Dietary glycaemic index and cognitive function: prospective associations in adults of the 1946 British birth cohort.
Evidence suggests that the rate of glucose release following consumption of carbohydrate-containing foods, defined as the glycaemic index (GI), is inversely associated with cognitive function. To date, most of the evidence stems from either single-meal studies or highly heterogeneous cohort studies. We aimed to study the prospective associations of diet GI at age 53 years with outcomes of verbal memory and letter search tests at age 69 years and rate of decline between 53 and 69 years. Longitudinal population-based birth cohort study. MRC National Survey for Health and Development. Cohort members (n 1252). Using multivariable linear and logistic regression, adjusted for potential confounders, associations of higher-GI diet with lower verbal memory, lower letter search speed and lower number of hits in a letter search test were attenuated after adjustments for cognitive ability at age 15 years, educational attainment, further training and occupational social class. No association was observed between diet GI at 53 years and letter search accuracy or speed-accuracy trade-off at 69 years, or between diet GI at 53 years and rate of decline between 53 and 69 years in any cognitive measure. Diet GI does not appear to predict cognitive function or decline, which was mainly explained by childhood cognitive ability, education and occupational social class. Our findings confirm the need for further research on the association between diet and cognition from a life-course perspective.


The minute brain of the copepod Tigriopus californicus supports a complex ancestral ground pattern of the tetraconate cerebral nervous systems.
Copepods are a diverse and ecologically crucial group of minute crustaceans that are relatively neglected in terms of studies on nervous system organization. Recently, morphological neural characters have helped clarify evolutionary relationships within Arthropoda, particularly among Tetraconata (i.e., crustaceans and hexapods), and indicate that copepods occupy an important phylogenetic position relating to both Malacostraca and Hexapoda. This taxon therefore provides the opportunity to evaluate those neural characters common to these two clades likely to be results of shared ancestry (homology) versus convergence (homoplasy). Here we present an anatomical characterization of the brain and central nervous system of the well-studied harpacticoid copepod species Tigriopus californicus. We show that this species is endowed with a complex brain possessing a central complex comprising a protocerebral bridge and central body. Deutocerebral glomeruli are supplied by the antennular nerves, and a lateral protocerebral olfactory neuropil corresponds to the malacostracan hemiellipsoid body. Glomeruli contain synaptic specializations comparable to the presynaptic "T-bars" typical of dipterous insects, including Drosophila melanogaster. Serotonin-like immunoreactivity pervades the brain and ventral nervous system, with distinctive deutocerebral distributions. The present observations suggest that a suite of morphological characters typifying the Tigriopus brain reflect a ground pattern organization of an ancestral Tetraconata, which possessed an elaborate and structurally differentiated nervous system.


Metacognition and cognition in inpatient MCT and CBT for comorbid anxiety disorders: A study of within-person effects.
Psychotherapists have long questioned what mediating processes are linked to outcome of psychotherapy. Few studies examining this question have assessed within-person changes in the process outcome relationship over time. The present study examined changes in cognition and metacognition over the course of therapy using a dataset from a randomized controlled trial comparing Metacognitive therapy (MCT) and Cognitive-behavioral therapy (CBT). The sample included 74 patients measured on process and symptom instruments weekly throughout therapy. Multilevel longitudinal models (sessions nested within patients) were used to examine the relationship between metacognition, cognition, and anxiety. Main effects of metacognition and cognition on anxiety and the interaction with treatment, as well as the reciprocal relationships, were investigated. The results indicate a main effect of both cognitions and metacognitions on predicting anxiety. However, there was no interaction with treatment condition. The reciprocal relationship of anxiety on metacognitions was larger in MCT compared with CBT. This is the first study documenting within-person effects of both cognitions and metacognitions on anxiety over the course of therapy. Implications for therapy are discussed. (PsycINFO Database Record


Chronic pancreatitis.
Summarize key clinical advances in chronic pancreatitis reported in 2016. Early diagnosis of chronic pancreatitis remains elusive. Recent studies suggest that endoscopic ultrasound may be less accurate than previously thought and new MRI techniques may be helpful. Genetic predisposition may independently affect the clinical course of chronic pancreatitis and the risk for pancreatic cancer. Cigarette smoking may have a greater negative impact on chronic pancreatitis than previously thought and moderate alcohol consumption may be protective. A multidisciplinary approach is necessary for the treatment of type 3 diabetes and nutritional deficiencies in chronic pancreatitis. Although endoscopic therapy remains a reasonable first-line option in treating chronic pancreatitis and its complications, early surgical intervention may be indicated for pain in select patients. Newer endoscopic ultrasound and MRI techniques are being evaluated to help with the early diagnosis of chronic pancreatitis. Both genetic predisposition and cigarette smoking are increasingly recognized as having a major impact in the course of the disease and the risk for pancreatic cancer. Endoscopic therapy is well tolerated and effective for the treatment of chronic pancreatitis and its complications although an early surgical approach for pain may be associated with improved clinical outcomes.


Gene expression signatures and small-molecule compounds link a protein kinase to Plasmodium falciparum motility.
Calcium-dependent protein kinases play a crucial role in intracellular calcium signaling in plants, some algae and protozoa. In Plasmodium falciparum, calcium-dependent protein kinase 1 (PfCDPK1) is expressed during schizogony in the erythrocytic stage as well as in the sporozoite stage. It is coexpressed with genes that encode the parasite motor complex, a cellular component required for parasite invasion of host cells, parasite motility and potentially cytokinesis. A targeted gene-disruption approach demonstrated that pfcdpk1 seems to be essential for parasite viability. An in vitro biochemical screen using recombinant PfCDPK1 against a library of 20,000 compounds resulted in the identification of a series of structurally related 2,6,9-trisubstituted purines. Compound treatment caused sudden developmental arrest at the late schizont stage in P. falciparum and a large reduction in intracellular parasites in Toxoplasma gondii, which suggests a possible role for PfCDPK1 in regulation of parasite motility during egress and invasion.


Assessment of Residual Pain and Dissatisfaction in Total Knee Arthroplasty: Methods Matter.
Residual pain after total knee arthroplasty (TKA) refers to knee pain after 3 to 6 months postoperatively. The estimates of the proportion of patients who experience residual pain after TKA vary widely. We hypothesized that the variation may stem from the range of methods used to assess residual pain. We analyzed data from 2 prospective studies to assess the proportion of subjects with residual pain as defined by several commonly used metrics and to examine the association of residual pain defined by each metric with participant dissatisfaction. We combined participant data from 2 prospective studies of TKA outcomes from subjects recruited between 2011 and 2014. Residual pain was defined using a range of metrics based on the WOMAC (Western Ontario and McMaster Universities Osteoarthritis Index) pain score (0 to 100, in which 100 indicates worst), including the minimal clinically important difference (MCID) and patient acceptable symptom state (PASS). We also examined combinations of MCID and PASS cutoffs. Subjects self-reported dissatisfaction following TKA, and we defined dissatisfied as somewhat or very dissatisfied at 12 months. We calculated the proportion of participants with residual pain, as defined by each metric, who reported dissatisfaction. We examined the association of each metric with dissatisfaction by calculating the sensitivity, specificity, positive predictive value, and Youden index. We analyzed data from 417 subjects with a mean age (and standard deviation) of 66.3 ± 8.3 years. Twenty-six participants (6.2%) were dissatisfied. The proportion of participants defined as having residual pain according to the various metrics ranged from 5.5% to >50%. The composite metric Improvement in WOMAC pain score ≥20 points or final WOMAC pain score ≤25 had the highest positive predictive value for identifying dissatisfied subjects (0.54 [95% confidence interval, 0.35 to 0.71]). No metric had a Youden index of ≥50%. Different metrics provided a wide range of estimates of residual pain following TKA. No estimate was both sensitive and specific for dissatisfaction in patients who underwent TKA, underscoring that measures of residual pain should be defined explicitly in reports of TKA outcomes. Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.


Laparoscopic cholecystectomy in a cardiac transplant candidate with an ejection fraction of less than 15%.
Laparoscopic procedures are becoming increasingly popular, even in the severely ill patient. We present a case of a cardiac transplant candidate with an ejection fraction of less than 15% who underwent a laparoscopic cholecystectomy. This is the first case in which intraoperative hemodynamic measurements were recorded in a patient with cardiopulmonary disease this severe. The patient underwent the procedure while measurements were made at crucial intervals (baseline, with incremental insufflation, reverse Trendelenberg, at desufflation) using a pulmonary artery catheter, transesophageal echocardiography with fractional area measurements, radial arterial line, as well as standard monitoring. This patient showed hemodynamic changes consistent with a person without severe heart disease. There was a rise in mean arterial and systemic vascular resistance with insufflation to 10 mm Hg, which was further exaggerated by reverse Trendelenberg. Heart rate and pulmonary artery wedge pressure increased slightly. The cardiac output and fractional area change declined minimally. We conclude that if a patient with congestive heart failure is medically optimized, and intra-abdominal pressures and surgical times are minimized, laparoscopic cholecystectomy may be performed with minimal risk to the patient.


Amyloid precursor protein in human breast cancer: an androgen-induced gene associated with cell proliferation.
Amyloid precursor protein (APP) is a transmembrane protein that is highly expressed in brain tissue. Recently, APP has been implicated in some human malignancies, and its regulation by androgens has also been demonstrated. Such findings suggest the importance of APP in hormone-dependent breast carcinoma, but APP has not yet been examined in breast carcinoma tissues. Therefore, in this study, we examined the biological and clinical significance of APP in breast carcinoma using immunohistochemistry and in vitro studies. APP immunoreactivity was detected in 57 out of 117 (49%) breast carcinoma tissues examined, and it was positively associated with androgen receptor (AR) expression. APP immunoreactivity was also significantly associated with Ki-67 LI and increased risk of recurrence in the estrogen receptor (ER)-positive cases, and was an independent prognostic factor in these patients. Subsequent in vitro experiments demonstrated that APP mRNA expression was significantly induced by biologically active androgen dihydrotestosterone in both a dose-dependent and a time-dependent manner in MCF-7 breast carcinoma cells, which was potently suppressed by an AR blocker hydroxyflutamide. Moreover, cell proliferation activity of MCF-7 and MDA-MB-231 cells was significantly associated with their APP expression level. These findings suggest that APP is an androgen-induced gene that promotes proliferation activity of breast carcinoma cells. Moreover, APP immunohistochemical status is considered a potent prognostic factor in ER-positive breast cancer patients.


Unrecognized sexually transmitted infections in rural South African women: a hidden epidemic.
Sexually transmitted infections (STIs) are of major public health concern in developing countries, not least because they facilitate transmission of human immunodeficiency virus (HIV). The present article presents estimates of the prevalence, on any given day, of STIs among women in rural South Africa and the proportion who are asymptomatic, symptomatic but not seeking care, and symptomatic and seeking care. The following data sources from Hlabisa district were used: clinical surveillance for STI syndromes treated in health facilities, microbiological studies among women attending antenatal and family planning clinics, and a community survey. Population census provided denominator data. Adequacy of drug treatment was determined through quality of care surveys. Of 55,974 women aged 15-49 years, a total of 13,943 (24.9%) were infected on any given day with at least one of Trichomonas vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, or Treponema pallidum. Of the women investigated, 6697 (48%) were asymptomatic, 6994 (50%) were symptomatic but not seeking care, 238 (1.7%) were symptomatic and would seek care, and 14 (0.3%) were seeking care on that day. Only 9 of the 14 women (65%) were adequately treated. STIs remained untreated because either women were asymptomatic or the symptoms were not recognized and acted upon. Improved case management alone is therefore unlikely to have a major public health impact. Improving partner treatment and women's awareness of symptoms is essential, while the potential of mass STI treatment needs to be explored.


Hollow MnCo2O4 submicrospheres with multilevel interiors: from mesoporous spheres to yolk-in-double-shell structures.
We present a general strategy to synthesize uniform MnCo2O4 submicrospheres with various hollow structures. By using MnCo-glycolate submicrospheres as the precursor with proper manipulation of ramping rates during the heating process, we have fabricated hollow MnCo2O4 submicrospheres with multilevel interiors, including mesoporous spheres, hollow spheres, yolk-shell spheres, shell-in-shell spheres, and yolk-in-double-shell spheres. Interestingly, when tested as anode materials in lithium ion batteries, the MnCo2O4 submicrospheres with a yolk-shell structure showed the best performance among these multilevel interior structures because these structures can not only supply a high contact area but also maintain a stable structure.


Impact of hormone replacement therapy on exercise training-induced improvements in insulin action in sedentary overweight adults.
Exercise training (ET) and hormone replacement therapy (HRT) are both recognized influences on insulin action, but the influence of HRT on responses to ET has not been examined. To determine if HRT use provided additive benefits for the response of insulin action to ET, we evaluated the impact of HRT use on changes in insulin during the course of a randomized, controlled, aerobic ET intervention. Subjects at baseline were sedentary, dyslipidemic, and overweight. These individuals were randomized to 6 months of one of 3 aerobic ET interventions or continued physical inactivity. In 206 subjects, an insulin sensitivity index (S(I)) was obtained with a frequently sampled intravenous glucose tolerance test pre- and post-ET. Baseline and postintervention fitness, regional adiposity, general adiposity, skeletal muscle biochemistry and histology, and serum lipoproteins were measured as other putative mediators influencing insulin action. Two-way analyses of variance were used to determine if sex or HRT use influenced responses to exercise training. Linear modeling was used to determine if predictors for response in S(I) differed by sex or HRT use(.) Women who used HRT (HRT+) demonstrated significantly greater improvements in S(I) with ET than women not using HRT (HRT-). In those HRT+ women, plasma triglyceride change best correlated with change in S(I). For HRT- women, capillary density change and, for men, subcutaneous adiposity change best correlated with change in S(I). In summary, in an ET intervention, HRT use appears to be associated with more robust responses in insulin action. Furthermore, relationships between ET-induced changes in insulin action and potential mediators of change in insulin action are different for men, and for women on or off HRT. These findings have implications for the relative utility of ET for improving insulin action in middle-aged men and women, particularly in the setting of differences in HRT use.


Sensory and nutritional quality of split peas (Pisum sativum) stored up to 34 y in residential storage.
The sensory and nutritional quality of split peas stored up to 34 y was determined. Nine samples of split peas representing 5 retail brands packaged in Nr 10 cans and stored at room temperature were obtained from donors. Duplicate cans of a fresh sample of split peas were purchased as controls. Can head space oxygen ranged from 0.255% to 20.1%. Water activity of the raw split peas ranged from 0.41 to 0.56. The green color of the raw split peas decreased over time as shown by increasing CIE a* values. Flavor, appearance, texture, and overall liking hedonic scores (9-point scale) of split-pea soup made from each sample ranged from 3.7 to 6.7 and decreased over time. Hedonic scores for appearance were correlated with the decrease in raw product green color (r(2)= 0.65). Hedonic scores for soup texture declined over time, which corresponded with increasing hardness of the cooked peas as measured by a TA.XT2 texture analyzer. All samples were judged to be acceptable in an emergency situation by over 75% of sensory panelists. Available thiamin was significantly lower in older samples while riboflavin remained unchanged. The results indicate that split pea quality declines over time, but the product maintains sufficient sensory acceptance to be considered for potential use in emergency storage and other applications where minimal stock rotation is a common practice.


A singlicate immunogenicity method to detect anti-polyethylene glycol antibodies: pre- and post-dose of PEGylated therapies.
Aim: Preexisting anti-polyethylene glycol (PEG) antibodies (APAs) may affect the efficacy and safety of PEGylated compounds. Omontys® and Krystexxa® withdrawal and SARS-CoV-2 RNA vaccine anaphylaxis have all been linked to APAs. This project aimed to develop and validate a method to detect total antibodies against PEG, pre- and post-dose. Materials & methods: The repetitive, linear PEG structure prevented the use of a bridging homogenous format, hence the requirement to use a solid-phase extraction and acid dissociation assay coupled with the Meso Scale Discovery® platform. Results & conclusion: Using singlicate analysis, the method was validated to successfully detect APA pre- and post-dose, with a crucial aspect of the method being the preparation of an appropriate negative control.


Immobility in the sedentary plant-parasitic nematode H. glycines is associated with remodeling of neuromuscular tissue.
The sedentary plant-parasitic nematodes are considered among the most economically damaging pathogens of plants. Following infection and the establishment of a feeding site, sedentary nematodes become immobile. Loss of mobility is reversed in adult males while females never regain mobility. The structural basis for this change in mobility is unknown. We used a combination of light and transmission electron microscopy to demonstrate cell-specific muscle atrophy and sex-specific renewal of neuromuscular tissue in the sedentary nematode Heterodera glycines. We found that both females and males undergo body wall muscle atrophy and loss of attachment to the underlying cuticle during immobile developmental stages. Male H. glycines undergo somatic muscle renewal prior to molting into a mobile adult. In addition, we found developmental changes to the organization and number of motor neurons in the ventral nerve cord correlated with changes in mobility. To further examine neuronal changes associated with immobility, we used a combination of immunohistochemistry and molecular biology to characterize the GABAergic nervous system of H. glycines during mobile and immobile stages. We cloned and confirmed the function of the putative H. glycines GABA synthesis-encoding gene hg-unc-25 using heterologous rescue in C. elegans. We found a reduction in gene expression of hg-unc-25 as well as a reduction in the number of GABA-immunoreactive neurons during immobile developmental stages. Finally, we found evidence of similar muscle atrophy in the phylogenetically diverged plant-parasitic nematode Meloidogyne incognita. Together, our data demonstrate remodeling of neuromuscular structure and function during sedentary plant-parasitic nematode development.


Surveillance and diagnosis of hepatocellular carcinoma in patients with cirrhosis.
Early identification of hepatocellular carcinoma (HCC) is more frequent because of surveillance programs for HCC worldwide. The optimal strategy of surveillance in cirrhosis is a current topical issue. In terms of diagnosis, recent advances in non-invasive imaging technology, including various techniques of harmonic ultrasound, new ultrasound contrast agents, multi-slice helical computed tomography and rapid high quality magnetic resonance, have all improved the accuracy of diagnosis. Consequently the role of liver biopsy in diagnosis of HCC has declined. The imaging diagnosis relies on the hallmark of arterial hypervascularity with portal venous washout. However, with recent advances in genomics and proteomics a great number of potential serum and tissue markers have been identified and are being developed as new candidate markers for both diagnosis and prognosis of hepatocellular carcinoma, and may increase the need for liver biopsy.


Effect of treatment delay on survival in patients with cervical cancer: a historical cohort study.
The objective of this study was to evaluate the effect of treatment delay on prognosis in patients with cervical cancer. The study group of this historic cohort study comprised 321 patients newly diagnosed with cervical cancer between 1999 and 2010. Time from diagnosis to treatment was analyzed both as a continuous variable and as a categorical variable in 3 groups that differed in waiting time between diagnosis and treatment initiation: 30 days or less (group 1, n = 134), 30 to 45 days (group 2, n = 86), and more than 45 days (group 3, n = 101). Associations between waiting time group, patients' characteristics, and disease outcome were investigated using t tests, analyses of variance and Cox regression analyses, Kaplan-Meier survival analysis, and log-rank (Mantel-Cox) tests. Time from diagnosis to treatment initiation, when analyzed as a continuous variable, was not a significant factor in survival. There were no between-group differences in age, smoking rate, marital status, gravidity, parity, tumor histology, or lymph node involvement. Early-stage disease and small tumor diameter were diagnosed most frequently in group 3. However, there was no significant between-group difference in 3-year survival rates (74.6%, 82.2%, and 80.8% in groups 1, 2, and 3, respectively; P = 0.38). On multivariate analysis, only stage, histology, and lymph node involvement were significant prognostic factors for survival. Before starting treatment, 28 patients underwent ovarian preservation procedures. Longer waiting time from diagnosis to treatment was not associated with worse survival. Our findings imply that if patients desire fertility or ovarian preservation procedures before starting treatment, it is acceptable to allow time for them.


A comparison of statistical approaches used for the optimization of soluble protein expression in Escherichia coli.
During a discovery project of potential inhibitors for three proteins, TNF-α, RANKL and HO-1, implicated in the pathogenesis of rheumatoid arthritis, significant amounts of purified proteins were required. The application of statistically designed experiments for screening and optimization of induction conditions allows rapid identification of the important factors and interactions between them. We have previously used response surface methodology (RSM) for the optimization of soluble expression of TNF-α and RANKL. In this work, we initially applied RSM for the optimization of recombinant HO-1 and a 91% increase of protein production was achieved. Subsequently, we slightly modified a published incomplete factorial approach (called IF1) in order to evaluate the effect of three expression variables (bacterial strains, induction temperatures and culture media) on soluble expression levels of the three tested proteins. However, soluble expression yields of TNF-α and RANKL obtained by the IF1 method were significantly lower (<50%) than those obtained by RSM. We further modified the IF1 approach by replacing the culture media with induction times and the resulted method called IF-STT (Incomplete Factorial-Stain/Temperature/Time) was validated using the three proteins. Interestingly, soluble expression levels of the three proteins obtained by IF-STT were only 1.2-fold lower than those obtained by RSM. Although RSM is probably the best approach for optimization of biological processes, the IF-STT is faster, it examines the most important factors (bacterial strain, temperature and time) influencing protein soluble expression in a single experiment, and can be used in any recombinant protein expression project as a starting point.


Analysis of social determinants of health and individual factors found in health equity frameworks: Applications to injury research.
This research evaluated existing health equity frameworks as they relate to social determinants of health (SDOHs) and individual factors that may impact injury outcomes and identify gaps in coverage using the Healthy People (HP) 2030 key domains. The study used a list of health equity frameworks sourced from previous literature. SDOHs and individual factors from each framework were identified and categorized into the Healthy People 2030 domains. Five injury topic areas were used as examples for how SDOHs and individual factors can be compared to injury topic-specific health disparities to identify health equity frameworks to apply to injury research. The study identified 59 SDOHs and individual factors from the list of 33 health equity frameworks. The number of SDOHs and individual factors identified varied by Healthy People 2030 domain: Neighborhood and Built Environment contained 16 (27.1%) SDOHs and individual actors, Social and Community Context contained 22 (37.3%), Economic Stability contained 10 (16.9%), Healthcare Access and Quality contained 10 (16.9%), and Education Access and Quality contained one (1.7%). Twenty-three (39.0%) SDOHs/individual factors related to traumatic brain injury, thirteen (22.0%) related to motor vehicle crashes and suicide, 11 (18.6%) related to drowning and older adult falls. Eight frameworks (24.2%) covered all HP 2030 key domains and may be applicable to injury topics. Incorporating health equity into research is critical. Health equity frameworks can provide a way to systematically incorporate health equity into research. The findings from this study may be useful to health equity research by providing a resource to injury and other public health fields. Health equity frameworks are a practical tool to guide injury research, translation, evaluation, and program implementation. The findings from this study can be used to guide the application of health equity frameworks in injury research for specific topic areas.


Variability in Perioperative Steroid Therapy Regimen for Cochlear Implantation as It Relates to Hearing Preservation.
We aimed to identify practice trends and association between physician training and administration of perioperative steroids for cochlear implantation (CI) as it relates to hearing preservation. Perioperative steroid therapy regimens are postulated to protect residual hearing and improve hearing preservation outcomes in CI. A 27-question online survey was developed by the senior authors using the Qualtrics Survey Tool, then distributed via email from September to November 2022 to otolaryngologists specializing in otology or neurotology and who practice in the United States or Canada. The survey was sent to 463 physicians, 162 (35.0%) of whom completed the survey. One hundred forty-four (31.1%) responses underwent analysis. All physicians administering preoperative steroids (n = 31) prefer preoperative oral prednisone. Of 143 physicians administering intraoperative steroids, 54.5% prefer intraoperative intravenous dexamethasone. More than half (77.6%) of 85 physicians administering postoperative steroids prefer postoperative oral prednisone. Postoperative steroid administration (p < 0.006) and taper utilization (p < 0.041) were greater among physicians who complete greater than 40 CIs annually (n = 47 [71.2%]; n = 30 [49.2%]) than physicians who complete up to 40 CIs annually (n = 37 [48.7%]; n = 20 [31.3%]), respectively. Physicians practicing for 5 to 20 years after residency are more prevalent in using postoperative steroid tapers than physicians practicing for fewer than 5 years after and more than 20 years after residency (n = 37 [51.4%] versus n = 14 [25.5%], p < 0.001). Consensus is needed about the optimal steroid treatment for CI patients. 4.


FUBP1: a new protagonist in splicing regulation of the DMD gene.
We investigated the molecular mechanisms for in-frame skipping of DMD exon 39 caused by the nonsense c.5480T>A mutation in a patient with Becker muscular dystrophy. RNase-assisted pull down assay coupled with mass spectrometry revealed that the mutant RNA probe specifically recruits hnRNPA1, hnRNPA2/B1 and DAZAP1. Functional studies in a human myoblast cell line transfected with DMD minigenes confirmed the splicing inhibitory activity of hnRNPA1 and hnRNPA2/B1, and showed that DAZAP1, also known to activate splicing, acts negatively in the context of the mutated exon 39. Furthermore, we uncovered that recognition of endogenous DMD exon 39 in muscle cells is promoted by FUSE binding protein 1 (FUBP1), a multifunctional DNA- and RNA-binding protein whose role in splicing is largely unknown. By serial deletion and mutagenesis studies in minigenes, we delineated a functional intronic splicing enhancer (ISE) in intron 38. FUBP1 recruitment to the RNA sequence containing the ISE was established by RNA pull down and RNA EMSA, and further confirmed by RNA-ChIP on endogenous DMD pre-mRNA. This study provides new insights about the splicing regulation of DMD exon 39, highlighting the emerging role of FUBP1 in splicing and describing the first ISE for constitutive exon inclusion in the mature DMD transcript.


Water balance across the cuticle of a soil insect.
1. The water regime in soil commonly approaches equilibrium of water potential with the insects living there. 2. Even under these conditions, non-equilibrium processes have a significant effect on water movement through the cuticle of soil insects. 3. Measurements of water potential on either side of the cuticle of Costelytra zealandica larvae showed that equilibrium is not reached while the insect is alive. There is an active outward flow of water by thermoosmosis associated with the flow of heat from the insect.


An innovative program to train health sciences researchers to be effective clinical and translational research mentors.
The creation of the Clinical Translational Science Awards for academic health sciences campuses in 2006 was implicitly accompanied by a call for a new paradigm of faculty development and mentoring to train the next generation of researchers and leaders in this new approach to research. Effective mentoring is critical to help early-career investigators become successful, independent researchers, and a new approach to mentoring is vital to recruit, advance, and retain fellows and junior faculty engaged in clinical and translational research. However, in addition to the many rewards of mentoring, there are numerous substantive barriers to effective mentoring. These barriers include a lack of training in how to be a mentor, lack of time and structural and financial support for mentoring, and competing personal, administrative, and clinical demands. The authors describe an innovative program, the University of California, San Francisco Mentor Development Program (MDP), established in 2006 and designed to train midcareer academic health sciences researchers to be more effective as clinical and translational research mentors. Using a framework for presenting innovations in academic research, they present the rationale, design, implementation, and mechanisms being used to evaluate and sustain the MDP. Specific details of the objectives and content of the MDP sessions are provided as well as evaluation criteria and a link to specific curriculum materials.


Utilizing melatonin to combat bacterial infections and septic injury.
Melatonin, also known as N-acetyl-5-methoxytryptamine, is a ubiquitously acting molecule that is produced by the pineal gland and other organs of animals, including humans. As melatonin and its metabolites are potent antioxidants and free radical scavengers, they are protective against a variety of disorders. Moreover, multiple molecular targets of melatonin have been identified, and its actions are both receptor-mediated and receptor-independent. Recent studies have shown that melatonin may be useful in fighting against sepsis and septic injury due to its antioxidative and anti-inflammatory actions; the results generally indicate a promising therapeutic application for melatonin in the treatment of sepsis. To provide a comprehensive understanding regarding the protective effects of melatonin against septic injury, in the present review we have evaluated the published literature in which melatonin has been used to treat experimental and clinical sepsis. Firstly, we present the evidence from studies that have used melatonin to resist bacterial pathogens. Secondly, we illustrate the protective effect of melatonin against septic injury and discuss the possible mechanisms. Finally, the potential directions for future melatonin research against sepsis are summarized.


Development and characterization of microsatellite loci in dioecious figs (Ficus, Moraceae).
Microsatellite markers for dioecious fig species, Ficus bernaysii King (section Sycocarpus), Ficus pachyrrhachis K. Schum. & Lauterb. (section Sycocarpus), and Ficus copiosa Steud. (section Sycidium) were developed to estimate gene flow among closely related fig species. Loci were developed from tri- and tetranucleotide-enriched genomic libraries. The six most repeatable and scorable loci for F. bernaysii and F. pachyrrhachis were characterized in 50 individuals each of five species from section Sycocarpus. Four primer pairs from F. copiosa were screened in 50 individuals each of two species from section Sycidium. Observed heterozygosity ranged from 0.53 to 0.93 in loci developed from F. bernaysii, 0.25 to 0.86 in loci developed from F. pachyrrhachis, and 0.68 to 0.87 in loci developed from F. copiosa. Amplification across the two Ficus sections was observed in seven of the ten total primer pairs.


Structural requirements of ligands for the oxysterol liver X receptors LXRalpha and LXRbeta.
LXRalpha and -beta are nuclear receptors that regulate the metabolism of several important lipids, including cholesterol and bile acids. Previously, we have proposed that LXRs regulate these pathways through their interaction with specific, naturally occurring oxysterols, including 22(R)-hydroxycholesterol, 24(S)-hydroxycholesterol, and 24(S),25-epoxycholesterol. Using a ligand binding assay that incorporates scintillation proximity technology to circumvent many of the problems associated with assaying extremely hydrophobic ligands, we now demonstrate that these oxysterols bind directly to LXRs at concentrations that occur in vivo. To characterize further the structural determinants required for potent LXR ligands, we synthesized and tested a series of related compounds for binding to LXRs and activation of transcription. These studies revealed that position-specific monooxidation of the sterol side chain is requisite for LXR high-affinity binding and activation. Enhanced binding and activation can also be achieved through the use of 24-oxo ligands that act as hydrogen bond acceptors in the side chain. In addition, introduction of an oxygen on the sterol B-ring results in a ligand with LXRalpha-subtype selectivity. These results support the hypothesis that naturally occurring oxysterols are physiological ligands for LXRs and show that a rational, structure-based approach can be used to design potent LXR ligands for pharmacologic use.


Isolation and characterization of myosin heavy chain isozymes of the bovine conduction system.
To determine the characteristics of cardiac myosin in the conduction system, a pure Purkinje fiber preparation, consisting of atrioventricular nodes and the ventricular conduction system, was obtained from bovine hearts. Two types of myosin heavy chain isozymes, alpha-type and beta-type, were fractionated by affinity chromotography using monoclonal antibodies CMA19 and HMC50, which are specific for the alpha-type heavy chain and beta-type heavy chain, respectively. Competitive enzyme-linked immunosorbent assay demonstrated that the content of beta-type in the atrioventricular node (30-40%) was higher than that in atrial ordinary myocardium (10-20%) and that of the alpha-type was 30-40% in the ventricular conduction system, which was much higher than that in the ventricular ordinary myocardium (less than 10%). By one- and two-dimensional electrophoresis of the peptides produced by partial and complete digestion, the peptide compositions of alpha-type and beta-type in the conduction system were shown to be very similar to those of alpha-type and beta-type in ordinary myocardium, respectively. The CA2+-activated ATPase activity of myosin of the atrioventricular nodes was lower than that of ordinary atrial myosin (0.46 +/- 0.03 versus 0.58 +/- 0.02 mumol Pi/mg/min, mean +/- SEM, p less than 0.05) and in contrast, that of ventricular specialized myocardium was higher than that of myosin in the ventricular ordinary working myocardium (0.32 +/- 0.03 versus 0.22 +/- 0.01 mumol Pi/mg/min, p less than 0.05). This was in good agreement with the relative proportion of myosin isozymes.(ABSTRACT TRUNCATED AT 250 WORDS)


The effect of submaxillariectomy on the uterine peroxidase activity and [14C]phenylalanine incorporation into uterine protein in rats.
Surgical removal of submaxillary glands in immature female rats caused an increase in size and about 3-fold increase in dry and wet weight of the uterus compared to that of the sham operated animals of the same age group. Histological examination reveal a significant increase in the diameter of the uterus with considerable elongation of the luminal epithelium from cubical to columnar in the experimental group. Biochemical studies showed that the uterine peroxidase, a marker enzyme for oestrogen action, increased (P less than 0.01) on submaxillariectomy. Incorporation of [14C]phenylalanine into the nuclear fraction of uterus was also enhanced significantly on removal of submaxillary glands. The elevation of peroxidase activity as well as of [14C]phenylalanine incorporation into the nuclear fraction due to removal of submaxillary glands were abolished in ovariectomized rats suggesting the involvement of ovarian hormones. The results show that oestrogen was responsible for all the above mentioned changes, whereas progesterone had little effect. Results further suggest the existence of a factor in the submaxillary glands through which they exert an effect on the uterus and ovary.


Informing decisions with disparate stakeholders: cross-sector evaluation of cash transfers in Malawi.
The Social Cash Transfer Programme (SCTP) in Malawi is a cross-sectoral policy with impacts on health, education, nutrition, agriculture and welfare. Implementation of the SCTP requires collaboration across sectors and across national and international stakeholders. Economic evaluation can inform investment by indicating whether benefits exceed costs, but economic evaluations that provide an overall benefit-cost ratio typically assume a common agreed objective and agreed set of value judgements. In reality, the various stakeholders involved in the delivery of the SCTP may have different remits and objectives and may differ in how they value the impacts of the programme. We use the SCTP as a case study to illustrate a cross-sectoral analytical framework that accounts for these differences. The stakeholders that contribute to the SCTP include the Ministry of Gender, Ministry of Finance, Ministry of Economic Planning and Development and Global Fund. We estimate how the SCTP changes outcomes in education, health, net production and poverty, and distinguish outcomes in three groups: SCTP recipients; population in Malawi not eligible for the SCTP and population in other countries. After estimating the direct effects and opportunity costs from investing in the SCTP, we summarize the results according to different perspectives. The SCTP is estimated to provide benefits in excess of costs from the perspective of national stakeholders. From the perspective of an international donor interested in health outcomes, its health benefits do not outweigh the opportunity costs unless health improvement in SCTP recipients is valued at 18 times that of other potential spending beneficiaries or the donor values broader outcomes than health alone. This work illustrates the potential of a cross-sectoral economic evaluation to guide debate about stakeholder contributions to the SCTP, and the value judgements required to favour the SCTP above other policy options.


The level of socioeconomic development of EU countries and the state of ISO 14001 certification.
This study examines the relationship between the level of socioeconomic development of the EU 28 countries and the adoption of International Organization for Standardization (ISO) 14001 environmental management system. First, through a multivariate comparative analysis of the secondary data obtained from the public statistics, a Hellwig's synthetic indicator is created to rank the level of socioeconomic development of the EU 28 countries. Then, using the total number of certificates issued in 2012 and the increase from 2011, this study has found a correlation between the level of national socioeconomic development and the adoption of ISO 14001 system in their businesses. Although there was no relationship between the number of ISO 14001 certificates in 2012 and the level of socioeconomic development at the national level, a weak negative correlation was observed between the increase of certification from 2011 to 2012 and the level of national socioeconomic development. The results suggest a higher interest in ISO 14001 adoption by the firms from the less developed countries than those from the more developed states.


Evaluation of the brainstem with high-resolution CT in cerebellar atrophic processes.
The authors studied the usefulness of computed tomography (CT) for evaluation of the brainstem in cerebellar atrophic processes. Twenty adult subjects without posterior fossa lesion were used for normal CT measurements of the brainstem. The measured values with CT corresponded to those with pneumotomography. Also reviewed were 49 patients with cerebellar atrophy which included spinocerebellar degeneration (25 patients), Shy-Drager syndrome (five), progressive supranuclear palsy (three), chronic phenytoin usage (10), and chronic alcoholism (six). All but the chronic alcoholism group showed atrophy of the brainstem at all locations of measurement when compared with normal controls (p less than 0.05). In addition, the patients with progressive supranuclear palsy had significantly more pronounced midbrain atrophy. In the chronic alcoholism group the measurements of the brachium pontis, the medulla, and the fourth ventricle differed significantly from those of normal controls (p less than 0.05).


RU 58841-myristate--prodrug development for topical treatment of acne and androgenetic alopecia.
Acne and androgenetic alopecia are linked to androgen effects and therefore should improve following topical application of antiandrogens. We present a new antiandrogen prodrug, RU 58841-myristate (RUM) for topical therapy. Almost devoid of affinity to the androgen receptor, as derived from investigations in the mouse fibroblast cell line 29 +/GR +, RUM is rapidly metabolised to the potent antiandrogen RU 58841 by cultured human foreskin fibroblasts and keratinocytes, male occipital scalp skin dermal papilla cells, and by cells of the sebaceous gland cell line SZ95. In order to improve a specific targeting of the hair follicle, RUM was loaded on solid lipid nanoparticles (SLN), which are already known to support dermal targeting effects. Physically stable RUM loaded SLN were produced by hot homogenization. Penetration/permeation studies carried out using the Franz diffusion cell proved only negligible permeation of reconstructed epidermis and excised porcine skin within 6 h, implying a more topical action of the drug. Targeting to the hair follicle using SLN was visualised by fluorescence microscopy, following the application of Nile Red labelled SLN to human scalp skin. Transmission electron microscopy (TEM) allowed to detect intact silver labelled SLN in porcine hair follicles of preparations applied to the skin for 24 h. RUM loaded SLN should be considered for topical antiandrogen therapy of acne and androgenetic alopecia.


A case study of bovine tuberculosis in an area of County Donegal, Ireland.
A descriptive analysis, to investigate the potential risk factors that might have contributed to the increased incidence of bovine tuberculosis (BTB) herd-breakdowns in the reference area of Co. Donegal during the fifth year of the four-area project (FAP), was performed. Seventy two different herds were restricted for BTB during the FAP; 10 of these herds were restricted twice, resulting in a total of 82 BTB breakdowns. During the first four years of the FAP, the number of BTB herd breakdowns in the area varied from a lowest of nine to a maximum of 18 per year, and were geographically dispersed. In the fifth year of the study a considerable increase in the number of BTB breakdowns (n = 32) was observed, and there was a spatial 'cluster' of infected herds in the eastern part of the study area. The increased number of BTB breakdowns during the fifth year most likely occurred because of the recrudescence of infection, herd-to-herd transmission and, to a lesser extent, purchase of infected cattle. Infected badgers remain as a possible but less likely source of infection, especially as an explanation for the cluster of infected herds. The analysis supports the hypothesis that BTB in herds is a problem that cannot be addressed successfully by dedicating our efforts to the elimination of single risk factors. Neither is it a problem that needs to be investigated only at the herd level, but rather at the area level, including groups of contiguous herds.


Impairment of coronary endothelial cell ET(B) receptor function after short-term inhalation exposure to whole diesel emissions.
Air pollutant levels positively correlate with increases in both acute and chronic cardiovascular disease. The pollutant diesel exhaust (DE) increases endothelin (ET) levels, suggesting that this peptide may contribute to DE-induced cardiovascular disease. We hypothesized that acute exposure to DE also enhances ET-1-mediated coronary artery constrictor sensitivity. Constrictor responses to KCl, U-46619, and ET-1 were recorded by videomicroscopy in pressurized intraseptal coronary arteries from rats exposed for 5 h to DE (300 microg/m(3)) or filtered air (Air). ET-1 constriction was augmented in arteries from DE-exposed rats. Nitric oxide synthase (NOS) inhibition [N(omega)-nitro-L-arginine (L-NNA), 100 microM] and endothelium inactivation augmented ET-1 responses in arteries from Air but not DE rats so that after either treatment responses were not different between groups. DE exposure did not affect KCl and U-46619 constrictor responses, while NOS inhibition augmented KCl constriction equally in both groups. Thus basal NOS activity does not appear to be affected by DE exposure. The endothelin type B (ET(B)) receptor antagonist BQ-788 (10 microM) inhibited ET-1 constriction in DE but not Air arteries, and constriction in the presence of the antagonist was not different between groups. Cytokine levels were not different in plasma from DE and AIR rats, suggesting that acute exposure to DE does not cause an immediate inflammatory response. In summary, a 5-h DE exposure selectively increases constrictor sensitivity to ET-1. This augmentation is endothelium-, NOS-, and ET(B) receptor dependent. These data suggest that DE exposure diminishes ET(B) receptor activation of endothelial NOS and augments ET(B)-dependent vasoconstriction. This augmented coronary vasoreactivity to ET-1 after DE, coupled with previous reports that DE induces production of ET-1, suggests that ET-1 may contribute to the increased incidence of cardiac events during acute increases in air pollution levels.


Characterization and expression of DNA sequences encoding putative type-II metallothioneins in the seagrass Posidonia oceanica.
Posidonia oceanica is a marine phanerogam, largely widespread in the Mediterranean sea, representing an important food substrate for many marine organisms. A progressive reduction of P. oceanica meadows has been reported, due to anthropogenic coastal activity. Studying mechanisms by which this species responds to environmental stresses, three DNA sequences putatively encoding metallothioneins (MTs) have been isolated, by PCR. Two sequences, Pomt2a (accession no. AJ249603) and Pomt2b (accession no. AJ249602), show high similarities with genes encoding type-II MTs and are interrupted by two and one intron, respectively. The third sequence, Pomt2c (accession no. AJ249604), is supposed to be a pseudogene, originated by retrotranscription of the Pomt2b mRNA. These sequences belong to a multigene family with at least five members. Northern hybridizations indicated that MT transcripts accumulation is constitutive and seasonally regulated. MT encoding RNAs increase after rhyzome harvesting and (at a lesser extent) after 15 d of cultivation in an aquarium. As for animal MTs, transcripts accumulation is observed also after exposure to trace metals such as copper and cadmium. In the case of copper, the effect depends on concentration. Finally, taking into consideration the great interest in studying the biogeochemical cycle of mercury in the Mediterranean basin and since P. oceanica is commonly considered a bioindicator of this metal, the effect of mercury treatments on the accumulation of MT transcripts has been analyzed: in only a few experiments a small increase in the level of transcripts was recorded, suggesting that MTs are not key elements in the mercury accumulation by this species.


[Basidiomycetes: A new source of secondary metabolites.].
The area of natural products research is the most rapidly growing field of organic chemistry, due to the great technical developments in the isolation and identification techniques. Today, near to one million natural products -isolated from the most diverse living things- are known. Microorganisms are among the least-studied of these. Nevertheless, they offer large possibilities for the discovery of new structures and biological activities. Among the microorganisms, the Basidiomycetes present a production capacity and a range of biologically active metabolites, which have scarcely been investigated. The wide spectrum of natural products with biological activity produced by Basidiomycetes includes antimicrobial agents, antifungal, antiviral and cytotoxic activities, enzymes, plant growth regulators and flavors. These metabolites are generally grouped by their chemical origin, and the relationship between chemical structure and the different biological activities reported. The main objective of this review is to bring an updated revision of the numerous and interesting biosynthetic pathways from basidiomycetes.


Codominant interference, antieffectors, and multitarget drugs.
The insufficient selectivity of drugs is a bane of present-day therapies. This problem is significant for antibacterial drugs, difficult for antivirals, and utterly unsolved for anticancer drugs, which remain ineffective against major cancers, and in addition cause severe side effects. The problem may be solved if a therapeutic agent could have a multitarget, combinatorial selectivity, killing, or otherwise modifying, a cell if and only if it contains a predetermined set of molecular targets and lacks another predetermined set of targets. An earlier design of multitarget drugs [Varshavsky, A. (1995) Proc. Natl. Acad. Sci. USA 92, 3663-3667] was confined to macromolecular reagents such as proteins, with the attendant difficulties of intracellular delivery and immunogenicity. I now propose a solution to the problem of drug selectivity that is applicable to small (</=1 kDa) drugs. Two ideas, codominant interference and antieffectors, should allow a therapeutic regimen to possess combinatorial selectivity, in which the number of positively and negatively sensed macromolecular targets can be two, three, or more. The nature of the effector and interference moieties in a multitarget drug determines its use: selective killing of cancer cells or, for example, the inhibition of a neurotransmitter-inactivating enzyme in a specific subset of the enzyme-containing cells. The in vivo effects of such drugs would be analogous to the outcomes of the Boolean operations "and," "or," and combinations thereof. I discuss the logic and applications of the antieffector and interference/codominance concepts, and the attendant problem of pharmacokinetics.


Adversarial Attack Type I: Cheat Classifiers by Significant Changes.
Despite the great success of deep neural networks, the adversarial attack can cheat some well-trained classifiers by small permutations. In this paper, we propose another type of adversarial attack that can cheat classifiers by significant changes. For example, we can significantly change a face but well-trained neural networks still recognize the adversarial and the original example as the same person. Statistically, the existing adversarial attack increases Type II error and the proposed one aims at Type I error, which are hence named as Type II and Type I adversarial attack, respectively. The two types of attack are equally important but are essentially different, which are intuitively explained and numerically evaluated. To implement the proposed attack, a supervised variation autoencoder is designed and then the classifier is attacked by updating the latent variables using gradient information. Besides, with pre-trained generative models, Type I attack on latent spaces is investigated as well. Experimental results show that our method is practical and effective to generate Type I adversarial examples on large-scale image datasets. Most of these generated examples can pass detectors designed for defending Type II attack and the strengthening strategy is only efficient with a specific type attack, both implying that the underlying reasons for Type I and Type II attack are different.


Pose estimates from online videos show that side-by-side walkers synchronize movement under naturalistic conditions.
Marker-less video-based pose estimation promises to allow us to do movement science on existing video databases. We revisited the old question of how people synchronize their walking using real world data. We thus applied pose estimation to 348 video segments extracted from YouTube videos of people walking in cities. As in previous, more constrained, research, we find a tendency for pairs of people to walk in phase or in anti-phase with each other. Large video databases, along with pose-estimation algorithms, promise answers to many movement questions without experimentally acquiring new data.


[Locoregional recurrence in breast cancer--what should be done?].
Recurrence is a quite common problem in breast cancer. Strictly local recurrence seems to be not much different from an early breast cancer situation with minimal tumor load, and surgical removal may be adequate. If recurrence with multiple local tumors and/or regional nodes is present, most patients will develop metastatic disease. Treatment of loco-regional recurrence depends on prognostic factors; where long survival may be expected, surgical excision and radiation therapy with normal fractionation schedules should be planned. Current controlled studies are expected to determine the value of adjuvant hormone/cytotoxic therapy. With poor prognostic signs only systemic treatment is expected to influence survival, whereas local treatment (surgery/radiation-therapy) should be given as palliative treatment.


Color Doppler imaging in thyroid eye disease and its correlation to disease activity.
Purpose: To evaluate alterations in orbital color Doppler imaging (CDI) parameters and their correlation to disease activity and severity in patients with thyroid eye disease (TED). Methods: Seventy-six orbits of 45 TED patients and 40 orbits of 40 normal controls were enrolled in this cross-sectional study. According to clinical activity score (CAS), patients were categorized to active (CAS ≥ 3) or inactive disease (CAS < 3). Patients were also classified as having mild, moderate or severe disease. Peak systolic velocity (PSV), end diastolic velocity (EDV), and resistance index (RI) in ophthalmic artery and central retinal artery, and maximum and minimum velocity in superior ophthalmic vein and central retinal vein were determined in all subjects. Results: There was a significant difference in maximum velocity of superior ophthalmic vein and EDV and RI of ophthalmic artery between patients with TED and normal subjects. Superior ophthalmic vein maximum and minimum velocity and ophthalmic artery RI were significantly higher in patients with active disease than inactive cases. Disease severity did not affect the blood flow parameters independently. A cutoff point of 3.99 cm/s in superior ophthalmic vein maximum velocity yielded a sensitivity of 91.2% and specificity of 81.2% in detecting active disease. Conclusion: Retrobulbar blood flow is altered in TED and is related to disease activity. Superior ophthalmic vein maximum velocity could be helpful in differentiation of active and inactive cases.


[The value of serum glucose, urine glucose and hbAIc for long term metabolic information in juvenile diabetics (author's transl)].
In 43 insuline dependent diabetics postprandial serum glucose concentrations as 24-hour glucose excretions have been determined. Their value for a long term metabolic information was compared to that of HbAIc concentrations. A correlation between postprandial serum glucose concentration of the partial correlations. Those excluded the influence of an interrelationship among the individual serum glucose concentrations, at the time of different presentations for metabolic control, on the serum glucose to HbAIc relationship. There was no correlation between the concentrations of HbAIc and the amount of glucose excreted into the urine. The study shows that single determinations of postprandial serum glucose concentrations, as the excretion of small amounts of glucose into the urine are of no use for a long term metabolic information in juvenile diabetics.


Halftime rotational atherectomy: a unique concept for diffuse long severely calcified lesions.
Rotational atherectomy (RA) is technically more difficult in a diffuse calcified lesion than in a focal calcified lesion. We hypothesized that taking a halftime can be another option for RA to the diffuse calcified lesions. Halftime was defined as at least one long break during RA, in which an operator pulled out the Rotablator system from the guide catheter before crossing the lesion. This study aimed to compare the complications between RA with and without halftime. We included 177 diffuse long severely calcified lesions (lesion lengths ≥ 30 mm) that required RA, and divided those lesions into a halftime group (n = 29) and a no-halftime group (n = 148). The primary outcome was periprocedural myocardial infarction (MI). The reference diameter was smaller in the halftime group than in the no-halftime group [1.82 (1.70-2.06) mm versus 2.17 (1.89-2.59) mm, p = 0.002]. The total run time was longer in the halftime group than in the non-halftime group [133.0 (102.0-223.0) seconds versus 71.5 (42.0-108.0) seconds, p < 0.001]. Although creatinine kinase (CK) and CK-myocardial band (MB) was significantly higher in the halftime group than in the no-halftime group [CK: 156 (97-308) U/L versus 99 (59-216) U/L, p = 0.021; CK-MB: 15 (8-24) U/L versus 5 (3-15) U/L, p < 0.001], periprocedural MI was not observed in the halftime group. In conclusion, periprocedural MI was not observed in RA with halftime. This preliminary study suggests that halftime RA may be a safe option for diffuse severely calcified lesions.


Tip-Enhanced Raman Excitation Spectroscopy (TERES): Direct Spectral Characterization of the Gap-Mode Plasmon.
The plasmonic properties of tip-substrate composite systems are of vital importance to near-field optical spectroscopy, in particular tip-enhanced Raman spectroscopy (TERS), which enables operando studies of nanoscale chemistry at a single molecule level. The nanocavities formed in the tip-substrate junction also offer a highly tunable platform for studying field-matter interactions at the nanoscale. While the coupled nanoparticle dimer model offers a correct qualitative description of gap-mode plasmon effects, it ignores the full spectrum of multipolar tip plasmon modes and their interaction with surface plasmon polariton (SPP) excitation in the substrate. Herein, we perform the first tip-enhanced Raman excitation spectroscopy (TERES) experiment and use the results, both in ambient and aqueous media, in combination with electrodynamics simulations, to explore the plasmonic response of a Au tip-Au substrate composite system. The gap-mode plasmon features a wide spectral window corresponding to a host of tip plasmon modes interacting with the plasmonic substrate. Simulations of the electric field confinement demonstrate that optimal spatial resolution is achieved when a hybrid plasmon mode that combines a multipolar tip plasmon and a substrate SPP is excited. Nevertheless, a wide spectral window over 1000 nm is available for exciting the tip plasmon with high spatial resolution, which enables the simultaneous resonant detection of different molecular species. This window is robust as a function of tip-substrate distance and tip radius of curvature, indicating that many choices of tips will work, but it is restricted to wavelengths longer than ∼600 nm for the Au tip-Au substrate combination. Other combinations, such as Ag tip-Ag substrate, can access wavelengths as low as 350 nm.


Fasting is neuroprotective following traumatic brain injury.
To determine the neuroprotective effect of fasting after traumatic brain injury (TBI) and to elucidate the potential underlying mechanisms, we used a controlled cortical impact (CCI) injury model to induce either a moderate or a severe injury to adult male Sprague Dawley rats. Tissue-sparing assessments were used to determine the level of neuroprotection of fasting, hypoglycemia (insulin 10 U), or ketone (1.66 mmoles/kg/day or 0.83 mmoles/kg/day; D-beta-hydroxtbutyrate) administration. Mitochondrial isolation and respiratory studies were utilized to determine the functionality of mitochondria at the site of injury. We also investigated biomarkers of oxidative stress, such as lipid/protein oxidation, reactive oxygen species (ROS) production, and intramitochondrial calcium load, as a secondary measure of mitochondrial homeostasis. We found that fasting animals for 24 hr, but not 48 hr, after a moderate (1.5 mm), but not severe (2.0 mm), CCI resulted in a significant increase in tissue sparing. This 24-hr fast also decreased biomarkers of oxidative stress and calcium loading and increased mitochondrial oxidative phosphorylation in mitochondria isolated from the site of injury. Insulin administration, designed to mimic the hypoglycemic effect seen during fasting did not result in significant tissue sparing after moderate CCI injury and in fact induced increased mortality at some injection time points. However, the administration of ketones resulted in increased tissue sparing after moderate injury. Fasting for 24 hr confers neuroprotection, maintains cognitive function, and improves mitochondrial function after moderate (1.5 mm) TBI. The underlying mechanism appears to involve ketosis rather than hypoglycemia.


The Occurrence of Ophidiomyces ophiodiicola in Northern Georgia Wild and Captive Snake Populations.
Ophidiomycosis, or snake fungal disease, is an emerging wildlife disease caused by the Ophidiomyces ophiodiicola fungus. The fungus can result in high mortality rates among infected snakes and has been documented across much of the eastern US, including southern Georgia. However, little is known about ophidiomycosis in northern Georgia. We surveyed wild snake populations in five counties of northern Georgia between March 2019 and March 2020 and swabbed captured snakes (n=27) for the presence of O. ophiodiicola DNA. We followed similar sampling protocols with a group of captive snakes (n=6) at the Elachee Nature Center in Hall County, Georgia. Quantitative PCR confirmed the presence of O. ophiodiicola DNA in 33% (11/33) of snakes. Eight of the confirmed positive samples were collected from wild snakes (30%, 8/27) across our sample region, while three were from our captive group (50%, 3/6). Our results indicated that O. ophiodiicola is present in wild snake populations in northern Georgia, and the pathogen is present in seemingly healthy captive snakes. This knowledge is critical for conservation and management efforts, but more research is needed to fully understand ophidiomycosis and its effect on snake populations in the region.


Temporal trends and epidemiology of Staphylococcus aureus surgical site infection in the Swiss surveillance network: a cohort study.
Staphylococcus aureus is the leading pathogen in surgical site infections (SSI). To explore trends and risk factors associated with S. aureus SSI. Risk factors for monomicrobial S. aureus SSI were identified from the Swiss multi-centre SSI surveillance system using multi-variate logistic regression. Both in-hospital and postdischarge SSI were identified using standardized definitions. Over a six-year period, data were collected on 229,765 surgical patients, of whom 499 (0.22%) developed monomicrobial S. aureus SSI; 459 (92.0%) and 40 (8.0%) were due to meticillin-susceptible S. aureus (MSSA) and meticillin-resistant S. aureus (MRSA), respectively. There was a significant decrease in the rate of MSSA SSI (P = 0.007), but not in the rate of MRSA SSI (P = 0.70). Independent protective factors for S. aureus SSI were older age [≥75 years vs <50 years: odds ratio (OR) 0.60, 95% confidence interval (CI) 0.44-0.83], laparoscopy/minimally invasive surgery (OR 0.68, 95% CI 0.50-0.92), non-clean surgery [OR 0.78 (per increase in wound contamination class), 95% CI 0.64-0.94] and correct timing of pre-operative antibiotic prophylaxis (OR 0.80, 95% CI 0.65-0.98). Independent risk factors were male sex (OR 1.38, 95% CI 1.14-1.66), higher American Society of Anesthesiologists' score (per one-point increment: OR 1.30, 95% CI 1.13-1.51), re-operation for non-infectious reasons (OR 4.59, 95% CI 3.59-5.87) and procedure type: cardiac surgery, laminectomy, and hip or knee arthroplasty had two-to nine-fold increased odds of S. aureus SSI compared with other procedures. SSI due to S. aureus are decreasing and becoming rare events in Switzerland. High-risk procedures that may benefit from specific preventive measures were identified. Unfortunately, many of the independent risk factors are not easily modifiable.


Engineering nanostructured polymer blends with controlled nanoparticle location using Janus particles.
Janus particles are used on a multigram scale for the blend compatibilization of two polymers in a twin screw mini-mixer. It is shown that the Janus particles can be located exclusively at the interface of the two polymer phases despite the high temperature and shear conditions. The domain sizes of the dispersed phase decrease with increasing content of Janus particles. The decrease is yet ongoing for high contents of Janus particles. Furthermore, the biphasic particles exhibit an ordered arrangement at the interface. Thus, the approach demonstrates that a nanoscopic structuring of the interface can be achieved under macroscopic processing conditions. The structural order occurs on two levels. The first is the complete adsorption at the interface and the second is the lateral ordering at the interface. The strong adsorption at the interface is explained in terms of the increased desorption energy of Janus particles. Secondly, the compatibilization efficiency is critically compared to state-of-the-art compatibilizers. The efficiency of the Janus particles is found to be superior as compared to block copolymer-based compatibilizers. The efficiency gap between Janus particles and block copolymer compatibilizers widens for larger amounts added.


Testing the efficacy of motivational strategies, empathic reflections, and lifelike features in a computerized intervention for alcohol use: A factorial trial.
Although computer delivered brief interventions (CDBIs) have been effective in reducing alcohol use, few studies have examined which components of CDBIs are most associated with drinking reductions. The current factorial trial deconstructed a brief alcohol intervention into component parts to identify main and interaction effects on drinking outcomes. Participants (N = 352) were randomly assigned to 1 of 16 possible combinations of four dichotomous (present vs. absent) CDBI components for which theoretical and empirical support was available: empathic reflections, motivational strategies, a spoken voice, and an animated narrator. We measured main and interaction effects of these components on the primary outcome of self-reported mean drinks per day at 1- and 3-month follow-up, as well as on secondary outcomes, including binge drinking, intentions to reduce drinking, and alcohol consequences. Participants reduced drinking across all alcohol use indices over the 3-month assessment period. These effects were stronger for participants who were exposed to motivational strategies, F = 7.7, p < .001. Empathic reflections, use of a spoken voice, and use of an animated narrator were not associated with reductions in alcohol use, either as main effects or in interaction with other factors. Results suggest that CDBIs using motivational strategies are effective in reducing alcohol use. However, empathic reflections and lifelike characteristics (e.g., narrator, voice) may operate differently in CDBIs than they do in person-delivered interventions. More research is needed to better understand how these (or other factors) may influence efficacy. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


GLUCOSE UTILIZATION BY CHICK EMBRYO HEART HOMOGENATES.
Homogenates of early chick embryos and homogenates of early chick embryonic hearts utilized the phosphogluconate pathway of glucose catabolism to a greater extent, relative to the glycolytic-Krebs cycle pathway, than did homogenates of hearts from older chick embryos or adult chicks. An abrupt drop in the relative participation by the phosphogluconate pathway in embryo heart homogenates occurs at about 5 to 7 days of incubation. Heart homogenates from adult chicks catabolize glucose almost entirely by the glycolytic-Krebs cycle pathway, with negligible participation by the phosphogluconate pathway.


miR-9-Mediated Inhibition of EFEMP1 Contributes to the Acquisition of Pro-Tumoral Properties in Normal Fibroblasts.
Tumor growth and invasion occurs through a dynamic interaction between cancer and stromal cells, which support an aggressive niche. MicroRNAs are thought to act as tumor messengers to "corrupt" stromal cells. We previously demonstrated that miR-9, a known metastamiR, is released by triple negative breast cancer (TNBC) cells to enhance the transition of normal fibroblasts (NFs) into cancer-associated fibroblast (CAF)-like cells. EGF containing fibulin extracellular matrix protein 1 (EFEMP1), which encodes for the ECM glycoprotein fibulin-3, emerged as a miR-9 putative target upon miRNA's exogenous upmodulation in NFs. Here we explored the impact of EFEMP1 downmodulation on fibroblast's acquisition of CAF-like features, and how this phenotype influences neoplastic cells to gain chemoresistance. Indeed, upon miR-9 overexpression in NFs, EFEMP1 resulted downmodulated, both at RNA and protein levels. The luciferase reporter assay showed that miR-9 directly targets EFEMP1 and its silencing recapitulates miR-9-induced pro-tumoral phenotype in fibroblasts. In particular, EFEMP1 siRNA-transfected (si-EFEMP1) fibroblasts have an increased ability to migrate and invade. Moreover, TNBC cells conditioned with the supernatant of NFs transfected with miR-9 or si-EFEMP1 became more resistant to cisplatin. Overall, our results demonstrate that miR-9/EFEMP1 axis is crucial for the conversion of NFs to CAF-like cells under TNBC signaling.


Hemochromatosis (HFE) gene mutations in Brazilian chronic hemodialysis patients.
Patients with chronic renal insufficiency (CRI) have reduced hemoglobin levels, mostly as a result of decreased kidney production of erythropoietin, but the relation between renal insufficiency and the magnitude of hemoglobin reduction has not been well defined. Hereditary hemochromatosis is an inherited disorder of iron metabolism. The importance of the association of hemochromatosis with treatment for anemia among patients with CRI has not been well described. We analyzed the frequency of the C282Y and H63D mutations in the HFE gene in 201 Brazilian individuals with CRI undergoing hemodialysis. The analysis of the effects of HFE mutations on iron metabolism and anemia with biochemical parameters was possible in 118 patients of this study (hemoglobin, hematocrit, ferritin levels, transferrin saturation, and serum iron). A C282Y heterozygous mutation was found in 7/201 (3.4%) and H63D homozygous and heterozygous mutation were found in 2/201 (1.0%) and 46/201 (22.9%), respectively. The allelic frequencies of the HFE mutations (0.017 for C282Y mutation and 0.124 for H63D mutation) did not differ between patients with CRI and healthy controls. Regarding the biochemical parameters, no differences were observed between HFE heterozygous and mutation-negative patients, although ferritin levels were not higher among patients with the H63D mutation (P = 0.08). From what we observed in our study, C282Y/H63D HFE gene mutations are not related to degrees of anemia or iron stores in CRI patients receiving intravenous iron supplementation (P > 0.10). Nevertheless, the present data suggest that the H63D mutation may have an important function as a modulating factor of iron overload in these patients.


COX19 mediates the transduction of a mitochondrial redox signal from SCO1 that regulates ATP7A-mediated cellular copper efflux.
SCO1 and SCO2 are metallochaperones whose principal function is to add two copper ions to the catalytic core of cytochrome c oxidase (COX). However, affected tissues of SCO1 and SCO2 patients exhibit a combined deficiency in COX activity and total copper content, suggesting additional roles for these proteins in the regulation of cellular copper homeostasis. Here we show that both the redox state of the copper-binding cysteines of SCO1 and the abundance of SCO2 correlate with cellular copper content and that these relationships are perturbed by mutations in SCO1 or SCO2, producing a state of apparent copper overload. The copper deficiency in SCO patient fibroblasts is rescued by knockdown of ATP7A, a trans-Golgi, copper-transporting ATPase that traffics to the plasma membrane during copper overload to promote efflux. To investigate how a signal from SCO1 could be relayed to ATP7A, we examined the abundance and subcellular distribution of several soluble COX assembly factors. We found that COX19 partitions between mitochondria and the cytosol in a copper-dependent manner and that its knockdown partially rescues the copper deficiency in patient cells. These results demonstrate that COX19 is necessary for the transduction of a SCO1-dependent mitochondrial redox signal that regulates ATP7A-mediated cellular copper efflux.


Rgt1, a glucose sensing transcription factor, is required for transcriptional repression of the HXK2 gene in Saccharomyces cerevisiae.
Expression of HXK2, a gene encoding a Saccharomyces cerevisiae bifunctional protein with catalytic and regulatory functions, is controlled by glucose availability, being activated in the presence of glucose and inhibited when the levels of the sugar are low. In the present study, we identified Rgt1 as a transcription factor that, together with the Med8 protein, is essential for repression of the HXK2 gene in the absence of glucose. Rgt1 represses HXK2 expression by binding specifically to the motif (CGGAAAA) located at -395 bp relative to the ATG translation start codon in the HXK2 promoter. Disruption of the RGT1 gene causes an 18-fold increase in the level of HXK2 transcript in the absence of glucose. Rgt1 binds to the RGT1 element of HXK2 promoter in a glucose-dependent manner, and the repression of target gene depends on binding of Rgt1 to DNA. The physiological significance of the connection between two glucose-signalling pathways, the Snf3/Rgt2 that causes glucose induction and the Mig1/Hxk2 that causes glucose repression, was also analysed.


The initial burst of impulses in responses of toad muscle spindles during stretch.
The responses of muscle spindles in the iliofibularis muscle of the cane toad Bufo marinus were examined during constant velocity stretch of the passive muscle. Spindles were found to show an 'initial burst' of high frequency impulses at the onset of stretch. Associated with the initial burst was a steep passive tension rise in the whole muscle, the short-range elastic component (Hill, 1968), called here the passive stiffness. The size of the initial burst was found to depend on muscle length in a similar way as whole-muscle tetanic tension. Repetitive stretch was found to reduce both the initial burst and passive stiffness. The time taken for both to return to their control values was 3 and 10 s respectively. If immediately following repetitive stretch the muscle, and hence the spindle, was held stretched for 3 s, the initial burst in response to a subsequent stretch from a shorter length remained reduced in size for 300 s. The depression could be reversed by a brief period of fusimotor stimulation. Hypertonic Ringer solutions were found to increase the initial burst and passive stiffness, while both were reduced in hypotonic solutions. Low concentrations of caffeine (1.5 mM) produced a similar decrease in both the initial burst and the passive stiffness. Calcium-free Ringer solution left the stiffness unchanged, and increased the whole dynamic response of the spindle. Metabolic exhaustion and poisoning of the muscle caused the initial burst to increase while decreasing the active tension. It is concluded that the initial burst is an intrafusal manifestation of the passive short-range stiffness of extrafusal muscle which is thought to be due to the formation of stable cross-bridges between the actin and myosin filaments of myofibrils.


Analysis of the proteome of intracellular Shigella flexneri reveals pathways important for intracellular growth.
Global proteomic analysis was performed with Shigella flexneri strain 2457T in association with three distinct growth environments: S. flexneri growing in broth (in vitro), S. flexneri growing within epithelial cell cytoplasm (intracellular), and S. flexneri that were cultured with, but did not invade, Henle cells (extracellular). Compared to in vitro and extracellular bacteria, intracellular bacteria had increased levels of proteins required for invasion and cell-to-cell spread, including Ipa, Mxi, and Ics proteins. Changes in metabolic pathways in response to the intracellular environment also were evident. There was an increase in glycogen biosynthesis enzymes, altered expression of sugar transporters, and a reduced amount of the carbon storage regulator CsrA. Mixed acid fermentation enzymes were highly expressed intracellularly, while tricarboxylic acid (TCA) cycle oxidoreductive enzymes and most electron transport chain proteins, except CydAB, were markedly decreased. This suggested that fermentation and the CydAB system primarily sustain energy generation intracellularly. Elevated levels of PntAB, which is responsible for NADPH regeneration, suggested a shortage of reducing factors for ATP synthesis. These metabolic changes likely reflect changes in available carbon sources, oxygen levels, and iron availability. Intracellular bacteria showed strong evidence of iron starvation. Iron acquisition systems (Iut, Sit, FhuA, and Feo) and the iron starvation, stress-associated Fe-S cluster assembly (Suf) protein were markedly increased in abundance. Mutational analysis confirmed that the mixed-acid fermentation pathway was required for wild-type intracellular growth and spread of S. flexneri. Thus, iron stress and changes in carbon metabolism may be key factors in the S. flexneri transition from the extra- to the intracellular milieu.


Opioid-sparing effects of 10 kHz spinal cord stimulation: a review of clinical evidence.
Chronic pain is a common condition that affects the physical, emotional, and mental well-being of patients and can significantly diminish their quality of life. Due to growing concerns about the substantial risks of long-term opioid use, both governmental agencies and professional societies have recommended prioritizing the use of nonpharmacologic treatments, when suitable, in order to reduce or eliminate the need for opioid use. The use of 10 kHz spinal cord stimulation (10 kHz SCS) is one such nonpharmacologic alternative for the treatment of chronic, intractable pain of the trunk and limbs. This review examines published clinical data regarding the efficacy of 10 kHz SCS for decreasing chronic pain in patients and its potential to reduce or eliminate opioid usage. Multiple prospective and retrospective studies in patients with intractable pain demonstrated that 10 kHz SCS treatment provided ≥50% pain relief in >70% patients after at least 1 year of treatment. Pain relief with 10 kHz SCS therapy ranged from 54% to 87% in the studies. More importantly, the mean daily dose of opioids required by patients in these studies was reduced after 10 kHz SCS treatment, and on average over 60% patients in studies either reduced or eliminated opioids at the last follow-up.


Functional display of bioactive peptides on the vGFP scaffold.
Grafting bioactive peptides into recipient protein scaffolds can often increase their activities by conferring enhanced stability and cellular longevity. Here, we describe use of vGFP as a novel scaffold to display peptides. vGFP comprises GFP fused to a bound high affinity Enhancer nanobody that potentiates its fluorescence. We show that peptides inserted into the linker region between GFP and the Enhancer are correctly displayed for on-target interaction, both in vitro and in live cells by pull-down, measurement of target inhibition and imaging analyses. This is further confirmed by structural studies highlighting the optimal display of a vGFP-displayed peptide bound to Mdm2, the key negative regulator of p53 that is often overexpressed in cancer. We also demonstrate a potential biosensing application of the vGFP scaffold by showing target-dependent modulation of intrinsic fluorescence. vGFP is relatively thermostable, well-expressed and inherently fluorescent. These properties make it a useful scaffold to add to the existing tool box for displaying peptides that can disrupt clinically relevant protein-protein interactions.


Aging dependent effect of nuclear tau.
Tau protein is characterized by a complex pattern of phosphorylation and is localized in the cytoplasm and nucleus in both neuronal and non-neuronal cells. Human AT100 nuclear tau, endowed by phosphorylation in Thr212/Ser214, was recently shown to decline in cornus ammonis 1 (CA1) and dentate gyrus (DG) in Alzheimer's disease (AD), but a defined function for this nuclear tau remains unclear. Here we show that AT100 progressively increases in the nuclei of neuronal and non-neuronal cells during aging, and decreases in the more severe AD stages, as recently shown, and in cancer cells (colorectal adenocarcinoma and breast cancer). AT100, in addition to a co-localization with the DAPI-positive heterochromatin, was detected in the nucleolus of pyramidal cells from the CA1 region, shown to be at its highest level in the more senescent cells and in the first stage of AD (ADI), and disappearing in the more severe AD cases (ADIV). Taking into account the nuclear distribution of AT100 during cell aging and its relation to the chromatin changes observed in degenerated neurons, as well as in cancerous cells, which are both cellular pathologies associated with age, we can consider the Thr212/Ser214 phosphorylated nuclear tau as a molecular marker of cell aging.


A brief measure of reactance to health warnings.
Reactance to persuasive messages involves perceived threat to freedom, anger, and counterarguing that may undermine the impact of health warnings. To understand reactance's effects, reliable and valid assessment is critical. We sought to develop and validate a brief Reactance to Health Warnings Scale (RHWS). Two independent samples of US adults completed the brief RHWS in studies that presented warnings on cigarette packs that smokers carried with them for 4 weeks (Study 1; n = 2149) or as digital images of cigarette packs that participants viewed briefly (Study 2; n = 1413). The three-item Brief RHWS had good internal consistency and test-retest reliability. The scale correlated with higher trait reactance and exposure to pictorial warnings, supporting its convergent validity. With respect to predictive validity, the Brief RHWS predicted perceived message effectiveness, quit intentions, avoidance of the warnings, and number of cigarettes smoked per day. The Brief RHWS can serve as an efficient adjunct to the development of persuasive messages.


Reconfigurable add-drop multiplexer for spatial modes.
We show how a spatial mode can be extracted from a light beam, leaving the other orthogonal modes undisturbed, and allowing a new signal to be retransmitted on that mode. The method is self-aligning, avoids fundamental splitting losses, and uses only local feedback loops on controllable beam splitters and phase shifters. It could be implemented with Mach-Zehnder interferometers in planar optics. The method can be extended to multiple simultaneous mode extractions. As a spatial reconfigurable optical add-drop multiplexer, it is hitless, allowing reconfiguration without interrupting the transmission of any channel.


Glasgow Coma Scale and outcomes after structural traumatic head injury in early childhood.
To assess the association of the Glasgow Coma Scale (GCS) with radiological evidence of head injury (the Abbreviated Injury Scale for the head region, AIS-HR) in young children hospitalized with traumatic head injury (THI), and the predictive value of GCS and AIS-HR scores for long-term impairment. Our study involved a 10-year retrospective review of a database encompassing all patients admitted to Starship Children's Hospital (Auckland, New Zealand, 2000-2010) with THI. We studied 619 children aged <5 years at the time of THI, with long-term outcome data available for 161 subjects. Both GCS and AIS-HR scores were predictive of length of intensive care unit and hospital stay (all p<0.001). GCS was correlated with AIS-HR (ρ=-0.46; p<0.001), although mild GCS scores (13-15) commonly under-estimated the severity of radiological injury: 42% of children with mild GCS scores had serious-critical THI (AIS-HR 3-5). Increasingly severe GCS or AIS-HR scores were both associated with a greater likelihood of long-term impairment (neurological disability, residual problems, and educational support). However, long-term impairment was also relatively common in children with mild GCS scores paired with structural THI more severe than a simple linear skull fracture. Severe GCS scores will identify most cases of severe radiological injury in early childhood, and are good predictors of poor long-term outcome. However, young children admitted to hospital with structural THI and mild GCS scores have an appreciable risk of long-term disability, and also warrant long-term follow-up.


Preparation and inoculation of mesophyll protoplasts from monocotyledenous and dicotyledenous hosts.
This unit describes a simple and efficient protocol for inoculating mesophyll protoplasts of either monocotyledonous or dicotyledenous host plants using polyethylene glycol. The unit covers isolation of biologically active protoplasts, their transfection with viral RNA, and conditions for the maintenance of transfected protoplasts. It also highlights critical aspects of these steps and discusses common problems and their solutions.


Can We Geographically Validate a Natural Language Processing Algorithm for Automated Detection of Incidental Durotomy Across Three Independent Cohorts From Two Continents?
Incidental durotomy is an intraoperative complication in spine surgery that can lead to postoperative complications, increased length of stay, and higher healthcare costs. Natural language processing (NLP) is an artificial intelligence method that assists in understanding free-text notes that may be useful in the automated surveillance of adverse events in orthopaedic surgery. A previously developed NLP algorithm is highly accurate in the detection of incidental durotomy on internal validation and external validation in an independent cohort from the same country. External validation in a cohort with linguistic differences is required to assess the transportability of the developed algorithm, referred to geographical validation. Ideally, the performance of a prediction model, the NLP algorithm, is constant across geographic regions to ensure reproducibility and model validity. Can we geographically validate an NLP algorithm for the automated detection of incidental durotomy across three independent cohorts from two continents? Patients 18 years or older undergoing a primary procedure of (thoraco)lumbar spine surgery were included. In Massachusetts, between January 2000 and June 2018, 1000 patients were included from two academic and three community medical centers. In Maryland, between July 2016 and November 2018, 1279 patients were included from one academic center, and in Australia, between January 2010 and December 2019, 944 patients were included from one academic center. The authors retrospectively studied the free-text operative notes of included patients for the primary outcome that was defined as intraoperative durotomy. Incidental durotomy occurred in 9% (93 of 1000), 8% (108 of 1279), and 6% (58 of 944) of the patients, respectively, in the Massachusetts, Maryland, and Australia cohorts. No missing reports were observed. Three datasets (Massachusetts, Australian, and combined Massachusetts and Australian) were divided into training and holdout test sets in an 80:20 ratio. An extreme gradient boosting (an efficient and flexible tree-based algorithm) NLP algorithm was individually trained on each training set, and the performance of the three NLP algorithms (respectively American, Australian, and combined) was assessed by discrimination via area under the receiver operating characteristic curves (AUC-ROC; this measures the model's ability to distinguish patients who obtained the outcomes from those who did not), calibration metrics (which plot the predicted and the observed probabilities) and Brier score (a composite of discrimination and calibration). In addition, the sensitivity (true positives, recall), specificity (true negatives), positive predictive value (also known as precision), negative predictive value, F1-score (composite of precision and recall), positive likelihood ratio, and negative likelihood ratio were calculated. The combined NLP algorithm (the combined Massachusetts and Australian data) achieved excellent performance on independent testing data from Australia (AUC-ROC 0.97 [95% confidence interval 0.87 to 0.99]), Massachusetts (AUC-ROC 0.99 [95% CI 0.80 to 0.99]) and Maryland (AUC-ROC 0.95 [95% CI 0.93 to 0.97]). The NLP developed based on the Massachusetts cohort had excellent performance in the Maryland cohort (AUC-ROC 0.97 [95% CI 0.95 to 0.99]) but worse performance in the Australian cohort (AUC-ROC 0.74 [95% CI 0.70 to 0.77]). We demonstrated the clinical utility and reproducibility of an NLP algorithm with combined datasets retaining excellent performance in individual countries relative to algorithms developed in the same country alone for detection of incidental durotomy. Further multi-institutional, international collaborations can facilitate the creation of universal NLP algorithms that improve the quality and safety of orthopaedic surgery globally. The combined NLP algorithm has been incorporated into a freely accessible web application that can be found at https://sorg-apps.shinyapps.io/nlp_incidental_durotomy/ . Clinicians and researchers can use the tool to help incorporate the model in evaluating spine registries or quality and safety departments to automate detection of incidental durotomy and optimize prevention efforts. Level III, diagnostic study.


Using data to plan public health programs: experience from state cancer prevention and control programs.
In 1989 the National Cancer Institute funded the second round of Data-Based Intervention Research (DBIR) cooperative agreements with state health agencies to implement a four-phase cancer prevention and control planning model that would establish ongoing cancer prevention and control programs. Activities included identifying and analyzing relevant data to develop a state cancer control plan. The authors reviewed the data analysis and planning activities of five DBIR projects to understand: how states use different types of available data to make public health planning decisions, in what ways available data were sufficient or insufficient for this planning, and perceived costs and benefits of a data-based planning approach. Many of the sources of and ways in which health statistics and behavioral data were used were consistent across states. Sources and use of data on the availability and utilization of health services and on cancer control policies were less consistent. Data were most useful in making decisions to address specific cancers, to target populations or regions, to identify general barriers, and to influence policy makers and the public. Data were less influential in identifying specific barriers within target populations and determining what proven intervention components should be implemented and how. The process of pulling this information together and involving working groups and coalitions was considered very beneficial in establishing the credibility of the state health agency in addressing the state's cancer problem. This process relied on a national infrastructure that provided financial resources, sources of data, and research results.


Vertebral body replacement with an expandable cage for reconstruction after spinal tumor resection.
The authors report their clinical experience with expandable cages used to stabilize the spine after vertebrectomy. The objectives of surgical treatment for spine tumors include a decrease in pain, decompression of the neural elements, mechanical stabilization of the spine, and wide resection to gain local control of certain primary tumors. Most of the lesions occur in the anterior column or vertebral body (VB). Anterior column defects following resection of VBs require surgical restoration of anterior column support. Recently, various expandable cages have been developed and used clinically for VB replacement (VBR). Between January 2001 and June 2003, the authors treated 15 patients who presented with primary spinal tumors and metastatic lesions from remote sites. All patients underwent vertebrectomy, VBR with an expandable cage, and anterior instrumentation with or without posterior instrumentation, depending on the stability of the involved segment. The correction of kyphotic angle was achieved at an average of 20 degrees. Pain scores according to the visual analog scale decreased from 8.4 to 5.2 at the last follow-up review. Patients whose Frankel neurological grade was below D attained at least a one-grade improvement after surgery. All patients achieved immediate stability postsurgery and there were no significant complications related to the expandable cage. The advantage of the expandable cage is that it is easy to use because it permits optimal fit and correction of the deformity by in vivo expansion of the device. These results are promising, but long-term follow up is required.