The expression of B-cell activating factor belonging to tumor necrosis factor superfamily (BAFF) significantly correlated with C4D in kidney allograft rejection.
B-cell activating factor, belonging to the tumor necrosis factor (TNF) superfamily (BAFF), plays an important role in the development and survival of B lymphocytes. The present study was to investigate the correlation of BAFF and complement residue C4d in kidney allograft biopsies. Transplant kidney biopsies as allograft were studied. 10 acute rejection (AR) cases, 3 interstitial fibrosis/tubular atrophy (IF/TA) cases, and 9 unknown etiology cases from January 1, 2001 to July 20, 2007 were analyzed. Transplanted kidneys were excised because of graft loss, and their pathological sections were studied. Ten other protocol biopsy specimens were studied. These recipients who received protocol biopsies had serum creatinine levels that were within normal range (normal value: < 144 micromol/L) from 2007. [corrected] BAFF, C4d, and immunoglobulin G (IgG) were detected by immunohistochemical stain methods. All were primary renal transplant recipients. In AR and IF/TA cases, BAFF was strongly stained, similar to C4d. And BAFF and C4d were distributed in the perinephric tubular epithelial cell cytoplasm and cytomembrane. In the unknown etiology sections and protocol biopsy sections, BAFF was weakly or not expressed, similar to C4d. In BAFF and C4d high expression sections, weak or moderate IgG deposition was also found. BAFF may be associated with the development of antibody-mediated allograft rejection in kidney transplantation.


A Community-Curated DokuWiki Resource on Diagnostics, Diversity, Pathogenicity and Genetic Control of Xanthomonads.
Xanthomonads, including Xanthomonas and Xylella species, constitute a large and significant group of economically and ecologically important plant pathogens. Up-to-date knowledge of these pathogens and their hosts is essential for the development of suitable control measures. Traditional review articles or book chapters have inherent limitations, including static content and rapid obsolescence. To address these challenges, we have developed a web-based knowledge platform dedicated to xanthomonads, inspired by the concept of living systematic reviews. This platform offers a dynamic resource that encompasses bacterial virulence factors, plant resistance genes, and tools for diagnostics and genetic diversity studies. Our goal is to facilitate access for newcomers to the field, provide continuing education opportunities for students, assist plant protection services with diagnostics, provide valuable information to breeders on sources of resistance and breeding targets, and offer comprehensive expert knowledge to other stakeholders interested in plant-pathogenic xanthomonads. This resource is available for queries and updates at https://euroxanth.ipn.pt.


Laparoscopic resection of a paraganglioma in the greater omentum mimicking a peripancreatic neoplasm: a case report.
A paraganglioma is a tumor originating in the sympathetic or parasympathetic nervous system. Its diagnosis may sometimes be confusing if it occurs in an atypical site. We described herein a case of a peripancreatic paraganglioma originating in the greater omentum. An asymptomatic, 61-year-old, female patient was referred to our hospital for detailed examination of a peripancreatic mass detected incidentally on computed tomography (CT). The differential diagnosis was a neuroendocrine neoplasm (NEN), and a biopsy using EUS-FNA was performed. Histologically, the tumor cells showed proliferation in solid cell nests and were positive for CD56, chromogranin A, and synaptophysin. These findings and the hypervascularity of the tumor on imaging studies were compatible with NEN. Since the imaging studies did not clearly demonstrate the continuity of the tumor with the pancreas, laparoscopic tumor resection without a pancreatectomy and sampling of the enlarged peripancreatic lymph nodes were planned as treatment. The absence of continuity with the pancreas was later confirmed by intraoperative observation, and the resection was carried out as planned. The resected tumor was pathologically considered as NEN at first in agreement with the preoperative diagnosis. However, several histological findings (such as a zelleballen-like growth pattern, pseudo-inclusion, and strong nuclear atypia compared with the cells' proliferative ability) were atypical for NEN, and paraganglioma was included in the differential diagnosis. Additional immunostainings of S-100 and AE1/AE3 were performed, leading to the final diagnosis of paraganglioma. Paragangliomas should be included in the differential diagnosis of an intraperitoneal mass of uncertain identity with hypervascularity.


Genomic organization of δ-guaiene synthase genes in Aquilaria crassna and its possible use for the identification of Aquilaria species.
The resinous portions of Aquilaria plants, called agarwood, have been used as medicines and incenses. Agarwood contains a great variety of sesquiterpenes, and a study using cultured cells of Aquilaria crassna showed that the production of sesquiterpenes (α-guaiene, α-humulene, and δ-guaiene) was induced by treatment with methyl jasmonate, which led to the cloning of δ-guaiene synthases. In the present study, analyses of genomic organization and Southern blotting of δ-guaiene synthase in A. crassna were performed in order to examine the genomic background of δ-guaiene synthases in Aquilaria plants. Genomic cloning and sequencing revealed five types of sequence in putative δ-guaiene synthases sharing more than 96% identity in exon regions, and that these enzymes belonged to the class III TPS subfamily with seven exons and six introns. Furthermore, Southern blotting revealed that at least five copies of δ-guaiene synthase existed in A. crassna. The hybridization of digested DNA of A. crassna and A. sinensis with probes made with a δ-guaiene synthase cDNA fragment resulted in different banding patterns for these two species. It may be possible to identify Aquilaria species by restriction fragment length polymorphism analyses with δ-guaiene synthase cDNA probes.


The influence of temperature and incubation time on deformability of human erythrocytes.
Human erythrocytes have been heated and stressed in a novel and controlled manner using rectangular microcapillaries. Heated cells attached to the capillary wall were stressed by liquid flow. Under particular conditions of stress, temperature and incubation time the body of the cell could be pulled in the flow, retaining a connection with the glass by means of a narrow process or tether. The tethers appear as: regularly beaded, irregularly beaded or without beads depending upon the incubation conditions. We have outlined the incubation regimes necessary to achieve these different responses in the temperature range 48--55 degrees C. The cells become less deformable as the incubation is continued beyond an optimum time. The behaviour of the tether is compared with that of a viscoelastic liquid. Circular dichroism studies of ghost membranes show that the denaturation of membrane proteins is partially reversible when incubation times are similar to those required to bring about a loss of deformability.


Mutation-selection equilibrium in games with multiple strategies.
In evolutionary games the fitness of individuals is not constant but depends on the relative abundance of the various strategies in the population. Here we study general games among n strategies in populations of large but finite size. We explore stochastic evolutionary dynamics under weak selection, but for any mutation rate. We analyze the frequency dependent Moran process in well-mixed populations, but almost identical results are found for the Wright-Fisher and Pairwise Comparison processes. Surprisingly simple conditions specify whether a strategy is more abundant on average than 1/n, or than another strategy, in the mutation-selection equilibrium. We find one condition that holds for low mutation rate and another condition that holds for high mutation rate. A linear combination of these two conditions holds for any mutation rate. Our results allow a complete characterization of nxn games in the limit of weak selection.


Migrant Pharmacopoeias: An Ethnobotanical Survey of Four Caribbean Communities in Amazonia (French Guiana).
French Guiana is an overseas French department in South America at the margin of the Amazon basin. Its population is characterized by an important number of cultural groups. Many inhabitants originate from the Caribbean (mostly Saint Lucia, Martinique, Guadeloupe, Haiti, and the Dominican Republic). The objectives of this study were to present an overview of the main uses of plants among the Caribbean populations in French Guiana, and how they contribute to the dynamics of plant-based practices, in order to provide insights into ethnobotanical convergences, divergences, and hybridizations (such as the importation of new species and associated practices, and the adoption of Amazonian species by Caribbean people). Interviews and botanical voucher collections were conducted throughout the coastal area of French Guiana. Sixteen Saint Lucian, nineteen Haitian, eighteen French Caribbean, and twelve Dominican informants were interviewed during the fieldwork. Altogether they use 212 botanical species. Some plants have recently been imported directly from the Caribbean, while adaptations have also taken place: some species that do not exist locally are abandoned while Amazonian species are integrated to form hybrid pharmacopoeias. The phytotherapies of these communities in French Guiana are still conserved as consistent sets of knowledge, although they tend to blend through an ongoing process of hybridization. The online version contains supplementary material available at 10.1007/s12231-021-09529-0.


The use of clinical, histologic, and serologic parameters to predict the intragastric extent of intestinal metaplasia: a recommendation for routine practice.
Surveillance of intestinal metaplasia (IM) of the gastric mucosa should be limited to patients at high risk of gastric cancer. Patients with extensive IM are at increased cancer risk; however, the intragastric extent of IM is usually unknown at the time of the initial diagnosis. To assess the predictive value of clinical, histologic, and serologic parameters for the intragastric extent of IM. Prospective, multicenter study. Eighty-eight patients with a previous diagnosis of IM of the gastric mucosa. Surveillance gastroscopy with extensive random biopsy sampling. Biopsy specimens were evaluated according to the Sydney classification system. In addition, serologic testing of Helicobacter pylori and cagA status, pepsinogens I and II, gastrin, and intrinsic factor antibodies was performed. The association between the available parameters and extensive IM was evaluated with logistic regression analysis. In 51 patients (58%), IM was present in the biopsy specimens from at least 2 intragastric locations. The most important predictors of extensive IM were a family history of gastric cancer, alcohol use > or = 1 unit/d (1 glass, approximately 10 mL or 8 g ethanol), moderate or marked IM of the index biopsy specimen, and a pepsinogen I to II ratio < 3.0. A simple risk score based on these factors could identify extensive IM in 24 of 25 patients (sensitivity 96%). A prospective cohort study should confirm the proposed risk stratification. A risk score of clinical, histologic, and serologic parameters can predict extensive intragastric IM and may serve as a practical tool to select patients for surveillance endoscopy in routine clinical practice.


Quinovic acid purified from medicinal plant Fagonia indica mediates anticancer effects via death receptor 5.
Plants are major source for discovery and development of anticancer drugs. Several plant-based anticancer drugs are currently in clinical use. Fagonia indica is a plant of medicinal value in the South Asian countries. Using mass spectrometry and NMR spectroscopy, several compounds were purified from the F. indica extract. We have used one of the purified compounds quinovic acid (QA) and found that QA strongly suppressed the growth and viability of human breast and lung cancer cells. QA did not inhibit growth and viability of non-tumorigenic breast cells. QA mediated its anticancer effects by inducing cell death. QA-induced cell death was associated with biochemical features of apoptosis such as activation of caspases 3 and 8 as well as PARP cleavage. QA also upregulated mRNA and protein levels of death receptor 5 (DR5). Further investigation revealed that QA did not alter DR5 gene promoter activity, but enhanced DR5 mRNA and protein stabilities. DR5 is one of the major components of the extrinsic pathway of apoptosis. Accordingly, Apo2L/TRAIL, the DR5 ligand, potentiated the anticancer effects of QA. Our results indicate that QA mediates its anticancer effects, at least in part, by engaging DR5-depentent pathway to induce apoptosis. Based on our results, we propose that QA in combination with Apo2L/TRAIL can be further investigated as a novel therapeutic approach for breast and lung cancers.


The Effect of the Establishment of Conditioned Reinforcement for Reading Content on Second-Graders' Reading Achievement.
There remains a gap in the current literature as to how to reliably measure and increase students' "voluntary reading," based on research suggesting a relation between reading amount and reading achievement. We tested the effect of the establishment of conditioned reinforcement for reading via a collaborative shared reading (CSR) conditioning procedure on eight 2nd-grade students with and without learning disabilities and developmental disorders. This conditioning procedure was composed of opportunities for reciprocal reading and collaboration on comprehension and vocabulary tasks related to the reading content, such that partners (teacher-participant or participant-participant) were required to work together. We utilized a combined small-n experimental-control simultaneous-treatment design with a single-case multiple-probe design nested within each small group in order to compare within- and between-group differences for participants in the CSR procedure with a teacher or peer. All participants for whom conditioned reinforcement for reading was established (n = 7) demonstrated gains in reading achievement after a maximum of nine sessions (412 min), with grade-level increases between 0.2 and 2.5 on measures of reading comprehension and between 0.3 and 3.1 on measures of vocabulary. The students in the teacher-yoked condition (n = 3) demonstrated more significant gains in their average increases in achievement, although the peer-yoked procedure was also effective and possibly more viable in a classroom setting. These results suggest that a CSR procedure with a teacher or peer should be considered as a means of increasing the reading achievement of early elementary students via increases in the reinforcement value of reading.


Heme oxygenase-1-derived carbon monoxide is an autocrine inhibitor of vascular smooth muscle cell growth.
Vascular smooth muscle cells (SMCs) generate carbon monoxide (CO) via the catabolism of heme by the enzyme heme oxygenase (HO). In the present study, we found that serum stimulated a time- and concentration-dependent increase in the levels of HO-1 messenger RNA (mRNA) and protein in vascular SMCs. The induction of HO-1 expression by serum was inhibited by actinomycin D or cycloheximide. In addition, serum stimulated HO activity, as reflected by an increase in the concentration of bilirubin in the culture media. Treatment of vascular SMCs with serum stimulated DNA synthesis and this was potentiated by the HO inhibitors, zinc and tin protoporphyrin-IX as well as by the CO scavenger, hemoglobin. The iron chelator desferrioxamine had no effect on DNA synthesis. However, exposure of vascular SMCs to exogenous CO inhibited serum-stimulated SMC proliferation and the phosphorylation of retinoblastoma protein. In addition, CO arrested SMCs at the G(1)/S transition phase of the cell cycle and selectively blocked the serum-stimulated expression of cyclin A mRNA and protein without affecting the expression of cyclin D1 and E. CO also inhibited the serum-stimulated activation of cyclin A-associated kinase activity and cyclin-dependent kinase 2 activity. These results demonstrate that serum stimulates HO-1 gene expression and CO synthesis. Furthermore, they show that CO acts in a negative feedback fashion to inhibit vascular SMC growth by regulating specific components of the cell cycle machinery. The capacity of vascular mitogens to induce CO synthesis may provide a novel mechanism by which these agents modulate cell growth.


A potential treatment of low intensity pulsed ultrasound on cavernous nerve injury for erectile dysfunction.
Erectile dysfunction after nerve injury is a common disease after radical prostatectomy. Brain-derived neurotrophic factor (BDNF) is a member of the neurotrophin family, which plays an important role in the survival of existing neurons, the differentiation of new neurons, and synaptic plasticity. It has been demonstrated that low-intensity pulsed ultrasound (LIPUS) accelerates bone healing and axonal regeneration after injury. LIPUS may also be able to stimulate neuronal activity and enhance the levels of neurotrophic factors. Evidence suggests that elevated levels of BDNF in the brain have protective effects against neurodegenerative diseases. Previous studies have shown that the treatment on cavernous nerve injury repair, and protective effect plus neuro-regeneration effect by low-intensity pulsed ultrasound. They shared the similar mechanism including several trophic factors stimulation, Pl3K/akt pathway activation, and anti-fibrosis mechanism. We hypothesized that due to its combined neuroregenerative and protective effects, the non-invasive and easy-to-use method of LIPUS stimulation could have a therapeutic effect on erectile dysfunction stemming from cavernous nerve injury.


The effect of pressure on the characteristics of supercritical carbon dioxide extracts from Calamintha nepeta subsp. nepeta.
Chemists and industrialists are continuously attempting to develop greener and more environmentally benign chemical processes to extract essential oils and bioactive metabolites of high purity, finding various applications in cosmetics, detergents, nutraceuticals and pharmaceuticals. An increase preferenced for natural products over synthetic ones has made supercritical fluid technology a primary alternative for the generation of high-value bioactive ingredients. This effective technique requires only moderate temperatures, eliminates clean-up steps and avoids the use of harmful organic solvents. In this context, our study was focused on the chemical analysis of Calamintha nepeta subsp. nepeta aromatic extracts obtained with supercritical carbon dioxide. The effect of different operating conditions on the capacity of the lipophilic solvent to extract the targeted volatile components was also studied. The process was carried out at a fairly low constant temperature of 40°C, and with varying the pressure from 90 to 300 bar. The chemical composition of the extracts was analyzed by gas chromatography-mass spectroscopy. The results showed that the composition pattern, the concentrations of individual components and the quality of the extractable analytes were affected by pressure increase. The extraction yields varied from 0.73 to 1.21 wt% at 90 and 300 bar, respectively.


Validation of a high throughput screening assay to identify small molecules that target the eukaryotic replicative helicase.
Mcm2-7 is the catalytic core of the eukaryotic replicative helicase, which together with CDC45 and the GINS complex unwind parental DNA to generate templates for DNA polymerase. Being a highly regulated and complex enzyme that operates via an incompletely understood multi-step mechanism, molecular probes of Mcm2-7 that interrogate specific mechanistic steps would be useful tools for research and potential future chemotherapy. Based upon a synthetic lethal approach, we previously developed a budding yeast multivariate cell-based high throughput screening (HTS) assay to identify putative Mcm inhibitors by their ability to specifically cause a growth defect in an mcm mutant relative to a wild-type strain[1]. Here, as proof of concept, we used this assay to screen a 1280-member compound library (LOPAC) for potential Mcm2-7 inhibitors. Primary screening and dose-dependent retesting identified twelve compounds from this library that specifically inhibited the growth of the Mcm mutant relative to the corresponding wild-type strain (0.9 % hit rate). Secondary assays were employed to rule out non-specific DNA damaging agents, establish direct protein-ligand interaction via biophysical methods, and verify in vivo DNA replication inhibition via fluorescence activated cell sorter analysis (FACS). We identified one agent (β-carboline-3-carboxylic acid N-methylamide, CMA) that physically bound to the purified Mcm2-7 complex (Kdapp119 µM), and at slightly higher concentrations specifically blocked S-phase cell cycle progression of the wild-type strain. In total, identification of Mcm2-7 as a CMA target validates our synthetic lethal HTS assay paradigm as a tool to identify chemical probes for the Mcm2-7 replicative helicase.


Biocompatible surface functionalization architecture for a diamond quantum sensor.
Quantum metrology enables some of the most precise measurements. In the life sciences, diamond-based quantum sensing has led to a new class of biophysical sensors and diagnostic devices that are being investigated as a platform for cancer screening and ultrasensitive immunoassays. However, a broader application in the life sciences based on nanoscale NMR spectroscopy has been hampered by the need to interface highly sensitive quantum bit (qubit) sensors with their biological targets. Here, we demonstrate an approach that combines quantum engineering with single-molecule biophysics to immobilize individual proteins and DNA molecules on the surface of a bulk diamond crystal that hosts coherent nitrogen vacancy qubit sensors. Our thin (sub-5 nm) functionalization architecture provides precise control over the biomolecule adsorption density and results in near-surface qubit coherence approaching 100 μs. The developed architecture remains chemically stable under physiological conditions for over 5 d, making our technique compatible with most biophysical and biomedical applications.


Threat and adaptation: The maternal lived experience of continuing pregnancy after receiving a prenatal diagnosis of agenesis of the corpus callosum.
One stated objective of prenatal screening and diagnosis is the preparation for delivering a baby with medical needs or disability, however, psychosocial outcomes of parents who received a prenatal diagnosis suggest that this objective is not yet realised. Preparation may be complicated by diagnostic and prognostic uncertainty. A prenatal diagnosis that includes significant uncertainty due to the heterogeneous presentations, classifications, causes and outcomes is agenesis of the corpus callosum. As a neuroanatomical anomaly identified in the second or third trimesters, the diagnosis is likely to cause distress for expectant mothers, yet there is limited guidance for holistic support. To begin to address the paucity of research, this hermeneutic phenomenological study sought to explore, and provide a telling of the maternal experience of continuing pregnancy after a prenatal diagnosis of agenesis of the corpus callosum. Through interviews and a series of online, asynchronous and facilitated focus groups, lived experiences during pregnancy from the time of diagnosis to birth were explored with 26 mothers who participated in this international study. Themes were constructed through reflexive thematic analysis to describe the experience of the lived phenomenon. The first theme, Under Threat, included subthemes of The Threat to the Life of the Baby and Threatened Image of the Expected Family. The second theme, Day to Day Toward Adaptation, included subthemes of Holding it Together and Falling Apart, and More Than Information, Searching for Meaning, Hope and Control. To realise the commonly stated objective of prenatal diagnosis, to support maternal preparation, healthcare professionals require awareness of the profound, yet individual experience of prenatal diagnosis to adequately respond and support mothers through their continued pregnancies. Healthcare services should be designed to flexibly respond in a woman- and family-centred manner to reduce the threat and support maternal adaptation after a prenatal diagnosis.


Costs and Benefits of Office Diagnostic Hysteroscopy
In Italy, the main location for performing diagnostic hysteroscopy is the office. The success of office hysteroscopy is related to savings of time and money and the convenience for the physician and patient. We evaluated 4000 diagnostic hysteroscopies performed between January 2, 1989, and March 1, 1996. In 17.8% of these patients we subsequently performed operative hysteroscopies (metroplasty, synechiolysis, myomectomy, polypectomy for polyps >2 cm), and in 5.2% a total abdominal hysterectomy for abnormal uterine bleeding or endometrial cancer. The low percentage of operations underlines the efficacy of office diagnostic hysteroscopy, especially if we evaluate costs and accuracy of this procedure in detecting uterine pathologies.


The real-world efficacy and safety of anlotinib in advanced non-small cell lung cancer.
Anlotinib is an anti-angiogenetic multi-targeted tyrosine kinase inhibitor. This study aimed to evaluate the efficacy and safety of anlotinib in advanced non-small cell lung cancer (aNSCLC) in the real world. Patients with aNSCLC receiving anlotinib were enrolled in two cohorts (treatment naive and previously treated). The endpoints included progression-free survival (PFS), overall survival (OS) and anlotinib-related adverse events (ar-AEs). 203 patients accrued in the study. In the treatment-naïve cohort (n = 80), the PFS was 7.4 (95% confidence interval [CI] 4.1-10.7) and OS was 10.8 (95% CI 5.8-15.8) months of monotherapy group (immature survival for combination group). In previously treated cohort (n = 123), the PFS was 8.0 months (95% CI 6.1-9.9) in the combination group and 4.3 months (95% CI 2.1-6.6) in the monotherapy group (hazard ratio [HR] 0.49; 95% CI 0.29-0.83; p = 0.007), respectively. The OS was 18.5 months (95% CI 10.5-26.6) in the combination group and 7.8 months (95% CI 7.1-8.4) in the monotherapy group (HR 0.38; 95% CI 0.22-0.66; p = 0.001), respectively. The ar-AEs of grade ≥ 3 in the monotherapy and the combination groups were hypertension (9.0 and 8.7%), fatigue (8.1 and 7.6%), hand-foot syndrome (8.1 and 6.5%), diarrhea (5.4 and 8.7%), proteinuria (5.4 and 5.4%), and mucositis oral (6.3 and 8.7%). In aNSCLC, anlotinib monotherapy has a promising efficacy in the first-line setting. It may be an option for those who are ineligible for chemotherapy; anlotinib combination therapy in a ≥ second-line setting showed manageable toxicities and encouraging efficacy, indicating a good application prospect. This study was retrospectively registered with ISRCTN Registry (ID ISRCTN35543977) on January 26th, 2021 and Chinese Clinical Trial Register (ChiCTR2000032265) on April 4th, 2020.


Salivary gland tumors exhibit distinct miRNA signatures involved in Wnt/β-Catenin signaling in formalin fixed paraffin embedded tissue samples.
Advances in high-throughput genomic technologies have enabled the identification of numerous selective tumor markers. However, adapting these newly identified markers to clinical practice is not always possible because most RNA molecules, including mRNAs of protein-coding genes and long non-coding RNAs, are not stable under laboratory conditions, making their testing a major challenge. In contrast to long RNA molecules, miRNAs offer a great advantage in that they are relatively stable due to their small size. Accordingly, herein we aimed to determine the expression levels of miRNAs that are involved in Wnt/β-catenin signaling pathway in formalin fixed paraffin embedded (FFPE) tissue samples of patients with salivary gland tumors. A total of 42 patients with salivary gland tumors were included in the study. The miRNA expression signatures were evaluated using the RT-qPCR. As a result, β-catenin positivity was observed in all salivary gland tumors without distinguishing between benign and malignant phenotypes. Remarkably, we found that miR-200a and miR-373 were significantly upregulated whereas miR-30c were downregulated in tissues of patients with salivary gland tumors, compared to adjacent healthy tissue samples. In addition, distinct expression signatures of these miRNAs were significantly associated with the clinicopathological findings of patients such as perineural invasion and lymph node metastasis. Additionally, miR-145 and miR-30a were found to be specifically downregulated in a mucoepidermoid carcinoma. Also, miR-26b was selectively increased in pleomorphic adenomas of the salivary gland. Collectively, our findings suggest that these miRNAs may play chief roles in the differential diagnosis of salivary gland tumors.


lncRNA-CR46018 positively regulates the Drosophila Toll immune response by interacting with Dif/Dorsal.
The Toll signaling pathway is highly conserved from insects to mammals. Drosophila is a model species that is commonly used to study innate immunity. Although many studies have assessed protein-coding genes that regulate the Toll pathway, it is unclear whether long noncoding RNAs (lncRNAs) play regulatory roles in the Toll pathway. Here, we evaluated the expression of the lncRNA CR46018 in Drosophila. Our results showed that this lncRNA was significantly overexpressed after infection of Drosophila with Micrococcus luteus. A CR46018-overexpressing Drosophila strain was then constructed; we expected that CR46018 overexpression would enhance the expression of various antimicrobial peptides downstream of the Toll pathway, regardless of infection with M. luteus. RNA-seq analysis of CR46018-overexpressing Drosophila after infection with M. luteus showed that upregulated genes were mainly enriched in Toll and Imd signaling pathways. Moreover, bioinformatics predictions and RNA-immunoprecipitation experiments showed that CR46018 interacted with the transcription factors Dif and Dorsal to enhance the Toll pathway. During gram-positive bacterial infection, flies overexpressing CR46018 showed favorable survival compared with flies in the control group. Overall, our current work not only reveals a new immune regulatory factor, lncRNA-CR46018, and explores its potential regulatory model, but also provides a new perspective for the effect of immune disorders on the survival of Drosophila melanogaster.


Hydatidiform mole in Jos, Nigeria.
Hydatidiform mole is a relatively common gynecological problem which could present like spontaneous abortion, one of the commonest gynecological emergencies. It has the propensity to become malignant but can easily be identified and treated. The aim of this study was to determine the demographics, clinical features, treatment options and outcome of patients with hydatidiform mole in our environment. This was a retrospective review of all the cases of hydatidiform mole seen at the Jos University Teaching Hospital (JUTH), Jos, Nigeria over a 5-year period. There were 34 cases of hydatidiform mole giving an incidence of 1 in 357 deliveries. However only 25 case notes were available for analysis and the mean age of patients was 28±3 years. Vaginal bleeding (92%), honeycomb appearance on ultrasound scan (84%), and passage of vesicles (60%) were the most common clinical findings while suction curettage was the mode of treatment for all the patients in this study. Twenty-eight percent of cases were confirmed by histology. No patient came for follow-up after the third month of diagnosis. Twenty percent of the patients booked for antenatal care within 9 months of diagnosis while 12% of patients presented as gynecological emergencies with features of malignant disease within six months of diagnosis. Hydatidiform mole is common in Jos, North Central Nigeria, and presents most commonly with vaginal bleeding with over 10% becoming malignant. Hence all patients who present with vaginal bleeding should be screened for HM. None of the patients completed the recommended duration of follow-up and only about ¼ had histology reports. Concerted efforts need to be made to address the challenges of patients adhering to recommended follow-up protocols and having to pay first before investigations are done.


Multiplex RT-PCR to simultaneously detect three viruses that infect peach.
Peach is a major crop in China, and like any other stone fruit, virus and virus-like diseases can reduce the yield and quality of the fruit. Herein, we developed a multiplex RT-PCR (mRT-PCR) assay for simultaneously detecting three viruses known to infect peach: peach-associated luteovirus (PaLV), peach virus D (PeVD) and nectarine stem-pitting-associated virus (NSPaV). Plant nad5 mRNA was used as the internal control. Field samples that were co-infected with PaLV, PeVD and NSPaV were used; we identified three primer pairs to be the most specific for detecting these viruses, followed by determining the ideal concentration of each primer pair and optimizing the annealing temperature for mRT-PCR. We also assessed the detection limit using serial dilutions of RNA and cDNA. The newly developed mRT-PCR assay could simultaneously detect PaLV, PeVD and NSPaV. To validate the reliability of mRT-PCR for virus detection, mRT-PCR was used to detect viruses in the leaves of 21 peach plants collected in Liaoning Province, China. The obtained results revealed the presence of single and co-infections. To conclude, the mRT-PCR assay developed herein is sensitive, reliable and economical, and we believe that it can thus be used for large-scale surveys of PaLV, PeVD and NSPaV. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, we developed a multiplex reverse transcriptase PCR (mRT-PCR) assay for simultaneously detecting three viruses that infect peach: peach-associated luteovirus (PaLV), peach virus D (PeVD) and nectarine stem-pitting-associated virus (NSPaV). This assay is simple, easy to perform, reliable and cost-effective, and can thus be applied for large-scale surveys of PaLV, PeVD and NSPaV.


A statistical approach to the classification of biological cells from their diffraction patterns.
An effective method of graphically representing complex biological data is presented, and a statistical classification technique is outlined including an example involving exfoliated cervical cells.


Carrageenan stimulates reduction of nitroblue tetrazolium by human neutrophils without membrane depolarization, myeloperoxidase secretion, or increased oxygen consumption.
Carrageenan, a sulfated polyanionic polysaccharide, is commonly used to induce inflammation in experimental animals, and this model is used to screen for the effectiveness of antiinflammatory drugs. Carrageenan-induced inflammation has been attributed to a variety of autocoids including histamine, bradykinin, complement, superoxide, and prostaglandins. This study examines the effects of carrageenan on human PMN in a serum-free system. Carrageenan was found to stimulate the reduction of NBT by PMNs without stimulating membrane depolarization, oxygen consumption, H2O2 production, or myeloperoxidase secretion. Carrageenan stimulates a heat-labile, NBT-reducing system which is unassociated with the usual stimulus-response coupling seen with other PMN activators such as PMA, FMLP, and zymosan.


Incorporation of amino acids by rat liver nucleoli in vitro.
Isolated rat liver nucleoli are shown to incorporate radioactive amino acids into proteins. The characteristics of this nucleolar protein-synthesizine system indicated that the incorporation observed was not due to cytoplasmic or bacterial contamination. The products synthesized by isolated nucleoli have been analyzed by polyacrylamide gel electrophoresis and subsequent autoradiography; this revealed a few radioactive bands suggesting that in vitro only a few proteins are labelled. The results presented are discussed in view of a possible nucleolar protein synthesis.


Prognosis of surgical hernia repair in cirrhotic patients with refractory ascites.
Abdominal wall hernias are common in patients with ascites. Elective surgical repair is recommended for the treatment of abdominal wall hernias. However, surgical hernia repair in cirrhotic patients with refractory ascites is controversial. In this study, we aimed to evaluate the outcomes of elective surgical hernia repair in patients with liver cirrhosis with and without refractory ascites. From January 2005 to June 2018, we retrospectively reviewed the records of consecutive patients with liver cirrhosis who underwent a surgical hernia repair. This study included 107 patients; 31 patients (29.0%) had refractory ascites. Preoperatively, cirrhotic patients with refractory ascites had a higher median model for end-stage liver disease (MELD) score (13.0 vs 11.0, P = 0.001) than those without refractory ascites. The 30-day mortality rate (3.2% vs 0%, P = 0.64) and the risk of recurrence (hazard ratio 0.410; 95% CI 0.050-3.220; P = 0.39) did not differ significantly between cirrhotic patients with refractory ascites and cirrhotic patients without refractory ascites. Among cirrhotic patients with refractory ascites, albumin (P = 0.23), bilirubin (P = 0.37), creatinine (P = 0.97), and sodium levels (P = 0.35) did not change significantly after surgery. In advanced liver cirrhosis patients with refractory ascites, hernias can be safely treated with elective surgical repair. Mortality rate within 30 days did not differ by the presence or absence of refractory ascites. Elective hernia repair might be beneficial for treatment of abdominal wall hernia in cirrhotic patients with refractory ascites.


Observation of the molecular response to light upon photoexcitation.
When a molecule interacts with light, its electrons can absorb energy from the electromagnetic field by rapidly rearranging their positions. This constitutes the first step of photochemical and photophysical processes that include primary events in human vision and photosynthesis. Here, we report the direct measurement of the initial redistribution of electron density when the molecule 1,3-cyclohexadiene (CHD) is optically excited. Our experiments exploit the intense, ultrashort hard x-ray pulses of the Linac Coherent Light Source (LCLS) to map the change in electron density using ultrafast x-ray scattering. The nature of the excited electronic state is identified with excellent spatial resolution and in good agreement with theoretical predictions. The excited state electron density distributions are thus amenable to direct experimental observation.


Efficient sensing of saccharin through interference synthesis of gum ghatti capped silver nanoparticles.
The presence of saccharin (SH) could be efficiently sensed (in the concentration range of 5 × 10-5 M to 5 × 10-1 M) through the interference synthesis of gum ghatti (GG) capped silver nanoparticles (GGAgNps). The synthesis used sodium borohydride and gum ghatti (GG) as the reducing and capping agents respectively. The strong hydrogen-bonding recognition between GG and SH was responsible for the interference. The intensity of the SPR peak of GGAgNps was found linearly dependent on [SH]. The SH detection was further enhanced when combo capping comprising of GG and chitosan (Ch) (in 1:1 weight ratio) was used while the use of gum acacia (GA) in place of Ch (in combo) decreased the detection sensitivity. The combo polysaccharide solutions had non-Newtonian behaviour and shear thinning property like GG. The method was also applied for the successful detection of SH in commercially available real juice samples.


Cloning, sequencing and characterization of the genome of a recombinant norovirus of the rare genotype GII.P7/GII.6 in China.
The genome sequence of a rare recombinant norovirus (NoV) genotype obtained from clinical samples in China was determined using one-step reverse transcription PCR. It was identified as the GII.P7/GII.6 genotype using both phylogenetic and SimPlot analyses. A high degree of variability was observed in the P2 subdomain, especially in the B-loop structure. The recombination breakpoints of all available GII.P7/GII.6 strains were mapped to two different positions within the RdRp region, both of which were at least 40 nt upstream of the overlap of ORF1 and 2. The GII.P7/GII.6 genotype appears to have been circulating in Asia for at least 10 years.


Lack of ERK activation and cell migration in FGF-2-deficient endothelial cells.
The formation of blood capillaries from preexisting vessels (angiogenesis) and vascular remodeling secondary to atherosclerosis or vessel injury are characterized by endothelial cell migration and proliferation. Numerous growth factors control these cell functions. Basic fibroblast growth factor (FGF-2), a potent angiogenesis inducer, stimulates endothelial cell proliferation, migration, and proteinase production in vitro and in vivo. However, mice genetically deficient in FGF-2 have no apparent vascular defects. We have observed that endothelial cell migration in response to mechanical damage in vitro is accompanied by activation of the extracellular signal-regulated kinase (ERK) pathway, which can be blocked by neutralizing anti-FGF-2 antibodies. Endothelial cells from mice that are genetically deficient in FGF-2 neither migrate nor activate ERK in response to mechanical wounding. Addition of exogenous FGF-2 restores a normal cell response, which shows that impaired migration results from the genetic deficiency of this growth factor. Injury-induced ERK activation in endothelial cells occurs only at the edge of the wound. In addition, FGF-2-induced ERK activation mediates endothelial cell migration in response to wounding without a significant effect on proliferation. These data show that FGF-2 is a key regulator of endothelial cell migration during wound repair.


Lipopolysaccharide increases gastric and circulating NUCB2/nesfatin-1 concentrations in rats.
Bacterial lipopolysaccharide (LPS) is an established animal model to study the innate immune response to Gram-negative bacteria mimicking symptoms of infection including reduction of food intake. LPS decreases acyl ghrelin associated with decreased concentrations of circulating ghrelin-O-acyltransferase (GOAT) likely contributing to the anorexigenic effect. We also recently described the prominent expression of the novel anorexigenic hormone, nucleobindin2 (NUCB2)/nesfatin-1 in gastric X/A-like cells co-localized with ghrelin in different pools of vesicles. To investigate whether LPS would affect gastric and circulating NUCB2/nesfatin-1 concentration, ad libitum fed rats were equipped with an intravenous (iv) catheter. LPS was injected intraperitoneally (ip, 100μg/kg) and blood was withdrawn before and at 2, 5, 7 and 24h post injection and processed for NUCB2/nesfatin-1 radioimmunoassay. Gastric corpus was collected to measure NUCB2 mRNA expression by RT-qPCR and NUCB2/nesfatin-1 protein concentration by Western blot. Injection of LPS increased plasma NUCB2/nesfatin-1 concentrations by 43%, 78% and 62% compared to vehicle at 2h, 5h and 7h post injection respectively (p<0.05) and returned to baseline at 24h. The plasma NUCB2/nesfatin-1 increase at 2h was associated with increased corpus NUCB2 mRNA expression (p<0.01), whereas NUCB2 mRNA was not detectable in white blood cells. Likewise, gastric NUCB2 protein concentration was increased by 62% after LPS compared to vehicle (p<0.01). These data show that gastric NUCB2 production and release are increased in response to LPS. These changes are opposite to those of ghrelin in response to LPS supporting a differential gastric regulation of NUCB2/nesfatin-1 and ghrelin expression derived from the same cell by immune challenge.


Septic Dislocation of Hip After Cephalomedullary Nail Fixation of Intertrochanteric Fractures of Femur: A Case Report.
We describe 3 cases of septic hip arthritis with a dislocation after failed, cephalomedullary nail (CMN) fixation following fragility intertrochanteric (IT) hip fractures that were treated with antibiotic spacer placement. Septic hip arthritis and compromised hip abductors can contribute to dislocation of the hip after CMN for IT fractures. This case report presents a treatment plan to address this rare complication, not yet reported after CMN for IT fractures in the literature, to control infection and optimize function.


Efficacy of oral brush cytology cell block immunocytochemistry in the diagnosis of oral leukoplakia and oral squamous cell carcinoma.
This study assessed the efficacy of using oral liquid-based brush cytology and cell block immunocytochemistry in the diagnosis of oral leukoplakia as minimally invasive diagnostic adjuncts. Seventy-two patients diagnosed clinically with either oral leukoplakia (OLK) or oral squamous cell carcinoma were included. Oral brush samples using Orcellex® brushes were obtained from all participants directly before undergoing surgical biopsy. Cell blocks were prepared for all samples and assessed for cytomorphology and immunocytochemistry of DNA mismatch repair proteins (MSH-6, MSH-2, MLH-1 and PMS-2). A combined index score of immunocytochemistry expression and cytology grading was compared against the gold standard (histopathological diagnosis). A significant association was observed between the cytological assessments of oral liquid-based brush cytology samples and the histopathological diagnosis (P < .005). In addition, there was a significant inverse correlation between the grade of oral epithelial dysplasia and the cumulative score of the studied DNA mismatch repair proteins (P < .005). Grading criteria for both oral liquid-based brush cytology and immunocytochemistry cumulative index scores are proposed based on the analysis of receiver operating characteristic curve coordinates. The diagnostic accuracy of this approach was outstanding in terms of discrimination between the presence or absence of oral epithelial dysplasia (0.961) and squamous cell carcinoma (0.977) separately. Oral liquid-based brush cytology cell block immunocytochemistry provides a reliable strategy to investigate oral mucosal epithelial disorders. This approach presents a minimally invasive, highly accurate and non-technically demanding method for the surveillance of oral potentially malignant disorders and squamous cell carcinoma.


Mast Cell Numbers in Primary Eosinophilic Colitis are Significantly Higher than in Secondary Tissue Eosinophilia and Normal Control: a Possible Link to Pathogenesis.
Primary Eosinophilic Colitis (PEC) is one of the rare eosinophilic gastrointestinal diseases with a poorly understood pathogenesis. Eosinophilic esophagitis (EE) is the most common and best-understood disease in this category. Activated mast cells (MCs) have a role to play in the tissue damage in EE. It is not known if PEC shares this mechanism. This cross-sectional study aimed to investigate the number of MCs in PEC and to compare them with cases of secondary colonic tissue eosinophilia (TE) and normal colon. The study included 19 PEC cases, 47 cases of secondary tissue eosinophilia and 50 normal colon tissues. Histopathological slides of all cases were reviewed to confirm the diagnosis and count the number of eosinophils. Glass slides for all cases were stained for C-kit (CD117) to highlight and count the MCs. The mean number of the MCs in normal controls was 9.7 MCs per HPF (SD= 4.6). The mean number of MCs in the PEC cases was 26.5 (SD=7.1) which was significantly higher than the normal counts (p-value <0.000). The mean number of MCs in the secondary TE group was 18.0 (SD=7.1), which was significantly higher than normal controls; p-value <0.000. Comparing MC counts in PEC and secondary TE also revealed a significant difference with a p-value of < 0.000. MCs in PEC are significantly higher than those in secondary TE and normal controls. This suggests the role of the MCs in the pathogenesis of Primary Eosinophilic Colitis.


1H NMR studies of the heme iron coordination in cytochrome c-552 from Euglena gracilis.
The coordination of the heme iron in cytochrome c-552 from Euglena gracilis was investigated by 1H NMR studies at 360 MHz. The data imply that the axial heme ligands are His-14 and Met-56 in both the oxidized and the reduced protein. Studies of mixed solutions of ferro- and ferricytochrome c-552, which provided much of the information on the heme structure, also showed that the intermolecular electron exchange is characterized by a bimolecular rate constant of 5-10(6) mol-1-s-1 at 29 degrees C, which is three orders of magnitude faster than the corresponding reaction in solutions of mammalian cytochromes c.


Optogenetic Methods to Control Tissue Mechanics in Drosophila.
Optogenetics is a powerful technique that allows the control of protein function with high spatiotemporal precision using light. Here, we describe the application of this method to control tissue mechanics during Drosophila embryonic development. We detail optogenetic protocols to either increase or decrease cell contractility and analyze the interplay between cell-cell interaction, tissue geometry, and force transmission during gastrulation.


Tolerance regeneration by T regulatory cells in autologous haematopoietic stem cell transplantation for autoimmune diseases.
Autologous haematopoietic stem cell transplantation shows increasing promise as a therapeutic option for patients with treatment-refractory autoimmune disease, particularly systemic sclerosis and multiple sclerosis. However, this intensive chemotherapy-based procedure is not always possible due to potential treatment toxicities and comorbidities. The biological mechanisms of how this procedure induces long-term remission in autoimmune disease are increasingly understood. The focus of this review is on recent research findings on the role of CD4+ T regulatory cells (Tregs) in resetting the immune system leading to the eradication of the autoimmune disease after transplantation. Discovery of the precise mechanisms of this process will allow development of novel Treg-based therapies and thus avoid the need for intensive chemotherapy-based treatment for these autoimmune diseases in the future.


Toxicity of sulfide-modified nanoscale zero-valent iron to Escherichia coli in aqueous solutions.
Sulfide-modified nanoscale zero-valent iron (S/nZVI) has been widely studied for groundwater remediation, but the potential environmental risks are poorly understood. This study examined the toxicity of S/nZVI to Escherichia coli in aqueous solutions. The sulfidation could reduce toxicity of nZVI, and S/nZVI exhibited a weaker toxicity at lower Fe/S molar ratio, resulting from the lower Fe0 content and higher sulfate and iron oxide. The toxicity of S/nZVI was significantly alleviated in the presence of N-Acetyl-L-cysteine (a scavenger for reactive oxygen species (ROS)), revealing that the ROS-induced oxidative stress was the principal mechanism. Moreover, Transmission Electron Microscopy images elucidated that the membranes of S/nZVI-treated cells were disrupted and S/nZVI existed on E. coli surface and in the cytoplasm. S/nZVI might have interacted with the amine, carboxyl, and ester groups on E. coli cell surface, as demonstrated by Fourier Transform Infrared Spectroscopy analysis. However, the presence of individual groundwater component (e.g., Ca2+, SO42-, HCO3- and humic acid) could more or less alleviate the toxicity of S/nZVI. Furthermore, S/nZVI only exhibited slight toxic effect (<0.15-log after 1 h) in the presence of the mixed components. The same faint toxicity was observed for the aged S/nZVI, indicating that S/nZVI could lose its toxicity over time.


Proteomic characterization of atopic dermatitis blood from infancy to adulthood.
Patients with atopic dermatitis (AD) have systemic biomarker dysregulation that differs by age group; however, the proteomic characteristics of these age-based changes are unknown. To profile blood proteins of patients with AD across different age groups versus age-appropriate controls. Using the Olink high-throughput proteomic platform, we profiled 375 serum proteins of 20 infants (age, 0-5 years), 39 children (age, 6-11 years), 21 adolescents (age, 12-17 years), and 20 adults (age, ≥18 years) with moderate-to-severe AD and 83 age-appropriate controls. Each group presented a distinct systemic proteomic signature. Th2-related proteins were increased in infant AD and further intensified with age through adolescence and adulthood (interleukin 4/CCL13/CCL17). In contrast, Th1 axis down-regulation was detected in infants with AD and gradually reversed to increased Th1 products (interferon γ/CXCL9/CXCL10/CCL2) in patients with AD from childhood to adulthood. Despite their short disease duration, infants already had evidence of systemic inflammation, with significant upregulation of innate immunity (interleukin 17C/ interleukin-1RN), T-cell activation/migration (CCL19), Th2 (CCL13/CCL17), and Th17 (PI3) proteins. Adults with AD present unique upregulation of cardiovascular proteins related to coagulation and diabetes. Cross-sectional observational study with a single time point. Systemic immune signatures of AD are age-specific beyond the shared Th2 immune activation. These data advocate for precision medicine approaches based on age-specific AD profiles.


Utilization of static and kinetic information for depth by young Malaŵians.
The utilization of static and kinetic information for depth by Malaŵian children and young adults in making monocular relative size judgments was investigated. Subjects viewed pairs of objects or photographic slides of the same pairs and judged which was the larger of each pair. The sizes and positions of the objects were manipulated such that the more distant object subtended a visual angle equal to, 80% of, or 70% of the nearer object. Motor parallax information was manipulated by allowing or preventing head movement. All subjects displayed sensitivity to static information for depth when the two objects subtended equal visual angles. When the more distant object was larger but subtended a smaller visual angle than the nearer object, subjects tended to base their judgments on retinal size. Motion parallax information increased accuracy of judgments of three-dimensional displays but reduced accuracy of judgments of pictorial displays. Comparisons are made between these results and those for American subjects.


Development of ratiometric carbohydrate sensor based on boron dipyrromethene (BODIPY) scaffold.
We designed a ratiometric carbohydrate sensor consisting of the boron dipyrromethene fluorophore substituted with boronic acid at the 2-position, based upon the strong substituent dependency of the absorbance/fluorescence wavelengths of BODIPY. The substituent is in equilibrium between the boronic acid B(OH)2 and boronate (B(OH)3-) forms, which have different absorbance/fluorescence wavelengths in the visible region. Reaction of the boronic acid moiety with hydroxy groups of carbohydrate affords a cyclic ester and shifts the equilibrium in favor of the boronate (B(OR)3-) form, resulting in a carbohydrate-concentration-dependent change of the fluorescence ratio. Thus, the sensor, BA-BODIPY, can ratiometrically detect carbohydrate at a pH near the pKa of cyclic ester formation.


A diacylglycerol photoswitching protocol for studying TRPC channel functions in mammalian cells and tissue slices.
Small molecular probes designed for photopharmacology and opto-chemogenetics are rapidly gaining widespread recognition for investigations of transient receptor potential canonical (TRPC) channels. This protocol describes the use of three photoswitchable diacylglycerol analogs-PhoDAG-1, PhoDAG-3, and OptoDArG-for ultrarapid activation and deactivation of native TRPC2 channels in mouse vomeronasal sensory neurons and olfactory type B cells, as well as heterologously expressed human TRPC6 channels. Photoconversion can be achieved in mammalian tissue slices and enables all-optical stimulation and shutoff of TRPC channels. For complete details on the use and execution of this protocol, please refer to Leinders-Zufall et al. (2018).


Aberrant ganglioside composition in glioblastoma multiforme and peritumoral tissue: A mass spectrometry characterization.
Tumor cells are characterized by aberrant glycosylation of the cell surface glycoconjugates. Gangliosides are sialylated glycosphingolipids highly abundant in neural tissue and considered as tumor markers and therapeutic targets. In this study, a detailed characterization of native ganglioside mixtures from glioblastoma multiforme, corresponding peritumoral tissue and healthy human brain was performed using mass spectrometry and high performance thin layer chromatography in order to elucidate their roles as tumor-associated antigens. Distinctive changes in ganglioside expression were determined in glioblastoma compared to healthy brain tissue showing 5 times lower total ganglioside content and higher abundance of simple gangliosides. Glioblastoma gangliosides were characterized by highly diverse ceramide composition with fatty acyl chains varying from 16 to 24 carbon atoms, while in normal and peritumoral tissue mostly C18 chains were found. The most abundant ganglioside in glioblastoma was GD3 (d18:1/18:0), followed by GD3 (d18:1/24:0) that was exclusively detected in glioblastoma tissue. Peritumoral tissue expressed higher abundance of GD3- and nLM1/GM1-species while lower GT1-species vs. normal brain. O-Ac-GD1, known as neurostatin, was detected in normal and peritumoral tissue, but not in glioblastoma. O-Ac-GD3 species were found exclusively in glioblastoma; MS structural characterization of the isomeric form possessing the O-acetylation at the inner sialic acid residue confirmed our previous finding that this isomer is glioma-associated. This, to our knowledge, the most detailed characterization of ganglioside composition in glioblastoma and peritumoral tissue, especially addressing the ceramide variability and O-acetylation of tumor-associated gangliosides, could contribute to recognition of new molecular targets for glioblastoma treatment and sub-classification.


The effects of stimulating the cerebellum on social sequences: A tDCS-fMRI pilot study: Los efectos de estimular el cerebelo en secuencias sociales: Un estudio piloto con tDCS y fMRI.
Research on the involvement of the cerebellum in social behavior and its relationship with social mentalizing has just begun. Social mentalizing is the ability to attribute mental states such as desires, intentions, and beliefs to others. This ability involves the use of social action sequences which are believed to be stored in the cerebellum. In order to better understand the neurobiology of social mentalizing, we applied cerebellar transcranial direct current stimulation (tDCS) on 23 healthy participants in the MRI scanner, immediately followed by measuring their brain activity during a task that required to generate the correct sequence of social actions involving false (i.e., outdated) and true beliefs, social routines and non-social (control) events. The results revealed that stimulation decreased task performance along with decreased brain activation in mentalizing areas, including the temporoparietal junction and the precuneus. This decrease was strongest for true belief sequences compared to the other sequences. These findings support the functional impact of the cerebellum on the mentalizing network and belief mentalizing, contributing to the understanding of the role of the cerebellum in social sequences.


The use of a caries detector dye during cavity preparation: a microbiological assessment.
During cavity preparation conventional tactile and optical criteria are used to assess the caries status of the enamel-dentine junction, cavity preparation being considered complete when this area is hard to a sharp probe and stain free. In the present study 201 cavities were prepared under rubber dam. When caries removal was considered complete using the conventional tactile and optical criteria, a caries detector dye (1% acid red in propylene glycol), which is claimed to stain 'infected' tissue red, was applied. Fifty-two per cent of cavities showed caries dye stain in some part of the enamel-dentine junction. Subsequent microbiological sampling of dye-stained and dye-unstained sites resulted in the recovery of low numbers of bacteria and revealed no difference in the level of infection of the two sites. It is concluded that the conventional tactile and optical criteria are satisfactory assessments of the caries status of tissue during cavity preparation and that subsequent use of a caries detector dye on hard and stain-free dentine will result in unnecessary tissue removal.


Systolic and diastolic cardiac dysfunction early after the initiation of doxorubicin therapy: significance of gender and concurrent mediastinal radiation.
Diastolic and systolic left ventricular (LV) function may be affected early after the initiation of doxorubicin therapy. However, the role of mediastinal radiation and other cytotoxic agents in the production of these early cardiac effects is unclear. In this study LV diastolic and systolic function were assessed before and after doxorubicin (223+/-122 mg.m-2; range, 40-618) in 33 patients. After doxorubicin, LV ejection fraction declined (0.61+/-0.08 to 0.56+/-0.08, P=0.0008), peak filling rate decreased (3.38+/-1.10 to 2.82+/-0.62 end diastolic volumes/s, P=0.006), and time to peak filling rate increased (162+/-39 to 182+/-45 ms, P=0.04). The changes in LV systolic and diastolic function were not related to doxorubicin dose and the use of other cytotoxic agents; the decrease in LV ejection fraction with doxorubicin was more notable in men and in patients who received mediastinal irradiation concurrently with doxorubicin. It is concluded that the use of doxorubicin was associated with the simultaneous early development of LV systolic and diastolic dysfunction. Male gender and concurrent mediastinal irradiation were independent influences, but doxorubicin dose and the use of other cytotoxic agents were not associated with worse cardiac dysfunction.


Pressures to adhere to treatment: observations on 'leverage' in English mental healthcare.
Coercion in psychiatric practice appears to be increasing. Is this in patients' best interest? Is it good medical practice?


A "complement-ary" AIDS vaccine.
The human immunodeficiency virus uses the human complement system to its advantage. Is it possible to turn the tables with a vaccine?


Identification and quantification of oligomeric proanthocyanidins, alkaloids, and flavonoids in lotus seeds: A potentially rich source of bioactive compounds.
Untargeted metabolomics was performed to study the profiles of 101 chemicals in lotus seeds using ultrahigh-performance liquid chromatography-photodiode array detection-high-resolution tandem mass spectrometry. Among them, 16 dimeric, 18 trimeric, and 4 tetrameric proanthocyanidins were theoretically identified based on the degree of polymerization, and the number of linkages and the presence of two dihydroflavonols and three glycosylated alkaloids were determined for the first time. The proanthocyanidin, flavonoid, amino acid, and total compound contents were quantified, revealing decreases in their levels during maturation as well as a polymerization process formation of polymers from monomers during seed maturation. Interestingly, glycosylated alkaloids were only detected in seed cotyledons being highest at green-brown stage, whereas proanthocyanidins were present at a concentration of 8,226.19 ± 249.96 μg/g (dry weight) in green-brown stage of seed coats. Our findings may provide insights into the utilization of lotus seeds as a functional food.


Ambulatory 24-hour fast using flexible insulin therapy in patients with type 1 diabetes.
Prolonged fasting may be necessary in life for religious, medical and other reasons. For this reason, our study investigated the feasibility and safety of a 24-h fast conducted at home for patients with type 1 diabetes. Thirty-four patients with type 1 diabetes performed a 24-h complete fast at home. Thirteen patients were treated with multiple insulin injections using either glargine (n=12) or NPH (n=1) as basal insulin. The remaining patients were treated with an insulin pump. All patients received their basal insulin only, which was adjusted to 40% of their total daily dose, and were monitored by either a Gold(®) or Guardian(®) continuous glucose monitoring (CGMS) device. Capillary glucose (SMBG) was targeted at 3.9-7.8 mmol/L, with a standardized protocol for correction of hyper- and hypoglycaemia. Interstitial glucose (IG) profiles were compared with the SMBG values; the IG profiles of patients using glargine or a pump and either of the two CGMS devices were also compared. All of the patients completed the 24-h fast with no major incident. At the end of the fast, 80% of the IG values were on target. The route by which insulin was delivered made no difference, but there were more IG values on target in patients monitored by the Guardian(®) device. IG was below target in 104 occurrences and above-target in 34. After a mean intake of 10 g of sucrose, below-target IG was corrected within 30 min [range: 15-40]. The mean insulin dose to correct above-target episodes was 1 U. Prolonged fasting is possible at home in patients with type 1 diabetes, provided the basal dose is adjusted. The use of CGMS is not necessary, but offers useful information on the patient's IG profile during the fast.


H+ and Cl- ion equilibrium across the red cell membrane in the carp.
H2O and electrolyte distribution were studied in carp erythrocytes at various pH values achieved by CO2/O2 equilibration in vitro. Intracellular pH was measured by means of glass electrode and by DMO-14C. rH+, rCl- and rDMO- varied linearly and in a comparable manner with pHe. At pHe = 7.8, rH+, rCl- and rDMO- were 0.21, 0.29 and 0.30. rCl- and rDMO- were closely correlated and exhibited only minor differences. rH+ was closely correlated with rCl- and rDMO-, but was, however, significantly lower than rCl- or rDMO-. This difference is considered to be due to a systematic error of the glass electrode when used in highly concentrated protein solutions. The coulometric determination of chloride in packed red cells is shown to be highly susceptible to protein. The results are consistent with the assumption that H+ and Cl- ions are passively distributed across the red cell membrane.


Identification of potential biomarkers and immune infiltration characteristics in recurrent implantation failure using bioinformatics analysis.
Recurrent implantation failure (RIF) is a frustrating challenge because the cause is unknown. The current study aims to identify differentially expressed genes (DEGs) in the endometrium on the basis of immune cell infiltration characteristics between RIF patients and healthy controls, as well as to investigate potential prognostic markers in RIF. GSE103465, and GSE111974 datasets from the Gene Expression Omnibus database were obtained to screen DEGs between RIF and control groups. Gene Ontology analysis, Kyoto Encyclopedia of Genes and Genomes Pathway analysis, Gene Set Enrichment Analysis, and Protein-protein interactions analysis were performed to investigate potential biological functions and signaling pathways. CIBERSORT was used to describe the level of immune infiltration in RIF, and flow cytometry was used to confirm the top two most abundant immune cells detected. 122 downregulated and 66 upregulated DEGs were obtained between RIF and control groups. Six immune-related hub genes were discovered, which were involved in Wnt/-catenin signaling and Notch signaling as a result of our research. The ROC curves revealed that three of the six identified genes (AKT1, PSMB8, and PSMD10) had potential diagnostic values for RIF. Finally, we used cMap analysis to identify potential therapeutic or induced compounds for RIF, among which fulvestrant (estrogen receptor antagonist), bisindolylmaleimide-ix (CDK and PKC inhibitor), and JNK-9L (JNK inhibitor) were thought to influence the pathogenic process of RIF. Furthermore, our findings revealed the level of immune infiltration in RIF by highlighting three signaling pathways (Wnt/-catenin signaling, Notch signaling, and immune response) and three potential diagnostic DEGs (AKT1, PSMB8, and PSMD10). Importantly, our findings may contribute to the scientific basis for several potential therapeutic agents to improve endometrial receptivity.


Functional and histologic imaging of urinary bladder wall after exposure to psychological stress and protamine sulfate.
To quantify the urinary bladder wall T1 relaxation time (T1) before and after the instillation contrast mixture in rats previously subjected to water avoidance stress (WAS) and/or acute exposure to protamine sulfate (PS). Female Wistar rats were randomized to receive either sham (control) or 1 h of WAS for ten consecutive days before the evaluation of nocturnal urination pattern in metabolic cages. T1 mapping of urinary bladder wall at 9.4 T was performed pre- and post- instillation of 4 mM Gadobutrol in a mixture with 5 mM Ferumoxytol. Subsequently, either T1 mapping was repeated after brief intravesical PS exposure or the animals were sacrificed for histology and analyzing the mucosal levels of mRNA. Compared to the control group, WAS exposure decreased the single void urine volume and shortened the post-contrast T1 relaxation time of mucosa- used to compute relatively higher ingress of instilled Gadobutrol. Compromised permeability in WAS group was corroborated by the urothelial denudation, edema and ZO-1 downregulation. PS exposure doubled the baseline ingress of Gadobutrol in both groups. These findings confirm that psychological stress compromises the paracellular permeability of bladder mucosa and its non-invasive assay with MRI was validated by PS exposure.


Characterization and evolution of napin-encoding genes in radish and related crucifers.
Three cDNA clones, encoding napin storage proteins from radish, were isolated and sequenced. They fall into two classes differing in the size of the primary translation product. Sequences of the two classes are very well conserved and they display an organization very similar to that of the homologous genes from rapeseed and Arabidopsis which have previously been described. On the basis of hybridization intensity and the number of restriction fragments, we estimate that the radish napin multigene family is represented by eight to twelve members. The use of probes specific to each subfamily demonstrates that they contribute to a similar extent to the production of napin mRNA. Analysis of the sequence data suggests that the napin ancestral genes are probably derived from successive duplication and divergence of a protogene. Comparing other available napin sequences with those of radish reveals intriguing features. Comparison of the coding sequences shows that the homology between the radish and rapeseed sequences is much higher than that between each of the four members of the Arabidopsis gene family. This would suggest that the duplications which gave rise to the different members occurred independently in the two groups of species after separation of Arabidopsis from the Brassica lineage. However, similar comparison carried out on the 3' -noncoding sequences does not support this hypothesis, but shows that slightly different duplicated genes probably already existed in the common ancestor to the three genera. This paradox can be resolved by assuming that, within each genus, coding sequences for napin-encoding genes have been considerably homogenized as a result of concerted evolution.


Lessons Learned from the Ottawa Regional STEMI Program.
Percutaneous coronary intervention (PCI) has become the dominant strategy for the treatment of ST-segment elevation myocardial infarction (STEMI) when rapid access to a catheterization facility is available. In communities where primary PCI is not feasible, a pharmacoinvasive strategy has become a recommended option. At the University of Ottawa Heart Institute, a care delivery model has been developed in which primary PCI and pharmacoinvasive strategies are applied for an entire region. This article reviews the lessons learned in setting up and maintaining a regional STEMI program.


Longitudinal Assessment of the Tiger Milk Medicinal Mushroom, Lignosus rhinocerus (Agaricomycetes), Sclerotium Cultivar: Uniformity of Bioactive Components.
Traditional use of the tiger milk medicinal mushroom, Lignosus rhinocerus, to treat various illnesses has been recorded for > 4 centuries. Successful cultivation of L. rhinocerus using proprietary solid-state fermentation (SSF) technology by LiGNO Biotech has enabled large-scale production of L. rhinocerus sclerotia (termed L. rhinocerus TM02) and further investigations into its medicinal properties. Pharmacological activities of L. rhinocerus TM02, including its antioxidant, anti-inflammatory, anticancer, and immunomodulatory effects and the bioactive components responsible, have been validated by various scientific studies. In this study, we assessed the consistency of the bioactive components in 11 batches of L. rhinocerus TM02 produced over a 9-year period. The different batches of L. rhinocerus TM02 consisted of stable protein, polysaccharide, and glycoprotein contents, and all tested samples were comparable to the wild type. L. rhinocerus TM02 had greater protein, carbohydrate, and glycoprotein contents, which were mostly bioactive compared to another cultivar from a different cultivation technology (TM-UN). Together with previous scientific validations, L. rhinocerus TM02 produced using SSF cultivation is of optimal quality with high consistent bioactive contents, which can be an appropriate indicator for quality validation of the much sought-after medicinal mushroom, L. rhinocerus.


[Framingham function estimates the risk of cardio vascular mortality more effectively than SCORE function in the population of the Canary Islands (Spain)].
To compare the performance of the Framingham and SCORE functions to estimate fatal cardiovascular events. In addition, we explored the ability of both functions to detect the risk contributed by factors not included in their equations: sedentariness, obesity, abdominal circumference, abdomen/height razón, abdomen/pelvis ratio, and excessive alcohol consumption. We performed a cross-sectional study of 5,289 individuals aged 30 to 69 years old, recruited by random sampling of the general population of the Canary Islands. We calibrated the Framingham and SCORE functions and estimated their concordance. The cardiovascular mortality rate for the population in this age range was compared with the risk predicted by the two functions. Among males, the mortality rate per 100,000 inhabitants was 67.4, while the Framingham, SCORE-low and SCORE-high estimations were 80, 140, and 270, respectively. Among females, the mortality rate was 19.3 while the estimations were 30, 50, and 70, respectively. Both functions detected the increased risk contributed by the factors studied, except for sedentariness among females with SCORE, and excessive alcohol consumption with both functions. Among males, taking cut points of > or = 12%, > or = 15%, and > or = 20% for Framingham, the concordance with SCORE-low yielded Kappa values of 0.6, 0.7, and 0.5, respectively. The Framingham function yielded the best estimate of cardiovascular mortality rates. Only Framingham detected the cardiovascular risk contributed by sedentariness in both genders. We recommend the use of the calibrated Framingham function for this population.


Drinking water treatment residue recycled to synchronously control the pollution of polycyclic aromatic hydrocarbons and phosphorus in sediment from aquatic ecosystems.
Great efforts have long been made to control sediment pollution from persistent organic pollutants and phosphorus for aquatic ecosystem restoration. This study proposed a novel recycling of drinking water treatment residue (DWTR) to synchronously control sediment polycyclic aromatic hydrocarbons (PAHs) and phosphorus pollution based on a 350-day incubation test. The results suggested that DWTR addition reduced approximately 88%- 96% of potential bioavailable PAHs and 76% of mobile phosphorus in sediment. The dominant mechanisms for both reductions by DWTR were immobilization, mainly through increasing sediment amorphous aluminum and iron. The tendency of enhanced PAHs degradation by DWTR was also observed, especially for high molecular weight PAHs (e.g., chrysene, indeno(1, 2, 3-cd)pyrene, and benzo(g, hi)perylene), which decreased by approximately 21.1%- 22.0% of the total. Additionally, accompanying a clear increase in the connections of microbial cooccurrence networks, the variations in bioavailable PAHs, amorphous aluminum and iron, and other properties (e.g., pH, nitrogen, and organic matter) significantly (p < 0.01) enhanced Flavobacterium enrichment, although the enrichment of many other microbes potentially related to PAHs degradation (e.g., C1-B045) decreased after DWTR addition. Therefore, DWTR could promote the construction of a "PAHs immobilization with microbial augmentation" system while immobilizing phosphorus in sediment, indicating the high feasibility of controlling multiple sediment pollution.


Salivary gland degeneration and ovarian development in the Rocky Mountain wood tick, Dermacentor andersoni Stiles (Acari: Ixodidae). I. Post-engorgement events.
It is well known that female ixodid ticks undergo salivary gland degeneration and the greater part of ovarian development following engorgement. The process has been particularly well studied in Amblyomma hebraeum, including the hormonal control of these processes. The purpose of this study, and the second one in this series, is to compare the processes in Dermacentor andersoni with those of A. hebraeum. A major difference between the two species is that virgin female D. andersoni feed to a much higher fed-to-unfed weight ratio than do virgin female A. hebraeum, the former achieving an average of 67× the unfed weight in this study, the latter normally achieving an average of only about 10-15× the unfed weight. We show here that although engorged virgins (defined here as those that exceed a fed-unfed weight ratio of 50, even though they do not detach from the host spontaneously) degenerate their salivary glands and develop their ovaries, these processes proceed more slowly in virgins that they do in mated females, as is the case for A. hebraeum.


Imaging-Negative Hepatocellular Carcinoma Presents as an Intrabiliary Mass.
Hepatocellular carcinoma (HCC) is the second most common cause of cancer-related death and one of the most prevalent cancers worldwide. HCC prognosis remains poor with an average survival rate between 6 and 12 months. Obstructive jaundice, as a main clinical feature, is uncommon in HCC. HCC with bile duct invasion is much rarer than HCC with vascular invasion. We present a case where a patient's HCC was diagnosed by endoscopic retrograde cholangiopancreatography and digital cholangioscopy because his HCC manifested as an obstructing lesion in the intrahepatic duct, but not in the liver.


Isolation and characterization of charge variants of infliximab biosimilar HS626.
Charge variants are the most commonly observed sources of heterogeneity in the routine manufacturing of monoclonal antibodies. To gain further insight into the structural foundation of charge heterogeneity and its influence on biological functions, an infliximab biosimilar HS626 from a biopharmaceutical facility was isolated by semipreparative cation exchange chromatography (CEX) to obtain fractions of acidic and basic charge variants and determine the main species. It was assessed again by CEX to ensure purities. Through a series of structural and physicochemical characterizations, we concluded that the acidic variants were caused by fragments, Met oxidation, Asn deamidation, higher levels of sialylation and galactosylation of N-linked glycans, and less high mannose. The basic variants resulted mainly from aggregates, fragments, and Met oxidation. Through further analysis of antigen binding affinity, cell death inhibitory activity, ADCC, and CDC, as well as FcRn, FcγRIIIa, and C1q affinity, we demonstrated that the charge heterogeneity did not affect biological functions. This research enhances the understanding of charge variants, which are usually effective components that should not be intentionally reduced unless biological functions are affected.


Design, synthesis and biological evaluation of new series of hexahydroquinoline and fused quinoline derivatives as potent inhibitors of wild-type EGFR and mutant EGFR (L858R and T790M).
New series of hexahydroquinoline and fused quinoline derivatives were designed and synthesized. The thirty seven new compounds were screened for in vitro antitumor activity against HepG2, HCT-116 and MCF-7 cancer cells. Results indicated that compounds 2e, 2h, 5b, 5c, 6a, 7d and 9b have the strongest potency against the three cancer cells, and they were further screened for in vitro cytotoxicity against A431 and H1975 cancer cells, as well as WI38 and WISH normal cells. Results revealed that 7d potently inhibited the growth of H1975 cells harboring EGFRT790M mutation (IC50 = 1.32 ± 0.2 µM) over A431 cells overexpressing EGFRWT (IC50 = 4.96 ± 0.3 µM). Moreover, the seven compounds displayed low cytotoxicity against the tested normal cells. The seven potent antitumor compounds were examined for their ability to inhibit the activity of EGFRWT. The attained data manifested that 7d has remarkable EGFRWT inhibitory activity (IC50 = 0.083 ± 0.002 μM) compared to erlotinib (IC50 = 0.067 ± 0.002 μM). Compound 7d was further studied for its enzymatic inhibitory activity against other eight human kinases, and it displayed outstanding inhibitory activity against EGFRL858R and EGFRT790M mutants (IC50 = 0.053 ± 0.002, 0.026 ± 0.001 μM, respectively), as well as JAK3 (IC50 = 0.069 ± 0.003 μM). Analysis of cell cycle evidenced that 7d induces cell cycle arrest in G2/M and pre-G1 phases in the tested cancer cells. In addition, cancer cell death induced by 7d was proved to take place via apoptosis supported by elevated Bax/Bcl-2 ratio in the tested cancer cells. Moreover, docking results confirmed the good binding interactions of 7d with EGFRWT, EGFRL858R, EGFRT790M and JAK3, which came in agreement with the results of in vitro enzyme assay. Further, 7d is predicted to have good oral absorption, good drug-likeness properties and low toxicity risks in human.


A case report of CBD and THC as analgesic therapy in a cat with chronic osteoarthritic pain.
A 10-year-old mixed breed male cat presented with clinical signs related to chronic orthopaedic pain. Upon physical examination, pain was noted, based on the feline Musculoskeletal Pain Index (FMPI). An analgesic treatment with a full spectrum cannabis oil (1.8% CBD and 0.8% THC) was proposed for 30 days (0,5 mg/kg based on CBD). The FMPI scale score decreased more than 50%. This case reported a satisfactory outcome for the patient and the owner, although this medication could increase ALT. Given the paucity of literature published to date on the treatment of veterinary species with cannabis-based medications, further clinical and pharmacokinetic studies are necessary to study the safety and efficacy of its use.


Prostaglandin D2 interacts at thromboxane receptor-sites on guinea-pig platelets.
The anti-aggregatory prostanoid, prostaglandin D2 (PGD2) does not completely inhibit ADP-induced aggregation of guinea-pig platelets and thus produces a bell-shaped dose-inhibition curve. The nature of this bell-shaped curve has now been investigated in guinea-pig platelet-rich plasma. Two selective thromboxane receptor antagonists, 13-aza-prostanoic acid (13-AZA; 16-64.4 microM) and BM 13.177 (5.9-29.8 microM), converted PGD2 to a full inhibitor of aggregation in a dose-related manner. The putative platelet PGD2 receptor antagonist, N-0164 (75 microM) also converted PGD2 to a full inhibitor of platelet aggregation. In contrast to 13-AZA and BM 13.177, higher concentrations of N-0164 (380 and 760 microM) caused a dose-related rightward shift of the PGD2 dose-inhibition curve. The thromboxane receptor antagonism of N-0164 was confirmed in studies in which the dose-aggregation curve to U-46619, a thromboxane mimetic, was competitively antagonized with a pA2 value of 4.67 and a slope of 1.13, comparable to that of 13-AZA. The results show that N-0164 acts as both a platelet PGD2 and thromboxane-receptor antagonist in both human and guinea-pig platelet-rich plasma. The results further indicate that PGD2 can interact at thromboxane receptors in guinea-pig platelets.


A novel tau-based rhesus monkey model of Alzheimer's pathogenesis.
Alzheimer's disease (AD) is a devastating condition with no effective treatments, with promising findings in rodents failing to translate into successful therapies for patients. Targeting the vulnerable entorhinal cortex (ERC), rhesus monkeys received two injections of an adeno-associated virus expressing a double tau mutation (AAV-P301L/S320F) in the left hemisphere, and control AAV-green fluorescent protein in the right ERC. Noninjected aged-matched monkeys served as additional controls. Within 3 months we observed evidence of misfolded tau propagation, similar to what is hypothesized to occur in humans. Viral delivery of human 4R-tau also coaptates monkey 3R-tau via permissive templating. Tau spreading is accompanied by robust neuroinflammatory response driven by TREM2+ microglia, with biomarkers of inflammation and neuronal loss in the cerebrospinal fluid and plasma. These results highlight the initial stages of tau seeding and propagation in a primate model, a more powerful translational approach for the development of new therapies for AD.


Stress free configuration of the human eye.
Numerical simulations of eye globes often rely on topographies that have been measured in vivo using devices such as the Pentacam or OCT. The topographies, which represent the form of the already stressed eye under the existing intraocular pressure, introduce approximations in the analysis. The accuracy of the simulations could be improved if either the stress state of the eye under the effect of intraocular pressure is determined, or the stress-free form of the eye estimated prior to conducting the analysis. This study reviews earlier attempts to address this problem and assesses the performance of an iterative technique proposed by Pandolfi and Holzapfel [1], which is both simple to implement and promises high accuracy in estimating the eye's stress-free form. A parametric study has been conducted and demonstrated reliance of the error level on the level of flexibility of the eye model, especially in the cornea region. However, in all cases considered 3-4 analysis iterations were sufficient to produce a stress-free form with average errors in node location <10(-6)mm and a maximal error <10(-4)mm. This error level, which is similar to what has been achieved with other methods and orders of magnitude lower than the accuracy of current clinical topography systems, justifies the use of the technique as a pre-processing step in ocular numerical simulations.


Healing of bony defects in the irradiated and unirradiated rat mandible.
A model system of the irradiated rat mandible has been developed and used in conjunction with a non-spontaneously healing mandibular defect. The contribution of the tissue components in the healing of bony defects was studied using demineralized bone powder (DBP) prepared from unirradiated or in vivo irradiated rat long bones. Better bony fill of the defects occurred in the irradiated beds filled with unirradiated DBP than in the unirradiated beds containing irradiated DBP. This suggests that, at least in the early postirradiation period, the bed is not the limiting factor in healing of bony defects and the osteogenic components of bone in the DBP may be most affected by irradiation. In the irradiated bed, the defects grafted 2 weeks after irradiation healed better than those grafted at 4 weeks. Thus, the timing of surgery after irradiation also plays a role in the healing process, with early surgery producing better results.


Decentralization for cost-effective conservation.
Since 1930, areas of state-managed forest in the central Himalayas of India have increasingly been devolved to management by local communities. This article studies the long-run effects of the devolution on the cost of forest management and on forest conservation. Village council-management costs an order of magnitude less per unit area and does no worse, and possibly better, at conservation than state management. Geographic proximity and historical and ecological information are used to separate the effects of management from those of possible confounding factors.


The contribution of cardiac myosin binding protein-c Ser282 phosphorylation to the rate of force generation and in vivo cardiac contractility.
Cardiac myosin binding protein-C phosphorylation plays an important role in modulating cardiac muscle function and accelerating contraction. It has been proposed that Ser282 phosphorylation may serve as a critical molecular switch that regulates the phosphorylation of neighbouring Ser273 and Ser302 residues, and thereby govern myofilament contractile acceleration in response to protein kinase A (PKA). Therefore, to determine the regulatory roles of Ser282 we generated a transgenic (TG) mouse model expressing cardiac myosin binding protein-C with a non-phosphorylatable Ser282 (i.e. serine to alanine substitution, TG(S282A)). Myofibrils isolated from TG(S282A) hearts displayed robust PKA-mediated phosphorylation of Ser273 and Ser302, and the increase in phosphorylation was identical to TG wild-type (TG(WT)) controls. No signs of pathological cardiac hypertrophy were detected in TG(S282A) hearts by either histological examination of cardiac sections or echocardiography. Baseline fractional shortening, ejection fraction, isovolumic relaxation time, rate of pressure development and rate of relaxation (τ) were unaltered in TG(S282A) mice. However, the increase in cardiac contractility as well as the acceleration of pressure development observed in response to β-adrenergic stimulation was attenuated in TG(S282A) mice. In agreement with our in vivo data, in vitro force measurements revealed that PKA-mediated acceleration of cross-bridge kinetics in TG(S282A) myocardium was significantly attenuated compared to TG(WT) myocardium. Taken together, our data suggest that while Ser282 phosphorylation does not regulate the phosphorylation of neighbouring Ser residues and basal cardiac function, full acceleration of cross-bridge kinetics and left ventricular pressure development cannot be achieved in its absence.


Dysphotopsia: a multifaceted optic phenomenon.
To summarize the literature on dysphotopsia, focusing on strategies for management and prevention. Clinical studies and ray-tracing diagrams have improved our understanding of the risk factors for positive and negative dysphotopsiae. Well known management and prevention strategies such as piggyback IOL, reverse optic capture, Nd:YAG capsulotomy, and IOL exchange are reviewed, as well as newer strategies such as the infertemporal approach to IOL implantation, 'in-the-bag nasal optic truncation' and new IOL design implantation, and new IOL design. Positive and negative dysphotopsiae are known to occur in a number of patients after cataract surgery. Although studying dysphotopsiae has provided many challenges given its subjective patient experience and its multifactorial cause, these same challenges have generated new theories and ideas regarding dysphotopsiae. A better understanding of the available methods for treatment and prevention can improve management of dysphotopsiae and patient satisfaction.


Family Presence and Sleep in the Intensive Care Unit.
Sleep is a vital component of health and healing. Hospitalized patients need sleep in order to overcome their illness, and family members of those patients also need restorative sleep. Nurses can assist with both patient's and family member's abilities to obtain nonfragmented sleep. Education is important for families to understand the importance of sleep, and nurses can also supply families with simple comfort measures such as ear plugs, eye masks, and a comfortable sleep location.


Long-Term Clinical Effectiveness of Ustekinumab in Patients With Crohn's Disease: A Retrospective Cohort Study.
This study clarifies the long-term effectiveness of ustekinumab based on real-life data from Japanese Crohn's disease (CD) patients. A total of 137 patients were included, and 124 patients (90.5%) were exposed to anti-tumor necrosis factor-α agents. The clinical remission rate at week 52 was 32.4% in moderate to severely active CD patients. The achievement of clinical remission for 8 weeks after ustekinumab therapy induction was associated with clinical remission at week 52. Ustekinumab persistence rate at week 104 was 81.4%. Ustekinumab is effective and persistent in CD patients with the previous treatment history of several biologics.


Accurate prediction of cis-regulatory modules reveals a prevalent regulatory genome of humans.
cis-regulatory modules(CRMs) formed by clusters of transcription factor (TF) binding sites (TFBSs) are as important as coding sequences in specifying phenotypes of humans. It is essential to categorize all CRMs and constituent TFBSs in the genome. In contrast to most existing methods that predict CRMs in specific cell types using epigenetic marks, we predict a largely cell type agonistic but more comprehensive map of CRMs and constituent TFBSs in the gnome by integrating all available TF ChIP-seq datasets. Our method is able to partition 77.47% of genome regions covered by available 6092 datasets into a CRM candidate (CRMC) set (56.84%) and a non-CRMC set (43.16%). Intriguingly, the predicted CRMCs are under strong evolutionary constraints, while the non-CRMCs are largely selectively neutral, strongly suggesting that the CRMCs are likely cis-regulatory, while the non-CRMCs are not. Our predicted CRMs are under stronger evolutionary constraints than three state-of-the-art predictions (GeneHancer, EnhancerAtlas and ENCODE phase 3) and substantially outperform them for recalling VISTA enhancers and non-coding ClinVar variants. We estimated that the human genome might encode about 1.47M CRMs and 68M TFBSs, comprising about 55% and 22% of the genome, respectively; for both of which, we predicted 80%. Therefore, the cis-regulatory genome appears to be more prevalent than originally thought.


Identification of various orthodontic materials as foreign bodies via panoramic radiography, cone beam computed tomography, magnetic resonance imaging, and ultrasonography: an in vitro study.
We evaluated the in vitro detection sensitivity of orthodontic materials (serving as foreign bodies) using panoramic radiography, cone beam computed tomography (CBCT), magnetic resonance imaging (MRI), and ultrasonography. Five different orthodontic materials served as foreign bodies: titanium-molybdenum alloy wire (TMA; ORMCO, Orange, CA, USA; 0.017 × 0.025 in in cross-sectional dimensions and 1 cm long); stainless steel bracket tooth #34 (American Orthodontics, Sheboygan, WI, USA); a monocrystalline, sapphire ceramic bracket tooth #34 (Skyortho Dental Supplies Medical, China); a polycrystalline alumina clear bracket, Damon clear bracket tooth #34 (ORMCO); and a 1 × 1 × 0.1 cm polyurethane-based thermoplastic material, Invisalign clear aligner (Align Technology, San Jose, CA, USA). Panoramic radiography, CBCT, MRI, and ultrasonography were used, and four observers scored all findings independently. The TMA and stainless steel bracket were visualised in all fields by panoramic radiography and CBCT. The sapphire and Damon brackets were very clear on CBCT. The Invisalign in air was evident only on CBCT. MRI was unable to identify any material in muscle. Ultrasonography detected the TMA, sapphire bracket, and the Invisalign in muscle but only the TMA on bone. Panoramic radiography does not reveal nonmetallic orthodontic equipment in air and reveals them only poorly in muscle. CBCT was the optimal imaging modality for all materials in all fields except for the Invisalign in muscle and bone. CBCT was the only method that revealed the Invisalign in air. MRI and ultrasonography should be used to detect orthodontic materials in muscle.


Compartmentalization of cold-stable and acetylated microtubules in the subsynaptic domain of chick skeletal muscle fibre.
The junction between motor nerve and skeletal muscle in vertebrates is characterized by a complex subneural structure in which the nicotinic acetylcholine receptor (AChR) accumulates at a surface density of up to 10,000-20,000 molecules per microns 2. Among the mechanisms involved in the maintenance of the accumulation of AChR molecules is the localized expression of AChR subunit genes in the sarcoplasmic nuclei underlying the motor endplate, referred to as 'fundamental' by Ranvier. Immunocytochemical analysis has revealed that in innervated muscle fibres, the Golgi apparatus is also localized to subjunctional domains. We now report an accumulation of cold-stable and acetylated microtubules in the region of the sarcoplasm underlying the endplate. Together, these observations indicate that the subsynaptic sarcoplasm represents a compartment that is specialized both for the transcription, post-translational processing and stabilization of proteins of the postsynaptic membrane, in particular AChR, and for their targeting to the motor endplate.