Adolescents with bulimia nervosa and eating disorder not otherwise specified-purging only.
The purpose of the study was to better understand the phenomenology of bulimic symptomatology in an adolescent clinic sample. Adolescents with bulimia nervosa (BN; n = 36) and eating disorders not otherwise specified-purging but no objective bulimic episodes (EDNOS-P; n = 20) were compared on the Eating Disorder Examination (EDE), the Beck Depression Inventory (BDI), the Rosenberg Self-Esteem Scale (RSES), and the Schedule for Affective Disorders and Schizophrenia for School-Age Children (K-SADS). Subjects with EDNOS-P and BN were equivalent in terms of age and weight, but were less likely to have intact families. Nearly one half of EDNOS-P subjects purged exclusively outside of eating episodes in which they experienced a sense of loss of control. Although still at clinically significant levels, EDNOS-P subjects reported less concerns regarding weight, shape, and eating relative to BN. Groups were not significantly different on psychiatric comorbidity, but differed on self-esteem. Results prompt reappraisal of current criteria of BN to encompass those who purge without binge eating.


Fetal human hematopoietic stem cells can differentiate sequentially into neural stem cells and then astrocytes in vitro.
In some rodent models, there is evidence that hematopoietic stem cells (HSC) can differentiate into neural cells. However, it is not known whether humans share this potential, and, if so, what conditions are sufficient for this transdifferentiation to occur. We addressed this question by assessing the ability of fetal human liver CD34(+)/CD133(+)/CD3(-) hematopoietic stem cells to generate neural cells and astrocytes in culture. We cultured fetal liver-derived hematopoietic stem cells in human astrocyte culture-conditioned medium or using a method wherein growing human astrocytes were separated from cultured, nonadherent hematopoietic stem cells by a semipermeable membrane in a double-chamber co-culture system. Hematopoietic stem cell cultures were probed for neural progenitor cell marker expression (nestin and bone morphogenic protein-2 [BMP-2]) during growth in both culture conditions. RT-PCR, western blotting, and immunocytochemistry assays showed that cells cultured in either condition expressed nestin mRNA and protein and BMP-2 mRNA. HSC similarly cultured in nonconditioned medium or in the absence of astrocytes did not express either marker. Cells expressing these neural markers were transferred and cultured on poly-D-lysine-coated dishes with nonconditioned growth medium for further study. Immunocytochemistry demonstrated that these cells differentiated into astrocytes after 8 days in culture as indicated by their morphology and expression of the astrocytic markers glial fibrillary acidic protein (GFAP) and S100, as well as by their rate of proliferation, which was identical to that of freshly isolated fetal brain astrocytes. These findings demonstrate that neural precursor gene expression can be induced when human hematopoietic stem cells are exposed to a suitable microenvironment. Furthermore, the neural stem cells generated in this environment can then differentiate into astrocytes. Therefore, human hematopoietic stem cells may be an alternative resource for generation of neural stem cells for therapy of central nervous system defects resulting from disease or trauma.


Violating the Weak Cosmic Censorship Conjecture in Four-Dimensional Anti-de Sitter Space.
We consider time-dependent solutions of the Einstein-Maxwell equations using anti-de Sitter (AdS) boundary conditions, and provide the first counterexample to the weak cosmic censorship conjecture in four spacetime dimensions. Our counterexample is entirely formulated in the Poincaré patch of AdS. We claim that our results have important consequences for quantum gravity, most notably to the weak gravity conjecture.


The Efficacy of "Care for Child Development" Intervention on the Improvement of the Development Skills of Orphanage Children.
Development refers to the progressive enhancement of skills and functional capacity, i.e., qualitative changes in the child's functions. The process of development begins before birth and continues throughout life. The present study aims to evaluate the effectiveness of the "Care for Child Development (CCD)" program on 4-42 months children's developmental skills in orphanages. In this study, two orphanages in the capitals of East and West Azerbaijan provinces were selected using the convenience sampling technique, and thirty children were included. Then, they were randomly divided into two intervention and control groups (each group, N=15). Next, after obtaining consent from the head of the orphanages, a group of volunteers from the healthcare center performed the CCD program, considering children's chronological ages (4 to 42 months), for three sessions a week, with each session lasting two hours and it lasted for three months. At the end of the intervention process, the Bayley Scale of Infant and Toddler Development 3rd version (BSID-III) and the Ages and Stages Questionnaire-II (ASQ) were completed for the two intervention and control groups to compare them in the cognitive, motor, communication, and personal-social domains. Comparing the two control and intervention groups using the T-test (difference in mean) indicates that except for the domain of cognitive skills (Bayley: P-value = 0.176), there was statistically a significant difference between the two groups in communication (ASQ: P-value = 0.001; Bayley: P-value = 0.003), motor (ASQ: P-value = 0.000; Bayley: P-value = 0.009), and personal-social (ASQ: P-value <0.000) skills. In the present study, it was concluded that it is required to apply interventions, including standard ones such as the CCD program in environments like orphanages, to enhance the developmental skills of those children living in them.


Rev and Rex proteins of human complex retroviruses function with the MMTV Rem-responsive element.
Mouse mammary tumor virus (MMTV) encodes the Rem protein, an HIV Rev-like protein that enhances nuclear export of unspliced viral RNA in rodent cells. We have shown that Rem is expressed from a doubly spliced RNA, typical of complex retroviruses. Several recent reports indicate that MMTV can infect human cells, suggesting that MMTV might interact with human retroviruses, such as human immunodeficiency virus (HIV), human T-cell leukemia virus (HTLV), and human endogenous retrovirus type K (HERV-K). In this report, we test whether the export/regulatory proteins of human complex retroviruses will increase expression from vectors containing the Rem-responsive element (RmRE). MMTV Rem, HIV Rev, and HTLV Rex proteins, but not HERV-K Rec, enhanced expression from an MMTV-based reporter plasmid in human T cells, and this activity was dependent on the RmRE. No RmRE-dependent reporter gene expression was detectable using Rev, Rex, or Rec in HC11 mouse mammary cells. Cell fractionation and RNA quantitation experiments suggested that the regulatory proteins did not affect RNA stability or nuclear export in the MMTV reporter system. Rem had no demonstrable activity on export elements from HIV, HTLV, or HERV-K. Similar to the Rem-specific activity in rodent cells, the RmRE-dependent functions of Rem, Rev, or Rex in human cells were inhibited by a dominant-negative truncated nucleoporin that acts in the Crm1 pathway of RNA and protein export. These data argue that many retroviral regulatory proteins recognize similar complex RNA structures, which may depend on the presence of cell-type specific proteins. Retroviral protein activity on the RmRE appears to affect a post-export function of the reporter RNA. Our results provide additional evidence that MMTV is a complex retrovirus with the potential for viral interactions in human cells.


In vitro anti-HIV-1 activity of salicylidene acylhydrazide compounds.
Salicylidene acylhydrazide compounds have been shown to inhibit bacterial pathogens, including Chlamydia and Neisseria gonorrhoeae. If such compounds could also target HIV-1, their potential use as topical microbicides to prevent sexually transmitted infections would be considerable. In this study, the in vitro anti-HIV-1 activity, cytotoxicity and mechanism of action of several salicylidene acylhydrazides were determined. Inhibitory activity was assessed using TZM-bl cells and primary peripheral blood mononuclear cells (PBMCs) as targets for HIV-1 infection. Antiviral activity was measured against cell-free and cell-associated virus and in vaginal fluid and semen simulants. Since the antibacterial activity of salicylidene acylhydrazides is reversible by Fe(2+), the ability of Fe(2+) and other cations to reverse the anti-HIV-1 activity of the compounds was determined. Real-time PCR was also employed to determine the stage affected in the HIV-1 replication cycle. Four compounds with 50% inhibitory concentrations against HIV-1 of 1-7 μM were identified. In vitro toxicity varied but was generally limited. Activity was similar against three R5 clade B primary isolates and whether the target for virus replication was TZM-bl cells or PBMCs. Compounds inhibited cell-free and cell-associated virus and were active in vaginal fluid and semen simulants. Fe(2+), but not other cations, reversed the anti-HIV-1 effect. Finally, the inhibitory effect of the compounds occurred at a post-integration step. In conclusion, salicylidene acylhydrazides were identified with in vitro anti-HIV-1 activity in the micromolar range. The activity of these compounds against other sexually transmitted pathogens makes them potential candidates to formulate for use as a broad-spectrum topical genital microbicide.


Iron(III) binding proteins of echidna (Tachyglossus aculeatus) and platypus (Ornithorhynchus anatinus).
Iron (III) binding proteins are isolated from echidna (Tachyglossus aculeatus multiaculeatus) and platypus (Ornithorhynchus anatinus) milk and blood. On the basis of several criteria it is shown that the milk proteins are not lactoferrins, but are transferrins similar to the corresponding transferrins from the blood. The heterogeneity of the proteins, particularly the echidna milk transferrin, is, at least in part, due to different levels of sialic acid. Their N-terminal sequences (30 residues) are determined and compared with those of other transferrins and lactoferrins. The role of the proteins is discussed.


[Pathogenesis of persistent and chronic forms of tick-borne encephalitis (experimental study)].
A long-term experiment was conducted to study various aspects of the pathogenesis of persistent and chronic tick-borne encephalitis (TBE). Virological, serological, pathomorphological, electron microscopic and immunofluorescent techniques have been utilized in this study. Persistent TBE infection of Syrian hamsters examined over the period from 40 days to 2 years was characterized by the presence of virus-specific antigens in the organs and of specific antibodies in the blood serum. The persisting TBE virus was found to be predominantly localized in the central nervous system and spleen. Nerve cells underwent ultrastructural changes which were characteristic of flavivirus infection and related to the morphogenesis of viral particles. The authors have developed an experimental model of a primary progressive form of TBE with early and late manifestations of clinical symptoms of the disease.


Endoscopic findings of upper gastrointestinal lesions in patients with pancreatic cancer.
Pancreatic cancer is often complicated with upper gastrointestinal lesions. However, there have been few endoscopic studies in pancreatic cancer patients. We retrospectively investigated the upper gastrointestinal lesions in patients with pancreatic cancer who underwent upper gastrointestinal endoscopy. Upper gastrointestinal endoscopy was performed in 75 patients with pancreatic cancer between 2003 and 2010. We examined upper gastrointestinal lesions, such as gastroduodenal invasion, ulcers, esophagogastric varices, radiation-induced gastroduodenal mucosal lesions, and portal hypertensive gastropathy. Among the 53 patients with pancreatic cancer who underwent upper gastrointestinal endoscopy at diagnosis, 23 gastrointestinal lesions were observed in 20 patients (38%) as follows: gastroduodenal invasion (n=11), esophagogastric varices (n=7), gastroduodenal ulcers (n=3), portal hypertensive gastropathy (n=1) and duodenal metastasis (n=1). Among the 75 patients with pancreatic cancer, 56 gastrointestinal lesions were identified in 46 patients (61%) during the clinical course as follows: gastroduodenal invasion (n=20), esophagogastric varices (n=14), radiation-induced gastroduodenal mucosal lesions (n=9), gastroduodenal ulcers (except radiation-induced ulcers) (n=8), portal hypertensive gastropathy (n=3), duodenal metastasis (n=1), and gastrointestinal bleeding from unknown primary site (n=1). Twenty-nine (52%) of the 56 gastrointestinal lesions showed symptoms related to the lesions. Fifteen (27%) lesions were accompanied by upper gastrointestinal bleeding. Fourteen (25%) lesions developed according to the progression of pancreatic cancer. We should pay attention to upper gastrointestinal lesions in patients with pancreatic cancer.


Sequentially surface modified hematite enables lower applied bias photoelectrochemical water splitting.
Hematite (α-Fe2O3) is a suitable candidate for photoelectrochemical water splitting due to its well-suited band structure, stability, and availability. However, water splitting using a low external potential is the major challenge that limits the practical application of hematite. Here, we achieve a very low onset potential using a sequential surface treatment approach to overcome two fundamental limiting factors, sluggish hole transfer, and interfacial recombination, independently. First, a heavily doped Fe2-xSnxO3 surface passivation layer was created by Sn4+ surface treatment which can robustly inhibit interfacial recombination. Then, an NiOOH catalyst layer was deposited that greatly enhances the charge transfer process across the passivated electrode/electrolyte interface. By exploiting this approach, the optimized sequentially treated photoanode (Fe2O3/Fe2-xSnxO3/NiOOH) exhibits a low photocurrent onset potential of 0.49 V vs. RHE and a saturated photocurrent density of 2.4 mA cm-2 V at 1.5 V vs. RHE. Transient photocurrent and impedance spectroscopy measurements further reveal that the combined Fe2-xSnxO3/NiOOH layers reduce interfacial recombination and enhance charge transfer across the electrode/electrolyte interface. The results provide convincing evidence that it is possible to address the problems of surface trap recombination and sluggish catalysis independently by employing surface passivation layers first and catalysts later sequentially.


Effects of anterior approach to quadratus lumborum block on postoperative cognitive function following hip surgery in older people: a randomized controlled clinical trial.
Peripheral nerve block, including the quadratus lumborum block (QLB), has been used for postoperative analgesia in hip surgery. However, the effects of QLB on cognitive function after hip surgery remain unknown. This study aimed to assess the effects of the anterior approach to QLB on postoperative cognitive function in older people undergoing hip surgery. Sixty older people who underwent hip surgery from May 2021 to May 2022 were randomly divided into the QLB (n = 30) and control groups (n = 30). The Montreal Cognitive Assessment (MoCA) score (mean ± SD) was measured one day preoperatively and seven and 30 days postoperatively. The frequency (%) of postoperative cognitive dysfunction (POCD) was examined seven and 30 days postoperatively. The visual analog scale (VAS) scores at rest and Bruggrmann comfort scale (BCS) scores [Median (IQR)] 6 h (t1), 12 h (t2), 24 h (t3), and 48 h (t4) after surgery were assessed. The plasma high mobility group box protein 1 (HMGB1) and levels of interleukin-6 (IL-6) (mean ± SD) were evaluated 1 h preoperatively (baseline) and 24 h postoperatively (day 1). The requirement for rescue analgesia [Median (IQR)], time to first off-bed activity (mean ± SD), and adverse effects after surgery were also recorded. Compared with the control group, the frequency of POCD was significantly lower in the QLB group seven days postoperatively (10.7% vs. 34.5%, P = 0.033), but no difference at 30 days postoperatively (3.6% vs. 10.3%, P = 0.319). There was no significant difference in MoCA scores between the two groups at one day preoperatively and 30 days postoperatively. However, the MoCA scores at seven days postoperatively were higher in the QLB group than in the control group (27.4 ± 1.81 vs. 26.4 ± 1.83, P = 0.043). In the QLB group, the VAS scores at t1, t2, and t3 were lower [3(2-4) vs. 4(3-4), P = 0.028; 3(2-3) vs. 4(3-5), P = 0.009; 2(1-3) vs. 2(2-3), P = 0.025], and the BCS scores at t1, t2, and t3 were higher than those in the control group [3(1-3) vs. 1(1-2), P = 0.006; 3(2-3) vs. 2(1-3), P = 0.011; 3(2-4) vs. 2(2-3), P = 0.041]. The patients in the QLB group reported significantly fewer requirements for rescue analgesia [0(0-1) vs. 1(0-2), P = 0.014]. The plasma levels of HMGB1 and IL-6 at 24 h postoperatively in the QLB group were significantly lower than in the control group (749.0 ± 185.7 vs. 842.1 ± 157.9, P = 0.046; 24.8 ± 8.1 vs. 31.9 ± 5.5, P < 0.001). The time to first off-bed activity from the end of surgery was shorter in the QLB group (25.3 ± 5.3 vs. 29.7 ± 6.9, P = 0.009). There was no significant difference in the incidence of postoperative complications between the two groups. Anterior QLB given to older people undergoing hip surgery could promote early postoperative cognitive function recovery, provide adequate postoperative analgesia, and inhibit the release of inflammatory factors. Chictr.org.cn identifier ChiCTR2000040724 (Date of registry: 08/12/2020, prospectively registered).


[Sports ability after patella dislocation].
A dislocation of the patella is a serious injury which can far affect the career of every athlete or even finish it. In a retrospective study we examined 24 athletes (mean age 19.3 years, post surgical observation period 39 (13 to 79) months) which suffered from a patellar dislocation and received arthroscopic surgical treatment. The diagnostics contained clinical investigation and X-rays of the knee joint in 2 plains as well as patella tangential, in pathological axial deformations of the leg completed with whole-leg-images and optional a torsion computed tomography. All 24 patients received an arthroscopical assisted suturing of the medial retinaculum, in 12 patients additional a lateral release was carried out. The postoperative course followed a standard pattern. In 24 operated patients the reoccurrence rate was 12.5 % (3 patients), which is within the range of the latest published figures (average 12.0 %). The Lysholm score was in 15 (62.4 %) of all patients higher than 80 demonstrating the good to very good results, the average Lysholm score was 83.4. Out of 24 patients with patellar dislocation 11 patients (45.8 %) were able to continue their sport career after surgical therapy at the pre-trauma level, 4 patients (16.6 %) had to continue at lower level, 9 patients (37.5 %) had to finish their sport career. The Tegner activity score decreased from 7.2 to 6.0 at about 1.2 points what means a clear decrease of the activity level. On average a full sportive level was reached 9.7 months after surgery. Especially sport athletes needs to be informed about the seriousness of the injury and the long process of rehabilitation.


Primary Open-Angle Glaucoma Diagnosis from Optic Disc Photographs Using a Siamese Network.
Primary open-angle glaucoma (POAG) is one of the leading causes of irreversible blindness in the United States and worldwide. Although deep learning methods have been proposed to diagnose POAG, these methods all used a single image as input. Contrastingly, glaucoma specialists typically compare the follow-up image with the baseline image to diagnose incident glaucoma. To simulate this process, we proposed a Siamese neural network, POAGNet, to detect POAG from optic disc photographs. The POAGNet, an algorithm for glaucoma diagnosis, is developed using optic disc photographs. The POAGNet was trained and evaluated on 2 data sets: (1) 37 339 optic disc photographs from 1636 Ocular Hypertension Treatment Study (OHTS) participants and (2) 3684 optic disc photographs from the Sequential fundus Images for Glaucoma (SIG) data set. Gold standard labels were obtained using reading center grades. We proposed a Siamese network model, POAGNet, to simulate the clinical process of identifying POAG from optic disc photographs. The POAGNet consists of 2 side outputs for deep supervision and uses convolution to measure the similarity between 2 networks. The main outcome measures are the area under the receiver operating characteristic curve, accuracy, sensitivity, and specificity. In POAG diagnosis, extensive experiments show that POAGNet performed better than the best state-of-the-art model on the OHTS test set (area under the curve [AUC] 0.9587 versus 0.8750). It also outperformed the baseline models on the SIG test set (AUC 0.7518 versus 0.6434). To assess the transferability of POAGNet, we also validated the impact of cross-data set variability on our model. The model trained on OHTS achieved an AUC of 0.7490 on SIG, comparable to the previous model trained on the same data set. When using the combination of SIG and OHTS for training, our model achieved superior AUC to the single-data model (AUC 0.8165 versus 0.7518). These demonstrate the relative generalizability of POAGNet. By simulating the clinical grading process, POAGNet demonstrated high accuracy in POAG diagnosis. These results highlight the potential of deep learning to assist and enhance clinical POAG diagnosis. The POAGNet is publicly available on https://github.com/bionlplab/poagnet.


Venous thrombosis: understanding the paradoxes of recurrence.
The decision to continue anticoagulant treatment in patients with a first venous thrombosis after the initial treatment period has strong, life-long implications. Both the risk of recurrence when treatment is stopped and the risk of bleeding when it is continued are high and will persist over a patient's lifetime. For clinicians, rational strategies to stratify their patients into levels of risk of recurrence are limited. To support in the decision to continue or not, it is of the utmost importance to understand why some people develop a second event and others do not and how these people can be identified. This is not easy as, contrary to intuition, the risk profile of a recurrent event is entirely different from that of a first: Some genetic factors that have a major effect on first thrombosis only marginally predict recurrence, while, for instance, the opposite is true for male sex. These paradoxes can be explained when we understand etiology of a first event, how rates for first and second event cannot be directly compared, and how fixed risk factors cannot be predictors, while factors that are not causes can yet be predictors. Integrating all knowledge and combining the best predicting variables will ultimately lead to ways to estimate an individual's recurrence risk and hence to decide on optimal further treatment.


Combination treatment with Fructus Ligustri Lucidi and Puerariae radix offsets their independent actions on bone and mineral metabolism in ovariectomized rats.
The aim of this study was to evaluate the efficacy of Fructus Ligustri Lucidi (FLL) and Puerariae radix (PR) combination treatment in bone and mineral metabolism in ovariectomized (OVX) rats in our search for an alternative regimen for the management of postmenopausal osteoporosis. Six-month-old OVX rats were used as postmenopausal osteoporotic models, and PR water extract (PR) and FLL water extract (WE) were added to commercial diets individually or in combination and administered to OVX rats for 12 weeks. Bone properties, calcium and phosphorus absorption, and bone biochemical markers were measured to investigate the potential interactions between the actions of PR and the actions of WE on bone and mineral metabolism in OVX rats. Long-term treatment with PR did not significantly improve bone properties but greatly ameliorated the secondary hyperparathyroidism induced by ovariectomy in the animals. WE significantly enhanced the intestinal calcium absorption rate and decreased the enlarged trabecular bone surface at the site of metaphysic tibia in OVX rats. However, the positive effects of WE or PR alone on bone and mineral metabolism were diminished when OVX rats were cotreated with WE and PR. The combination of these two herbs offsets their independent actions on bone and mineral metabolism in vivo. The results of the present study could provide insights to medical professionals to further their understanding of the potential negative impact of herb-herb interactions when a combination of herbal mixtures is used for the management of osteoporosis.


Mitochondrial-Based Therapeutics for the Treatment of Spinal Cord Injury: Mitochondrial Biogenesis as a Potential Pharmacological Target.
Spinal cord injury (SCI) is characterized by an initial trauma followed by a progressive cascade of damage referred to as secondary injury. A hallmark of secondary injury is vascular disruption leading to vasoconstriction and decreased oxygen delivery, which directly reduces the ability of mitochondria to maintain homeostasis and leads to loss of ATP-dependent cellular functions, calcium overload, excitotoxicity, and oxidative stress, further exacerbating injury. Restoration of mitochondria dysfunction during the acute phases of secondary injury after SCI represents a potentially effective therapeutic strategy. This review discusses the past and present pharmacological options for the treatment of SCI as well as current research on mitochondria-targeted approaches. Increased antioxidant activity, inhibition of the mitochondrial permeability transition, alternate energy sources, and manipulation of mitochondrial morphology are among the strategies under investigation. Unfortunately, many of these tactics address single aspects of mitochondrial dysfunction, ultimately proving largely ineffective. Therefore, this review also examines the unexplored therapeutic efficacy of pharmacological enhancement of mitochondrial biogenesis, which has the potential to more comprehensively improve mitochondrial function after SCI.


Evolution of amino acid metabolism inferred through cladistic analysis.
Because free amino acids were most probably available in primitive abiotic environments, their metabolism is likely to have provided some of the very first metabolic pathways of life. What were the first enzymatic reactions to emerge? A cladistic analysis of metabolic pathways of the 16 aliphatic amino acids and 2 portions of the Krebs cycle was performed using four criteria of homology. The analysis is not based on sequence comparisons but, rather, on coding similarities in enzyme properties. The properties used are shared specific enzymatic activity, shared enzymatic function without substrate specificity, shared coenzymes, and shared functional family. The tree shows that the earliest pathways to emerge are not portions of the Krebs cycle but metabolisms of aspartate, asparagine, glutamate, and glutamine. The views of Horowitz (Horowitz, N. H. (1945) Proc. Natl. Acad. Sci. U. S. A. 31, 153-157) and Cordón (Cordón, F. (1990) Tratado Evolucionista de Biologia, Aguilar, Madrid, Spain), according to which the upstream reactions in the catabolic pathways and the downstream reactions in the anabolic pathways are the earliest in evolution, are globally corroborated; however, with some exceptions. These are due to later opportunistic connections of pathways (actually already suggested by these authors). Earliest enzymatic functions are mostly catabolic; they were deaminations, transaminations, and decarboxylations. From the consensus tree we extracted four time spans for amino acid metabolism development. For some amino acids catabolism and biosynthesis occurred at the same time (Asp, Glu, Lys, Leu, Ala, Val, Ile, Pro, Arg). For others ultimate reactions that use amino acids as a substrate or as a product are distinct in time, with catabolism preceding anabolism for Asn, Gln, and Cys and anabolism preceding catabolism for Ser, Met, and Thr. Cladistic analysis of the structure of biochemical pathways makes hypotheses in biochemical evolution explicit and parsimonious.


Formation of indigo by recombinant mammalian cytochrome P450.
The development of bicistronic systems for coexpression of recombinant human cytochrome P450 enzymes (P450s) with their redox partner, NADPH-cytochrome P450 reductase (NPR), has enabled P450 activity to be reconstituted within bacterial cells. During expression of recombinant P450 2E1 and some other forms, we observed the formation of a blue pigment in bacterial cultures. The pigment was extracted from cultures and shown to comigrate with standard indigo on TLC. UV-visible spectroscopy and mass spectrometric analysis provided further support for identification of the pigment as indigo. Indigo is known to form following the spontaneous oxidation of 3-hydroxyindole. Accordingly, we speculated that indole, formed as a breakdown product of tryptophan in bacteria, was hydroxylated by the P450 system, leading to indigo formation. Bacterial membranes containing recombinant P450 2E1 and human NPR were incubated in vitro with indole and shown to catalyze formation of a blue pigment in a time- and cofactor-dependent manner. These studies suggest potential applications of mammalian P450 enzymes in industrial indigo production or in the development of novel colorimetric assays based on indole hydroxylation.


The Alaska Haemophilus influenzae type b experience: lessons in controlling a vaccine-preventable disease.
Before 1991, Alaska Native children experienced one of the highest rates of invasive Haemophilus influenzae type b disease. H influenzae type b vaccine has led to a near-elimination of invasive H influenzae type b disease in the United States. We describe challenges encountered in controlling H influenzae type b disease in Alaska and update the current status of H influenzae disease and carriage in Alaska as lessons to other populations. We reviewed data from statewide H influenzae disease surveillance conducted during 1980-2004. Vaccine coverage data were based on audits from tribal facilities and the National Immunization Survey. H influenzae type b colonization data were based on 6 carriage studies. After universal infant vaccination in 1991, H influenzae type b disease among Alaska Native and non-Native children < 5 years of age decreased by 94% and 96%, respectively. After a 1996 change in H influenzae type b vaccine from polyribosylribitol phosphate-outer membrane protein conjugate vaccine to H influenzae type b oligosaccharide-CRM197 vaccine, the incidence of H influenzae type b disease increased in rural Alaska Natives from 19.8 to 91.1 cases per 100000 per year < 5 years of age. During 2001-2004, with use of polyribosylribitol phosphate-outer membrane protein conjugate vaccine, the rate of H influenzae type b disease in Alaska Native and non-Native children aged < 5 years decreased to 5.4 and 0 per 100000 per year, respectively. In postvaccine studies, H influenzae type b carriage has decreased in Alaska Native children < 5 years of age. H influenzae type b vaccination has resulted in a dramatic decrease in invasive H influenzae type b disease in Alaska; however, despite high rates of H influenzae type b vaccine coverage, H influenzae type b disease rates among rural Alaska Native children < 5 years of age remain higher than the rates among non-Native Alaska and other US children. Equity in disease rates may not be achieved in indigenous populations with the current vaccines unless other environmental and household factors contributing to disease transmission are addressed.


Pharmacokinetics of metformin during pregnancy.
Our objective was to evaluate the pharmacokinetics of metformin during pregnancy. Serial blood and urine samples were collected over one steady-state dosing interval in women treated with metformin during early to late pregnancy (n = 35) and postpartum (n = 16). Maternal and umbilical cord blood samples were obtained at delivery from 12 women. Metformin concentrations were also determined in breast milk samples obtained over one dosing interval in 6 women. Metformin renal clearance increased significantly in mid (723 +/- 243 ml/min, P < 0.01) and late pregnancy (625 +/- 130 ml/min, P < 0.01) compared with postpartum (477 +/- 132 ml/min). These changes reflected significant increases in creatinine clearance (240 +/- 70 ml/min, P < 0.01 and 207 +/- 56 ml/min, P < 0.05 versus 165 +/- 44 ml/min) and in metformin net secretion clearance (480 +/- 190 ml/min, P < 0.01 and 419 +/- 78 ml/min, P < 0.01 versus 313 +/- 98 ml/min) in mid and late pregnancy versus postpartum, respectively. Metformin concentrations at the time of delivery in umbilical cord plasma ranged between nondetectable (<5 ng/ml) and 1263 ng/ml. The daily infant intake of metformin through breast milk was 0.13 to 0.28 mg, and the relative infant dose was <0.5% of the mother's weight-adjusted dose. Our results indicate that metformin pharmacokinetics are affected by pregnancy-related changes in renal filtration and net tubular transport and can be roughly estimated by the use of creatinine clearance. At the time of delivery, the fetus is exposed to metformin concentrations from negligible to as high as maternal concentrations. In contrast, infant exposure to metformin through the breast milk is low.


jMorp: Japanese Multi-Omics Reference Panel update report 2023.
Modern medicine is increasingly focused on personalized medicine, and multi-omics data is crucial in understanding biological phenomena and disease mechanisms. Each ethnic group has its unique genetic background with specific genomic variations influencing disease risk and drug response. Therefore, multi-omics data from specific ethnic populations are essential for the effective implementation of personalized medicine. Various prospective cohort studies, such as the UK Biobank, All of Us and Lifelines, have been conducted worldwide. The Tohoku Medical Megabank project was initiated after the Great East Japan Earthquake in 2011. It collects biological specimens and conducts genome and omics analyses to build a basis for personalized medicine. Summary statistical data from these analyses are available in the jMorp web database (https://jmorp.megabank.tohoku.ac.jp), which provides a multidimensional approach to the diversity of the Japanese population. jMorp was launched in 2015 as a public database for plasma metabolome and proteome analyses and has been continuously updated. The current update will significantly expand the scale of the data (metabolome, genome, transcriptome, and metagenome). In addition, the user interface and backend server implementations were rewritten to improve the connectivity between the items stored in jMorp. This paper provides an overview of the new version of the jMorp.


The Nordic Maintenance Care Program: Does psychological profile modify the treatment effect of a preventive manual therapy intervention? A secondary analysis of a pragmatic randomized controlled trial.
Chiropractic maintenance care is effective as secondary/tertiary prevention of non-specific low back pain (LBP), but the potential effect moderation by psychological characteristics is unknown. The objective was to investigate whether patients in specific psychological sub-groups had different responses to MC with regard to the total number of days with bothersome pain and the number of treatments. Data from a two-arm randomized pragmatic multicenter trial with a 12-month follow up, designed to investigate the effectiveness of maintenance care, was used. Consecutive patients, 18-65 years of age, with recurrent and persistent LBP seeking chiropractic care with a good effect of the initial treatment were included. Eligible subjects were randomized to either maintenance care (prescheduled care) or to the control intervention, symptom-guided care. The primary outcome of the trial was the total number of days with bothersome LBP collected weekly for 12 months using an automated SMS system. Data used to classify patients according to psychological subgroups defined by the West Haven-Yale Multidimensional Pain Inventory (adaptive copers, interpersonally distressed and dysfunctional) were collected at the screening visit. A total of 252 subjects were analyzed using a generalized estimating equations linear regression framework. Patients in the dysfunctional subgroup who received maintenance care reported fewer days with pain (-30.0; 95% CI: -36.6, -23.4) and equal number of treatments compared to the control intervention. In the adaptive coper subgroup, patients who received maintenance care reported more days with pain (10.7; 95% CI: 4.0, 17.5) and more treatments (3.9; 95% CI: 3.5, 4.2). Patients in the interpersonally distressed subgroup reported equal number of days with pain (-0.3; 95% CI: -8.7, 8.1) and more treatments (1.5; 95% CI: 0.9, 2.1) on maintenance care. Psychological and behavioral characteristics modify the effect of MC and should be considered when recommending long-term preventive management of patients with recurrent and persistent LBP.


Usefulness of the bleeding time to predict the risk of clinical bleeding in patients with uraemia.
In a prospective study of haemostatic status of 15 uraemic Nigerians a highly significant prolongation of bleeding time (p less than 0.001) and significantly lower haematocrit levels (p less than 0.001) were detected in comparison to healthy controls. A highly significant positive correlation (r = +0.778) between prolongation of bleeding time and blood urea was present. Six patients had overt clinical bleeding with significantly prolonged bleeding time (p less than 0.01) compared to uraemics without overt bleeding. Blood urea was significantly different in both groups (p greater than 0.1). Platelet count, prothrombin time, partial thromboplastin time and Hess tourniquet test did not differ between the uraemic patients and controls. Haemostatic dysfunction has been noted in Nigerian uraemics we studied. Bleeding time is a useful but simple means of assessing such dysfunction.


Association of nascent polypeptide and transfer ribonucleic acid with 30S ribosomal subunits.
1. Crude extracts of Escherichia coli programmed in protein synthesis by endogenous mRNA have incorporated amino acids into protein. Analysis of such extracts by sucrose-gradient centrifugation in low Mg(2+) concentration has revealed that 30S ribosomal subunits carry associated radioactive material of which a considerable proportion can be removed from ribosomes by treatment of pre-labelled extracts with puromycin. 2. Gradient analyses of incorporations carried out in the additional presence of added (32)P-labelled tRNA have indicated that tRNA sediments in the regions of the newly synthesized nascent protein and that both labels are associated with all ribosomal components detected on the gradients under the experimental conditions employed. 3. 30S ribosomal subunits carrying both (32)P and (14)C labels have been isolated, disrupted with sodium dodecyl sulphate, and analysed by chromatography on Sephadex G-200 columns. Both labels elute closely together and well away from a tRNA marker analysed under identical conditions. 4. It is proposed that 30S ribosomal subunits, isolated from extracts which have synthesized nascent peptides under the direction of endogenous mRNA, carry associated peptidyl-tRNA.


Deep Convolutional Neural Networks Help Scoring Tandem Mass Spectrometry Data in Database-Searching Approaches.
Spectrum annotation is a challenging task due to the presence of unexpected peptide fragmentation ions as well as the inaccuracy of the detectors of the spectrometers. We present a deep convolutional neural network, called Slider, which learns an optimal feature extraction in its kernels for scoring mass spectrometry (MS)/MS spectra to increase the number of spectrum annotations with high confidence. Experimental results using publicly available data sets show that Slider can annotate slightly more spectra than the state-of-the-art methods (BoltzMatch, Res-EV, Prosit), albeit 2-10 times faster. More interestingly, Slider provides only 2-4% fewer spectrum annotations with low-resolution fragmentation information than other methods with high-resolution information. This means that Slider can exploit nearly as much information from the context of low-resolution spectrum peaks as the high-resolution fragmentation information can provide for other scoring methods. Thus, Slider can be an optimal choice for practitioners using old spectrometers with low-resolution detectors.


[Diagnostic significance of the type of in vitro interaction between blood lymphocytes and serotonin in multiple sclerosis].
The method for laboratory diagnosis of multiple sclerosis (MS)--the loading serotonin test in vitro based on a specific type of interaction between MS patient's blood lymphocytes and serotonin--is suggested. An examination of 56 MS patients, 12 patients with vertebrale nervous system pathology, 33 patients with MS phenocopies and 19 healthy subjects revealed an initial low serotonin rosette-forming cell (RFC) content in MS patients, comparing to all the subjects studied. In contrast to healthy subjects and patients with other neurological disorders, serotonin receptor lymphocyte expression in MS patients does not change during serotonin stimulation in vitro and serotonin RFC number does not increase. The type of interaction is specific and selective, being confirmed in reaction of rosette-formation by serotonin excess and by the results of rosette-formation reaction with other biologically active components (histamine, nor adrenaline). A serotonin reception inhibition, specific for MS, can be determined already in the disease debut, regardless of patient's gender and age, clinical type, disease stage and duration, which proves to be of most importance. The loading serotonin test indexes are significant for differential diagnosis. The method patented in 1999 is realized in vitro, excludes a double blood collection that reduces allergy and infection danger for patients is easy to make and cost-effective.


Electromyography activation of shoulder and trunk muscles is greater during closed chain compared to open chain exercises.
To compare the activation of shoulder and trunk muscles between six pairs of closed (CC) and open chain (OC) exercises for the upper extremity, matched for performance characteristics. The secondary aims were to compare shoulder and trunk muscle activation and shoulder activation ratios during each pair of CC and OC exercise. Twenty-two healthy young adults were recruited. During visit 1, the 5-repetition maximum resistance was established for each CC and OC exercise. During visit 2, electromyography activation from the infraspinatus (INF), deltoid (DEL), serratus anterior (SA), upper, middle and lower trapezius (UT, MT, LT), erector spinae (ES) and external oblique (EO) muscles was collected during 5-repetition max of each exercise. Average activation was calculated during the concentric and eccentric phases of each exercises. Activation ratios (DEL/INF, UT/LT, UT/MT, UT/SA) were also calculated. Linear mixed models compared the activation by muscle collapsed across CC and OC exercises. A paired t-test compared the activation of each muscle and the activation ratios (DEL/INF, UT/LT, UT/MT, UT/SA) between each pair of CC and OC exercises. The INF, LT, ES, and EO had greater activation during both concentric (p = 0.03) and eccentric (p < 0.01) phases of CC versus OC exercises. Activation ratios were lower in CC exercises compared to OC exercises (DEL/INF, 3 pairs; UT/LT, 2 pairs; UT/MT, 1 pair; UT/SA, 3 pairs). Upper extremity CC exercises generated greater activation of shoulder and trunk muscles compared to OC exercises. Some of the CC exercises produced lower activation ratios compared to OC exercises.


Associations between family factors and premarital heterosexual relationships among female college students in Tehran.
Although premarital heterosexual relationships, especially those involving sexual contact, are discouraged in Iran, particularly for females, a considerable minority of young people are involved in such relationships. However, the determinants of such relationships have not been identified, especially those pertaining to family. In 2005-2006, a random sample of 1,378 unmarried female college students from four universities in Tehran completed anonymous, self-administered surveys that asked whether the respondent had ever had a premarital heterosexual relationship. Bivariate and multivariate regression analyses were conducted to examine associations between such relationships and family factors, including socioeconomic measures, parent-child communication and closeness, family values and atmosphere, and parental control. Having had a boyfriend was positively associated with paternal income (odds ratio, 1.3), maternal educational attainment (1.3) and more liberal family values (1.3), and negatively associated with parent-child closeness (0.6). Very strict and very relaxed parental control during adolescence were both associated with having had a boyfriend, but only the former was associated with having had premarital sex. In addition, respondents were more likely to have had premarital intercourse if they did not live with both parents (2.0) or if their family had more liberal values (1.3); they had reduced odds of having had sex if they had a closer relationship with their parents (0.7). Good family relationships may reduce the likelihood that youth will engage in premarital sex, possibly by fostering parent-child closeness. Moderate parental control may discourage premarital relationships more effectively than lesser or greater degrees of control.


Mutation extraction tools can be combined for robust recognition of genetic variants in the literature.
As the cost of genomic sequencing continues to fall, the amount of data being collected and studied for the purpose of understanding the genetic basis of disease is increasing dramatically. Much of the source information relevant to such efforts is available only from unstructured sources such as the scientific literature, and significant resources are expended in manually curating and structuring the information in the literature. As such, there have been a number of systems developed to target automatic extraction of mutations and other genetic variation from the literature using text mining tools. We have performed a broad survey of the existing publicly available tools for extraction of genetic variants from the scientific literature. We consider not just one tool but a number of different tools, individually and in combination, and apply the tools in two scenarios. First, they are compared in an intrinsic evaluation context, where the tools are tested for their ability to identify specific mentions of genetic variants in a corpus of manually annotated papers, the Variome corpus. Second, they are compared in an extrinsic evaluation context based on our previous study of text mining support for curation of the COSMIC and InSiGHT databases. Our results demonstrate that no single tool covers the full range of genetic variants mentioned in the literature. Rather, several tools have complementary coverage and can be used together effectively. In the intrinsic evaluation on the Variome corpus, the combined performance is above 0.95 in F-measure, while in the extrinsic evaluation the combined recall performance is above 0.71 for COSMIC and above 0.62 for InSiGHT, a substantial improvement over the performance of any individual tool. Based on the analysis of these results, we suggest several directions for the improvement of text mining tools for genetic variant extraction from the literature.


Aeroallergen sensitization patterns among patients with chronic rhinitis with or without concomitant asthma.
The aim of this study was to explore the differences in the pattern of allergen sensitization in CR individuals without or with asthma, according to asthma severity. A total of 1066 adults were evaluated. Asthma and chronic⁄allergic rhinits were identified by specialists, questionnaries and skin-prick test. The phenotypic characterization was avaliable from skin-prick test to an aeroallergen extended panel, total IgE and pulmonary function. Using questionnaires and clinical evaluation, participants were classified into the groups: chronic rhinitis alone (CRA) and chronic rhinitis + asthma, the latter subdivided into CR + mild asthma (CRMA) and CR + moderate to severe asthma (CRMSA). Aerollergen sensitization was defined by a positive prick test to one or more allergens associated with nasal symptoms and/or asthma. The association between CR and asthma was evaluated by multivariable logistic regression. The evidence of effect modification of pattern of sensitization in CR on the association with asthma severity and outcomes was examined by introducing interactions terms in the logistic regression models adjusting for confounders. Frequency of sensitization to aeroallergens was higher in association with asthma in comparison to CRA (CRMA 70.4%; CRMSA 65.0%; CRA 47.0%; p = 0.000). Similarly, the presence of asthma was associated to aeroallergen multiple sensitization (51.5%) (OR = 2.10, 95% CI 1.27-3.50). Additionally, the sensitization to mites, cockroaches, animal epithelium, grasses, and molds, were higher in asthma (56.8%, 24.3%, 12%, 7.13% and 10.3%, respectively). Sensitization to Alternaria alternata, Cladosporium herbarum and dog epithelium was exclusive in asthma groups. A concomitant asthma diagnosis was directly associated with a positive allergen sensitization at least one allergen (62.7%, OR = 2.45, 95% CI 1.80-3.34) and polissensitization (51.5%, OR = 2.10, 95% CI 1.27-3.50). Asthma is associated with multiple allergen sensitization among patients with CR. Some unique profiles of aeroallergen sensitization were observed in patients with CR and asthma. Nevertheless, no difference was found in the sensitization in relation to asthma severity, which suggest atopy is not the main underlying mechanism for asthma severity among patients with CR. Level 3.


Distinct cord blood C-peptide, adipokine, and lipidomic signatures by in utero HIV exposure.
Early-life metabolic derangements in HIV-exposed uninfected (HEU) infants have been reported. Pregnant women with HIV and HIV-uninfected pregnant women were enrolled with their newborns in a US cohort from 2011 to 2015. We measured cord insulin, C-peptide, and metabolic cytokines of HEU and HIV-unexposed uninfected (HUU) newborns using ELISA and metabolites, lipid subspecies, and eicosanoids via liquid chromatography/mass spectrometry. Linear regression was employed to assess the association of intrauterine HIV/ART with insulin and C-peptide. Graphical lasso regression was used to identify differences between metabolite/lipid subspecies networks associated with C-peptide. Of 118 infants, 56 were HEU, ART exposed. In adjusted analyses, mean cord insulin (β = 0.295, p = 0.03) and C-peptide (β = 0.522, p < 0.01) were significantly higher in HEU vs. HUU newborns. HEU neonates exhibited primarily positive associations between complex lipids and C-peptide, indicative of fuel storage, and augmented associations between cord eicosanoids and cytokines. HUU neonates exhibited negative associations with lipids and C-peptide indicative of increased fuel utilization. Higher cord insulin and C-peptide in HEU vs. HUU newborns as well as differences in cord metabolites, metabolic-related cytokines, and eicosanoids may reflect a propensity for fuel storage and an inflammatory milieu suggestive of fetal metabolic changes associated with in utero HIV/ART exposure. There is a paucity of studies assessing cord blood and neonatal metabolic health in HIV-exposed uninfected (HEU) newborns, an increasing population worldwide. Compared to HIV-unexposed uninfected (HUU) newborns, HEU newborns exhibit alterations in fuel homeostasis and an inflammatory milieu associated with in utero HIV/antiretroviral therapy (ART) exposure. The long-term implications of these neonatal findings are as yet unknown, but merit continued evaluation as this important and growing population ages into adulthood.


Investigating the importance of sediment resuspension in Alexandrium fundyense cyst population dynamics in the Gulf of Maine.
Cysts of Alexandrium fundyense, a dinoflagellate that causes toxic algal blooms in the Gulf of Maine, spend the winter as dormant cells in the upper layer of bottom sediment or the bottom nepheloid layer and germinate in spring to initiate new blooms. Erosion measurements were made on sediment cores collected at seven stations in the Gulf of Maine in the autumn of 2011 to explore if resuspension (by waves and currents) could change the distribution of over-wintering cysts from patterns observed in the previous autumn; or if resuspension could contribute cysts to the water column during spring when cysts are viable. The mass of sediment eroded from the core surface at 0.4 Pa ranged from 0.05 kg m-2 near Grand Manan Island, to 0.35 kg m-2 in northern Wilkinson Basin. The depth of sediment eroded ranged from about 0.05 mm at a station with sandy sediment at 70 m water depth on the western Maine shelf, to about 1.2 mm in clayey-silt sediment at 250 m water depth in northern Wilkinson Basin. The sediment erodibility measurements were used in a sediment-transport model forced with modeled waves and currents for the period October 1, 2010 to May 31, 2011 to predict resuspension and bed erosion. The simulated spatial distribution and variation of bottom shear stress was controlled by the strength of the semi-diurnal tidal currents, which decrease from east to west along the Maine coast, and oscillatory wave-induced currents, which are strongest in shallow water. Simulations showed occasional sediment resuspension along the central and western Maine coast associated with storms, steady resuspension on the eastern Maine shelf and in the Bay of Fundy associated with tidal currents, no resuspension in northern Wilkinson Basin, and very small resuspension in western Jordan Basin. The sediment response in the model depended primarily on the profile of sediment erodibility, strength and time history of bottom stress, consolidation time scale, and the current in the water column. Based on analysis of wave data from offshore buoys from 1996 to 2012, the number of wave events inducing a bottom shear stress large enough to resuspend sediment at 80 m ranged from 0 to 2 in spring (April and May) and 0 to 10 in winter (October through March). Wave-induced resuspension is unlikely in water greater than about 100 m deep. The observations and model results suggest that a millimeter or so of sediment and associated cysts may be mobilized in both winter and spring, and that the frequency of resuspension will vary interannually. Depending on cyst concentration in the sediment and the vertical distribution in the water column, these events could result in a concentration in the water column of at least 104 cysts m-3. In some years, resuspension events could episodically introduce cysts into the water column in spring, where germination is likely to be facilitated at the time of bloom formation. An assessment of the quantitative effects of cyst resuspension on bloom dynamics in any particular year requires more detailed investigation.


Seviteronel, a Novel CYP17 Lyase Inhibitor and Androgen Receptor Antagonist, Radiosensitizes AR-Positive Triple Negative Breast Cancer Cells.
Increased rates of locoregional recurrence (LR) have been observed in triple negative breast cancer (TNBC) despite multimodality therapy, including radiation (RT). Recent data suggest inhibiting the androgen receptor (AR) may be an effective radiosensitizing strategy, and AR is expressed in 15-35% of TNBC tumors. The aim of this study was to determine whether seviteronel (INO-464), a novel CYP17 lyase inhibitor and AR antagonist, is able to radiosensitize AR-positive (AR+) TNBC models. In cell viability assays, seviteronel and enzalutamide exhibited limited effect as a single agent (IC50 > 10 μM). Using clonogenic survival assays, however, AR knockdown and AR inhibition with seviteronel were effective at radiosensitizing cells with radiation enhancement ratios of 1.20-1.89 in models of TNBC with high AR expression. AR-negative (AR-) models, regardless of their estrogen receptor expression, were not radiosensitized with seviteronel treatment at concentrations up to 5 μM. Radiosensitization of AR+ TNBC models was at least partially dependent on impaired dsDNA break repair with significant delays in repair at 6, 16, and 24 h as measured by immunofluorescent staining of γH2AX foci. Similar effects were observed in an in vivo AR+ TNBC xenograft model where there was a significant reduction in tumor volume and a delay to tumor doubling and tripling times in mice treated with seviteronel and radiation. Following combination treatment with seviteronel and radiation, increased binding of AR occurred at DNA damage response genes, including genes involved both in homologous recombination and non-homologous end joining. This trend was not observed with combination treatment of enzalutamide and RT, suggesting that seviteronel may have a different mechanism of radiosensitization compared to other AR inhibitors. Enzalutamide and seviteronel treatment also had different effects on AR and AR target genes as measured by immunoblot and qPCR. These results implicate AR as a mediator of radioresistance in AR+ TNBC models and support the use of seviteronel as a radiosensitizing agent in AR+ TNBC.


Kinetic analysis of [11C]befloxatone in the human brain, a selective radioligand to image monoamine oxidase A.
[11C]Befloxatone measures the density of the enzyme monoamine oxidase A (MAO-A) in the brain. MAO-A is responsible for the degradation of different neurotransmitters and is implicated in several neurologic and psychiatric illnesses. This study sought to estimate the distribution volume (VT) values of [11C]befloxatone in humans using an arterial input function. Seven healthy volunteers were imaged with positron emission tomography (PET) after [11C]befloxatone injection. Kinetic analysis was performed using an arterial input function in association with compartmental modeling and with the Logan plot, multilinear analysis (MA1), and standard spectral analysis (SA) at both the regional and voxel level. Arterialized venous samples were drawn as an alternative and less invasive input function. An unconstrained two-compartment model reliably quantified VT values in large brain regions. A constrained model did not significantly improve VT identifiability. Similar VT results were obtained using SA; however, the Logan plot and MA1 slightly underestimated VT values (about -10%). At the voxel level, SA showed a very small bias (+2%) compared to compartmental modeling, Logan severely underestimated VT values, and voxel-wise images obtained with MA1 were too noisy to be reliably quantified. Arterialized venous blood samples did not provide a satisfactory alternative input function as the Logan-VT regional values were not comparable to those obtained with arterial sampling in all subjects. Binding of [11C]befloxatone to MAO-A can be quantified using an arterial input function and a two-compartment model or, in parametric images, with SA.


PKCdelta associates with and is involved in the phosphorylation of RasGRP3 in response to phorbol esters.
RasGRP is a family of guanine nucleotide exchange factors that activate small GTPases and contain a C1 domain similar to the one present in protein kinase C (PKC). In this study, we examined the interaction of RasGRP3 and PKC in response to the phorbol ester PMA. In Chinese hamster ovary or LN-229 cells heterologously expressing RasGRP3, phorbol 12-myristate 13-acetate (PMA) induced translocation of RasGRP3 to the perinuclear region and a decrease in the electrophoretic mobility of RasGRP3. The mobility shift was associated with phosphorylation of RasGRP3 on serine residues and seemed to be PKCdelta-dependent because it was blocked by the PKCdelta inhibitor rottlerin as well as by a PKCdelta kinase-dead mutant. Using coimmunoprecipitation, we found that PMA induced the physical association of RasGRP3 with PKCdelta and, using in situ methods, we showed colocalization of PKCdelta and RasGRP3 in the perinuclear region. PKCdelta phosphorylated RasGRP3 in vitro. Previous studies suggest that ectopic expression of RasGRP3 increases activation of Erk1/2. We found that overexpression of either PKCdelta or RasGRP3 increased the activation of Erk1/2 by PMA. In contrast, coexpression of PKCdelta and RasGRP3 yielded a level of phosphorylation of Erk1/2 similar to that of control vector cells. Our results suggest that PKCdelta may act as an upstream kinase associating with and phosphorylating RasGRP3 in response to PMA. The interaction between RasGRP3 and PKCdelta points to the existence of complex cross-talk between various members of the phorbol ester receptors which can have important impact on major signal transduction pathways and cellular processes induced by phorbol esters or DAG


Intermediate in the O-O bond cleavage reaction of an extradiol dioxygenase.
The reactive oxy intermediate of the catalytic cycle of extradiol aromatic ring-cleaving dioxygenases is formed by binding the catecholic substrate and O2 in adjacent ligand positions of the active site metal [usually Fe(II)]. This intermediate and the following Fe(II)-alkylperoxo intermediate resulting from oxygen attack on the substrate have been previously characterized in a crystal of homoprotocatechuate 2,3-dioxygenase (HPCD). Here a subsequent intermediate in which the O-O bond is broken to yield a gem diol species is structurally characterized. This new intermediate is stabilized in the crystal by using the alternative substrate, 4-sulfonylcatechol, and the Glu323Leu variant of HPCD, which alters the crystal packing.


High level of transgene expression in cell cultures and in the mouse by replication-incompetent adenoviruses harboring modified VAI genes.
Replication-incompetent adenoviruses are currently used in gene therapy trials. Most of the work designed to increase the expression from these vectors concerns the modification of cis sequences of the foreign transcription unit, so as to improve the transcription level or the stability of the mRNA. In this report, we show that an alternative strategy based on the coexpression of modified VAI genes can efficiently increase gene expression both in cell cultures and in animals. The VAI RNA is synthesized mainly during the late phase of the adenovirus cycle and increases the translation of late adenovirus gene products by counteracting the effect of an interferon-induced kinase, the PKR. We have constructed several modified VAI genes in which the central domain was deleted or substituted by exogenous sequences. These modified VAI genes, or the native VAI gene, were cloned into the left part of adenovirus type 5 genomes harboring their own endogenous VAI gene. One of the resulting viruses (Ad-VAr) increased 12.5- to 502-fold the expression level of reporter genes, either expressed as a constitutive cell line from an extrachromosomal DNA or introduced into cells by coinfection with another adenovirus vector. This effect was independent of the promoter, the coding sequence, and the 5' untranslated mRNA sequence and was obvious in the two non-E1-complementing cell lines tested (HeLa and Vero). Coinfection of Ad-VAr with adenoviruses expressing the luciferase gene from the major late promoter or Rous sarcoma virus (RSV) promoter by the intravenous route in mice increased by more than 33 (MLP)- to 128 (RSV)- and 4,700 (MLP)- to 30,000 (RSV)-fold the expression level of the reporter gene in the lungs and liver, respectively. The intramuscular coinoculation of Ad-VAr and Ad-MLP-gD (a recombinant adenovirus vaccine expressing gD from the pseudorabies herpes virus) led to a 10-fold decrease in the protective dose of Ad-gD in mice. Ad-VAfull, a similar adenovirus in which the native VAI gene was cloned at the left part of the genome, showed no evidence of efficacy in cell culture and in mice. These results suggest that the use of modified VAI genes expressed at the early phase of the cycle can be helpful in the design of potent adenovirus vectors.


Compact broadband spectrometer based on upconversion and downconversion luminescence.
In this Letter, a compact spectrometer based on upconversion and downconversion luminescence for operation in the infrared, visible, and ultraviolet bands is presented. The proposed spectrometer has three components that are used for dispersion, frequency conversion, and detection. The conversion component converts the incident signal beam into a spectral window appropriate for the detection component. The detection component images the speckle pattern generated by scattering or diffraction in the random structure of the dispersion component. With the two-dimensional intensity data captured from both the speckle pattern and a calibration measurement process, one can reconstruct the spectra of the signal beam by solving a matrix equation. A smoothing simulated annealing algorithm has been implemented to improve the accuracy of the spectral reconstruction. We have analyzed possible sources of error in the algorithm and the corresponding limits of operation. The reported broadband, compact, high-resolution, luminescence-based spectrometer is well suited for portable spectroscopy applications.


Improvement of predictive models of risk of disease progression in chronic hepatitis C by incorporating longitudinal data.
Existing predictive models of risk of disease progression in chronic hepatitis C have limited accuracy. The aim of this study was to improve upon existing models by applying novel statistical methods that incorporate longitudinal data. Patients in the Hepatitis C Antiviral Long-term Treatment Against Cirrhosis trial were analyzed. Outcomes of interest were (1) fibrosis progression (increase of two or more Ishak stages) and (2) liver-related clinical outcomes (liver-related death, hepatic decompensation, hepatocellular carcinoma, liver transplant, or increase in Child-Turcotte-Pugh score to ≥7). Predictors included longitudinal clinical, laboratory, and histologic data. Models were constructed using logistic regression and two machine learning methods (random forest and boosting) to predict an outcome in the next 12 months. The control arm was used as the training data set (n = 349 clinical, n = 184 fibrosis) and the interferon arm, for internal validation. The area under the receiver operating characteristic curve for longitudinal models of fibrosis progression was 0.78 (95% confidence interval [CI] 0.74-0.83) using logistic regression, 0.79 (95% CI 0.77-0.81) using random forest, and 0.79 (95% CI 0.77-0.82) using boosting. The area under the receiver operating characteristic curve for longitudinal models of clinical progression was 0.79 (95% CI 0.77-0.82) using logistic regression, 0.86 (95% CI 0.85-0.87) using random forest, and 0.84 (95% CI 0.82-0.86) using boosting. Longitudinal models outperformed baseline models for both outcomes (P < 0.0001). Longitudinal machine learning models had negative predictive values of 94% for both outcomes. Prediction models that incorporate longitudinal data can capture nonlinear disease progression in chronic hepatitis C and thus outperform baseline models. Machine learning methods can capture complex relationships between predictors and outcomes, yielding more accurate predictions; our models can help target costly therapies to patients with the most urgent need, guide the intensity of clinical monitoring required, and provide prognostic information to patients.


I get paid fast; what about you?
The process of billing an insurance company for health care services has changed radically. In the past few years, the emphasis has been on automation. The change is fueled by the opinion of cost containment experts who claim that automation will help reduce costs in the U.S. health care delivery system. Key to success for the provider in adapting to this change will be understanding the coding used in the billing process and following standards of accuracy and fairness. This article is not intended to represent the adjudication rules of any particular insurance company. It is the result of experience as a practicing surgeon and as a consultant in the health care field.


Universal dental adhesives: Current status, laboratory testing, and clinical performance.
Increasing demand for simplified and user-friendly adhesive systems has led to the development of a new class of adhesives termed as Universal Adhesives (UAs). The term "Universal" reflects manufacturers' claims that these adhesives can be applied with any adhesion strategy and offer the versatility of use with a variety of direct and indirect restorative materials. The aim of this review was to synthesize the literature regarding the current status of UAs, their adhesion potential to various substrates and their performance in different restorative situations. In vitro studies, clinical trials and systematic reviews were identified utilizing controlled vocabulary and keyword searches in Medline and EMBASE databases. About 282 studies (272 in vitro studies; 11 clinical studies) were included. Available laboratory and clinical evidence does not support the claim that UAs can be used with any adhesive strategy. Although, they can chemically bond to various tooth and direct/indirect restorative substrates, the stability of this bond is material-dependent and subject to hydrolytic degradation. Hence, additional measures are still needed to ensure long-term durability. which undermines the versatility of UAs. The lack of long-term data regarding the clinical performance of UAs further complicates clinical decision-making. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2121-2131, 2019.


[Facial angioedema after filler injections. Description of five cases].
The use of fillers for cosmetic purposes is becoming increasingly frequent. Although initially considered inert, these products produce adverse reactions around the injection site. We present 5 cases of women with a history of filler injections who presented a hard and persistent angioedema followed by local subcutaneous nodules. They were referred to the allergist for suspected allergy related angioedema without response to usual antihistamine treatment. The angioedema episodes initiated 27.6 months (range 1 to 48) after the fillers treatment. The patients underwent exacerbations and remissions of angioedema, partially relieved with oral steroids and, in 2 cases, local triamcinolone injections. Mean time from onset of symptoms to remission of angioedema was 8.75 months (range 1 to 24). Until October 2009 four patients continued into remission after 24.5 months (range 7 to 36) free of symptoms. One patient continued with exacerbations 11 months after the initial symptoms. Fillers may cause angioedema as an adverse event and should be considered in the differential diagnosis of persistent angioedema. They are only sensitive to steroid treatment and in some steroid dependent cases they respond to ciclosporin. The frequency of angioedema after filler injections among patients with angioedema in the Unit of Asthma Allergy and Clinical Immunology was 0.5%.


Familial and herd-level associations with paratuberculosis enzyme-linked immunosorbent assay status in beef cattle.
A cross-sectional study was performed to determine the odds of having a positive paratuberculosis ELISA result if the dam was ELISA positive in Texas beef cattle, adjusted for individual and herd-level risk factors for seropositivity. Texas beef cattle (n = 2,621) were tested for paratuberculosis by using a commercial ELISA and microbiologic culture of feces for Mycobacterium avium subsp. paratuberculosis (MAP). Pedigree data were collected to identify dam-and sire-offspring pairs. Bayesian mixed-effects logistic regression was used to estimate the odds of seropositivity associated with age, dam ELISA status, sire ELISA status, herd size, herd history of clinical paratuberculosis, within-herd seroprevalence, within-herd fecal MAP prevalence, and within-herd fecal non-MAP Mycobacterium spp. prevalence. Herd of residence was included as a random effect to account for the correlation of observations within the same herd. Statistically probable associations were observed between ELISA status and herd fecal MAP prevalence [OR (odds ratio) 1.28 per 1% increase; P < 0.001] and herd seroprevalence (OR 1.21 per 1% increase; P < 0.001). The association with dam ELISA status was small (OR 1.35) and not highly probable (P = 0.69). Results indicate that use of dam ELISA status to make culling decisions in beef cattle may not improve the success of paratuberculosis control programs. Alternative strategies may be more effective for reducing the odds of seropositivity.


Nonparametric motion characterization using causal probabilistic models for video indexing and retrieval.
This paper describes an original approach for content-based video indexing and retrieval. We aim at providing a global interpretation of the dynamic content of video shots without any prior motion segmentation and without any use of dense optic flow fields. To this end, we exploit the spatio-temporal distribution, within a shot, of appropriate local motion-related measurements derived from the spatio-temporal derivatives of the intensity function. These distributions are then represented by causal Gibbs models. To be independent of camera movement, the motion-related measurements are computed in the image sequence generated by compensating the estimated dominant image motion in the original sequence. The statistical modeling framework considered makes the exact computation of the conditional likelihood of a video shot belonging to a given motion or more generally to an activity class feasible. This property allows us to develop a general statistical framework for video indexing and retrieval with query-by-example. We build a hierarchical structure of the processed video database according to motion content similarity. This results in a binary tree where each node is associated to an estimated causal Gibbs model. We consider a similarity measure inspired from Kullback-Leibler divergence. Then, retrieval with query-by-example is performed through this binary tree using the maximum a posteriori (MAP) criterion. We have obtained promising results on a set of various real image sequences.


Whole tobacco smoke extracts to model tobacco dependence in animals.
Smoking tobacco is highly addictive and a leading preventable cause of death. The main addictive constituent is nicotine; consequently it has been administered to laboratory animals to model tobacco dependence. Despite extensive use, this model might not best reflect the powerful nature of tobacco dependence because nicotine is a weak reinforcer, the pharmacology of smoke is complex and non-pharmacological factors have a critical role. These limitations have led researchers to expose animals to smoke via the inhalative route, or to administer aqueous smoke extracts to produce more representative models. The aim was to review the findings from molecular/behavioural studies comparing the effects of nicotine to tobacco/smoke extracts to determine whether the extracts produce a distinct model. Indeed, nicotine and tobacco extracts yielded differential effects, supporting the initiative to use extracts as a complement to nicotine. Of the behavioural tests, intravenous self-administration experiments most clearly revealed behavioural differences between nicotine and extracts. Thus, future applications for use of this behavioural model were proposed that could offer new insights into tobacco dependence.


Human rotavirus antigen detection by enzyme-immunoassay with antisera against Nebraska calf diarrhoea virus.
A four-layer solid phase enzyme-immunoassay (EIA) with antisera against Nebraska calf diarrhoea virus (NCDV) as immunoreagents was developed to detect human rotavirus antigens from stool specimens of patients with acute rotavirus gastroenteritis. Polystyrene beads were used as the solid phase, guinea-pig and rabbit anti-NCDV immunoglobulin as the catching and secondary antibody, and peroxidase-conjugated swine anti-rabbit immunoglobulin as the indicator antibody. A comparison of the developed NCDV-EIA with an identical EIA, using antisera against human rotavirus (HRV-EIA) instead of NCDV antisera, was made with 216 stool specimens positive or negative for rotavirus. A complete agreement was obtained between the two methods provided that appropriate confirmatory tests were included. The developed NCDV-EIA was as sensitive and specific for rotavirus as the HRV-EIA, and it allowed the detection of both established rotavirus types 1 and 2 from stools with equal sensitivity. The difficulties in cultivating human rotavirus in vitro for immunisation and the relative ease of growing NCDV in widely-used continuous cell lines make NCDV a good alternative in the preparation of the highly specific and sensitive rotavirus antisera required in immunoassays, and facilitate the setting-up methods for the routine diagnosis of rotavirus gastroenteritis by EIA or RIA in diagnostic virus laboratories.


Impact of an antimicrobial stewardship program on patients with acute bacterial skin and skin structure infections.
The impact of an antimicrobial stewardship program (ASP) on the management of therapy and hospital resources for patients with acute bacterial skin and skin structure infections (ABSSSIs) at a community teaching hospital was evaluated. A retrospective, observational chart review was performed to evaluate the impact of the ASP on patients admitted to Akron City Hospital with a diagnosis of ABSSSI between February 1 and August 20, 2012. Information on patient demographic characteristics, comorbidities, ABSSSI subtype, antibiotic therapy, microbiology, surgical interventions, and ASP recommendations was collected from medical records and the ASP intervention log. ASP recommendations were organized into five categories: dosage changes, de-escalation, antibiotic regimen change (i.e., change antibiotic regimen to a broad-spectrum antimicrobial or target a pathogen not being covered), infectious diseases (ID) formal consultation, and other. A total of 62 patients were included in the study. A total of 85 recommendations were made to attending physicians for these 62 patients, with an acceptance rate of 95%. The most common interventions included dosage changes, de-escalation, antibiotic regimen change, and ID consultation. When compared with historical data for 1149 patients, the intervention group had a significantly lower mean length of stay (LOS). The 30-day all-cause readmission rate was also significantly lower in the intervention group; however, the 30-day ABSSSI readmission rate did not differ significantly between groups. Interventions made by an ASP including a clinical pharmacist were associated with significant reductions in the mean LOS and 30-day all-cause readmission rate for patients with an ABSSSI compared with historical data.


Etiology of acute undifferentiated fever in patients presenting to the emergency department of a tertiary care center in Karachi, Pakistan.
Our study aimed at identifying the characteristics and etiology of various causes of acute undifferentiated fever in patients presenting to the emergency department of a tertiary care hospital. This was a retrospective study conducted at the department of emergency medicine, Aga Khan University Hospital from January to June 2016. Adult patients presenting to Emergency department with acute undifferentiated fever were enrolled. Descriptive statistics were calculated in terms of mean±SD for continuous variables like age of the patients and duration of fever, whereas frequency and percentage were computed for categorical variables like gender and causes of fever. A total of one hundred and fifty five patients were included. Out of these 97 (62.6%) were males and 58 (37.4%) were females. Most patients (25.2%, n= 39) were diagnosed as malaria followed closely by dengue fever (n=33, 21.3%) and then enteric fever (n= 10, 6.5%). while 41.9% (n=65) were diagnosed as suspected viral fever based on clinical judgment and inconclusive laboratory results. Malaria was found to be the most common confirmed cause of acute undifferentiated fever followed by dengue and enteric fever. The provision of accurate epidemiological data will enable resources to be directed towards key areas and will be of practical importance to clinicians.


New molecular targets for hepatocellular carcinoma: the ErbB1 signaling system.
Hepatocellular carcinoma (HCC) is a major cause of cancer-related deaths. This malignancy is often diagnosed at an advanced state, when most potentially curative therapies are of limited efficacy. In addition, HCC is a type of tumor highly resistant to available chemotherapeutic agents, which leaves HCC patients with no effective therapeutic options and a poor prognosis. From a molecular perspective, HCC is a heterogeneous type of tumor. However, in most cases, HCC emerges on a background of persistent liver injury, inflammation and hepatocellular proliferation, which is characteristic of chronic hepatitis and cirrhosis. Recent studies have revealed that dysregulation of a limited number of growth and survival-related pathways can play a key role in HCC development. The epidermal growth factor receptor (ErbB1) can be bound and activated by a broad family of ligands, and can also engage in extensive cross talk with other signaling pathways. This system is considered as an important defense mechanism for the liver during acute tissue injury; however, accumulating evidences suggest that its chronic stimulation can participate in the neoplastic conversion of the liver. Agents that target the ErbB1 receptor have shown antineoplastic activity in other types of tumors, but their efficacy either alone or in combination with other compounds has just started to be tested in experimental and human HCC. Here, we review the evidences that support the involvement of the ErbB1 in HCC development and that provide a rationale for ErbB1 targeting in HCC prevention and treatment.


Association between ozone and respiratory admissions among children and the elderly in Vancouver, Canada.
In this study, we examine the impact of ozone on daily respiratory admissions in both young children and the elderly in greater Vancouver, British Columbia. Study subjects included children less than 3 yr of age and adults 65 yr of age or over living in greater Vancouver who had acute hospital admissions for any respiratory diseases (ICD-9 codes 460-519) during the 13-yr period 1986-1998. Bidirectional case-crossover analysis was used to investigate associations between ambient ozone and respiratory hospitalizations after adjustment for other pollutants, including carbon monoxide, nitrogen dioxide, sulfur dioxide, and coefficient of haze. Potential effect modification by socioeconomic status as measured by household income was also examined. Respiratory admissions were associated with ozone levels 2, 3, 4, and 5 days prior to admission in both children and the elderly, with the strongest association observed at a lag of 4 days. Odds ratios for hospital admission of 1.22 (95% CI: 1.15-1.30) for children and 1.13 (1.09-1.18) for the elderly, respectively, were found, based on an increment in exposure corresponding to the interquartile range for ozone. Adjusting for other pollutants did not attenuate the ozone effect on respiratory admissions. Nor did socioeconomic status appear to modify the association between ozone and respiratory admissions in either children or the elderly. We concluded that ambient ozone is positively associated with respiratory hospital admission among young children and the elderly in Vancouver, British Columbia. These associations persisted after adjustment for both copollutant exposures and socioeconomic status.


Is High-Intensity Speech Intervention Better? A Comparison of High-Intensity Intervention Versus Low-Intensity Intervention in Children With a Cleft Palate.
Purpose The purpose of this study was to compare the effect of speech intervention provided with a low intensity with speech intervention provided with a high intensity on the speech and health-related quality of life (HRQoL) in Dutch-speaking children with a cleft palate with or without a cleft lip (CP ± L) between 4 and 12 years. Method A longitudinal, prospective, randomized controlled trial with a multiple baseline design was used. Twelve children with a CP ± L (M age = 8.0 years, SD = 1.54) were divided into two groups using block randomization stratified by age and gender: One group received low-intensity speech intervention (LISI; n = 6) and one group received high-intensity speech intervention (HISI; n = 6). Children in the LISI group received intervention with a session duration of 1 hr, a dose frequency of 1 session per week, and a total intervention duration of 10 weeks. Children in the HISI group received intervention with a session duration of 1 hr, a dose frequency of 5 sessions per week, and a total intervention duration of 2 weeks. The cumulative intervention intensity was kept constant. Both groups received identical therapy programs provided by the same experienced speech therapist. Perceptual speech assessments were performed on baseline and posttreatment data points. Changes in HRQoL were assessed using the Velopharyngeal Insufficiency Effects on Life Outcomes (VELO) questionnaire. Both groups were compared over time using (generalized) linear mixed models. Results No significant Time × Group interactions were observed for the percentage of correctly produced consonants at the word and sentence levels, indicating no differences in evolution over time among the two groups. The variables speech understandability, speech acceptability, and the total VELO scores significantly improved following HISI, but not following LISI. Conclusions Children in the HISI group made equal and, for some variables, even superior progress in only 2 weeks of therapy compared to children in the LISI group who received 10 weeks of therapy. HISI is a promising strategy to improve speech outcomes and HRQoL in a shorter time period.


Candy Wrapper for the Earth's inner core.
Recent global expansion of seismic data motivated a number of seismological studies of the Earth's inner core that proposed the existence of increasingly complex structure and anisotropy. In the meantime, new hypotheses of dynamic mechanisms have been put forward to interpret seismological results. Here, the nature of hemispherical dichotomy and anisotropy is re-investigated by bridging the observations of PKP(bc-df) differential travel-times with the iron bcc/hcp elastic properties computed from first-principles methods.The Candy Wrapper velocity model introduced here accounts for a dynamic picture of the inner core (i.e., the eastward drift of material), where different iron crystal shapes can be stabilized at the two hemispheres. We show that seismological data are best explained by a rather complicated, mosaic-like, structure of the inner core, where well-separated patches of different iron crystals compose the anisotropic western hemispherical region, and a conglomerate of almost indistinguishable iron phases builds-up the weakly anisotropic eastern side.


The debate continues on the best age to stop breastfeeding.
The ongoing debate on the optimal duration of breastfeeding in high-income countries is challenging. The decision to continue breastfeeding beyond the introduction of solids according to WHO indications involves multifaceted considerations, such as nutritional needs, psychological factors and cultural influences, given the growing number of immigrants. The impact on maternal health, sleep routines and family dynamics should be carefully weighed too. CONCLUSION: Next studies should investigate associations of different modalities and periods of breastfeeding with physical and neurodevelopmental outcomes, including psychological aspects. A personalised and gradual approach is suggested, guided by informed decisions and a supportive network.


Ultrastructural and temporal observations of the potyvirus cylindrical inclusions (Cls) show that the Cl protein acts transiently in aiding virus movement.
A systematic ultrastructural study across the edge of an advancing infection in pea seed-borne mosaic potyvirus-infected pea cotyledons showed the cylindrical inclusion (CI) protein to exist in transient functional states. Initially, the characteristic CI pinwheel inclusion bodies were positioned centrally over the plasmodesmal apertures (including those of plasmodesmata connected to the previously infected cell), in agreement with a proposed role in virus movement (Carrington et al., 1998, Plant J., 13, in press). The viral coat protein was associated with these structures and was seen within the modified plasmodesma, most notably in a continuous channel that passed along the axis of the pinwheel and through the plasmodesma. The CI protein was not detected within the plasmodesmal cavities. Later in the infection (i.e., behind the zone of active virus replication) the CI was no longer associated with cell walls, or with coat protein, and showed signs of structural degeneration. In contrast, the coat protein remained within plasmodesmal cavities. The role of the CI in assisting virus movement is not known but the presence of the CI was linked with an apparent transient reduction in callose in the vicinity of the plasmodesmata.


Neurobehavioral dynamics following chronic sleep restriction: dose-response effects of one night for recovery.
Establish the dose-response relationship between increasing sleep durations in a single night and recovery of neurobehavioral functions following chronic sleep restriction. Intent-to-treat design in which subjects were randomized to 1 of 6 recovery sleep doses (0, 2, 4, 6, 8, or 10 h TIB) for 1 night following 5 nights of sleep restriction to 4 h TIB. Twelve consecutive days in a controlled laboratory environment. N = 159 healthy adults (aged 22-45 y), median = 29 y). Following a week of home monitoring with actigraphy and 2 baseline nights of 10 h TIB, subjects were randomized to either sleep restriction to 4 h TIB per night for 5 nights followed by randomization to 1 of 6 nocturnal acute recovery sleep conditions (N = 142), or to a control condition involving 10 h TIB on all nights (N = 17). Primary neurobehavioral outcomes included lapses on the Psychomotor Vigilance Test (PVT), subjective sleepiness from the Karolinska Sleepiness Scale (KSS), and physiological sleepiness from a modified Maintenance of Wakefulness Test (MWT). Secondary outcomes included psychomotor and cognitive speed as measured by PVT fastest RTs and number correct on the Digit Symbol Substitution Task (DSST), respectively, and subjective fatigue from the Profile of Mood States (POMS). The dynamics of neurobehavioral outcomes following acute recovery sleep were statistically modeled across the 0 h-10 h recovery sleep doses. While TST, stage 2, REM sleep and NREM slow wave energy (SWE) increased linearly across recovery sleep doses, best-fitting neurobehavioral recovery functions were exponential across recovery sleep doses for PVT and KSS outcomes, and linear for the MWT. Analyses based on return to baseline and on estimated intersection with control condition means revealed recovery was incomplete at the 10 h TIB (8.96 h TST) for PVT performance, KSS sleepiness, and POMS fatigue. Both TST and SWE were elevated above baseline at the maximum recovery dose of 10 h TIB. Neurobehavioral deficits induced by 5 nights of sleep restricted to 4 h improved monotonically as acute recovery sleep dose increased, but some deficits remained after 10 h TIB for recovery. Complete recovery from such sleep restriction may require a longer sleep period during 1 night, and/or multiple nights of recovery sleep. It appears that acute recovery from chronic sleep restriction occurs as a result of elevated sleep pressure evident in both increased SWE and TST.


Medicare Star excludes diabetes patients with poor CVD risk factor control.
CMS recently added medication adherence to antihypertensives, antihyperlipidemics, and oral antihyperglycemics to its Medicare Star quality measures. These CMS metrics exclude patients with <2 medication fills (ie, "early nonadherence") and patients concurrently taking insulin. This study examined the proportion of patients with diabetes prescribed cardiovascular disease (CVD) medications excluded from Star adherence metrics and assessed the relationship of both Star-defined adherence and exclusion from Star metrics with CVD risk factor control. Cross-sectional, population-based analysis of 129,040 patients with diabetes aged ≥65 years in 2010 from 3 Kaiser Permanente regions. We estimated adjusted risk ratios to assess the relationship between achieving Star adherence and being excluded from Star adherence metrics, with CVD risk factor control (glycated hemoglobin [A1C]<8.0%, low-density lipoprotein cholesterol [LDL-C]<100 mg/dL, and systolic blood pressure [SBP]<130 mm Hg) in patients with diabetes. Star metrics excluded 27% of patients with diabetes prescribed oral medications. Star-defined nonadherence was negatively associated with CVD risk factor control (risk ratio [RR], 0.95, 0.84, 0.96 for A1C, LDL-C, and SBP control, respectively; P<.001). Exclusion from Star metrics due to early nonadherence was also strongly associated with poor control (RR, 0.83, 0.56, 0.87 for A1C, LDL-C, and SBP control, respectively; P<.001). Exclusion for insulin use was negatively associated with A1C control (RR, 0.78; P<.0001). Medicare Star adherence measures underestimate the prevalence of medication nonadherence in diabetes and exclude patients at high risk for poor CVD outcomes. Up to 3 million elderly patients with diabetes may be excluded from these measures nationally. Quality measures designed to encourage effective medication use should focus on all patients treated for CVD risk.


Potential impact of natural conditions on the rostral oral health: a study of horses in Iceland.
In the context of a study of Equine Odontoclastic Tooth Resorption and Hypercementosis (EOTRH), elderly Icelandic horses were examined in their originating country. The subject of this survey was the impact of dental treatment and keeping and feeding management on the health of the rostral oral cavity. A total of 170 horses (aged ≥ 15) located in different areas of Northwest, West, and South Iceland. The horse owners completed a questionnaire about keeping, feeding, and preventative dental care. The horses were clinically examined with particular attention to gingival and dental disorders of the rostral mouth, such as gingivitis, fistula, gingiva retraction, enlargement of the roots and dental calculus as well as integrity of incisors, bite angle, and tooth mobility. Most of the horses were kept on pasture all year with little supplementary feeding. The frequency of dental treatment in Iceland was low. Only a few horses were affected by tooth loss, gingivitis, gingival recession, and incisor calculus. Calculus occurred more frequently and more severely in canines than in incisors. The findings did not depend on the location of the horse. Assuming that the remarkable health of the rostral oral cavity in elderly Icelandic horses was not caused by preventive dental care, it is likely that it was due to living conditions in Iceland. Continuity, intensity of chewing, and nativeness of forage might promote the health of incisors and tissue surrounding the tooth even in elderly horses.


Functional MRI evidence of an abnormal neural network for pitch processing in congenital amusia.
Congenital amusia (tone deafness) is a lifelong disorder that prevents typically developing individuals from acquiring basic musical skills. Electrophysiological evidence indicates that congenital amusia is related to a musical pitch deficit that does not seem to arise from a dysfunction of the auditory cortex but rather from an anomaly along a frontotemporal auditory pathway. In order to better localize the neural basis of this pitch disorder, here we conducted a functional magnetic resonance imaging (fMRI) study. Congenital amusic adults and "musically intact" controls were scanned while passively listening to pure-tone melodic-like sequences in which the pitch distance between consecutive tones was varied parametrically. In both amusics and controls, brain activity increased as a function of increasing pitch distance, even for fine pitch changes, in both the left and right auditory cortices. These results support prior electrophysiological work showing that the auditory cortex of amusic individuals responds normally to pitch. In contrast, the right inferior frontal gyrus showed an abnormal deactivation in the amusic group, as well as reduced connectivity with the auditory cortex as compared with controls. These fMRI data are highly consistent with previous gray and white matter anomalies found in amusics in the auditory and inferior frontal cortices, as well as reduced white matter connections between these 2 regions.


Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells.
Expansion microscopy (ExM) is a technique that physically expands preserved cells and tissues before microscope imaging, so that conventional diffraction-limited microscopes can perform nanoscale-resolution imaging. In ExM, biomolecules or their markers are linked to a dense, swellable gel network synthesized throughout a specimen. Mechanical homogenization of the sample (e.g., by protease digestion) and the addition of water enable isotropic swelling of the gel, so that the relative positions of biomolecules are preserved. We previously presented ExM protocols for analyzing proteins and RNAs in cells and tissues. Here we describe a cookbook-style ExM protocol for expanding cultured HeLa cells with immunostained microtubules, aimed to help newcomers familiarize themselves with the experimental setups and skills required to successfully perform ExM. Our aim is to help beginners, or students in a wet-lab classroom setting, learn all the key steps of ExM. © 2020 The Authors.