, diabetes, and hypertension. Argentina is one of the major consumers of SSBs per capita worldwide. Determining the impact of SSB reduction on health will inform policy debates. We used the Cardiovascular Disease Policy Model-Argentina (CVD Policy Model-Argentina), a local adaptation of a well-established computer simulation model that projects cardiovascular and mortality events for the population 35-94 years old, to estimate the impact of reducing SSB consumption on diabetes incidence, cardiovascular events, and mortality in Argentina during the period 2015-2024, using local demographic and consumption data. Given uncertainty regarding the exact amount of SSBs consumed by different age groups, we modeled 2 estimates of baseline consumption (low and high) under 2 different scenarios: a 10% and a 20% decrease in SSB consumption. We also included a range of caloric compensation in the model (0%, 39%, and 100%). We used Monte Carlo simulations to generate 95% uncertainty intervals (UIs) around our primary outcome measures for each intervention scenario. Over the 2015-2024 period, a 10% reduction in SSBs with a caloric compensation of 39% is projected to reduce incident diabetes cases by 13,300 (95% UI 10,800-15,600 [low SSB consumption estimate]) to 27,700 cases (95% UI 22,400-32,400 [high SSB consumption estimate]), i.e., 1.7% and 3.6% fewer cases, respectively, compared to a scenario of no change in SSB consumption. It would also reduce myocardial infarctions by 2,500 (95% UI 2,200-2,800) to 5,100 (95% UI 4,500-5,700) events and all-cause deaths by 2,700 (95% UI 2,200-3,200) to 5,600 (95% UI 4,600-6,600) for "low" and "high" estimates of SSB intake, respectively. A 20% reduction in SSB consumption with 39% caloric compensation is projected to result in 26,200 (95% UI 21,200-30,600) to 53,800 (95% UI 43,900-62,700) fewer cases of diabetes, 4,800 (95% UI 4,200-5,300) to 10,000 (95% UI 8,800-11,200) fewer myocardial infarctions, and 5,200 (95% UI 4,300-6,200) to 11,000 (95% UI 9,100-13,100) fewer deaths. The largest reductions in diabetes and cardiovascular events were observed in the youngest age group modeled (35-44 years) for both men and women; additionally, more events could be avoided in men compared to women in all age groups. The main limitations of our study are the limited availability of SSB consumption data in Argentina and the fact that we were only able to model the possible benefits of the interventions for the population older than 34 years. Our study finds that, even under conservative assumptions, a relatively small reduction in SSB consumption could lead to a substantial decrease in diabetes incidence, cardiovascular events, and mortality in Argentina.


A yeast gene important for protein assembly into the endoplasmic reticulum and the nucleus has homology to DnaJ, an Escherichia coli heat shock protein.
When nuclear localization sequences (termed NLS) are placed at the N terminus of cytochrome c1, a mitochondrial inner membrane protein, the resulting hybrid proteins do not assemble into mitochondria when synthesized in the yeast Saccharomyces cerevisiae. Cells lacking mitochondrial cytochrome c1, but expressing the hybrid NLS-cytochrome c1 proteins, are unable to grow on glycerol since the hybrid proteins are associated primarily with the nucleus. A similar hybrid protein with a mutant NLS is transported to and assembled into the mitochondria. To identify proteins that might be involved in recognition of nuclear localization signals, we isolated conditional-lethal mutants (npl, for nuclear protein localization) that missorted NLS-cytochrome c1 to the mitochondria, allowing growth on glycerol. The gene corresponding to one complementation group (NPL1) encodes a protein with homology to DnaJ, an Escherichia coli heat shock protein. npl1-1 is allelic to sec63, a gene that affects transit of nascent secretory proteins across the endoplasmic reticulum. Rothblatt, J. A., R. J. Deshaies, S. L. Sanders, G. Daum, and R. Schekman. 1989. J. Cell Biol. 109:2641-2652. The npl1 mutants reported here also weakly affect translocation of preprocarboxypeptidaseY across the ER membrane. A normally nuclear hybrid protein containing a NLS fused to invertase and a nucleolar protein are not localized to the nucleus in npl1/sec63 cells at the nonpermissive temperature. Thus, NPL1/SEC63 may act at a very early common step in localization of proteins to the nucleus and the ER. Alternatively, by affecting ER and nuclear envelope assembly, npl1 may indirectly alter assembly of proteins into the nucleus.


Environmental engineering of navigation infrastructure: a survey of existing practices, challenges, and potential opportunities.
Navigation infrastructure such as channels, jetties, river training structures, and lock-and-dam facilities are primary components of a safe and efficient water transportation system. Planning for such infrastructure has until recently involved efforts to minimize impacts on the environment through a standardized environmental assessment process. More recently, consistent with environmental sustainability concepts, planners have begun to consider how such projects can also be constructed with environmental enhancements. This study examined the existing institutional conditions within the US Army Corps of Engineers and cooperating federal agencies relative to incorporating environmental enhancements into navigation infrastructure projects. The study sought to (1) investigate institutional attitudes towards the environmental enhancement of navigation infrastructure (EENI) concept, (2) identify potential impediments to implementation and solutions to such impediments, (3) identify existing navigation projects designed with the express intent of enhancing environmental benefit in addition to the primary project purpose, (4) identify innovative ideas for increasing environmental benefits for navigation projects, (5) identify needs for additional technical information or research, and (6) identify laws, regulations, and policies that both support and hinder such design features. The principal investigation tool was an Internet-based survey with 53 questions. The survey captured a wide range of perspectives on the EENI concept including ideas, concerns, research needs, and relevant laws and policies. Study recommendations included further promotion of the concept of EENI to planners and designers, documentation of existing projects, initiation of pilot studies on some of the innovative ideas provided through the survey, and development of national goals and interagency agreements to facilitate implementation.


Non-O ABO blood group genotypes differ in their associations with Plasmodium falciparum rosetting and severe malaria.
Blood group O is associated with protection against severe malaria and reduced size and stability of P. falciparum-host red blood cell (RBC) rosettes compared to non-O blood groups. Whether the non-O blood groups encoded by the specific ABO genotypes AO, BO, AA, BB and AB differ in their associations with severe malaria and rosetting is unknown. The A and B antigens are host RBC receptors for rosetting, hence we hypothesized that the higher levels of A and/or B antigen on RBCs from AA, BB and AB genotypes compared to AO/BO genotypes could lead to larger rosettes, increased microvascular obstruction and higher risk of malaria pathology. We used a case-control study of Kenyan children and in vitro adhesion assays to test the hypothesis that "double dose" non-O genotypes (AA, BB, AB) are associated with increased risk of severe malaria and larger rosettes than "single dose" heterozygotes (AO, BO). In the case-control study, compared to OO, the double dose genotypes consistently had higher odds ratios (OR) for severe malaria than single dose genotypes, with AB (OR 1.93) and AO (OR 1.27) showing most marked difference (p = 0.02, Wald test). In vitro experiments with blood group A-preferring P. falciparum parasites showed that significantly larger rosettes were formed with AA and AB host RBCs compared to OO, whereas AO and BO genotypes rosettes were indistinguishable from OO. Overall, the data show that ABO genotype influences P. falciparum rosetting and support the hypothesis that double dose non-O genotypes confer a greater risk of severe malaria than AO/BO heterozygosity.


Bleeding events and mortality in SCT patients: a retrospective study of hematopoietic SCT patients with organ dysfunctions due to severe sepsis or GVHD.
In autologous and allogeneic hematopoietic SCT (HSCT) neutropenia may be associated with severe infection. Immunodeficiency associated with GVHD and its treatment in allogeneic HSCT is also a risk for severe infection. In both periods, patients may develop severe sepsis with organ failure. To gain insights into treatment possibilities, HISTORY, a multicenter retrospective study reviewed HSCT patient records on mortality, organ dysfunction, platelet count and bleeding events. All transplantation records from 16 European centers were reviewed for 1.5 years. Of 2092 patients screened, 160 were documented for HSCT with respiratory and/or cardiovascular organ dysfunction because of sepsis and/or GVHD. Mortality was 53.1% at 28 days and 65.6% at 100 days. HSCT patients with sepsis and organ dysfunction are at highest risk of death (49.5%). Death from refractory septic shock was 15.2%, and it was 20% from respiratory failure and 64.7% from sepsis. Fewer than 3% of HSCT patients died from bleeding complications; however, individuals at increased risk of bleeding were excluded. Despite low platelet counts, an increased risk of bleeding could be established only if thrombocytopenia dropped below 13 x 10(9)/l. Thus, there might be a therapeutic window for treatment strategies for severe sepsis in HSCT, such as drotrecogin alpha (activated).


Desfechos em fetos e recém-nascidos expostos a infecções na gravidez.
To analyze outcomes in fetuses and newborns exposed to infections during pregnancy. Cross-sectional, quantitative study, carried out in a public maternity hospital in Maceió, Alagoas, Brazil. The sample consisted of 145 medical records of pregnant women admitted between 2015 and 2018 with possible vertically transmitted infections. Incomplete medical records or those that did not make it possible to describe fetal/neonatal exposure were excluded. The chi-squared test was used to verify the association between variables. A greater occurrence of congenital syphilis was observed (28.8%). There was more than one outcome in the same individual, such as low birth weight (39%), respiratory distress (20.5%), oligohydramnios (20%), congenital malformation and small size for gestational age (10.8%). Maternal infections and the number of prenatal visits revealed an association with fetal/neonatal outcome (p ≤ 0.05). The data obtained indicate the occurrence of unfavorable fetal/neonatal outcomes when related to neonatal infections and indicate the need for strategies that strengthen the coping with vertical transmissions.


DATA MINING APPROACH FOR IN-HOSPITAL TREATMENT OUTCOME IN PATIENTS WITH ACUTE CORONARY SYNDROME.
Risk stratification is nowadays crucial when estimating the patient's prognosis in terms of treatment outcome and it also helps in clinical decision making. Several risk assessment models have been developed to predict short-term outcomes in patients with acute coronary syndrome. This study was aimed at developing an outcome prediction model for patients with acute coronary syndrome submitted to percutaneus coronary intervention using data mining approach. A total of 2030 patients hospitalized for acute coronary syndrome and treated with percutaneous coronary intervention from December 2008 to December 2011 were assigned to a derivation cohort. Demographic and anamnestic data, clinical characteristics on admission, biochemical analysis of blood parameters on admission, and left ventricular ejection fraction formed the basis ofthe study. A number of machine learning algorithms available within Waikato Environment for Knowledge Discovery had been evaluated and the most successful was chosen. The predictive model was subsequently validated in a different population of 931 patients (validation cohort), hospitalized during 2012. The best prediction results were achieved using Alternating Decision Tree classifier, which was able to predict in-hospital mortality with 89% accuracy, and preserved good performance on validation cohort with 87% accuracy. Alternating Decision Tree classifier identified a subset of 6 attributes most relevant to mortality prediction: systolic and diastolic blood pressure, heart rate, left ventricular ejection fraction, age, and troponin value. Data mining approach enabled the authors to develop a model capable of predicting the in-hospital outcome following percutaneous coronary intervention. The model showed excellent sensitivity and specificity during internal validation.


Dual spring force couples yield multifunctionality and ultrafast, precision rotation in tiny biomechanical systems.
Small organisms use propulsive springs rather than muscles to repeatedly actuate high acceleration movements, even when constrained to tiny displacements and limited by inertial forces. Through integration of a large kinematic dataset, measurements of elastic recoil, energetic math modeling and dynamic math modeling, we tested how trap-jaw ants (Odontomachus brunneus) utilize multiple elastic structures to develop ultrafast and precise mandible rotations at small scales. We found that O. brunneus develops torque on each mandible using an intriguing configuration of two springs: their elastic head capsule recoils to push and the recoiling muscle-apodeme unit tugs on each mandible. Mandibles achieved precise, planar, circular trajectories up to 49,100 rad s-1 (470,000 rpm) when powered by spring propulsion. Once spring propulsion ended, the mandibles moved with unconstrained and oscillatory rotation. We term this mechanism a 'dual spring force couple', meaning that two springs deliver energy at two locations to develop torque. Dynamic modeling revealed that dual spring force couples reduce the need for joint constraints and thereby reduce dissipative joint losses, which is essential to the repeated use of ultrafast, small systems. Dual spring force couples enable multifunctionality: trap-jaw ants use the same mechanical system to produce ultrafast, planar strikes driven by propulsive springs and for generating slow, multi-degrees of freedom mandible manipulations using muscles, rather than springs, to directly actuate the movement. Dual spring force couples are found in other systems and are likely widespread in biology. These principles can be incorporated into microrobotics to improve multifunctionality, precision and longevity of ultrafast systems.


Electrospun Filtering Membrane Designed as Component of Self-Decontaminating Protective Masks.
The 2019 coronavirus outbreak and worsening air pollution have triggered the search for manufacturing effective protective masks preventing both particulate matter and biohazard absorption through the respiratory tract. Therefore, the design of advanced filtering textiles combining efficient physical barrier properties with antimicrobial properties is more newsworthy than ever. The objective of this work was to produce a filtering electrospun membrane incorporating a biocidal agent that would offer both optimal filtration efficiency and fast deactivation of entrapped viruses and bacteria. After the eco-friendly electrospinning process, polyvinyl alcohol (PVA) nanofibers were stabilized by crosslinking with 1,2,3,4-butanetetracarboxylic acid (BTCA). To compensate their low mechanical properties, nanofiber membranes with variable grammages were directly electrospun on a meltblown polypropylene (PP) support of 30 g/m2. The results demonstrated that nanofibers supported on PP with a grammage of around only 2 g/m2 presented the best compromise between filtration efficiencies of PM0.3, PM0.5, and PM3.0 and the pressure drop. The filtering electrospun membranes loaded with benzalkonium chloride (ADBAC) as a biocidal agent were successfully tested against E. coli and S. aureus and against human coronavirus strain HCoV-229E. This new biocidal filter based on electrospun nanofibers supported on PP nonwoven fabric could be a promising solution for personal and collective protection in a pandemic context.


Neuronal activity in cortical motor areas related to ipsilateral, contralateral, and bilateral digit movements of the monkey.
1. Single cell activity was studied in the precentral (PCM), premotor (PM), and supplementary (SMA) motor cortex of the monkey to compare magnitudes of activity changes in relation to ipsilateral, contralateral, and bilateral digit movements. 2. Three Japanese monkeys were trained to press a small key with the right or left hand, or with both hands, in accordance with visual instruction signals given 2.6-5.4 s before a visual movement-trigger signal. Great care was taken to train the animal to use only the required part of the limb. As a result of extensive training, electromyographic (EMG) studies revealed that muscle activities before the key press were limited to the digit and hand muscles of the limb instructed to move. No overt increase or decrease in activity was detectable in the proximal limb or body muscles in relation to the key-press movements or instructions. 3. Even though the movement was thus limited to distal forelimb, distinct ipsilateral relationships were observed in 8.2% of the task-related PCM neurons. They changed their activity before ipsilateral and bilateral (but not before contralateral) key press. 4. A majority of the neurons recorded from the digit area of PCM (mostly limited to the anterior bank of the central sulcus) exhibited a contralateral relationship; namely the activity increased or decreased before the onset of the contralateral and bilateral key-press movements. In most of them, the magnitudes of the activity changes before the contralateral and bilateral movements were similar. 5. In proximal limb and trunk areas of PCM and also in the somatosensory cortex, no neurons were found to exhibit distinct relations to any of the key-press movements. 6. In both SMA and PM, a number of neurons exhibited relationships of the type never or only rarely observed in the primary motor cortex. Thirty-seven percent of SMA and 62% of PM neurons exhibited premovement activity changes before all of the key-press movements. The movement-specific type of activity was observed in 28% of SMA and 16% of PM neurons. In these neurons, the activity changes were observed in relation to only one of the right or left key-press movements or exclusively in relation to the bilateral key press. Neuronal activity resembling the majority of the PCM neurons (contralateral type) was observed in 31% of SMA and 13% of PM neurons. 7. Instruction-induced changes in activity were more often found in the secondary than in the primary motor area.(ABSTRACT TRUNCATED AT 400 WORDS)


New initiator system for bonding to dentin using methylcyclohexanedione.
As a part of a series of studies to examine the role of the initiator system in dentin bonding, the effectiveness of a new polymerization initiator system consisting of 2-methyl-1,3-cyclohexanedione (MCHD) and CuCl2 in dentin bonding was investigated in terms of the effects of dentin conditioners and CuCl2 concentration formulated in MMA/PMMA resins. Curing time and tensile bond strength to bovine dentin were measured using resins composed of MMA containing 0.001-0.03 wt% CuCl2 and PMMA powder containing 2 wt% MCHD. The dentin surfaces were treated with six conditioners consisting of 10 wt% phosphoric acid or citric acid containing 0-3 wt% FeCl3 or CuCl2. The bond strengths of about 10 MPa obtained under optimal conditions were comparable to those reported for tributylborane-initiated MMA resin which is one of the most effective bonding resins available. Thus, the resin using MCHD/CuCl2 system was concluded to be very effective for dentin bonding, especially when dentin was treated with acidic conditioners containing ferric or copper chloride.


Differential trajectories of hypometabolism across cognitively-defined Alzheimer's disease subgroups.
Disentangling biologically distinct subgroups of Alzheimer's disease (AD) may facilitate a deeper understanding of the neurobiology underlying clinical heterogeneity. We employed longitudinal [18F]FDG-PET standardized uptake value ratios (SUVRs) to map hypometabolism across cognitively-defined AD subgroups. Participants were 384 amyloid-positive individuals with an AD dementia diagnosis from ADNI who had a total of 1028 FDG-scans (mean time between first and last scan: 1.6 ± 1.8 years). These participants were categorized into subgroups on the basis of substantial impairment at time of dementia diagnosis in a specific cognitive domain relative to the average across domains. This approach resulted in groups of AD-Memory (n = 135), AD-Executive (n = 8), AD-Language (n = 22), AD-Visuospatial (n = 44), AD-Multiple Domains (n = 15) and AD-No Domains (for whom no domain showed substantial relative impairment; n = 160). Voxelwise contrasts against controls revealed that all AD-subgroups showed progressive hypometabolism compared to controls across temporoparietal regions at time of AD diagnosis. Voxelwise and regions-of-interest (ROI)-based linear mixed model analyses revealed there were also subgroup-specific hypometabolism patterns and trajectories. The AD-Memory group had more pronounced hypometabolism compared to all other groups in the medial temporal lobe and posterior cingulate, and faster decline in metabolism in the medial temporal lobe compared to AD-Visuospatial. The AD-Language group had pronounced lateral temporal hypometabolism compared to all other groups, and the pattern of metabolism was also more asymmetrical (left < right) than all other groups. The AD-Visuospatial group had faster decline in metabolism in parietal regions compared to all other groups, as well as faster decline in the precuneus compared to AD-Memory and AD-No Domains. Taken together, in addition to a common pattern, cognitively-defined subgroups of people with AD dementia show subgroup-specific hypometabolism patterns, as well as differences in trajectories of metabolism over time. These findings provide support to the notion that cognitively-defined subgroups are biologically distinct.


Increase in bone marrow lesions associated with cartilage loss: a longitudinal magnetic resonance imaging study of knee osteoarthritis.
Although bone marrow lesions (BMLs) are powerful predictors of joint space loss as visualized on radiographs, the natural history of these lesions, their relationship to cartilage loss, and the association between change in these lesions and cartilage loss are unknown. These questions were tested using longitudinal magnetic resonance imaging (MRI) data in a natural history study of symptomatic knee osteoarthritis (OA). MRI of the knee was performed at baseline, 15 months, and 30 months in 217 patients with primary knee OA (122 men, 95 women; mean +/- SD age 66.4 +/- 9.4 years). To assess mechanical alignment, long-limb films were obtained at 15 months. Subchondral bone marrow abnormalities, graded in the medial and lateral tibiofemoral joints, were defined as poorly marginated areas of increased signal intensity in the marrow on fat-suppressed, T2-weighted images. Cartilage morphologic features in the medial and lateral tibiofemoral joints were scored at all time points using a semiquantitative scale. For each of the medial and lateral compartments, generalized estimating equations were used to evaluate the longitudinal relationship of tibiofemoral BMLs to the tibiofemoral cartilage score, with adjustment for malalignment. Fifty-seven percent of knees had BMLs at baseline, of which 99% remained the same or increased in size at followup. Knee compartments with a higher baseline BML score had greater cartilage loss. An increase in BMLs was strongly associated with further worsening of the cartilage score. Enlarging or new BMLs occurred mostly in malaligned limbs, on the side of the malalignment (e.g., new medial BMLs in varus-aligned knees). The association of BML change with medial tibiofemoral cartilage loss was not significant after adjusting for alignment. Lesions of the bone marrow are unlikely to resolve and often get larger over time. Compared with BMLs that stay the same, enlarging BMLs are strongly associated with more cartilage loss. Furthermore, any change in BML is mediated by limb alignment.


Qualitative assessment of healthy volunteer experience receiving subcutaneous infusions of high-dose benzathine penicillin G (SCIP) provides insights into design of late phase clinical studies.
Secondary prophylaxis to prevent rheumatic heart disease (RHD) progression, in the form of four-weekly intramuscular benzathine benzylpenicillin G (BPG) injections, has remained unchanged since 1955. Qualitative investigations into patient preference have highlighted the need for long-acting penicillins to be delivered less frequently, ideally with reduced pain. We describe the experience of healthy volunteers participating in a phase-I safety, tolerability and pharmacokinetic trial of subcutaneous infusions of high-dose benzathine penicillin G (BPG)-the SCIP study (Australian New Zealand Clinical Trials Registry ACTRN12622000916741). Participants (n = 24) received between 6.9 mL to 20.7 mL (3-9 times the standard dose) of BPG as a single infusion into the abdominal subcutaneous tissues via a spring-driven syringe pump over approximately 20 minutes. Semi-structured interviews at four time points were recorded, transcribed verbatim and thematically analysed. Tolerability and specific descriptors of the experience were explored, alongside thoughts on how the intervention could be improved for future trials in children and young adults receiving monthly BPG intramuscular injections for RHD. Participants tolerated the infusion well and were able describe their experiences throughout. Most reported minimal pain, substantiated via quantitative pain scores. Abdominal bruising at the infusion site did not concern participants nor impair normal activities. Insight into how SCIP could be improved for children included the use of topical analgesia, distractions via television or personal devices, a drawn-out infusion time with reduced delivery speed, and alternative infusion sites. Trust in the trial team was high. Qualitative research is an important adjunct for early-phase clinical trials, particularly when adherence to the planned intervention is a key driver of success. These results will inform later-phase SCIP trials in people living with RHD and other indications.


Maternal verbal responsiveness and directiveness: consistency, stability, and relations to child early linguistic development.
Maternal responsive and directive speech to children at ages 0;10 and 2;0 was investigated by applying a procedure first introduced by Flynn and Masur (2007) to a new language community (Finnish). The issues examined were consistency and stability over time, and also the role of responsiveness and directiveness in child linguistic development at 1;0 and 2;6. The measures of maternal speech from each age were used to predict the results of the subsequent linguistic assessment. Negative correlations between responsive and directive utterances were found at both ages. The frequencies of responsive utterances and supportive directives increased over time. Responsiveness was positively, and intrusive directiveness negatively, related to child early comprehensive skills and the use of symbolic actions and communicative gestures. By contrast, no relations were found between responsiveness and directiveness and children's later linguistic capacities.


Correspondence of coral holobiont metabolome with symbiotic bacteria, archaea and Symbiodinium communities.
Microbial symbiotic partners, such as those associated with Scleractinian corals, mediate biochemical transformations that influence host performance and survival. While evidence suggests microbial community composition partly accounts for differences in coral physiology, how these symbionts affect metabolic pathways remains underexplored. We aimed to assess functional implications of variation among coral-associated microbial partners in hospite. To this end, we characterized and compared metabolomic profiles and microbial community composition from nine reef-building coral species. These data demonstrate metabolite profiles and microbial communities are species-specific and are correlated to one another. Using Porites spp. as a case study, we present evidence that the relative abundance of different sub-clades of Symbiodinium and bacterial/archaeal families are linked to positive and negative metabolomic signatures. Our data suggest that while some microbial partners benefit the union, others are more opportunistic with potential detriment to the host. Consequently, coral partner choice likely influences cellular metabolic activities and, therefore, holobiont nutrition.


Redox-active ligand assisted electrocatalytic water oxidation by a mononuclear cobalt complex.
In this report, the synthesis, characterization and electrocatalytic oxidation of water by a mononuclear cobalt(iii) complex, [CoIII(dpaq)(Cl)]Cl (1) featuring a redox-active pentadentate amidate ligand (H-dpaq = 2-[bis(pyridin-2-ylmethyl)]amino-N-quinolin-8-yl-acetamidate) is reported. Complex 1 has been found to be a stable and homogeneous water oxidation catalyst (WOC) in 0.1 M phosphate buffer (pH 8.0). A series of experiments (rinse test, SEM, EDX spectroscopy) confirm that this complex acts as a molecular electrocatalyst, and not a precursor of CoOx. The electrocatalytic water oxidation proceeds with high faradaic efficiency (81%) and fast rate (85 s-1). Analysis of the electrochemical reaction kinetics by foot-of-the-wave (FOWA) methodology reveals a high turnover frequency of 1.6 × 104 s-1 which is comparable to the best performing Ru-based WOCs.


Hypercalcemia in thyrotoxicosis.
Hypercalcemia occurs in approximately one of every five patients with thyrotoxicosis, and one of seven patients with hypercalcemia and thyrotoxicosis will have hyperparathyroidism as the cause of the serum calcium elevation. While there are no clinical features which permit easy identification of patients with hyperparathyroidism and thyrotoxicosis, determination of serum parathyroid hormone levels may help. Parathyroid hormone levels may be normal or suppressed if hypercalcemia is due to hyperthyroidism alone, and an elevated parathyroid hormone level suggest coexisting hyperparathyroidism.


[Probiotics: innocuousness, prevention and risks].
Probiotics have been defined as live microorganisms which, when ingested in adequate numbers, confer health benefits to the host. They are currently consumed without any age restrictions and adverse effects such as sepsis, a marker of the risk of invasion of the bloodstream, are extremely infrequent. However, some health professionals express doubts about probiotics being truly innocuous. This review discusses the incidence of sepsis secondary to probiotics use, mainly lactobacilli and bifidobacteria, evaluated through molecular biology or classic culture techniques, showing that sepsis in large numbers of individuals along decennia is extremely low, of the order of 0,02% en some centers or as low as 1 case/million population in France. These data are important considering the use different species and strains of these microorganisms. Few studies which have reported other adverse effects but many of these have problems with their design that cast doubt about the validity of their results. On the contrary, it has been shown that probiotic microorganisms exert positive stimulatory effects on innate and acquired immunity, with decrease of the manifestations of atopy and eczema. These positive effects are further evidenced by the beneficial effects of many species of probiotics in preventing necrotizing enterocolitis in patients as functionally labile as premature-born babies.


THE PLACENTAL TRANSMISSION OF ANTIBODIES IN THE SKIN SENSITIVE TYPE OF HUMAN ALLERGY.
1. Quantitative studies of the skin-sensitizing antibodies, blocking antibodies, and hemagglutinins in sera of allergic human beings have been made. 2. A comparison of 12 maternal and the 12 corresponding cord sera by the method of passive transfer showed the human placenta to be impermeable to skin-sensitizing antibodies. 3. Direct skin tests and passive transfer studies of 6 infants at the ages of 3 to 6 months showed negative reactions to the antigens to which their mothers were sensitive. 4. The blocking antibody present in the sera of hay fever patients after treatment with pollen extract injections was also demonstrated in the cord sera. The apparent placental transmission of this antibody gave further evidence that it was distinct from the skin-sensitizing antibody. Infant sera obtained at the ages of 3 to 6 months showed no evidence of this immune antibody. 5. The cord sera from 4 of these cases were shown to contain the same isoagglutinins as the maternal sera, showing that the placentas were permeable to these antibodies. The mothers and their offspring reacted alike to Schick testing. 6. Typhoid agglutinins were demonstrated in maternal and cord sera of an adult who previously received injections of triple typhoid vaccine, whereas the serum of the corresponding infant at the age of 3 months failed to show agglutinins.


Delayed Management of an Orbital Floor Blow-out Fracture.
A bony fracture in the orbital floor, the most common site, can lead to tissue herniation, enophthalmos, hypoglobus, or strabismic diplopia. Several surgical approaches for repair have been described in the literature. This report is a description of an illustrative case and a brief summary of the literature related to the transconjunctival approach to orbital floor fracture repair as performed by ophthalmologists. A 19-year-old female patient had fallen from a 5-meter-high fence and sustained panfacial fractures, including both orbits and the surrounding sinuses. An acute repair was performed by a maxillofacial team to stabilize the facial structure . Following neurosurgical stabilization, she was referred to ophthalmology with pronounced hypoglobus and enophthalmos, diplopia, relative afferent pupillary defect, and a slightly pale right optic nerve head. Surgery was performed under general anesthesia using the transconjunctival approach and an alloplastic implant. This approach was effective, providing excellent exposure while reducing the risks of lower eyelid retraction and surgical scars associated with the transcutaneous approach.


Human Milk Fortification in Very Preterm Infants in China: A Multicenter Survey.
To investigate the use of human milk fortifier (HMF) for very preterm infants (VPIs) and complications and nutritional status of VPIs due to various breast milk enhancement strategies among the Chinese population. VPIs with birth weight < 1,800 g and wholly or predominantly breastfed were assigned to the following fortification groups: no HMF, early HMF (adding HMF at an enteral volume of ≤ 80 ml·kg-1·day-1), middle HMF (adding HMF at an enteral volume of 80-100 ml·kg-1·day-1), and late HMF (adding HMF at an enteral volume of ≥100 ml·kg-1·day-1). The growth status and complications for various groups were evaluated. We enrolled 985 VPIs, of which 847 VPIs (86.0%) received HMF, whereas 138 VPIs (14.0%) did not. The number of VPIs in the early, middle, and late fortification groups were 89 (9.0%), 252 (25.6%), and 506 (51.4%), respectively. The complete fortification of the early, middle, and late fortification groups was achieved in 13.2 ± 11.0, 13.8 ± 11.7, and 12.3 ± 13.0 days, respectively, without significant differences (p > 0.05). The groups did not exhibit significant differences in the incidence of feeding intolerance, necrotizing enterocolitis (Bell stage ≥ 2), late-onset sepsis, and metabolic bone diseases (p > 0.05). The middle fortification groups exhibited the fastest growth velocity and the least dramatic decrease in the Z-score of weight and length, and the lowest incidence of EUGR (35.7%), whereas the "no HMF" groups exhibited the slowest growth velocity and the largest decline in the Z-score, and the highest incidence of EUGR (61.6%). The usage rate of HMF was relatively low among Chinese VPIs, fortification often occurred in the late feeding stage, and the time to reach complete fortification was long. Adding HMF and different breast milk enhancement strategies did not increase the incidence of feeding intolerance and necrotizing enterocolitis. The enteral volume of 80-100 ml·kg-1·day-1 with HMF addition led to increased growth in the weight and length and lower EUGR incidence, indicating that the addition of HMF at the specific feeding volume might be the best practice for promoting growth.


Subarachnoid hemorrhage in sickle-cell disease.
The neurological complications of sickle-cell disease include cerebral intracerebral hemorrhage; subarachnoid hemorrhage (SAH) has been infrequently reported. Among 325 patients with sickle-cell disease followed at the University of Illinois between 1975 and 1989, 11 cases of SAH were identified. Aneurysms were found in 10 of these patients, three of whom had multiple aneurysms. All of the patients had some degree of anemia and nine underwent craniotomy without hematological or neurological complications. From this review it appears that SAH is not uncommon in sickle-cell disease patients and tends to occur at a younger age and with smaller aneurysm size than in the general population. With proper perioperative management, including exchange transfusions to reduce the proportion of hemoglobin S to less than 30%, these patients can undergo angiography and craniotomy without an increased incidence of complications. The techniques used in managing sickle-cell disease patients with SAH are discussed.


[W(CN)6(L)](1-/2-) (L = bidentate ligand) as a useful building unit to construct molecule-based magnetic systems.
Metal ions heavier than the 3d metals have attracted increasing attention due to their strong magnetic exchange coupling, arising from the diffuse nature of the extended valence orbitals of the 4d and 5d elements and the involvement of a large magnetic anisotropy contingent on their strong spin-orbit coupling. [W(CN)6(L)](1-/2-) (L = 2,2'-bipyridine (bpy) and 1,10-phenanthroline (phen)) serves as a suitable building block for constructing molecule-based magnetic systems. Discrete molecular entities and one-dimensional chains are preferably obtained when the restricted paramagnetic precursor [W(CN)6(bpy)](-) encounters metal counterparts with the available coordination sites. The bimetallic magnetic assemblies exhibit a variety of magnetic features, along with a magnetostructural correlation. These include slow relaxation of magnetization, typical for single-molecule magnets and single-chain magnets, spin-canting, and metamagnetism. The two-dimensional structure is stabilized by using the reduced [W(CN)6(bpy)](2-) complex. In this perspective article, detailed discussions of molecular materials based on the W(V) ion are reviewed.


Evaluation of a feline viral rhinotracheitis-feline calicivirus disease vaccine.
An attenuated respiratory disease vaccine against feline viral rhinotracheitis (FVR) and feline calicivirus (FCV) disease was evaluated for safety and efficacy in specific-pathogen-free cats. Twenty cats were vaccinated twice intramuscularly, with 28 days between vaccinations. Ten unvaccinated cats were used as contact controls. Adverse effects were not noticed after vaccination, and the vaccinal virus did not spread to contact controls. Arithmetical mean serum-neutralizing titers against vaccinal FCV strain F9 and challenge FCV strain 255 were 1:13 and 1:15 at 28 days after the 1st inoculation. These titers increased to 1:45 and 1:196 after the 2nd inoculation. After challenge exposure of vaccinated cats to virulent FCV 255 virus, mean titers increased to 1:129 and 1:865, respectively for F9 and 255 viruses. The F9 postchallenge mean titer for vaccinated cats was 21.5 times higher than that for the 8 contact controls that survived challenge exposure. The arithmetical mean serum neutralizing titer for FVR was low (1:2) after the 1st vaccination, but increased to 1:35 after the 2nd vaccination. Challenge exposure to virulent FVR virus resulted in a marked anamnestic immune response (mean titer of 1:207, compared with 1:12 for contact controls). In general, vaccinated cats remained alert and healthy after challenge exposure with FCV-255, whereas unvaccinated contact control cats developed definite signs of FCV disease, including central nervous system (CNS) depression (6 of 10) and dyspnea indicative of pneumonia (5 of 10). Two controls died of severe pneumonia. A mild fibrile response was detected in 28% of vaccinated cats, compared with a more severe febrile response in 78% of control cats. Some vaccinated cats developed minute lingual ulcers that did not appear to be detrimental to the health of the cat. After FVR challenge exposure, vaccinated cats were free of serious clinical signs. Five of 18 vaccinated cats had mild signs of FVR, including an occasional sneeze, low temperature, and mild serous lacrimation for 1 or 2 days. Contact controls developed definite clinical signs of FVR. The combined FVR-FCV vaccine appears to be safe and reasonably efficacious. Vaccination against FCV disease and FVR should be part of the routine feline immunization program.


Feasibility of Multiparametric Imaging with PET/MR in Head and Neck Squamous Cell Carcinoma.
The purpose of this study was to investigate and assess the correlation and reproducibility of multiparametric imaging in head and neck cancer patients. Twenty-one patients were included in this prospective scan-rescan study. All patients were scanned twice on an integrated PET and MRI scanner. Gross tumor volumes were defined on T2-weighted MR images, and volumes of interest were defined on diffusion-weighted MRI and 18F-FDG PET (VOIDWI, VOIPET). Overlap between volumes was assessed as a percentwise overlap. 18F-FDG uptake and diffusion were measured using SUV and apparent diffusion coefficient, and correlation was tested across and within patients and as a voxel-by-voxel analysis. Seventeen patients were available for correlation analysis, and 12 patients were available for assessment of tumor overlap. The median tumor overlap between VOIDWI and VOIPET was 82% (VOIDWI in VOIPET) and 62% (VOIPET in VOIDWI) on scan 1 and scan 2, respectively. Across patients, the correlation between SUV and apparent diffusion coefficient was weak and nonsignificant. However, in individual patients a weak but significant correlation was identified on a voxel-by-voxel basis. In multiparametric imaging with the integrated PET/MR scanner, the VOIs from DWI and 18F-FDG PET were both within the target volume for radiotherapy and overlapped substantially although not completely. No correlation between 18F-FDG uptake and DWI could be found across patients, but within individual patients a statistically significant, but weak, voxel-by-voxel correlation was found. The findings suggest that information on glucose uptake and diffusion coefficient carries complementary information of interest that may be relevant for radiotherapy treatment planning.


Propensity score applied to survival data analysis through proportional hazards models: a Monte Carlo study.
Propensity score methods are increasingly used in medical literature to estimate treatment effect using data from observational studies. Despite many papers on propensity score analysis, few have focused on the analysis of survival data. Even within the framework of the popular proportional hazard model, the choice among marginal, stratified or adjusted models remains unclear. A Monte Carlo simulation study was used to compare the performance of several survival models to estimate both marginal and conditional treatment effects. The impact of accounting or not for pairing when analysing propensity-score-matched survival data was assessed. In addition, the influence of unmeasured confounders was investigated. After matching on the propensity score, both marginal and conditional treatment effects could be reliably estimated. Ignoring the paired structure of the data led to an increased test size due to an overestimated variance of the treatment effect. Among the various survival models considered, stratified models systematically showed poorer performance. Omitting a covariate in the propensity score model led to a biased estimation of treatment effect, but replacement of the unmeasured confounder by a correlated one allowed a marked decrease in this bias. Our study showed that propensity scores applied to survival data can lead to unbiased estimation of both marginal and conditional treatment effect, when marginal and adjusted Cox models are used. In all cases, it is necessary to account for pairing when analysing propensity-score-matched data, using a robust estimator of the variance.


Endocrine factors in juvenile essential hypertension.
Of 103 borderline essential hypertensives below the age of 36 years, 11 (10.7%) showed low renin, 57 (55.4%) showed normal renin, and 35 (33.9%) showed high renin. Various hormones such as plasma catecholamines, PA, DOC, B, cortisol 18-OH-DOC, 18-OH-B, ACTH, AVP and PRL tended to be high in the high renin group and tended to be low in the low renin group. Furthermore, urinary excretion of kallikrein was also increased in the high renin group and reduced in the low renin group. There was no hormone which was increased in the low renin group. Regardless of renin levels, pressor responses to exogenous angiotensin II were increased in these juvenile essential hypertensives. From these studies on endocrine factors it is concluded that essential hypertensives are largely divided into 3 groups; the high hormone group, the normal hormone group and the low hormone group, and increased hormones may play an important role in the development of hypertension in the high hormone group. However, in the low hormone group and a part of the normal hormone group the mechanism of hypertension cannot be revealed by the study on endocrine factors.


[Mechanisms of "warm" tachycardia and "cold" bradycardia in cats].
The infusion of a small volume of blood with temperature higher ("warm") or below ("cold") the temperature of atrial blood into the right atrium induced "warm" tachycardia and "cold" bradycardia, resp., in anesthetized and in alert cats. Infusion of the blood with temperature equal to that of atrial blood induced no changes of the heart rate. Infusion of the same volume of "cold" or "warm" blood after adreno- and cholinoreceptor-blocking agents was accompanied by a less severe tachycardia or bradycardia. There seem to be two mechanisms in the initiation of the above effects: the neural and the myogenic ones.


Anisotropic dynamics of optical vortex-beam propagating in biaxial crystals: a numerical method based on asymptotic expansion.
One difficulty of angular spectrum representation method in studying optical propagation inside anisotropic crystals is to calculate the double integrals containing highly oscillating functions. In this paper, we introduce an accuracy and numerically cheap method based on asymptotic expansion theory to overcome this difficulty, which therefore allows to compute optical fields with arbitrary incident beam and is not restricted to the paraxial limit. This numerical method is benchmarked against the analytical solutions in uniaxial crystals and excellent agreements between them are obtained. As an application, we adopt it to investigate the propagation of a Gaussian vortex-beam in a biaxial crystal. The general features of anisotropic dynamics and power conversion between field components are revealed. The numerical results is interpreted by making appropriate analytical approximation to the wave equations.


Distinct amplification of an untranslated regulatory sequence in the egfr gene contributes to early steps in breast cancer development.
Overexpression of the epidermal growth factor receptor (egfr) gene is a common feature in breast cancer. We demonstrated recently that the expression of EGFR in breast cancer strongly correlates with the length of a CA simple sequence repeat within the first 2000 bases in intron 1 of the egfr gene [CA simple sequence repeat (CA-SSR) I; H. Buerger et al., Cancer Res., 60: 854-857, 2000]. Using a standardized semiautomated method of microsatellite analysis for loss of heterozygosity detection, we identified an allelic imbalance (AI) at the egfr locus in 55 of 163 primary breast cancer cases. Fine mapping of the chromosomal region at 7p12-15 around the egfr gene using 10 CA-SSR markers showed that mutations of egfr in breast cancer are frequently restricted to the first intron of egfr. Thereby, the simple sequence repeat CA-SSR I in intron 1 was affected in 84% of the patients with AI. Reverse transcription-PCR analysis of 23 breast cancer tissues with AI excluded the presence of in-frame deletions between exon 2 and exon 7. For additional characterization of the underlying phenomenon leading to the detection of an AI in microsatellite analysis, a quantitative 5'-nuclease assay for the first CA-SSR I in intron 1 was established. In breast cancer cases with AI the presence of amplifications of this sequence was shown. Kaplan-Meier analysis revealed a statistically significant worse prognosis for patients with AI in the cancer tissue at the egfr locus compared with patients without AI. Interestingly, 75% of the patients bearing AI of CA-SSR I in the tumor also showed AI at normal, nontumorous breast tissue. Our data strongly support the assumption that distinct amplifications in intronic sequences of the egfr gene, which enhance the basic transcription activity of the gene, represent one of the first steps in breast carcinogenesis. Furthermore, they point to the presence of prognosis-associated markers for breast cancer already in morphological normal breast tissue.


Internal jugular vein reconstruction: application of conventional type A and novel type K methods.
During neck dissection, the current practice is to preserve the internal jugular vein in the majority of cases. However, sacrifice of bilateral internal jugular veins is required in rare cases. Simultaneous excision of both internal jugular veins is known to frequently cause fatal complications. Even if staged, bilateral internal jugular vein sacrifice still occasionally leads to fatal complications (in 2 per cent). We report two different methods of unilateral internal jugular vein reconstruction, in two cases requiring excision of bilateral internal jugular veins, and we review the significance of this reconstruction procedure. The first patient underwent conventional type A reconstruction (using Katsuno's classification): end-to-end anastomosis of the internal jugular vein to the external jugular vein. For the second patient, we anastomosed the internal jugular vein to the anterior jugular vein, preserving the flow of the external jugular vein. This method, termed type K, had two main expected benefits: facial drainage via the preserved external jugular vein; and provision of a built-in safeguard in the case of occlusion (via the preserved venous networks between the internal jugular vein and the external jugular vein, e.g. the facial vein). In both cases, the reconstructed internal jugular vein was patent and the post-operative course was uneventful, with no severe complications. The current and previous findings strongly indicate that the reconstruction of at least one internal jugular vein is highly recommended for patients requiring bilateral internal jugular vein sacrifice. Our type K method may represent a useful technique for this procedure.


Antihypertensive medication use in the Department of Veterans Affairs: a national analysis of prescribing patterns from 2000 to 2002.
Studies describe differences between recommendations for hypertension treatment and actual drug use. Antihypertensive use data from the Department of Veterans Affairs (VA) for 1995 to 1999 showed a downward trend for calcium antagonist (CA) use and increased use of beta-blockers (BB) and thiazide diuretics (TD). This study evaluates national VA antihypertensive treatment for 2000 to 2002 and compares these data to treatment data for 1995 to 1999. National VA pharmacy data were used to determine use of BB, TD, CA, angiotensin converting enzyme inhibitors (ACEI), angiotensin receptor blockers (ARB), and combinations of antihypertensive drugs for 2000 to 2002. Dispensing data were converted to treatment days. In addition to national trends, data were analyzed regionally to examine geographic differences. Pharmacoeconomic analysis estimated the financial impact of medication changes. Antihypertensive drug use in the VA represented more than 1 billion days in 2002. The ACEI were most commonly used, representing 33.4% and 33.5% of treatment days in 2000 and 2002, respectively. Changes from 2000 to 2002 were 21.9% to 24.2% for BB, 29.3% to 24.4% for CA, and 13.2% to 14.2% for TD. Use of ARB increased from 2.1% to 3.7% of treatment days. Analysis of the 21 VA regions showed geographic variation. For example, the proportion of BB treatment days is highest in a northeast VA region (28.6%) and lowest in a southeast region (19.9%). In 2002 the VA has saved an estimated US 8.5 million dollars because of changes in medication use. As a proportion of antihypertensive agent use, CA continues to fall in the VA, whereas BB, TD, and ARB use have increased. However, TD use remains low, despite national guidelines that promote use of this class of agent.


The integrated structure of care: evidence for the efficacy of models of clinical governance in the prevention of fragility fractures after recent sentinel fracture after the age of 50 years.
Randomized clinical trials and observational studies on the implementation of clinical governance models, in patients who had experienced a fragility fracture, were examined. Literature was systematically reviewed and summarized by a panel of experts who formulated recommendations for the Italian guideline. After experiencing a fracture, several strategies may be adopted to reduce the risk of recurrent fragility fractures and associated morbidity and mortality. Clinical governance models, such as the fracture liaison service (FLS), have been introduced for the identification, treatment, and monitoring of patients with secondary fragility fractures. A systematic review was conducted to evaluate the association between multidisciplinary care systems and several outcomes in patients with a fragility fracture in the context of the development of the Italian Guidelines. PubMed, Embase, and the Cochrane Library were investigated up to December 2020 to update the search of the Scottish Intercollegiate Guidelines Network. Randomized clinical trials (RCTs) and observational studies that analyzed clinical governance models in patients who had experienced a fragility fracture were eligible. Three authors independently extracted data and appraised the risk of bias in the included studies. The quality of evidence was assessed using the Grading of Recommendations Assessment, Development, and Evaluation methodology. Effect sizes were pooled in a meta-analysis using random-effects models. Primary outcomes were bone mineral density values, antiosteoporotic therapy initiation, adherence to antiosteoporotic medications, subsequent fracture, and mortality risk, while secondary outcomes were quality of life and physical performance. Fifteen RCTs and 62 observational studies, ranging from very low to low quality for bone mineral density values, antiosteoporotic initiation, adherence to antiosteoporotic medications, subsequent fracture, mortality, met our inclusion criteria. The implementation of clinical governance models compared to their pre-implementation or standard care/non-attenders significantly improved BMD testing rate, and increased the number of patients who initiated antiosteoporotic therapy and enhanced their adherence to the medications. Moreover, the treatment by clinical governance model respect to standard care/non-attenders significantly reduced the risk of subsequent fracture and mortality. The integrated structure of care enhanced the quality of life and physical function among patients with fragility fractures. Based on our findings, clinicians should promote the management of patients experiencing a fragility fracture through structured and integrated models of care. The task force has formulated appropriate recommendations on the implementation of multidisciplinary care systems in patients with, or at risk of, fragility fractures.


Mortality, In-Hospital Morbidity, Care Practices, and 2-Year Outcomes for Extremely Preterm Infants in the US, 2013-2018.
Despite improvement during recent decades, extremely preterm infants continue to contribute disproportionately to neonatal mortality and childhood morbidity. To review survival, in-hospital morbidities, care practices, and neurodevelopmental and functional outcomes at 22-26 months' corrected age for extremely preterm infants. Prospective registry for extremely preterm infants born at 19 US academic centers that are part of the Eunice Kennedy Shriver National Institute of Child Health and Human Development Neonatal Research Network. The study included 10 877 infants born at 22-28 weeks' gestational age between January 1, 2013, and December 31, 2018, including 2566 infants born before 27 weeks between January 1, 2013, and December 31, 2016, who completed follow-up assessments at 22-26 months' corrected age. The last assessment was completed on August 13, 2019. Outcomes were compared with a similar cohort of infants born in 2008-2012 adjusting for gestational age. Extremely preterm birth. Survival and 12 in-hospital morbidities were assessed, including necrotizing enterocolitis, infection, intracranial hemorrhage, retinopathy of prematurity, and bronchopulmonary dysplasia. Infants were assessed at 22-26 months' corrected age for 12 health and functional outcomes, including neurodevelopment, cerebral palsy, vision, hearing, rehospitalizations, and need for assistive devices. The 10 877 infants were 49.0% female and 51.0% male; 78.3% (8495/10848) survived to discharge, an increase from 76.0% in 2008-2012 (adjusted difference, 2.0%; 95% CI, 1.0%-2.9%). Survival to discharge was 10.9% (60/549) for live-born infants at 22 weeks and 94.0% (2267/2412) at 28 weeks. Survival among actively treated infants was 30.0% (60/200) at 22 weeks and 55.8% (535/958) at 23 weeks. All in-hospital morbidities were more likely among infants born at earlier gestational ages. Overall, 8.9% (890/9956) of infants had necrotizing enterocolitis, 2.4% (238/9957) had early-onset infection, 19.9% (1911/9610) had late-onset infection, 14.3% (1386/9705) had severe intracranial hemorrhage, 12.8% (1099/8585) had severe retinopathy of prematurity, and 8.0% (666/8305) had severe bronchopulmonary dysplasia. Among 2930 surviving infants with gestational ages of 22-26 weeks eligible for follow-up, 2566 (87.6%) were examined. By 2-year follow-up, 8.4% (214/2555) of children had moderate to severe cerebral palsy, 1.5% (38/2555) had bilateral blindness, 2.5% (64/2527) required hearing aids or cochlear implants, 49.9% (1277/2561) had been rehospitalized, and 15.4% (393/2560) required mobility aids or other supportive devices. Among 2458 fully evaluated infants, 48.7% (1198/2458) had no or mild neurodevelopmental impairment at follow-up, 29.3% (709/2419) had moderate neurodevelopmental impairment, and 21.2% (512/2419) had severe neurodevelopmental impairment. Among extremely preterm infants born in 2013-2018 and treated at 19 US academic medical centers, 78.3% survived to discharge, a significantly higher rate than for infants born in 2008-2012. Among infants born at less than 27 weeks' gestational age, rehospitalization and neurodevelopmental impairment were common at 2 years of age.


A New Zebrafish Model to Measure Neuronal α-Synuclein Clearance In Vivo.
The accumulation and aggregation of α-synuclein (α-SYN) is a common characteristic of synucleinopathies, such as Parkinson's Disease (PD), Dementia with Lewy Bodies (DLB) or Multiple System Atrophy (MSA). Multiplications of the wildtype gene of α-SYN (SNCA) and most point mutations make α-SYN more aggregate-prone, and are associated with mitochondrial defects, trafficking obstruction, and impaired proteostasis, which contribute to elevated neuronal death. Here, we present new zebrafish models expressing either human wildtype (wt), or A53T mutant, α-SYN that recapitulate the above-mentioned hallmarks of synucleinopathies. The appropriate clearance of toxic α-SYN has been previously shown to play a key role in maintaining cell homeostasis and survival. However, the paucity of models to investigate α-SYN degradation in vivo limits our understanding of this process. Based on our recently described imaging method for measuring tau protein clearance in neurons in living zebrafish, we fused human SNCA to the photoconvertible protein Dendra2 which enabled analyses of wt and A53T α-SYN clearance kinetics in vivo. Moreover, these zebrafish models can be used to investigate the kinetics of α-SYN aggregation and to study the mechanisms, and potential new targets, controlling the clearance of both soluble and aggregated α-SYN.


Mother-daughter correlation of central obesity and other noncommunicable disease risk factors: Tehran Lipid and Glucose Study.
This study aimed to investigate the mother-daughter correlation for central obesity and other noncommunicable disease risk factors. The authors used metabolic and anthropometric data from the Tehran Lipid and Glucose Study, enrolling 1041 mother-daughter pairs for the current study. Three age strata were defined: 3 to 9 years for childhood (146 mother-daughter pairs), 10 to 17 years for adolescence (395 mother-daughter pairs), and 18 to 25 years for early adulthood (500 mother-daughter pairs). Familial associations for central obesity and other noncommunicable disease risk factors were assessed. The prevalence of central obesity was 44.7% in mothers and 11.2% in daughters (6.2% in the 3-9, 19.2% in the 10-17, and 6.4% in the 18-25 years groups). Mothers with central obesity were more likely than nonobese mothers to have daughters with central obesity (10.5% and 1.7%, respectively; P = .0001). Central obesity indices among daughters were positively correlated with those of their mothers in all 3 age strata. Correlations for other noncommunicable disease risk factors were analyzed before and after adjusting the risk factor levels for mothers' and daughters' waist circumferences (WCs) within each group to determine whether risk factor correlations were, in part, a result of the central obesity correlations. After the non-communicable disease risk factor levels of participants were adjusted for their WCs, the mother-daughter correlations remained significant. The consistent association of central obesity between mothers and daughters may indicate the key role that could be played by the mother in the primary prevention of central obesity, particularly in high-risk families.


Comparative study on antibody immobilization strategies for efficient circulating tumor cell capture.
Methods for isolation and quantification of circulating tumor cells (CTCs) are attracting more attention every day, as the data for their unprecedented clinical utility continue to grow. However, the challenge is that CTCs are extremely rare (as low as 1 in a billion of blood cells) and a highly sensitive and specific technology is required to isolate CTCs from blood cells. Methods utilizing microfluidic systems for immunoaffinity-based CTC capture are preferred, especially when purity is the prime requirement. However, antibody immobilization strategy significantly affects the efficiency of such systems. In this study, two covalent and two bioaffinity antibody immobilization methods were assessed with respect to their CTC capture efficiency and selectivity, using an anti-epithelial cell adhesion molecule (EpCAM) as the capture antibody. Surface functionalization was realized on plain SiO2 surfaces, as well as in microfluidic channels. Surfaces functionalized with different antibody immobilization methods are physically and chemically characterized at each step of functionalization. MCF-7 breast cancer and CCRF-CEM acute lymphoblastic leukemia cell lines were used as EpCAM positive and negative cell models, respectively, to assess CTC capture efficiency and selectivity. Comparisons reveal that bioaffinity based antibody immobilization involving streptavidin attachment with glutaraldehyde linker gave the highest cell capture efficiency. On the other hand, a covalent antibody immobilization method involving direct antibody binding by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC)-N-hydroxysuccinimide (NHS) reaction was found to be more time and cost efficient with a similar cell capture efficiency. All methods provided very high selectivity for CTCs with EpCAM expression. It was also demonstrated that antibody immobilization via EDC-NHS reaction in a microfluidic channel leads to high capture efficiency and selectivity.


A developmentally controlled competitive STAT5-PU.1 DNA binding mechanism regulates activity of the Ig κ E3' enhancer.
Stage-specific rearrangement of Ig H and L chain genes poses an enigma because both processes use the same recombinatorial machinery, but the H chain locus is accessible at the pro-B cell stage, whereas the L chain loci become accessible at the pre-B cell stage. Transcription factor STAT5 is a positive-acting factor for rearrangement of distal V(H) genes, but attenuation of IL-7 signaling and loss of activated STAT5 at the pre-B cell stage corresponds with Igκ locus accessibility and rearrangement, suggesting that STAT5 plays an inhibitory role at this locus. Indeed, loss of IL-7 signaling correlates with increased activity at the Igκ intron enhancer. However, the κE3' enhancer must also be regulated as this enhancer plays a role in Igκ rearrangement. We show in this study that STAT5 can repress κE3' enhancer activity. We find that STAT5 binds to a site that overlaps the κE3' PU.1 binding site. We observed reciprocal binding by STAT5 and PU.1 to the κE3' enhancer in primary bone marrow cells, STAT5 and PU.1 retrovirally transduced pro-B cell lines, or embryonic stem cells induced to differentiate into B lineage cells. Binding by STAT5 corresponded with low occupancy of other enhancer binding proteins, whereas PU.1 binding corresponded with recruitment of IRF4 and E2A to the κE3' enhancer. We also find that IRF4 expression can override the repressive activity of STAT5. We propose a novel PU.1/STAT5 displacement model during B cell development, and this, coupled with increased IRF4 and E2A activity, regulates κE3' enhancer function.


Non-swarming grasshoppers exhibit density-dependent phenotypic plasticity reminiscent of swarming locusts.
Locusts are well known for exhibiting an extreme form of density-dependent phenotypic plasticity known as locust phase polyphenism. At low density, locust nymphs are cryptically colored and shy, but at high density they transform into conspicuously colored and gregarious individuals. Most of what we know about locust phase polyphenism come from the study of the desert locust Schistocerca gregaria (Forskål), which is a devastating pest species affecting many countries in North Africa and the Middle East. The desert locust belongs to the grasshopper genus Schistocerca Stål, which includes mostly non-swarming, sedentary species. Recent phylogenetic studies suggest that the desert locust is the earliest branching lineage within Schistocerca, which raises a possibility that the presence of density-dependent phenotypic plasticity may be a plesiomorphic trait for the whole genus. In order to test this idea, we have quantified the effect of rearing density in terms of the resulting behavior, color, and morphology in two non-swarming Schistocerca species native to Florida. When reared in both isolated and crowded conditions, the two non-swarming species, Schistocerca americana (Drury) and Schistocerca serialis cubense (Saussure) clearly exhibited plastic reaction norms in all traits measured, which were reminiscent of the desert locust. Specifically, we found that both species were more active and more attracted to each other when reared in a crowded condition than in isolation. They were mainly bright green in color when isolated, but developed strong black patterns and conspicuous background colors when crowded. We found a strong effect of rearing density in terms of size. There were also more mechanoreceptor hairs on the outer face of the hind femora in the crowded nymphs in both species. Although both species responded similarly, there were some clear species-specific differences in terms of color and behavior. Furthermore, we compare and contrast our findings with those on the desert locust and other relevant studies. We attribute the presence of density-dependent phenotypic plasticity in the non-swarming Schistocerca species to phylogenetic conservatism, but there may be a possible role of local adaptation in further shaping the ultimate expressions of plasticity.


[Intrageneric O-antigenic relations of Providencia bacteria].
The authors present literature and personal data on the intragenera O-antigenic relations of the strains of a collection of bacteria belonging to Providencia genus, including standard strains of 53 serological O-groups whose antigenic interrelations were studied serologically in the tests of agglutination and adsorption of agglutinins. Experimental results confirmed the presence of unilateral relations by the O-antigen between the serological groups 01-021, 03-021 and bilateral relations between the serological groups 01-03; there were also revealed relations by the O-antigens between the serological groups 09-036, 014-023, 018-035, 039-053, and 039-027 of a unilateral character. The data obtained added information to the antigenic structure of Providencia and were of significance for the preparation of diagnostic O-sera of Providencia.


Increased sympathetic activity in rat white adipose tissue during prolonged fasting.
The effect of prolonged fasting on sympathetic activity was examined in rat white adipose tissue (WAT) and, for comparison purposes, in interscapular brown adipose tissue (IBAT). Preliminary experiments showed that 6-hydroxydopamine or tyramine administration to fed animals produced similar reductions in norepinephrine (NE) content of WAT and IBAT. Fasting for 48 h did not affect tissue NE content significantly, but induced a threefold increase in [3H]NE uptake by retroperitoneal and epididymal adipose tissue, contrasting with a 50% reduction in IBAT. Measured with DL-alpha-methyl-p-tyrosine, NE fractional rates of turnover were faster and calculated turnover rates were three times higher in retroperitoneal and epididymal tissue from fasted rats than in tissues from fed controls. In experiments with [3H]NE, although fractional rates did not change significantly, calculated NE turnover also increased in retroperitoneal and epididymal tissue after food deprivation. In contrast, in IBAT, NE turnover either did not change (measured with DL-alpha-methyl-p-tyrosine) or, in the experiments with [3H]NE, decreased significantly after fasting. These and other data suggest that a centrally controlled selective activation of WAT sympathetic fibers contributes to fasting lipolysis.


Association of HIV infection with the development of severe and complicated malaria cases at a rural hospital in Zimbabwe.
To determine the association between HIV infection and progression of Plasmodium falciparum malaria illness in Hurungwe district, Zimbabwe. Prospective cohort. Hurungwe Rural Hospital in Mashonaland West Province, Zimbabwe. Blood slide positive P. falciparum malaria patients. Development of severe and complicated malaria. A total of 659 clinical malaria cases were investigated and 237 (36.0%) confirmed cases entered the study. The total HIV positive malaria patients were 82 (34.6%) of confirmed cases or 12.4% of the total clinical cases. The case fatality rate was 5.9% (14 deaths) in the confirmed cases and 11 of these deaths were HIV positive. The commonly reported complications were high parasite count of 2% or more (38.5%), anaemia (29.0%), cerebral malaria (23.1%), low blood pressure (8.3%) and renal failure (1.2%). The HIV positive cases which developed severe and complicated malaria were 72, 30.4% of the sample studied or 55.8% of the total severe and complicated cases. The mode of transport to the nearest health centre was the only confounding factor identified during the analysis. After adjusting for this confounding factor, the risk of developing severe and complicated malaria was 2.35 (95% CI 1.85 to 2.98) times more in the HIV positive malaria patients than in HIV negative patients. We conclude that HIV infection is significantly associated with the development of severe and complicated malaria. There is need for future studies to determine whether HIV positive malaria patients require different management protocol from HIV negative malaria patients.


APOE alleles are not associated with calcific aortic stenosis.
To analyse the association of APOE alleles with aortic stenosis (AS) in a large study population. Patients with AS (n = 538) and a control group of the same age without heart disease (n = 536) were recruited. Left heart catheterisation was performed and mean gradient, aortic valve area, presence of stenotic coronary artery disease (CAD) and cardiovascular risk factors (hypercholesterolaemia, hypertension, smoking, diabetes mellitus and family history of CAD) were assessed. The frequency of the APOE major alleles e2, e3 and e4 was assessed by genotyping the polymorphisms APOE334 and APOE472 with a 5' exonuclease assay (TaqMan). Mean gradient across the aortic valve in cases was 50 (SD 20) mm Hg corresponding to a mean aortic valve area of 0.84 (SD 0.34) cm(2). 270 patients with AS had stenotic CAD. Among patients with AS, the prevalence of hypercholesterolaemia (64% v 40%, p < 0.001), smoking (43% v 27%, p < 0.001), diabetes (27% v 17%, p < 0.01), family history of CAD (30% v 21%, p </= 0.05), and male sex (65% v 44%, p < 0.001) was higher in those with than in those without CAD. The frequency of the major alleles was not different between cases and controls (APOE e2: 104 (19.3%) v 94 (17.5%); APOE e3: 319 (59.3%) v 332 (61.9%); APOE e4: 115 (21.3%) v 110 (20.5%); all p > 0.10). APOE e4 is not associated with AS, reflecting the different genetic backgrounds of CAD and AS.


Number agreement processing in adolescents with and without developmental language disorder (DLD): evidence from event-related brain potentials.
In morphologically richer languages, including French, one must learn the specific properties of number agreement in order to understand the language, and this learning process continues into adolescence. This study examined similarities and differences between French-speaking adolescents with and without developmental language disorder (DLD) when processing number agreement, and investigated how morpho-syntactic regularity affected language processing. Using event-related potentials (ERP) and only grammatical sentences with audio-visual mismatches, we studied ERP correlates to three types of number agreement: (1) regular determiner agreement in noun phrases, (2) regular subject-verb plural liaison, and (3) irregular subject-verb agreement. We also included a lexico-semantic mismatch condition to investigate lexico-semantic processing in our participants. 17 adolescents with DLD (M = 14.1 years) and 20 (pre)teens with typical language (TL, M = 12.2 years) participated in the study. Our results suggest three patterns. First, French-speaking teenagers without DLD are still consolidating their neurocognitive processing of morpho-syntactic number agreement and generally display ERP profiles typical of lower language proficiency than adult native speakers. Second, differences in morphosyntactic processing between teenagers with and without DLD seem to be limited to rule-based (regular) number agreement. Third, there is little evidence for corresponding differences in lexico-semantic processing.


Inter- and intrahemispheric dissociations in ideomotor apraxia: a large-scale lesion-symptom mapping study in subacute brain-damaged patients.
Pantomimes of object use require accurate representations of movements and a selection of the most task-relevant gestures. Prominent models of praxis, corroborated by functional neuroimaging studies, predict a critical role for left parietal cortices in pantomime and advance that these areas store representations of tool use. In contrast, lesion data points to the involvement of left inferior frontal areas, suggesting that defective selection of movement features is the cause of pantomime errors. We conducted a large-scale voxel-based lesion-symptom mapping analyses with configural/spatial (CS) and body-part-as-object (BPO) pantomime errors of 150 left and right brain-damaged patients. Our results confirm the left hemisphere dominance in pantomime. Both types of error were associated with damage to left inferior frontal regions in tumor and stroke patients. While CS pantomime errors were associated with left temporoparietal lesions in both stroke and tumor patients, these errors appeared less associated with parietal areas in stroke than in tumor patients and less associated with temporal in tumor than stroke patients. BPO errors were associated with left inferior frontal lesions in both tumor and stroke patients. Collectively, our results reveal a left intrahemispheric dissociation for various aspects of pantomime, but with an unspecific role for inferior frontal regions.


Mechanisms of blood pressure alterations in response to the Valsalva maneuver in postural tachycardia syndrome.
The postural tachycardia syndrome (POTS) is characterized clinically by orthostatic lightheadedness and tachycardia. When these patients perform a Valsalva maneuver, there is an excessive blood pressure increment after cessation of the maneuver (phase IV) that is sometimes associated with headaches. It is not known whether excessive phase IV is due to excessive peripheral vascular tone (an alpha-adrenergic mechanism) or is a manifestation of increased beta-adrenergic tone (hyperadrenergic state). The authors undertook a pharmacologic study evaluating the effect of intravenous phentolamine (alpha-adrenergic antagonist) and propranolol (beta-adrenergic antagonist) on the different phases of the Valsalva maneuver in a group of patients with POTS and age-matched normal control subjects. Patients with POTS had mean phases, when compared with controls, that were characterized by more negative II_E (p = 0.07), smaller II_L (p = 0.04), and significantly larger phase IV (p = 0.001). The effect of phentolamine was qualitatively and quantitatively different in POTS when compared with controls. Ten mg phentolamine in controls resulted in a significant accentuation of phase II_E (p = 0.001), attenuation of phase II_L (p = 0.002), and increase of phase IV (57.6 vs 30.7 mm Hg; p = 0.025). These changes resembled those of patients with POTS at baseline. In patients with POTS, the phase II abnormalities, already present, were further accentuated (p <0.001), and phase IV became smaller (50.6 vs 73.8 mm Hg; p = 0.09). Propranolol had no significant effect on phases II_E and II_L, but significantly reduced phase IV in both controls (p <0.05) and in patients with POTS (p <0.001) and improved the headache symptoms, when present, during and after phase IV. The authors conclude that phase IV is mainly under beta-adrenergic regulation and that the exaggerated phase IV in POTS is a result of a hyperadrenergic state.


Calcitonin gene-related peptide in vivo positive inotropy is attributable to regional sympatho-stimulation and is blunted in congestive heart failure.
Calcitonin gene-related peptide (CGRP) is a nonadrenergic/noncholinergic (NANC) peptide with vasodilatative/inotropic action that may benefit the failing heart. However, precise mechanisms for its in vivo inotropic action remain unclear. To assess this, dogs with normal or failing (sustained tachypacing) hearts were instrumented for pressure-dimension analysis. In control hearts, CGRP (20 pmol/kg per minute) enhanced cardiac contractility (eg, +33+/-4.2% in end-systolic elastance) and lowered afterload (-14.2+/-2% in systemic resistance, both P<0.001). The inotropic response was markedly blunted by heart failure (+6.5+/-2%; P<0.001 versus control), whereas arterial dilation remained unaltered (-19.3+/-5%). CGRP-positive inotropy was not attributable to reflex activation because similar changes were observed in the presence of a ganglionic blocker. However, it was fully prevented by the beta-receptor antagonist (timolol), identifying a dominant role of sympatho-stimulatory signaling. In control hearts, myocardial interstitial norepinephrine assessed by microdialysis almost doubled in response to CGRP infusion, whereas systemic plasma levels were unchanged. In addition, CGRP receptors were not observed in ventricular myocardium but were prominent in coronary arteries and the stellate ganglia. Ventricular myocytes isolated from normal and failing hearts displayed no inotropic response to CGRP, further supporting indirect sympatho-stimulation as the primary in vivo mechanism. In contrast, the peripheral vasodilatative capacity of CGRP was similar in femoral vascular rings from normal and failing hearts in dogs. Thus, CGRP-mediated positive inotropy is load-independent but indirect and attributable to myocardial sympathetic activation rather than receptor-coupled stimulation in canine hearts. This mechanism is suppressed in heart failure, so that afterload reduction accounts for CGRP-enhanced function in this setting.


Circulating tumor cell telomerase activity as a prognostic marker for overall survival in SWOG 0421: a phase III metastatic castration resistant prostate cancer trial.
Circulating tumor cells (CTC) are promising biomarkers in metastatic castration resistant prostate cancer (mCRPC), and telomerase activity (TA) is a recognized cancer marker. Therefore, we hypothesized that CTC TA may be prognostic of overall survival (OS) in mCRPC. To test this, we used a novel Parylene-C slot microfilter to measure live CTC TA in S0421, a phase III SWOG-led therapeutic trial. Blood samples underwent CTC capture and TA measurement by microfilter, as well as parallel enumeration by CellSearch (Janssen/J&J). Cox regression was used to assess baseline (pre-treatment) TA versus OS, and recursive partitioning was used to explore potential prognostic subgroups and to generate Kaplan-Meier (KM) OS curves. Samples were obtained from 263 patients and generated 215 TA measures. In patients with baseline CTC count ≥5 (47% of patients), higher CTC TA was associated with hazard ratio 1.14 (p = 0.001) for OS after adjusting for other clinical covariates including CTC counts and serum PSA at study entry. Recursive partitioning identified new candidate risk groups with KM OS curve separation based on CTC counts and TA. Notably, in men with an intermediate range baseline CTC count (6-54 CTCs/7.5 ml), low versus high CTC TA was associated with median survival of 19 versus 12 months, respectively (p = 0.009). Baseline telomerase activity from CTCs live-captured on a new slot microfilter is the first CTC-derived candidate biomarker prognostic of OS in a large patient subgroup in a prospective clinical trial. CTC telomerase activity thus merits further study and validation as a step towards molecular CTC-based precision cancer management.


Studies on the replication of KRV single-stranded linear DNA.
The autonomous parvovirus, Kilham rat virus (KRV), is composed of three structural proteins (Salzman & White, 1970) and a single molecule of DNA. The DNA molecule is linear and single-stranded (ss). It is believed to have both 3' and 5'-terminal palindromic sequences (Salzman, 1977). Replication of the ss DNA may involve double-stranded (ds) DNA intermediates of one unit length as found in the virion or multiple length concatamers (Salzman & White, 1973; Gunther & May, 1976; Lavelle & Li, 1977). We have attempted to determine when the synthesis of KRV ds DNA can be determined in synchronized infected cells and the structure of the ds DNA molecules.


Successful implementation of an appendectomy process improvement protocol.
A key component of a process improvement program is the institution of hospital-specific protocols to address certain disparities and streamline patient care. In that regard, we evaluated the implementation of an outpatient laparoscopic appendectomy (OLA) protocol at a tertiary military hospital. We hypothesized that OLA would reduce length of stay (LOS) without increasing complications. In August 2016, our institution implemented an OLA protocol-defined as discharge within 24 hours of surgery. Exclusion criteria included age <18 years old, grade 4 or 5 appendicitis, immunosuppression, current pregnancy, and no supervision during the first 24 hours postdischarge. To determine OLA's effect on LOS, analysis of variance was used to perform a comparison between the years 2014 and 2017. Successful outpatient appendectomies were recorded preprotocol and postprotocol, as well as readmission complications. In 2017, the first full year of protocol implementation, 44 of 59 (75%) patients met the inclusion criteria, and all but 2 (42 of 44, 95%) stayed for less than 24 hours. Of the two outliers, one developed acute on chronic kidney disease and one had a slow return of bowel function following grade 3 appendicitis. Complications were low across all years (one per year). In 2017, the readmission was for percutaneous drainage of an abscess. Overall, protocol implementation produced a significant decrease in LOS. OLA protocol decreased LOS at a military hospital and should be expanded to other department of defense (DoD) facilities. Further research is needed to identify cost benefit to the military health system. III.


Pregnancy in patients with congenital heart disease.
Pregnancy and delivery in women with congenital heart disease remain inadvisable for patients at high risk due to the following congenital diseases: (1.) Severe left ventricular outflow tract obstruction (increase of pressure gradient); (2.) Eisenmenger syndrome (increase of right to left shunt with worsening of cyanosis and fetal growth retardation); (3.) Marfan syndrome with enlarged aortic root (risk of aortic dissection). In women suffering from cyanotic congenital heart disease the main risk is fetal complications. These are correlated to oxygen saturation and to the type of maternal disease, particularly if a palliative shunt operation has never been performed. Women with left to right shunt, right outflow tract obstruction or previous correction of the disease can go through pregnancy with low risk of mortality. The complications (mainly heart failure and arrhythmias) can be well managed with medical treatment, and the fetal outcome is similar to that in the general population.


Mathematical modeling and finite element simulation of slow release of drugs using hydrogels as carriers with various drug concentration distributions.
In drug release systems using hydrogels as carriers, the presence of the polymer network will reduce the drug release rate, which can extend the release period. For a controlled-release process of drug, usually the ideal situation is to get a zero-order drug release rate. In this paper, the mathematical model of hydrogel swelling processes is constructed on the basis of a biphasic theory, and then an integrated equation that considers both water convection and drug diffusion phenomena is used to describe the drug release process. The effects of the initial drug concentration with nonuniform distributions along the radial direction of hydrogel carriers on the release of drugs are studied through simulating two-dimensional hydrogel swelling processes by means of the COMSOL Multiphysics software. The simulation results show that along with the hydrogel swelling, the drug release rate is changing, and the major influencing factors of the drug release rate are water convection and drug diffusion coefficient, which are affected by water volume fraction, drug concentration distribution in matrix, and carrier radius. The results also indicate that the initial drug concentration distribution following a sine curve can result in an ideal zero-order release process.


Heterogeneity in cellular and antibody responses against thyrotropin receptor in patients with Graves' disease detected using synthetic peptides.
Graves' disease (GD) is characterized by the presence of autoantibodies to the thyrotropin receptor (TSHr). These antibodies bind to the TSHr and stimulate thyroid cells, thus causing hyperthyroidism. To understand the regulation of TSHr-specific immune responses in Graves' disease, it is important to evaluate the T-cell response in patients with GD against TSHr. In this study we used 11 different peptides that were derived from two regions (i.e. amino acid, AA 12-46 and 316-397) unique to the TSHr when compared to other glycoprotein hormone receptors, and which also have the highest predicted immunogenicity. We evaluated both lymphocyte proliferation as a measure of T-cell response and antibody binding to each of these peptides in nine patients with GD and eight healthy subjects. Patients with GD showed considerable lymphocyte proliferative and antibody responses against several of these peptides. There was considerable heterogeneity in immune responses amongst the patients. Moreover, our data suggested that several peptides contained both T cell and antibody reactive epitopes and might represent some of the highly immunogenic regions of the TSHr.


Ganfort, a blinding drug to the physician.
Ganfort eye drops are indicated for adult patients with open-angle glaucoma who have poor response to topical β-blockers or prostaglandin analogues. They contain 0.3 mg of bimatoprost (a prostaglandin) and 5 mg of timolol (β-blocker). The authors present a case of a 45-year-old man with glaucoma presenting with shortness of breath. On admission, he had a normal heart rate, pulse oximetry and examination. Despite being of low risk stratification using the Wells score and the Modified Geneva Score for pulmonary emboli, CT pulmonary angiogram scan subsequently showed extensive bilateral multiple large pulmonary emboli. This case demonstrates that the systemic absorption of ocular β blockers is extremely high and it bypasses the first pass metabolism. Therefore, even one drop of Ganfort into each eye once a day was sufficient to disguise the tachycardia of a large pulmonary emboli.


Cloning and characterization of a cAMP-specific cyclic nucleotide phosphodiesterase.
Cyclic nucleotide phosphodiesterases (PDEs) regulate intracellular levels of cAMP and cGMP by hydrolyzing them to their corresponding 5' monophosphates. We report here the cloning and characterization of a novel cAMP-specific PDE from mouse testis. This unique phosphodiesterase contains a catalytic domain that overall shares <40% sequence identity to the catalytic domain of all other known PDEs. Based on this limited homology, this new PDE clearly represents a previously unknown PDE gene family designated as PDE8. The cDNA for PDE8 is 3,678 nucleotides in length and is predicted to encode an 823 amino acid enzyme. The cDNA includes a full ORF as it contains an in-frame stop codon before the start methionine. PDE8 is specific for the hydrolysis of cAMP and has a Km of 0.15 microM. Most common PDE inhibitors are ineffective antagonists of PDE8, including the nonspecific PDE inhibitor 3-isobutyl-1-methylxanthine. Dipyridamole, however, an inhibitor that is generally considered to be relatively specific for the cGMP selective PDEs, does inhibit PDE8 with an IC50 of 4.5 microM. Tissue distribution studies of 22 different mouse tissues indicates that PDE8 has highest expression in testis, followed by eye, liver, skeletal muscle, heart, 7-day embryo, kidney, ovary, and brain in decreasing order. In situ hybridizations in testis, the tissue of highest expression, shows that PDE8 is expressed in the seminiferous epithelium in a stage-specific manner. Highest levels of expression are seen in stages 7-12, with little or no expression in stages 1-6.


Evaluation of Malnutrition and Quality of Life in Patients Treated for Oral and Oropharyngeal Cancer.
Oral and oropharyngeal cancer is a debilitating disease with high morbidity and mortality. Depending on the site and extent of the involvement of the cancer and the type of treatment modality, these patients can develop pain, trismus, xerostomia, dysphagia, and taste disturbances, compromising them socially and nutritionally. The aim of the study was to evaluate malnutrition and quality of life in patients treated for oral and oropharyngeal cancer. Methodology. A cross-sectional study was conducted which included 97 patients treated for oral and oropharyngeal cancer. The quality of life of the selected patients was assessed by using a validated European Organization for the Research and Treatment of Cancer's Quality of Life Questionnaire, Head and Neck and Mandibular Function Impairment Questionnaire. Pre- and posttreatment weight of the patients were assessed, and weight loss of ≥10% of pretreatment weight was considered as malnutrition. The chi-square test was used to correlate the symptoms with the quality of life. A paired t test was used to assess the differences in weight before and after treatment, and a p value of <0.005 was considered as significant. The most commonly reported symptoms were xerostomia (93.81%), pain (81.44%), and dysphagia (76.3%). A total of 40.2% of the individuals in the study had malnutrition. Malnutrition was comparatively lower in the group who had nutritional supplements. The quality of life in patients treated for oral and oropharyngeal cancer deteriorates immediately after the treatment; however, it significantly improves over time.