Use and Utility of Skeletal Surveys to Evaluate for Occult Fractures in Young Injured Children.
To describe the percentage and characteristics of children aged <24 months with non-motor vehicle crash (MVC)-related injuries who undergo a skeletal survey and have occult fractures. We performed a retrospective chart review of a stratified, systematic random sample of 1769 children aged<24 months with non-MVC-related bruises, burns, fractures, abdominal injuries, and head injuries at 4 children's hospitals between 2008 and 2012. Sampling weights were assigned to each child to allow for representative hospital-level population estimates. Logistic regression models were used to test for associations between patient characteristics with outcomes of skeletal survey completion and occult fracture identification. Skeletal surveys were performed in 46.3% of children aged 0 to 5 months, in 21.1% of those aged 6 to 11 months, in 8.0% of those aged 12 to 17 months, and in 6.2% of those aged 18 to 24 months. Skeletal surveys were performed most frequently in children with traumatic brain injuries (64.7%) and rib fractures (100%) and least frequently in those with burns (2.1%) and minor head injuries (4.4%). In adjusted analyses, older age, private insurance, and reported history of accidental trauma were associated with decreased skeletal survey use (P ≤ .001 for all). The prevalence of occult fractures on skeletal surveys ranged from 24.6% in children aged 0 to 5months to 3.6% in those aged 18 to 24 months, and varied within age categories based on the presenting injury (P < .001). The high rate of occult fractures in infants aged 0 to 5 months underscores the importance of increasing the use of skeletal surveys in this population. Further research is needed to identify the injury characteristics of older infants and toddlers most at risk for occult fractures.


Inactivation of enoyl-CoA reductase in pigeon liver fatty acid synthetase by pyridoxal 5'-phosphate: evidence for the presence of one lysine residue at the active site.
Pigeon liver fatty acid synthetase (FAS) was rapidly inactivated by pyridoxal 5'-phosphate (PLP). Assays of the partial activities of the PLP-treated synthetase showed that only the enoyl-CoA reductase was decreased significantly. The inactivation of both the overall activity and enoyl-CoA reductase activity of FAS by PLP could be reversed by dialysis or dilution but not by reduction with sodium borohydride. Malonyl-CoA and acetyl-CoA did not protect the enzyme, whereas NADPH provided 68% protection against PLP-inactivation indicating that PLP modified lysine residues present at or near the co-enzyme binding site. PLP-treated enzyme after reduction with sodium borohydride exhibited fluorescence with a maximum at 397 nm (irradiation at 325 nm). Stoichiometric analysis showed that modification of four lysine residues per enzyme molecule resulted in complete inactivation of the overall and enoyl-CoA reductase activities of FAS. NADPH prevented the inactivation by protecting two of these lysine residues from modification, suggesting the presence of two essential lysine residues per enzyme molecule. These results are consistent with the hypothesis that each subunit of the enzyme contains an enoyl-CoA reductase domain in which a lysine residue, at or near the active site, interacts with NADPH.


Inbreeding intensifies sex- and age-dependent disease in a wild mammal.
The mutation accumulation theory of senescence predicts that age-related deterioration of fitness can be exaggerated when inbreeding causes homozygosity for deleterious alleles. A vital component of fitness, in natural populations, is the incidence and progression of disease. Evidence is growing for natural links between inbreeding and ageing; between inbreeding and disease; between sex and ageing; and between sex and disease. However, there is scant evidence, to date, for links among age, disease, inbreeding and sex in a single natural population. Using ecological and epidemiological data from a long-term longitudinal field study, we show that in wild European badgers (Meles meles) exposed naturally to bovine tuberculosis (bTB), inbreeding (measured as multilocus homozygosity) intensifies a positive correlation between age and evidence of progressed infection (measured as an antibody response to bTB), but only among females. Male badgers suffer a steeper relationship between age and progressed infection than females, with no influence of inbred status. We found no link between inbreeding and the incidence of progressed infection during early life in either sex. Our findings highlight an age-related increase in the impact of inbreeding on a fitness-relevant trait (disease state) among females. This relationship is consistent with the predictions of the mutation accumulation theory of senescence, but other mechanisms could also play a role. For example, late-life declines in condition, arising through mechanisms other than mutation accumulation might have increased the magnitude of inbreeding depression in late life. Whichever mechanism causes the observed patterns, we have shown that inbreeding can influence age-dependent patterns of disease and, by extension, is likely to affect the magnitude and timing of the late-life declines in components of fitness that characterise senescence. Better understanding of sex-specific links between inbreeding, disease and ageing provides insights into population-level pathogen dynamics and could influence management strategies for wildlife reservoirs of zoonotic disease.


Increased hospital length of stay attributable to Clostridium difficile infection in patients with four co-morbidities: an analysis of hospital episode statistics in four European countries.
Hospital-onset Clostridium difficile infection (CDI) places a significant burden on health care systems throughout Europe, estimated at around €3 billion per annum. This burden is shared between national payers and hospitals that support additional bed days for patients diagnosed with CDI while in hospital or patients re-admitted from a previous hospitalisation. This study was performed to quantify additional hospital stay attributable to CDI in four countries, England, Germany, Spain, and The Netherlands, by analysing nationwide hospital-episode data. We focused upon patients at increased risk of CDI: with chronic obstructive pulmonary disease, heart failure, diabetes, or chronic kidney disease, and aged 50 years or over. Multivariate regression and propensity score matching models were developed to investigate the impact of CDI on additional length of hospital stay, controlling for confounding factors such as underlying disease severity. Patients in England had the longest additional hospital stay attributable to CDI at 16.09 days, followed by Germany at 15.47 days, Spain at 13.56 days, and The Netherlands at 12.58 days, derived using regression analysis. Propensity score matching indicated a higher attributable length of stay of 32.42 days in England, 15.31 days in Spain, and 18.64 days in The Netherlands. Outputs from this study consistently demonstrate that in European countries, for patients whose hospitalisation is complicated by CDI, the infection causes a statistically significant increase in hospital length of stay. This has implications for optimising resource allocation and budget setting at both the national and hospital level to ensure that levels of CDI-complicated hospitalisations are minimised.


Case Report: Paroxysmal nocturnal hemoglobinuria in a woman heterozygous for G6PD A-.
We describe a case of paroxysmal nocturnal hemoglobinuria (PNH) in a woman who is heterozygous for the glucose-6-phosphate dehydrogenase A- ( G6PDA-) allele. PNH is associated with one or more clones of cells that lack complement inhibition due to loss of function somatic mutations in the PIGA gene. PIGA encodes the enzyme phosphatidylinositol glycan anchor biosynthesis, class A, which catalyses the first step of glycosylphosphatidylinisotol ( GPI) anchor synthesis. Two GPI anchored red cell surface antigens regulate complement lysis. G6PD catalyses the first step of the pentose phosphate pathway and enzyme variants, frequent in some populations have been selected because they confer resistance to malaria, are associated with hemolysis in the presence of oxidizing agents including several drugs. The patient had suffered a hemolytic attack after taking co-trimoxazole, a drug that precipitates hemolysis in G6PD deficient individuals. Since both G6PD and PIGA are X-linked we hypothesized that the PIGA mutation was on the X-chromosome carrying the G6PDA- allele. Investigations showed that in fact the PIGA mutation was on the X-chromosome carrying the normal G6PD B allele. We speculate that complement activation on G6PD A- red cells exposed to Bactrim might have triggered complement activation inducing the lysis of G6PD B PNH Type II red blood cells or that the patient may have had a PNH clone expressing G6PDA- at the time of the hemolytic episode.


Oral administration of recombinant cholera toxin subunit B inhibits IL-12-mediated murine experimental (trinitrobenzene sulfonic acid) colitis.
Trinitrobenzene sulfonic acid (TNBS)-induced colitis is an IL-12-driven, Th1 T cell-mediated colitis that resembles human Crohn's disease. In the present study, we showed initially that the oral administration of recombinant subunit B of cholera toxin (rCT-B) at the time of TNBS-induced colitis by intrarectal TNBS instillation inhibits the development of colitis or, at later time when TNBS-induced colitis is well established, brings about resolution of the colitis. Dose-response studies showed that a majority of mice (68%) treated with rCT-B at a dose of 100 microg (times four daily doses) exhibited complete inhibition of the development of colitis, whereas a minority (30%) treated with rCT-B at a dose of 10 microg (times four daily doses) exhibited complete inhibition; in both cases, however, the remaining mice exhibited some reduction in the severity of inflammation. In further studies, we showed that rCT-B administration is accompanied by prevention/reversal of increased IFN-gamma secretion (the hallmark of a Th1 response) without at the same time causing an increase in IL-4 secretion. This decreased IFN-gamma secretion was not associated with the up-regulation of the secretion of counterregulatory cytokines (IL-10 or TGF-beta), but was associated with a marked inhibition of IL-12 secretion, i.e., the secretion of the cytokine driving the Th1 response. Finally, we showed that rCT-B administration results in increased apoptosis of lamina propria cells, an effect previously shown to be indicative of IL-12 deprivation. From these studies, rCT-B emerges as a powerful inhibitor of Th1 T cell-driven inflammation that can conceivably be applied to the treatment of Crohn's disease.


Colonoscopy and Colorectal Cancer Mortality in the Veterans Affairs Health Care System: A Case-Control Study.
Colonoscopy is widely used in the Veterans Affairs (VA) health care system for colorectal cancer (CRC) prevention, but its effect on CRC mortality is unknown. To determine whether colonoscopy is associated with decreased CRC mortality in veterans and whether its effect differs by anatomical location of CRC. Case-control study. VA-Medicare administrative data. Case patients were veterans aged 52 years or older who were diagnosed with CRC between 2002 and 2008 and died of the disease by the end of 2010. Case patients were matched to 4 control patients without prior CRC on the basis of age, sex, and facility. Conditional logistic regression was performed to calculate odds ratios (ORs) for exposure to colonoscopy, with adjustment for race, Charlson Comorbidity Index score, selected chronic conditions, nonsteroidal anti-inflammatory drug use, and family history of CRC. Exposure to colonoscopy was determined from 1997 to 6 months before CRC diagnosis in case patients and to a corresponding date in control patients. Subgroup analysis was performed for patients who had undergone screening colonoscopy. A total of 4964 case patients and 19 856 control patients were identified. Case patients were significantly less likely to have undergone any colonoscopy (OR, 0.39 [95% CI, 0.35 to 0.43]). Colonoscopy was associated with reduced mortality for left-sided cancer (OR, 0.28 [CI, 0.24 to 0.32]) and right-sided cancer (OR, 0.54 [CI, 0.47 to 0.63]). The results were similar for patients who had undergone screening colonoscopy (overall OR, 0.30 [CI, 0.24 to 0.38]). Sensitivity analyses that varied the interval between CRC diagnosis and colonoscopy exposure did not affect the primary findings. Unmeasured confounding. In this study using national VA-Medicare data, colonoscopy was associated with significant reductions in CRC mortality among veterans and was associated with greater benefit for left-sided cancer than right-sided cancer. U.S. Department of Veterans Affairs.


[Orthodontic treatment failure: the advice of a forensic expert].
If patients consider that their orthodontic treatment is a failure, they can claim compensation from their practitioner. Most often, discontented patients call on their third-party liability insurance when they are covered for "legal expenses protection". The patient's insurance company will then get in touch with the practitioner's insurance firm. Three-quarters of all claims are dealt with by the insurance companies. However, if an agreement cannot be reached or if the practitioner's insurance company manages to establish that he/she is not at fault, the patient can apply to the Tribunal de Grande Instance, the French Regional Court. The judge appoints a forensic expert who will examine the patient and follow a very precise procedure involving a dozen different questions. The advice of the forensic expert is therefore very valuable to orthodontists, in particular regarding their duty to inform patients and the importance of the medical file in order to avoid being held accountable following a course of treatment.


A quantitative three-dimensional assessment of abnormal variations in facial soft tissues of adult patients with cleft lip and palate.
To supply quantitative information about the facial soft tissues of adult operated patients with cleft lip and palate (CLP). The three-dimensional coordinates of soft tissue facial landmarks were obtained using an electromagnetic digitizer in 18 Caucasian patients with CLP (11 males and 7 females aged 19 to 27 years) and 162 healthy controls (73 females and 89 males aged 18 to 30 years). From the landmarks, 15 facial dimensions and two angles were calculated. Data were compared with those collected in healthy individuals by computing z-scores. Two summary anthropometric measurements for quantifying craniofacial variations were assessed in both the patients and reference subjects: the mean z-score (an index of overall facial size), and its SD, called the craniofacial variability index (an index of facial harmony). In treated patients with CLP, facial size was somewhat smaller than in normal individuals, but in all occasions the mean z-score fell inside the normal interval (mean +/- 2 SD). Almost all patients had a craniofacial variability index larger than the normal interval, indicating a global disharmonious appearance. Overall, in patients pronasale, subnasale, and pogonion were more posterior, the nose was shorter and larger, the face was narrower, and the soft tissue profile and upper lip were flatter than in the reference population. The facial soft tissue structures of adult operated patients with CLP differed from those of normal controls of the same age, sex, and ethnic group. In this patient group, surgical corrections of CLP failed to provide a completely harmonious appearance, even if the deviations from the reference were limited. Further analyses of larger groups of patients are needed.


Optimal patient positioning for ligamentotaxis during balloon kyphoplasty of the thoracolumbar and lumbar spine.
Percutaneous balloon kyphoplasty aims to restore vertebral height, correct angular deformity and stabilize the spine in the setting of vertebral compression fractures. The patient is positioned prone with supports under the iliac crests and upper thorax to allow gravity to extend the spine. In the treatment of lumbar fractures, we evaluated patient positioning with the contribution of hip extension to increase anterior ligamentotaxis, thus facilitating restoration of vertebral height. Our positioning technique created a mean anterior height increase from 72% to 78% of the average height of the cranial and caudal vertebrae (p=0.037). Balloon inflation did not significantly further increase anterior or posterior vertebral height, or Cobb angle.


Acute stimulation of white adipocyte respiration by PKA-induced lipolysis.
We examined the effect of β-adrenergic receptor (βAR) activation and cAMP-elevating agents on respiration and mitochondrial uncoupling in human adipocytes and probed the underlying molecular mechanisms. Oxygen consumption rate (OCR, aerobic respiration) and extracellular acidification rate (ECAR, anaerobic respiration) were examined in response to isoproterenol (ISO), forskolin (FSK), and dibutyryl-cAMP (DB), coupled with measurements of mitochondrial depolarization, lipolysis, kinase activities, and gene targeting or knock-down approaches. ISO, FSK, or DB rapidly increased oxidative and glycolytic respiration together with mitochondrial depolarization in human and mouse white adipocytes. The increase in OCR was oligomycin-insensitive and contingent on cAMP-dependent protein kinase A (PKA)-induced lipolysis. This increased respiration and the uncoupling were blocked by inhibiting the mitochondrial permeability transition pore (PTP) and its regulator, BAX. Interestingly, compared with lean individuals, adipocytes from obese subjects exhibited reduced OCR and uncoupling capacity in response to ISO. Lipolysis stimulated by βAR activation or other maneuvers that increase cAMP levels in white adipocytes acutely induces mitochondrial uncoupling and cellular energetics, which are amplified in the absence of scavenging BSA. The increase in OCR is dependent on PKA-induced lipolysis and is mediated by the PTP and BAX. Because this effect is reduced with obesity, further exploration of this uncoupling mechanism will be needed to determine its cause and consequences.


Comparison of 2 blood culture media shows significant differences in bacterial recovery for patients on antimicrobial therapy.
Antimicrobial removal devices in blood culture media are designed to remove antibiotics from the blood culture solution, thereby facilitating bacterial growth. How well these devices function clinically has not been established. All blood drawn for culture from adult inpatients and emergency department visitors in a level I trauma center was placed in paired BACTEC Plus and BacT/Alert FAN culture media and studied simultaneously, consecutively, and prospectively between 1 February and 30 September 2011. All cultures were processed per standard laboratory protocols. Of 9395 total cultures collected, 1219 (13%) were positive, 831 were included, and 524 (33%) contained pathogens. BACTEC had a 4.5-hour faster detection time (P < .0001), and isolated exclusively 182 of 524 (35%; P < .001) pathogens, 136 of 345 (39%) of the gram-positive cocci (P < .001), 48 of 175 (27%; P = .02) of the gram-negative rods, 101 of 195 (52%) of Staphylococcus aureus (P < .001), and 59 of 120 (49%; P = .004) septic events. If active antibiotics had been dosed 0-4 or 4-48 hours prior to culture collection, the odds of that culture growing in BACTEC were 4.8- and 5.2-fold greater, respectively, than of growing in BacT/Alert (P < .0001). Both were equivalent in the recovery of yeast and when no antimicrobials were dosed. BACTEC media has faster time to detection and increased bacterial recovery over the BacT/Alert media in the following categories: overall growth, pathogens, septic events, gram-positive cocci, gram-negative rods, Staphylococcus aureus, and cultures where antimicrobials were dosed up to 48 hours before culture collection.


Outcome evaluation of 'sharing the gift of life': an organ and tissue donation educational program for American Indians.
Culturally focused education about deceased donation is needed for American Indians (AIs). This study tested a program designed to impact intention to serve as a deceased donor for reservation dwelling AIs. A pre/posttest design and a community-based participatory research approach were used. The study was based upon the Transtheoretical Model. Adult participants (N = 1580, 58% women) were from four Northern Plains reservations. An outreach coordinator delivered the program using print and video materials. The outcome was stage of motivational readiness (SMR) to serve as a deceased donor. McNemar's test was used to compare pre- to postintervention changes in SMR. At baseline, 55% of participants were not thinking about being a donor (precontemplation stage) and 45% were thinking about it (contemplation stage). Postintervention, 43.1% of participants were unchanged in SMR and 56.9% progressed in SMR. Of those who progressed, 26.5% (n = 418) changed to the contemplation stage, 19.4% (n= 306) changed to the preparation stage (signed a donor card or joined a registry), and 11.1% (n = 175) confirmed a discussion of the decision with family (action stage). Progression in SMR from pre/post was significant, chi(2)(1) = 18.32, p < 0.05. The intervention resulted in important changes in deceased donor intentions for reservation dwelling AIs.


Sewers in the city: a case study of individual-level mortality and public health initiatives in Northampton, Massachusetts, at the turn of the century.
Emerging industrial communities of nineteenth-century New England experienced both rapid population growth and lagging development of public health infrastructures. In turn, high mortality in these newly urban cities contributed to a delay in the regional mortality transition of the late nineteenth century. Analyzing death records and a file of linked cause-specific death and manuscript census records for the industrializing community of Northampton, Massachusetts, we show that early in the city's development, mortality clustered near industrial activities and open sewers. When industrial areas were sewered, clustering of mortality abated, and differences between industrial and commercial areas of the town were no longer significant. These findings illustrate Szreter's emphasis on considering both the benefits and costs of development. (1) Initial development contributed to high mortality in newly emergent urban-industrial centers like Northampton and was abated only when lagging public health infrastructures caught up with rapid growth and development near the turn of the century.


Myeloid-Derived Suppressor Cells and Radiotherapy.
Myeloid-derived suppressor cells (MDSC) are a heterogeneous population of pathologically activated, mostly immature, myeloid cells that exert robust immunosuppressive functions. MDSCs expand during oncogenesis and have been linked to accelerated disease progression and resistance to treatment in both preclinical tumor models and patients with cancer. Thus, MDSCs stand out as promising targets for the development of novel immunotherapeutic regimens with superior efficacy. Here, we summarize accumulating preclinical and clinical evidence indicating that MDSCs also hamper the efficacy of radiotherapy (RT), as we critically discuss the potential of MDSC-targeting strategies as tools to achieve superior immunotherapeutic tumor control by RT in the clinic.


Indirect revascularization with the dural inversion technique for pediatric moyamoya disease: 20-year experience.
OBJECTIVENumerous surgical procedures facilitate revascularization of the ischemic brain in patients with moyamoya disease. Dural inversion is a technique in which flaps of dura mater centered around the middle meningeal artery are inverted, encouraging the formation of a rich collateral blood supply. This procedure has been used in combination with encephaloduroarteriosynangiosis for more than 20 years at the authors' institution for the treatment of pediatric moyamoya disease. The objective of this study was to describe the clinical and radiographic outcomes for a cohort of consecutive pediatric moyamoya patients undergoing dural inversion.METHODSClinical and radiographic data on patients who had undergone dural inversion in the period from 1997 to 2016 were reviewed. Univariate and multivariate logistic regression and Kaplan-Meier analyses were performed to assess the risk of postoperative stroke, functional outcome, and the angiographic degree of revascularization.RESULTSDural inversion was performed on 169 hemispheres in 102 patients. Median follow-up was 4.3 years. Six patients (3.6% of hemispheres) suffered postoperative ischemic or hemorrhagic stroke. Overall mortality was 1.0%. Good postoperative neurological status (modified Rankin Scale [mRS] score ≤ 2) was observed in 90 patients (88%); preoperative and postoperative mRS scores showed significant improvement (p < 0.001). Eighty-six percent of hemispheres had Matsushima grade A or B revascularization. Younger age was associated with postoperative stroke and poor functional outcome. Patients with secondary moyamoya syndrome had a significantly worse radiographic outcome. The cumulative 5-year Kaplan-Meier risk for stroke was 6.4%.CONCLUSIONSDural inversion is a useful technique of cerebral revascularization in pediatric moyamoya disease. A 20-year experience demonstrates the safety and efficacy of this technique with a relatively low rate of postoperative stroke, good functional outcomes, and favorable angiographic results.


Altered hypothalamus-pituitary-adrenal gland axis regulation in the expanded CGG-repeat mouse model for fragile X-associated tremor/ataxia syndrome.
The human FMR1 gene contains an unstable CGG-repeat in its 5' untranslated region. The repeat length in the normal population is polymorphic (5-54 CGG-repeats). Individuals carrying lengths beyond 200 CGGs (i.e. the full mutation) show hypermethylation and as a consequence gene silencing of the FMR1 gene. The absence of the gene product FMRP causes the fragile X syndrome, the most common inherited form of mental retardation. Elderly carriers of the premutation (PM), which is defined as a repeat length between 55 and 200 CGGs, can develop a progressive neurodegenerative syndrome: fragile X-associated tremor/ataxia syndrome (FXTAS). The high FMR1 mRNA levels observed in cells from PM carriers have led to the hypothesis that FXTAS is caused by a pathogenic RNA gain-of-function mechanism. Apart from tremor/ataxia, specific psychiatric symptoms have been described in PM carriers with or without FXTAS. Since these symptoms could arise from elevated stress hormone levels, we investigated hypothalamic-pituitary-adrenal (HPA) axis regulation using a knock-in mouse model with an expanded CGG-repeat in the PM range (>98 repeats) in the Fmr1 gene, which shows repeat instability, and displays biochemical, phenotypic and neuropathological characteristics of FXTAS. We show elevated levels of corticosterone in serum and ubiquitin-positive inclusions in both the pituitary and adrenal gland of 100-week-old animals. In addition, we demonstrate ubiquitin-positive inclusions in the amygdala from aged expanded CGG-repeat mice. We hypothesize that altered regulation of the HPA axis and the amygdala and higher stress hormone levels in the mouse model for FXTAS may explain associated psychological symptoms in humans.


Rapid efficient synthesis and characterization of silver, gold, and bimetallic nanoparticles from the medicinal plant Plumbago zeylanica and their application in biofilm control.
Nanoparticles (NPs) have gained significance in medical fields due to their high surface-area-to-volume ratio. In this study, we synthesized NPs from a medicinally important plant - Plumbago zeylanica. Aqueous root extract of P. zeylanica (PZRE) was analyzed for the presence of flavonoids, sugars, and organic acids using high-performance thin-layer chromatography (HPTLC), gas chromatography-time of flight-mass spectrometry (GC-TOF-MS), and biochemical methods. The silver NPs (AgNPs), gold NPs (AuNPs), and bimetallic NPs (AgAuNPs) were synthesized from root extract and characterized using ultraviolet-visible spectra, X-ray diffraction (XRD), energy-dispersive spectrometry (EDS), transmission electron microscopy (TEM), and dynamic light scattering (DLS). The effects of these NPs on Acinetobacter baumannii, Staphylococcus aureus, and Escherichia coli biofilms were studied using quantitative biofilm inhibition and disruption assays, as well as using fluorescence, scanning electron microscopy, and atomic force microscopy. PZRE showed the presence of phenolics, such as plumbagin, and flavonoids, in addition to citric acid, sucrose, glucose, fructose, and starch, using HPTLC, GC-TOF-MS, and quantitative analysis. Bioreduction of silver nitrate (AgNO₃) and chloroauric acid (HAuCl₄) were confirmed at absorbances of 440 nm (AgNPs), 570 nm (AuNPs), and 540 nm (AgAuNPs), respectively. The maximum rate of synthesis at 50°C was achieved with 5 mM AgNO₃ within 4.5 hours for AgNPs; and with 0.7 mM HAuCl4 within 5 hours for AuNPs. The synthesis of AgAuNPs, which completed within 90 minutes with 0.7 mM AgNO₃ and HAuCl₄, was found to be the fastest. Fourier-transform infrared spectroscopy confirmed bioreduction, while EDS and XRD patterns confirmed purity and the crystalline nature of the NPs, respectively. TEM micrographs and DLS showed about 60 nm monodispersed Ag nanospheres, 20-30 nm Au nanospheres adhering to form Au nanotriangles, and about 90 nm hexagonal blunt-ended AgAuNPs. These NPs also showed antimicrobial and antibiofilm activity against E. coli, A. baumannii, S. aureus, and a mixed culture of A. baumannii and S. aureus. AgNPs inhibited biofilm in the range of 96%-99% and AgAuNPs from 93% to 98% in single-culture biofilms. AuNPs also showed biofilm inhibition, with the highest of 98% in S. aureus. AgNPs also showed good biofilm disruption, with the highest of 88% in A. baumannii. This is the first report on rapid and efficient synthesis of AgNPs, AuNPs and AgAuNPs from P. zeylanica and their effect on quantitative inhibition and disruption of bacterial biofilms.


Seasonal and pandemic human influenza viruses attach better to human upper respiratory tract epithelium than avian influenza viruses.
Influenza viruses vary markedly in their efficiency of human-to-human transmission. This variation has been speculated to be determined in part by the tropism of influenza virus for the human upper respiratory tract. To study this tropism, we determined the pattern of virus attachment by virus histochemistry of three human and three avian influenza viruses in human nasal septum, conchae, nasopharynx, paranasal sinuses, and larynx. We found that the human influenza viruses-two seasonal influenza viruses and pandemic H1N1 virus-attached abundantly to ciliated epithelial cells and goblet cells throughout the upper respiratory tract. In contrast, the avian influenza viruses, including the highly pathogenic H5N1 virus, attached only rarely to epithelial cells or goblet cells. Both human and avian viruses attached occasionally to cells of the submucosal glands. The pattern of virus attachment was similar among the different sites of the human upper respiratory tract for each virus tested. We conclude that influenza viruses that are transmitted efficiently among humans attach abundantly to human upper respiratory tract, whereas inefficiently transmitted influenza viruses attach rarely. These results suggest that the ability of an influenza virus to attach to human upper respiratory tract is a critical factor for efficient transmission in the human population.


[Ross procedure in treatment of aortic disease: clinical experience of 15 cases].
To summarize the clinical experience of Ross procedure in treatment of aorta valve diseases. The clinical data of 15 patients with aorta valve diseases, 12 men and 3 women, aged 30 +/- 14, including 13 cases of congenital aorta valve disease, 1 case of senile degenerative aortic valve disease, and 1 case of subacte bacterial endocarditis complicated by aortic stenosis (AS), with the heart function (NYHA) of class II in 11 cases and class III in 4 cases, underwent Ross procedure from October 1994 to September 2002 in Anzhen Hospital. Before operation, ultrasound cardiography showed moderate to severe AS and/or aortic insufficiency (AI) with an average aortic valve annulus diameter (AVD) of 2.4 +/- 0.4 cm and normal pulmonary valve. Operation was performed on all patients under cardiopulmonary bypass and moderate hypothermia. The operation procedure was as follows: (1) to take off the auto-pulmonary artery valve; (2) to remove the dysfunctional aortic valve and auto-transplant the pulmonary valve on the aortic root; and (3) to put a pulmonary homograft so as to re-establish on the right ventricular outflow tract. The perieoperative mortality is 0. After the operation, the mean pressure gradient of aortic valve was in the normal limitation (7.23 +/- 0.14 mm Hg), the left ventricular diastolic diameter decreased significantly (P < 0.001), the left ventricular ejection fraction was 0.48 +/- 0.22, and the heart function (NYHA) was at the classes I - II in all the patients. All cases received follow-up of 1 - 9 years, their heart function was all in Class I, and the function of their aortic and pulmonary valves remained well. Ross procedure is a kind of effective alterative operation in treating patients with aortic valve disease.


Atlantoaxial rotatory fixation in the setting of associated congenital malformations: a modified classification system.
A case report. To raise awareness of the development of atlantoaxial rotatory fixation (AARF) in the setting of congenital vertebral anomalies/malformations. Klippel-Feil Syndrome (KFS) is a complex, heterogeneous condition noted as congenital fusion of 2 or more cervical vertebrae with or without spinal or extraspinal manifestations. Although believed to be a rare occurrence in the population, KFS may be underreported. Proper diagnosis of KFS and other congenital conditions affecting the spine is imperative to devise proper management protocols and avoid potential complications resulting from the altered biomechanics associated with such conditions and their abnormal vertebral morphology. Craniovertebral dislocation and AARF may cause severe cervicomedullary and spinal cord compression and could thereby be potentially fatal, especially in patients with KFS who present with congenitally-associated comorbidities. A 13-year-old boy with Chiari type I malformation, craniofacial abnormalities, and other irregularities underwent thoracolumbar spine surgery for his scoliosis curve correction at another institution, which immediately following surgery he became a quadriparetic. The initial preoperative assessment of his cervical spine was limited and the associated KFS was initially undiagnosed. At 14 years of age, he presented to our clinic with an ASIA-C spinal cord injury. Plain radiographs, normal and 3-dimensional reformatted computed tomographs (CT), and magnetic resonance imaging (MRI) noted assimilation of the patient's occiput to the atlas (occipitalization) with congenital fusion of C2-C3, indicative of KFS, and the presence of anterior craniovertebral dislocation with a Fielding and Hawkins type II AARF. Closed reduction of the craniovertebral dislocation was noted, but his atlantoaxial rotatory subluxation was nonresponsive and fixed (AARF). As such, at the age of 14, the patient underwent posterior instrumentation and fusion from the occiput to C4 to maintain reduction of thecraniovertebral dislocation and reduce his AARF. At 9 months postoperative follow-up of his craniovertebral surgery, the instrumentation remained intact, reduction of the atlantoaxial rotatory subluxation was maintained, and posterior bone fusion was noted. Neurologically, he remained an ASIA-C without any substantial return of function. This report raises awareness for the need of a thorough evaluation of the cervical spine to determine patients at high risk for craniovertebral dislocation and atlantoaxial rotatory subluxation, primarily in the context of KFS or other congenital conditions. Three-dimensional CT and MR imaging are ideal radiographic methods to determine the presence and extent of craniovertebral dislocation, AARF, and of abnormal vertebral anatomy/malformations. In addition, the authors propose a modification to the Fielding and Hawkins classification of AARF to include variants and subtypes that account for abnormal anatomy and congenital anomalies/malformations.


Understanding conceptus-maternal interactions: what tools do we need to develop?
Communication between the maternal endometrium and developing embryo/conceptus is critical to support successful pregnancy to term. Studying the peri-implantation period of pregnancy is critical as this is when most pregnancy loss occurs in cattle. Our current understanding of these interactions is limited, due to the lack of appropriate in vitro models to assess these interactions. The endometrium is a complex and heterogeneous tissue that is regulated in a transcriptional and translational manner throughout the oestrous cycle. While there are in vitro models to study endometrial function, they are static and 2D in nature or explant models and are limited in how well they recapitulate the in vivo endometrium. Recent developments in organoid systems, microfluidic approaches, extracellular matrix biology, and in silico approaches provide a new opportunity to develop in vitro systems that better model the in vivo scenario. This will allow us to investigate in a more high-throughput manner the fundamental molecular interactions that are required for successful pregnancy in cattle.


[Diabetes mellitus and pregnancy: screening and therapy].
There are two entities to differentiate: 1. Diabetes in pregnancy (pre-existent type 1 or type 2 diabetes with tendency of deterioration during pregnancy or first occurrence of a type 1 or type 2 diabetes with persistence after pregnancy. 2. Impaired Glucose Tolerance = IGT (Diagnosis of an impairment of glucose tolerance during pregnancy) Aims of the screening are the avoidance of complications for mother and child. There is no uniform consent regarding screening during pregnancy. With pregnant women without risk factors, the blood sugar is determined without eating and drinking in plasma between 24 and 28 weeks of gestation. If values are > 4.8 mmol/l, an oral 75 g glucose tolerance test is carried out. Alternatively, a 50 g oral glucose tolerance test can be made. With pregnant women with risk factors, we already carry out an oral 75 g glucose tolerance test in the first trimester. If the values are normal, we repeat the 75 g glucose tolerance test between 24 and 28 weeks of gestation. Main pillars of therapy are diet and movement. In gestational diabetes, an insulin therapy is indicated if blood sugars are too high, fetal growth is sonographically accelerated or a polyhydramnion is present. Oral antidiabetic medicaments are contraindicated during pregnancy. Pregnancies with diabetes in pregnancy or gestational diabetes are risk pregnancies, which are controlled more intensively. Delivery is ideally at term or with confirmed lung maturity. If an impaired glucose tolerance is diagnosed during pregnancy, an oral 75 g glucose tolerance test as well as annually blood sugar controls are carried out because of the augmented risk of developing a type 2 diabetes later in life.


Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and -independent approaches.
Epithelial circulating tumor cells (CTCs) are detectable in patients with non-small cell lung cancer (NSCLC). However, epithelial to mesenchymal transition, a widely reported prerequisite for metastasis, may lead to an underestimation of CTC number. We compared directly an epithelial marker-dependent (CellSearch) and a marker-independent (isolation by size of epithelial tumor cells [ISET]) technology platform for the ability to identify CTCs. Molecular characteristics of CTCs were also explored. Paired peripheral blood samples were collected from 40 chemonäive, stages IIIA to IV NSCLC patients. CTCs were enumerated by Epithelial Cell Adhesion Molecule-based immunomagnetic capture (CellSearch, Veridex) and by filtration (ISET, RareCell Diagnostics). CTCs isolated by filtration were assessed by immunohistochemistry for epithelial marker expression (cytokeratins, Epithelial Cell Adhesion Molecule, epidermal growth factor receptor) and for proliferation status (Ki67). CTCs were detected using ISET in 32 of 40 (80%) patients compared with 9 of 40 (23%) patients using CellSearch. A subpopulation of CTCs isolated by ISET did not express epithelial markers. Circulating tumor microemboli (CTM, clusters of ≥ 3 CTCs) were observed in 43% patients using ISET but were undetectable by CellSearch. Up to 62% of single CTCs were positive for the proliferation marker Ki67, whereas cells within CTM were nonproliferative. Both technology platforms detected NSCLC CTCs. ISET detected higher numbers of CTCs including epithelial marker negative tumor cells. ISET also isolated CTM and permitted molecular characterization. Combined with our previous CellSearch data confirming CTC number as an independent prognostic biomarker for NSCLC, we propose that this complementary dual technology approach to CTC analysis allows more complete exploration of CTCs in patients with NSCLC.


Determination of blood folate using acid extraction and internally standardized gas chromatography-mass spectrometry detection.
Whole blood folate level is a superior indicator of folate nutritional status than serum/plasma level. Problems with and lack of confidence in results of current whole blood folate assays have limited its popularity for assessing folate nutritional status. Here, an acid extraction GCMS detection method that measures total folate whole blood is presented. Folates are released from the matrix of whole blood and cleaved to para-aminobenzoic acid (pABA) by acid hydrolysis in the presence of [(13)C(6)]pABA as internal standard (IS). The hydrolysate is passed over a C18 resin to remove heme. The pABA isotopomers are ethyl esterified, isolated on C18 resin, and trifluoroacetylated. Following normal-phase HPLC separation, the isotopomers are silylated to their tBDMS derivatives. The abundance of these derivatives are measured at m/z 324 for [(13)C(6)]pABA as IS and m/z 318 for pABA from whole blood folate. Our method uses readily available chemicals and our results agree well with those using Lactobacillus casei, the current gold standard reference assay. The presence of folate analogs (methotrexate) or antibacterials (sulfonamines) does not affect our method. This feature makes it useful in monitoring folate status of patients undergoing chemotherapy. Before using our method, pABA supplements must be discontinued for a few days.


Follow-up plasma apolipoprotein E levels in the Australian Imaging, Biomarkers and Lifestyle Flagship Study of Ageing (AIBL) cohort.
Alzheimer's disease (AD) is a growing socioeconomic problem worldwide. Early diagnosis and prevention of this devastating disease have become a research priority. Consequently, the identification of clinically significant and sensitive blood biomarkers for its early detection is very important. Apolipoprotein E (APOE) is a well-known and established genetic risk factor for late-onset AD; however, the impact of the protein level on AD risk is unclear. We assessed the utility of plasma ApoE protein as a potential biomarker of AD in the large, well-characterised Australian Imaging, Biomarkers and Lifestyle Study of Ageing (AIBL) cohort. Total plasma ApoE levels were measured at 18-month follow-up using a commercial bead-based enzyme-linked immunosorbent assay: the Luminex xMAP human apolipoprotein kit. ApoE levels were then analysed between clinical classifications (healthy controls, mild cognitive impairment (MCI) and AD) and correlated with the data available from the AIBL cohort, including but not limited to APOE genotype and cerebral amyloid burden. A significant decrease in ApoE levels was found in the AD group compared with the healthy controls. These results validate previously published ApoE protein levels at baseline obtained using different methodology. ApoE protein levels were also significantly affected, depending on APOE genotypes, with ε2/ε2 having the highest protein levels and ε4/ε4 having the lowest. Plasma ApoE levels were significantly negatively correlated with cerebral amyloid burden as measured by neuroimaging. ApoE is decreased in individuals with AD compared with healthy controls at 18-month follow-up, and this trend is consistent with our results published at baseline. The influence of APOE genotype and sex on the protein levels are also explored. It is clear that ApoE is a strong player in the aetiology of this disease at both the protein and genetic levels.


The ultrastructure of myocardial hypertrophy: why does the compensated heart fail?
Several qualitative features of the ultrastructure of pressure overload and thyrotoxic myocardium are unique markers of the type and quantity of increased work the heart has been required to perform. Furthermore, they are reminiscent of features of normally growing myocytes, implying that the changes in the hypertrophied cell are the consequence of normally present capacities for adaptation to a demand for increased myocardial work. Thyrotoxic myocardium has two features which distinguish it from normal and pressure overloaded hearts: the mitochondria are large and have a peculiar fragile or lacey appearance. Many myocytes show considerable disorganization of sarcomeric myofilaments. Pressure overloaded hearts have smaller and more numerous mitochondria than the normal myocyte. Their sarcomeres have thicker Z bands than controls. Double intercalated discs are also a feature of these myocytes. Several features of hypertrophied myocytes are seen in both types of hypertrophy: RER and ribosomes on the external nuclear membrane. There are polyribosomes aligned along the long axes of thick filaments, presumably involved in myosin synthesis or transformation within the cell. There are areas of sarcomerogenesis both under the sarcolemma and within the cell at the intercalated disc. These are characterized by fragments of myofilaments, polyribosomes and rough endoplasmic reticulum. Quantitatively, myocyte composition is transiently disturbed but, like that of normally growing hearts, returns to control values as the adaptation to stress is negotiated.


Bcl3 prevents acute inflammatory lung injury in mice by restraining emergency granulopoiesis.
Granulocytes are pivotal regulators of tissue injury. However, the transcriptional mechanisms that regulate granulopoiesis under inflammatory conditions are poorly understood. Here we show that the transcriptional coregulator B cell leukemia/lymphoma 3 (Bcl3) limits granulopoiesis under emergency (i.e., inflammatory) conditions, but not homeostatic conditions. Treatment of mouse myeloid progenitors with G-CSF--serum concentrations of which rise under inflammatory conditions--rapidly increased Bcl3 transcript accumulation in a STAT3-dependent manner. Bcl3-deficient myeloid progenitors demonstrated an enhanced capacity to proliferate and differentiate into granulocytes following G-CSF stimulation, whereas the accumulation of Bcl3 protein attenuated granulopoiesis in an NF-κB p50-dependent manner. In a clinically relevant model of transplant-mediated lung ischemia reperfusion injury, expression of Bcl3 in recipients inhibited emergency granulopoiesis and limited acute graft damage. These data demonstrate a critical role for Bcl3 in regulating emergency granulopoiesis and suggest that targeting the differentiation of myeloid progenitors may be a therapeutic strategy for preventing inflammatory lung injury.


Wideband optical vector network analyzer based on optical single-sideband modulation and optical frequency comb.
A novel approach to increase the measurement range of the optical vector network analyzer (OVNA) based on optical single-sideband (OSSB) modulation is proposed and experimentally demonstrated. In the proposed system, each comb line in an optical frequency comb (OFC) is selected by an optical filter and used as the optical carrier for the OSSB-based OVNA. The frequency responses of an optical device-under-test (ODUT) are thus measured channel by channel. Because the comb lines in the OFC have fixed frequency spacing, by fitting the responses measured in all channels together, the magnitude and phase responses of the ODUT can be accurately achieved in a large range. A proof-of-concept experiment is performed. A measurement range of 105 GHz and a resolution of 1 MHz is achieved when a five-comb-line OFC with a frequency spacing of 20 GHz is applied to measure the magnitude and phase responses of a fiber Bragg grating.


The Caenorhabditis elegans unc-93 gene encodes a putative transmembrane protein that regulates muscle contraction.
unc-93 is one of a set of five interacting genes involved in the regulation or coordination of muscle contraction in Caenorhabditis elegans. Rare altered-function alleles of unc-93 result in sluggish movement and a characteristic "rubber band" uncoordinated phenotype. By contrast, null alleles cause no visibly abnormal phenotype, presumably as a consequence of the functional redundancy of unc-93. To understand better the role of unc-93 in regulating muscle contraction, we have cloned and molecularly characterized this gene. We isolated transposon-insertion alleles and used them to identify the region of DNA encoding the unc-93 protein. Two unc-93 proteins differing at their NH2 termini are potentially encoded by transcripts that differ at their 5' ends. The putative unc-93 proteins are 700 and 705 amino acids in length and have two distinct regions: the NH2 terminal portion of 240 or 245 amino acids is extremely hydrophilic, whereas the rest of the protein has multiple potential membrane-spanning domains. The unc-93 transcripts are low in abundance and the unc-93 gene displays weak codon usage bias, suggesting that the unc-93 protein is relatively rare. The unc-93 protein has no sequence similarity to proteins listed in current data-bases. Thus, unc-93 is likely to encode a novel membrane-associated muscle protein. We discuss possible roles for the unc-93 protein either as a component of an ion transport system involved in excitation-contraction coupling in muscle or in coordinating muscle contraction between muscle cells by affecting the functioning of gap junctions.


Integrin alphavbeta3 binding to human alpha5-laminins facilitates FGF-2- and VEGF-induced proliferation of human ECV304 carcinoma cells.
Human ECV304 cells respond reproducibly by tube formation to complex basement membrane matrices. Laminins are major glycoproteins of basement membranes. We therefore studied the ability of ECV304 cells to attach to defined laminin isoforms and to fibronectin, and identified the involved laminin receptors. The cells bound poorly to fibronectin, to some extent to laminin-1, whereas laminin-2/4 and -10/11 were strong adhesive substrates. Antibody perturbation assays showed that adhesion to laminin-1 was mediated by integrin alpha6beta1, and adhesion to laminin-2/4 by cooperative activity of integrins alpha3beta1 and alpha6beta1. Adhesion of ECV 304 cells to laminin-10/11 was mainly mediated by integrins alpha3beta1, with minor involvement of alpha6beta1/4 and alphavbeta3. Solid-phase binding assays confirmed that integrin alphavbeta3 binds human laminin-10/11 and -10, in an RGD-dependent fashion. Although integrin alphavbeta3 played a very minor role in cell adhesion to laminin-10/11, this interaction facilitated growth factor-induced proliferation of ECV304 cells. In response to FGF-2 or VEGF, the cells proliferated better when attached on laminin-10/11 than on laminin-1, -2/4, or gelatin. The proliferation induced by the joint application of laminin-10/11 and either one of the growth factors could be blocked by antibodies against integrin alphavbeta3. Fragments of several other basement membrane components are known to interact with alphavbeta3. The current data show that that integrin alphavbeta3 can bind intact alpha5-containing laminin trimers. Since the laminin alpha5 chain is broadly expressed in adult basement membranes, this interaction could be physiologically important. Our data suggest that this interaction is involved in the regulation of cellular responses to growth factors known to be involved in epithelial and endothelial development.


Risk factors of postoperative coronal malalignment following long-segment spinal fusion surgery in which multilevel lateral lumbar interbody fusion was used for degenerative lumbar kyphoscoliosis.
In patients with adult spinal deformity, especially degenerative lumbar kyphoscoliosis (DLKS), preoperative sagittal malalignment and coronal malalignment (CM) often coexist. Lateral lumbar interbody fusion (LLIF) has recently been widely chosen for DLKS treatment due to its minimal invasiveness and excellent sagittal alignment correction. However, postoperative CM may remain or occur due to an oblique takeoff phenomenon following multilevel LLIF, resulting in poor clinical outcomes. The authors investigated the risk factors for postoperative CM after long-segment fusion corrective surgery in which multilevel LLIF was used in patients with DLKS. Fifty-four consecutive patients with DLKS, main Cobb angle ≥ 20°, and lumbar lordosis ≤ 20° who underwent corrective spinal fusion surgery, including extreme lateral interbody fusion at ≥ 3 segments, were included at the authors' institute between April 2014 and October 2019. Patients who underwent suitable 3-column osteotomy, classified as grade 3-6 per the Scoliosis Research Society-Schwab criteria, were excluded. Patients were divided into CM and non-CM groups based on postoperative CM evaluated using standard standing-position radiographs obtained 2 years postoperatively. CM was defined as an absolute C7-CSVL (deviation of C7 plumb line off central sacral vertical line; calculated by defining the convex side of the CSVL as positive numerical values) value of ≥ 3.0 cm. Patient demographics and preoperative sagittal alignment parameters were evaluated. The following variables were measured to assess coronal alignment: main Cobb angle; preoperative C7-CSVL; amount of lateral listhesis; L4, L5, and sacral coronal tilt angles; coronal vertebral deformity angles; and coronal spine rigidity. Regarding risk factors for postoperative CM, patient characteristics, preoperative sagittal parameters, and coronal parameters did not significantly differ between the 2 groups, except for preoperative C7-CSVL (p = 0.016). Multivariate logistic regression analysis revealed that preoperative C7-CSVL (+1 cm; OR 1.23, 95% CI 1.05-1.50; p = 0.007) was a significant predictor of postoperative CM. Receiver operating characteristic curve analysis demonstrated that the cutoff value for preoperative C7-CSVL was +0.3 cm, the sensitivity was 85.7%, the specificity was 60.6%, and the area under the curve was 0.70. In corrective fusion surgery for DLKS in which multilevel LLIF was used, the occurrence of postoperative CM was associated with preoperative C7-CSVL. According to the C7-CSVL, which was evaluated by defining the convex side of the CSVL as positive numerical values and the concave side as negative numerical values, the CM group had a significantly higher value of preoperative C7-CSVL than did the non-CM group. Alternative corrective fusion methods, other than multiple LLIFs, may be considered in DLKS cases with a C7-CSVL of +0.3 cm or greater.


Diverse Transcriptome Responses to Salinity Change in Atlantic Cod Subpopulations.
Adaptation to environmental variation caused by global climate change is a significant aspect of fisheries management and ecology. A reduction in ocean salinity is visible in near-shore areas, especially in the Baltic Sea, where it is affecting the Atlantic cod population. Cod is one of the most significant teleost species, with high ecological and economical value worldwide. The population of cod in the Baltic Sea has been traditionally divided into two subpopulations (western and eastern) existing in higher- and lower-salinity waters, respectively. In recent decades, both Baltic cod subpopulations have declined massively. One of the reasons for the poor condition of cod in the Baltic Sea is environmental factors, including salinity. Thus, in this study, an oligonucleotide microarray was applied to explore differences between Baltic cod subpopulations in response to salinity fluctuations. For this purpose, an exposure experiment was conducted consisting of salinity elevation and reduction, and gene expression was measured in gill tissue. We found 400 differentially expressed genes (DEGs) involved in the immune response, metabolism, programmed cell death, cytoskeleton, and extracellular matrix that showed a subpopulation-dependent pattern. These findings indicate that osmoregulation in Baltic cod is a complex process, and that western and eastern Baltic cod subpopulations respond differently to salinity changes.


Standards-based, open-source electronic health record systems: a desirable future for the U.S. health industry.
Many healthcare systems are moving toward a fully electronic health record (EHR) in order to better manage patient care. Unfortunately, in the United States, many current EHR systems leave much to be desired. Among well-documented criticisms are that they tend to be inflexible, proprietary, nonintuitive, expensive, difficult to maintain and rarely interoperable across health systems. From the clinician's perspective, these flaws sometimes make having an EHR system seem no better than retaining a paper-based system. Open-source software, a great success in other information-intensive industries, is one possible solution to these problems, and may help integrate a functional EHR system into, and across, more health systems and clinics because of the greater potential for local customization. We believe that the advantages of an open-source EHR system outweigh the costs of a more traditional, proprietary EHR system, and recommend that more work be done to advance an interoperable open-source EHR system in the United States. Open-source EHR systems have the potential to improve healthcare in the United States as they have done in many other areas around the world.


Mead Johnson Critical Care Symposium for the Practising Surgeon. 2. Complications of monitoring systems.
Current invasive monitoring techniques, although valuable in the care of critically ill patients, also have disadvantages. Complications of these techniques include errors in interpretation of measurements and complications of venous access and indwelling lines. To prevent these, the author recommends a standard routine for inserting and changing catheters, regular calibration of transducers and insertion of Swan-Ganz catheters with the balloon inflated with 1.0 to 1.5 ml of air.


Depression Prediction by Using Ecological Momentary Assessment, Actiwatch Data, and Machine Learning: Observational Study on Older Adults Living Alone.
Although geriatric depression is prevalent, diagnosis using self-reporting instruments has limitations when measuring the depressed mood of older adults in a community setting. Ecological momentary assessment (EMA) by using wearable devices could be used to collect data to classify older adults into depression groups. The objective of this study was to develop a machine learning algorithm to predict the classification of depression groups among older adults living alone. We focused on utilizing diverse data collected through a survey, an Actiwatch, and an EMA report related to depression. The prediction model using machine learning was developed in 4 steps: (1) data collection, (2) data processing and representation, (3) data modeling (feature engineering and selection), and (4) training and validation to test the prediction model. Older adults (N=47), living alone in community settings, completed an EMA to report depressed moods 4 times a day for 2 weeks between May 2017 and January 2018. Participants wore an Actiwatch that measured their activity and ambient light exposure every 30 seconds for 2 weeks. At baseline and the end of the 2-week observation, depressive symptoms were assessed using the Korean versions of the Short Geriatric Depression Scale (SGDS-K) and the Hamilton Depression Rating Scale (K-HDRS). Conventional classification based on binary logistic regression was built and compared with 4 machine learning models (the logit, decision tree, boosted trees, and random forest models). On the basis of the SGDS-K and K-HDRS, 38% (18/47) of the participants were classified into the probable depression group. They reported significantly lower scores of normal mood and physical activity and higher levels of white and red, green, and blue (RGB) light exposures at different degrees of various 4-hour time frames (all P<.05). Sleep efficiency was chosen for modeling through feature selection. Comparing diverse combinations of the selected variables, daily mean EMA score, daily mean activity level, white and RGB light at 4:00 pm to 8:00 pm exposure, and daily sleep efficiency were selected for modeling. Conventional classification based on binary logistic regression had a good model fit (accuracy: 0.705; precision: 0.770; specificity: 0.859; and area under receiver operating characteristic curve or AUC: 0.754). Among the 4 machine learning models, the logit model had the best fit compared with the others (accuracy: 0.910; precision: 0.929; specificity: 0.940; and AUC: 0.960). This study provides preliminary evidence for developing a machine learning program to predict the classification of depression groups in older adults living alone. Clinicians should consider using this method to identify underdiagnosed subgroups and monitor daily progression regarding treatment or therapeutic intervention in the community setting. Furthermore, more efforts are needed for researchers and clinicians to diversify data collection methods by using a survey, EMA, and a sensor.


Setting safe acute exposure limits for halon replacement chemicals using physiologically based pharmacokinetic modeling.
Most proposed replacements for Halon 1301 as a fire suppressant are halogenated hydrocarbons. The acute toxic endpoint of concern for these agents is cardiac sensitization. An approach is described that links the cardiac endpoint as assessed in dogs to a target arterial concentration in humans. Linkage was made using a physiologically based pharmacokinetic (PBPK) model. Monte Carlo simulations, which account for population variability, were used to establish safe exposure times at different exposure concentrations for Halon 1301 (bromotrifluoromethane), CF(3)I (trifluoroiodomethane), HFC-125 (pentafluoroethane), HFC-227ea (1,1,1,2,3,3,3-heptafluoropropane), and HFC-236fa (1,1,1,3,3,3-hexafluoropropane). Application of the modeling technique described here not only makes use of the conservative cardiac sensitization endpoint, but also uses an understanding of the pharmacokinetics of the chemical agents to better establish standards for safe exposure. The combined application of cardiac sensitization data and physiologically based modeling provides a quantitative approach, which can facilitate the selection and effective use of halon replacement candidates.


Rational combination therapy in refractory migraine.
Refractory migraine (RM) headaches pose important treatment challenges to the patients who live with them and the clinicians who try to treat them. Defined based on the lack of response to acute, preventive, and nonpharmacologic treatment, RM is often treated with a combination of treatments. Although combination therapy for RM has not been systematically studied in randomized trials, clinical experience suggests that a rational approach to RM treatment, utilizing a combination of treatments, may be effective where monotherapy has failed. In this article we briefly identify patient populations appropriate for more aggressive migraine prevention with combination therapy. We then discuss modifiable risk factors and comorbidities in migraine and then focus on the use of rational combination therapy, as well as the duration migraine preventatives should be considered for use. Future research is needed to evaluate the full potential of rational combination treatment as a strategy for treating and ultimately preventing RM.


Comparison of MRI techniques for detecting microadenomas in Cushing's disease.
OBJECTIVE Many centers use conventional and dynamic contrast-enhanced MRI (DMRI) sequences in patients with Cushing's disease. The authors assessed the utility of the 3D volumetric interpolated breath-hold examination, a spoiled-gradient echo 3D T1 sequence (SGE) characterized by superior soft tissue contrast and improved resolution, compared with DMRI and conventional MRI (CMRI) for detecting microadenomas in patients with Cushing's disease. METHODS This study was a blinded assessment of pituitary MRI in patients with proven Cushing's disease. Fifty-seven patients who had undergone surgery for Cushing's disease (10 male, 47 female; age range 13-69 years), whose surgical findings were considered to represent a microadenoma, and who had been examined with all 3 imaging techniques were included. Thus, selection emphasized patients with prior negative or equivocal MRI on referral. The MRI annotations were anonymized and 4 separate imaging sets were independently read by 3 blinded, experienced clinicians: a neuroradiologist and 2 pituitary surgeons. RESULTS Forty-eight surgical specimens contained an adenoma (46 ACTH-staining adenomas, 1 prolactinoma, and 1 nonfunctioning microadenoma). DMRI detected 5 adenomas that were not evident on CMRI, SGE detected 8 adenomas not evident on CMRI, including 3 that were not evident on DMRI. One adenoma was detected on DMRI that was not detected on SGE. McNemar's test for efficacy between the different MRI sets for tumor detection showed that the addition of SGE to CMRI increased the number of tumors detected from 18 to 26 (p = 0.02) based on agreement of at least 2 of 3 readers. CONCLUSIONS SGE shows higher sensitivity than DMRI for detecting and localizing pituitary microadenomas, although rarely an adenoma is detected exclusively by DMRI. SGE should be part of the standard MRI protocol for patients with Cushing's disease.


Neonectria and Cylindrocarpon: the Nectria mammoidea group and species lacking microconidia.
Neonectria (Hypocreales: Nectriaceae) species having Cylindrocarpon anamorphs that lack microconidia and chlamydospores include: Neo. discophora var. discophora, Neo. discophora var. rubi, stat nov. et comb. nov., Neo. lucida, comb. nov., Neo. viridispora, sp. nov. and Neo. westlandica, comb. nov. Perithecia of these species are red and perithecial anatomyis of the N. mammoidea type, with a palisade of hypha-like cells in the outer perithecial wall. These species occur on recently dead or dying trees. Perithecia of Neo. betulae, sp. nov and Neo. dumontii, sp. nov. are anatomically and biologically similar to those of Neo. discophora. The only known culture of Neo. betulae remained sterile, while Neo. dumontii has not been cultured; their anamorphs are presumed to be Cylindrocarpon. Analyses of mit ssu rDNA sequences indicate that Neonectria/Cylindrocarpon is monophyletic. Within the genus, species having N. mammoidea type perithecia are paraphyletic. Most species cluster with Neo. discophora, but Neo. westlandica and Neo. trachosa are basal to a clade that includes species that do not have a N. mammoidea-type perithecium. Nectria fuckeliana clusters independently of Neonectria and Nectria. Although reported to have a Cylindrocarpon anamorph, fresh ascospore isolates of N. fuckeliana did not produce Cylindrocarpon macroconidia but produced acremonium- or verticillium-like anamorphs. A key to nectriaceous species of Neonectria that have Cylindrocarpon anamorphs that lack microconidia and chlamydospores and/or that have a N. mammoidea type perithecial wall anatomy is presented. New combinations are proposed for other species formerly included in Nectria that have non-microconidial Cylindrocarpon anamorphs: Neonectria cinnamomea, Neo. jungneri, Neo. platycephala, Neo. phaeodisca and Neo. verrucospora.


[The variants of changes in pro- and antioxidative processes in viral hepatitis and the necessity of differential approach in their antioxidative therapy].
There was investigated the changes in pro- and antioxidative processes in viral hepatitis of different etiology. It was revealed that the mechanisms mentioned above essentially differ from each other and for their treatment, accordingly, different groups of antioxidative drugs should be used. The data obtained indicates the impossibility of "universal" antioxidative therapy. The role of oxidative stress in the mechanisms of different types of hepatitis and unforeseen negative effect of antioxidative therapy requires special caution and attention in selection of mitochondrial respiratory chain stabilizing antioxidants. In hepatitis A, which is characterized by relatively less intensity of peroxidation natural antiradical remedies should be used (e.g. tocopherol). In viral hepatitis B and especially in hepatitis C, it is desirable to use the selenium-containing antioxidants. In hepatitis B, which is characterized by relatively less content of nitric oxide, it is recommended to use the stimulants of NO synthesis.


Early implant placement with simultaneous guided bone regeneration following single-tooth extraction in the esthetic zone: a cross-sectional, retrospective study in 45 subjects with a 2- to 4-year follow-up.
The concept of early implant placement is a treatment option in postextraction sites of single teeth in the anterior maxilla. Implant placement is performed after a soft tissue healing period of 4 to 8 weeks. Implant placement in a correct three-dimensional position is combined with a simultaneous guided bone regeneration procedure to rebuild esthetic facial hard and soft tissue contours. In this retrospective, cross-sectional study, 45 patients with an implant-borne single crown in function for 2 to 4 years were recalled for examination. Clinical and radiologic parameters, routinely used in implant studies, were assessed. All 45 implants were clinically successful according to strict success criteria. The implants demonstrated ankylotic stability without signs of a peri-implant infection. The peri-implant soft tissues were clinically healthy as indicated by low mean plaque (0.42) and sulcus bleeding index (0.51) values. None of the implants revealed a mucosal recession on the facial aspect as confirmed by a clearly submucosal position of all implant shoulders. The mean distance from the mucosal margin to the implant shoulder was -1.93 mm on the facial aspect. The periapical radiographs showed stable peri-implant bone levels, with a mean distance between the implant shoulder and the first bone-implant contact of 2.18 mm. This retrospective study demonstrated successful treatment outcomes for all 45 implants examined. The mid-term follow-up of 2 to 4 years also showed that the risk for mucosal recession was low with this treatment concept. Prospective clinical studies are required to confirm these encouraging results.


Microglia is associated with p-Tau aggregates in the olfactory bulb of patients with neurodegenerative diseases.
The olfactory bulb (OB) seems to be the first affected structure in neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and Lewy body dementia (LBD). Deposits of protein aggregates, increased dopaminergic neurons, and decreased cholinergic inputs have all been described in the OB of these diseases. We investigated here the contribution of the activated microglial cells to the increased deposits of protein aggregates. We quantified the number of activated microglial cells and astrocytes in the OB of patients with histological diagnosis of PD (n = 5), AD (n = 13), and LBD (n = 7) and aged-matched controls (n = 8). Specific consensus diagnostic criteria were applied for AD, LBD, and PD. Protein aggregates were scored in the OB as grade 0, none; grade 1, mild; grade 2, moderate; and grade 3, severe. OB sections from the 33 subjects were stained with specific antibodies markers for reactive astrocytes (GFAP) and microglial cells (Iba1 and HLA-DR). The total number of Iba1-ir (Iba-immunoreactive) and HLAD-DR cells was estimated by stereological analysis, while quantification of astrocytes was performed by GFAP optical density. Statistical analysis was done using the Stata 12.0 software. The number of microglia and activated microglia cells (HLA-RD-ir) was increased in patients with neurodegenerative diseases (p < 0.05). Moreover, the density of GFAP-ir cells was higher in the OB of patients. Neither the number of microglia cells nor the density of astrocytes correlated with the number of b-amyloid and alpha-synuclein deposits, but the density of Iba1-ir cells correlated with the number of p-Tau aggregates. Activated microglial cells and reactive astrocytes are present in the OB of patients with neurodegenerative diseases. The lack of correlation between the number of activated microglia cells and protein deposits indicate that they might independently contribute to the degenerative process. The presence of microglia is related to phosphorylated Tau deposits in neurodegenerative diseases.


[Physiopathologic implications of telomerase].
Telomerase is a specialized RNA-directed DNA polymerase that extends telomeres of eukaryotic chromosomes. Telomerase gene expression is active in germinal, stem and tumoral cells and repressed in normal human somatic cells. Activation of telomerase involves immortalization, while repression of this enzyme involves the shortening of telomeres in each round of chromosomal replication. Telomere shortening constitutes a mechanism for cells to control cell division and the entrance in the replicative senescence state. If the expression of this enzyme is forced, senescent human cells will extend their life span reaching the immortalization and malignization of the cells. Activation or depression of telomerase activity has important implications in aging and cancer therapy. Telomerase regulation is a multifactorial process in which telomerase expression, post-translational protein-protein interactions and protein phosphorylation are involved.


Active transcutaneous bone conduction hearing implants: Systematic review and meta-analysis.
In July 2018 the active transcutaneous bone conduction hearing implant received FDA approval in the US (for patients 12 years and older with conductive and/or mixed hearing loss or single-sided deafness), reflecting the current trend of moving away from percutaneous hearing solutions towards intact skin systems. To critically assess the current literature on safety, efficacy and subjective benefit after implantation with an active transcutaneous bone conduction hearing device. Literature investigation was performed by electronic database search including PubMed and Cochrane Central Register of Controlled Trials, and manual search of relevant journals and reference lists of included studies. Randomized controlled trials, clinical controlled trials and cohort studies, case series and case reports investigating subjective and objective outcomes. Retrieved literature was screened and extracted by two reviewers independently. Subgroup analysis of indications (conductive and/or mixed hearing loss, single-sided deafness) and participant ages (pediatric vs. adults) was conducted on patients with active transcutaneous bone conduction devices. Sensitivity analysis was performed to test the stability of the results in meta-analysis. 39 citations reporting on pre- and postoperative audiological results, speech performance in quiet and in noise, localization testing as well as subjective outcomes were included in this systematic review. Functional gain as well as word recognition score outcomes could be further investigated via meta-analysis. All outcomes reported and summarized here reflect beneficial audiological performance and high patient satisfaction, accompanied with a low complications rate (minor event incidence rate: 9.9 person-years; major incidence rate: 148.9 person-years) for the indications of conductive and mixed hearing loss as well as in individuals suffering from single-sided deafness for all age groups of subjects who underwent active transcutaneous bone conduction hearing device implantation. A limiting factor of this systematic review was the Level of Evidence of the reviewed literature, comprising 2a/3a studies (cohort studies and case-control studies). Furthermore, the reporting standards, especially in outcomes such as word recognition scores in quiet and in noise, vary across study cites from various countries, which impedes comparisons. Last but not least, no other comparable other device was retrieved as the active transcutaneous bone conduction hearing device is the only available at the moment. The device's transcutaneous technology results in a minor event incidence rate of one in 9.9 person-years and a major incidence rate of one in 148.9 person-years. Based on the audiological outcomes, high patient satisfaction as well as the low complication rate, the authors recommend the active transcutaneous bone conduction hearing device as a safe and effective treatment for patients suffering from hearing loss within the device's indication criteria (conductive and/or mixed hearing loss or single-sided deafness).


What is missing from the sturgeon jaw: Developmental morphology of the upper jaw in Acipenser.
Sturgeons belong to the family Acipenseridae, the most species-rich extant family of Acipenseriformes, a basal actinopterygian group of key importance in assessing the early radiations of the actinopterygians. At the same time, acipenseriforms display unique specializations in the morphology of the snout and jaws which make them a valuable model for studying evolutionary novelties. However, despite a long history of research, the homologies of the snout and the mandibular arch of acipenseriforms remain uncertain preventing further studies on the evolutionary origin of their unique snout and jaw structure, and in particular, of the upper jaw symphysis, the key apomorphy of the group and the preoral snout. In the present study, a detailed description of the upper jaw morphology and development in sturgeons is provided in order to address its composition in terms of the common actinopterygian archetype. Based on the obtained results, the upper jaw of acipenseriforms is assumed to have lost the autopalatine portion, which most likely is represented by the separate cartilages supporting the tentacles. Also, the conventional interpretation of the sturgeon's maxilla as dermopalatine is rejected on the grounds of this bone structure and development. Paedomorphosis is proposed to be the most likely mechanism explaining the evolutionary origin of the upper jaw symphysis and supposed modifications of the snout in sturgeons.


Segmentation, Binding, and Illusory Conjunctions.
We investigate binding within the framework of a model of excitatory and inhibitory cell assemblies that form an oscillating neural network. Our model is composed of two such networks that are connected through their inhibitory neurons. The excitatory cell assemblies represent memory patterns. The latter have different meanings in the two networks, representing two different attributes of an object, such as shape and color. The networks segment an input that contains mixtures of such pairs into staggered oscillations of the relevant activities. Moreover, the phases of the oscillating activities representing the two attributes in each pair lock with each other to demonstrate binding. The system works very well for two inputs, but displays faulty correlations when the number of objects is larger than two. In other words, the network conjoins attributes of different objects, thus showing the phenomenon of "illusory conjunctions," as in human vision.


Three-dimensional imaging of living and dying neurons with atomic force microscopy.
Atomic Force Microscopy (AFM) has been used to image the morphology of developing neurons and their processes. Additionally, AFM can physically interact with the cell under investigation in numerous ways. Here we use the AFM to both three-dimensionally image the neuron and to inflict a nano/micro-puncture to its membrane. Thus, the same instrument used as a tool to precisely penetrate/cut the membrane at the nanoscale level is employed to image the morphological responses to damage. These first high resolution AFM images of living chick dorsal root ganglion cells and cells of sympathetic ganglion and their growing processes provide confirmation of familiar morphologies. The increased resolution of the AFM revealed these structures to be significantly more complex and variable than anticipated. Moreover we describe novel, dynamic, and unreported architectures, particularly large dorsally projecting ridges, spines, and ribbons of cytoplasm that appear and disappear on the order of minutes. In addition, minute (ca. 100 nm) hair-like extensions of membrane along the walls of nerve processes that also shift in shape and density, appearing and disappearing over periods of minutes were seen. We also provide "real time" images of the death of the neuron cell body after nano/micro scale damage to its membrane. These somas excreted their degraded cytoplasm, revealed as an enlarging pool beneath and around the cell. Conversely, identical injury, even repeated perforations and nanoslices, to the neurite's membrane do not lead to demise of the process. This experimental study not only provides unreported neurobiology and neurotrauma, but also emphasizes the unique versatility of AFM as an instrument that can (1) physically manipulate cells, (2) provide precise quantitative measurements of distance, surface area and volume at the nanoscale if required, (3) derive physiologically significant data such as membrane pressure and compliance, and (4) during the same period of study--provide unexcelled imaging of living samples.


Feline faces: Unraveling the social function of domestic cat facial signals.
Lately, there has been a growing interest in studying domestic cat facial signals, but most of this research has centered on signals produced during human-cat interactions or pain. The available research on intraspecific facial signaling with domesticated cats has largely focused on non-affiliative social interactions. However, the transition to intraspecific sociality through domestication could have resulted in a greater reliance on affiliative facial signals that aid with social bonding. Our study aimed to document the various facial signals that cats produce during affiliative and non-affiliative intraspecific interactions. Given the close relationship between the physical form and social function of mammalian facial signals, we predicted that affiliative and non-affiliative facial signals would have noticeable differences in their physical morphology. We observed the behavior of 53 adult domestic shorthair cats at CatCafé Lounge in Los Angeles, CA. Using Facial Action Coding Systems designed for cats, we compared the complexity and compositionality of facial signals produced in affiliative and non-affiliative contexts. To measure complexity and compositionality, we examined the number and types of facial muscle movements (AUs) observed in each signal. We found that compositionality, rather than complexity, was significantly associated with the social function of intraspecific facial signals. Our findings indicate that domestication likely had a significant impact on the development of intraspecific facial signaling repertoires in cats.


Relationship between Connective Tissue Morphology and Lower-Limb Stiffness in Endurance Runners. A Prospective Study.
The lower limb behaves like a spring compressing and decompressing during running, where lower-limb stiffness is one of the most influential factors. This prospective observational study is aimed at examining the relationship between the connective tissue morphology and lower-limb stiffness and investigating whether the barefoot/shod condition influences on such relationship. 14 male amateur runners (10-km time trial <50') were included. Data were recorded over one session, where participants ran 2 trials (i.e., barefoot and shod conditions) of 3 minutes at 12 km/h, where running spatiotemporal parameters and vertical (Kvert) and leg stiffness (Kleg) were obtained. Prior to testing trials, thickness and cross-sectional area (CSA) were recorded for Achilles (AT) and patellar tendons (PT) and plantar fascia (PF) with ultrasound. Under barefoot condition, a positive correlation was found between Kleg and AT-thickness and CSA and PF-thickness; and between Kvert and AT-thickness and PF thickness. Under shod condition, a positive correlation was found between Kleg and PT-CSA and PT-thickness, and between Kvert and PT-CSA and PT-thickness. The results reveal a specificity of the relationship between the lower-limb stiffness and the morphology of the connective tissue. Greater tendon shows higher lower-limb stiffness when that tendon is specially demanded by the function.


Conjunctivorhinostomy without osteal perforation.
The present techniques of conjunctivorhinostomy require perforation of the frontal process of the maxillary bone to open a passageway between the conjunctival sac and the nasal fossa. A relatively complex operation is required, and an almost horizontal conduit is left, whose flow is rarely good. A new technique of conjunctivorhinostomy obviates the necessity for osseous perforation by passing a tube beneath the soft tissues of the face just superficial to the maxillary bone, between the lacrimal lake and the nasal atrium. The external diameter of the tube should not exceed 2 mm to avoid producing a visible elevation on the surface of the face. The almost vertical position of the tube and the placement of its inferior opening in the zone of maximal respiratory flow ensure good drainage of tears. The operation can be performed in a few minutes, with local anesthesia, in the surgeon's office.


An analysis of movement and discomfort of the female breast during exercise and the effects of breast support in three cases.
Breast pain is common during exercise, occurring in up to 56% of subjects in some surveys. This pain is mainly associated with the movement of breast tissue. In an attempt to analyse this movement and the resulting pain, the movement of the female breast tissue was quantified in four conditions of breast support ("sports bra", "fashion bra", "crop top" and bare breasted) during four different activities (running, jogging, aerobics march and walking). These activities represented general patterns of exercise for adult females in Australian society. All three subjects were healthy, active, young women of varying breast size (12B, 14B & 14C) that were typical of young women. Two of the subjects had noted breast pain whilst exercising. The results showed that wearing external support for the breast tissue reduced absolute vertical movement and maximum downward deceleration force on the breast. Support also reduced perceived pain. When compared to other forms of breast support a "sports bra" (brassiere designed for breast support whilst exercising) provided superior support for the breast in relation to the amplitude of movement, deceleration forces on the breast, and perceived pain. The data indicates that adult females should wear appropriate breast support to reduce perceived breast discomfort or pain. Of the three garments examined in this study, the fitted sports bra provided superior support and pain reduction.


Atorvastatin reduces proteinuria in non-diabetic chronic kidney disease patients partly via lowering serum levels of advanced glycation end products (AGEs).
There is accumulating evidence that advanced glycation end products (AGEs) play a role in the development and progression of chronic kidney disease (CKD). We have previously found that atorvastatin treatment significantly reduces serum levels of AGEs in type 2 diabetic patients and subjects with non-alcoholic steatohepatitis in a cholesterol lowering-independent manner. In this study, we examined whether atorvastatin could reduce proteinuria partly via reduction of serum levels of AGEs in non-diabetic CKD patients. Ten non-diabetic normotensive stage I or II CKD patients with dyslipidemia were enrolled. Patients were treated with atorvastatin (10 mg/day) for 1 year. All subjects underwent determination of blood chemistries, proteinuria and serum levels of AGEs at baseline and after 1 year. Atorvastatin treatment for 1 year significantly decreased circulating levels of total cholesterol, LDL-cholesterol, triglycerides, and AGEs, while it increased HDL-cholesterol levels. Further, although atorvastatin treatment did not affect estimated glomerular filtration rate, it significantly reduced proteinuria. In univariate analyses, proteinuria levels were correlated with total cholesterol, LDL-cholesterol, triglycerides, HDL-cholesterol (inversely) and AGEs. Multiple stepwise regression analysis revealed that AGE level was a sole independent correlate of proteinuria. In this initial examination of the patients in this study, our present study suggests that atorvastatin could decrease proteinuria in non-diabetic CKD patients with dyslipidemia partly via reduction of serum levels of AGEs. Atorvastatin may have AGE-lowering effects in CKD patients as well that could contribute to renoprotective properties of this agent.


Deciphering the Allosteric Process of the Phaeodactylum tricornutum Aureochrome 1a LOV Domain.
The conformational-driven allosteric protein diatom Phaeodactylum tricornutum aureochrome 1a (PtAu1a) differs from other light-oxygen-voltage (LOV) proteins for its uncommon structural topology. The mechanism of signaling transduction in the PtAu1a LOV domain (AuLOV) including flanking helices remains unclear because of this dissimilarity, which hinders the study of PtAu1a as an optogenetic tool. To clarify this mechanism, we employed a combination of tree-based machine learning models, Markov state models, machine-learning-based community analysis, and transition path theory to quantitatively analyze the allosteric process. Our results are in good agreement with the reported experimental findings and reveal a previously overlooked Cα helix and protein linkers as important in promoting the protein conformational changes. This integrated approach can be considered as a general workflow and applied on other allosteric proteins to provide detailed information about their allosteric mechanisms.


Identification of Species by Combining Molecular and Morphological Data Using Convolutional Neural Networks.
Integrative taxonomy is central to modern taxonomy and systematic biology, including behavior, niche preference, distribution, morphological analysis, and DNA barcoding. However, decades of use demonstrate that these methods can face challenges when used in isolation, for instance, potential misidentifications due to phenotypic plasticity for morphological methods, and incorrect identifications because of introgression, incomplete lineage sorting, and horizontal gene transfer for DNA barcoding. Although researchers have advocated the use of integrative taxonomy, few detailed algorithms have been proposed. Here, we develop a convolutional neural network method (morphology-molecule network [MMNet]) that integrates morphological and molecular data for species identification. The newly proposed method (MMNet) worked better than four currently available alternative methods when tested with 10 independent data sets representing varying genetic diversity from different taxa. High accuracies were achieved for all groups, including beetles (98.1% of 123 species), butterflies (98.8% of 24 species), fishes (96.3% of 214 species), and moths (96.4% of 150 total species). Further, MMNet demonstrated a high degree of accuracy ($>$98%) in four data sets including closely related species from the same genus. The average accuracy of two modest subgenomic (single nucleotide polymorphism) data sets, comprising eight putative subspecies respectively, is 90%. Additional tests show that the success rate of species identification under this method most strongly depends on the amount of training data, and is robust to sequence length and image size. Analyses on the contribution of different data types (image vs. gene) indicate that both morphological and genetic data are important to the model, and that genetic data contribute slightly more. The approaches developed here serve as a foundation for the future integration of multimodal information for integrative taxonomy, such as image, audio, video, 3D scanning, and biosensor data, to characterize organisms more comprehensively as a basis for improved investigation, monitoring, and conservation of biodiversity. [Convolutional neural network; deep learning; integrative taxonomy; single nucleotide polymorphism; species identification.].


Expression of the voltage-gated chloride channel ClC-2 in rod bipolar cells of the rat retina.
Voltage-gated chloride channels (ClC) are highly conserved during evolution and appear to participate in a variety of physiological functions. Recently, ClC-2 was proposed to play a role in stabilizing the chloride equilibrium potential near or below the resting membrane potential in neurons expressing ligand-gated chloride channels. Because rod bipolar cells in mammalian retina express three forms of inhibitory ligand-gated chloride channels, we decided to study ClC-2 localization and function in the rat retina. RNA encoding ClC-1, -2, -3, -4, and -5 was detected by reverse transcription-PCR in the rat retina. ClC-2-specific antibodies identified protein on cell bodies and in synaptic layers. Double-immunofluorescence staining revealed that intense ClC-2 immunoreactivity colocalized with PKC-stained rod bipolar cells. Patch-clamp experiments performed with individual rod bipolar cells demonstrated the presence of a time-dependent, inwardly rectified current activated at hyperpolarizing membrane potentials. This current demonstrated selectivity for different anions (Cl(-) > I(-) > gluconate), was inhibited by Cd(2+), and was minimally reduced by 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid. These features are consistent with currents generated by ClC-2 channels. Our data indicate that functional ClC-2 channels are present in retinal rod bipolar cells and support a role for ClC-2 in maintaining Cl(-) homeostasis in neurons with ligand-gated chloride channels.


Functional connectivity of amygdala subnuclei in PTSD: a narrative review.
While the amygdala is often implicated in the neurobiology of posttraumatic stress disorder (PTSD), the pattern of results remains mixed. One reason for this may be the heterogeneity of amygdala subnuclei and their functional connections. This review used PRISMA guidelines to synthesize research exploring the functional connectivity of three primary amygdala subnuclei, basolateral (BLA), centromedial (CMA), and superficial nuclei (SFA), in PTSD (N = 331) relative to trauma-exposed (N = 155) and non-trauma-exposed controls (N = 210). Although studies were limited (N = 11), preliminary evidence suggests that in PTSD compared to trauma-exposed controls, the BLA shows greater connectivity with the dorsal anterior cingulate, an area involved in salience detection. In PTSD compared to non-trauma-exposed controls, the BLA shows greater connectivity with the middle frontal gyrus, an area involved in attention. No other connections were replicated across studies. A secondary aim of this review was to outline the limitations of this field to better shape future research. Importantly, the results from this review indicate the need to consider potential mediators of amygdala subnuclei connectivity, such as trauma type and sex, when conducting such studies. They also highlight the need to be aware of the limited inferences we can make with such small samples that investigate small subcortical structures on low field strength magnetic resonance imaging scanners. Collectively, this review demonstrates the importance of exploring the differential connectivity of amygdala subnuclei to understand the pathophysiology of PTSD and stresses the need for future research to harness the strength of ultra-high field imaging to gain a more sensitive picture of the neural connectivity underlying PTSD.