MNK1, a new MAP kinase-activated protein kinase, isolated by a novel expression screening method for identifying protein kinase substrates.
We have developed a novel expression screening method for identifying protein kinase substrates. In this method, a lambda phage cDNA expression library is screened by in situ, solid-phase phosphorylation using purified protein kinase and [gamma-32P]ATP. Screening a HeLa cDNA library with ERK1 MAP kinase yielded cDNAs of previously characterized ERK substrates, c-Myc and p90RSK, demonstrating the utility of this method for identifying physiological protein kinase substrates. A novel clone isolated in this screen, designated MNK1, encodes a protein-serine/threonine kinase, which is most similar to MAP kinase-activated protein kinase 2 (MAPKAP-K2), 3pK/MAPKAP-K3 and p90RSK. Bacterially expressed MNK1 was phosphorylated and activated in vitro by ERK1 and p38 MAP kinases but not by JNK/SAPK. Further, MNK1 was activated upon stimulation of HeLa cells with 12-O-tetradecanoylphorbol-13-acetate, fetal calf serum, anisomycin, UV irradiation, tumor necrosis factor-alpha, interleukin-1beta, or osmotic shock, and the activation by these stimuli was differentially inhibited by the MEK inhibitor PD098059 or the p38 MAP kinase inhibitor SB202190. Together, these results indicate that MNK1 is a novel class of protein kinase that is activated through both the ERK and p38 MAP kinase signaling pathways.


AXL and MET receptor tyrosine kinases are essential for lung cancer metastasis.
The AXL and MET receptors regulate key processes in tumor growth, metastasis, and drug resistance; thus, they have recently been implicated as promising therapeutic targets in various tumors. We investigated the metastatic potential and crosstalk between these receptors in non‑small cell lung cancer (NSCLC). We found that the treatment of NSCLC cells with hepatocyte growth factor (HGF) and growth arrest-specific 6 (Gas6), as ligands for MET and AXL, respectively, promoted their migration and invasion ability. However, treatment with inhibitors of each of these receptors significantly reduced the migratory and invasiveness of the cells, although their inhibitory rates varied according to the inhibition of each receptor. In addition, the suppression of each receptor by shRNA resulted in reduced migration and invasiveness. Notably, the suppression of AXL was more effective than the suppression of MET in the inhibition of migration and invasion. In accordance with in vitro results, when the cells were transferred via tail vein injection, AXL inhibition was more efficient in attenuating metastasis than MET inhibition. Clinically, AXL or MET expression is associated with a poor prognosis in primary tumors of NSCLC. In summary, AXL and MET can regulate tumor metastasis, but AXL was shown to be more potent than MET in lung metastasis. Thus, we conclude that AXL might be a suitable therapeutic target for the inhibition of lung metastasis.


[Role of bacteriochlorophyll in stabilization of the structure of the near-central and peripheral light-harvesting complexes from purple photosynthetic bacteria].
Pheophytinization of bacteriochlorophyll (BChl) at low pH was investigated in the core (LH1) and peripheral (LH2) light-harvesting complexes, as well as in the ensemble of the reaction center (RC) with the LH1 complex. The stages in disintegration of the native BChl forms in the LH1 complex and in its ensemble with RC were revealed. They were observed as emergence of the absorption band of monomeric BChl and an increase in its intensity, followed by its transformation into the band of monomeric bacteriopheophytin (BPh) and then into the band of aggregated BPh. Unlike the LH1 complex, in the case of the LH2 complex monomeric BChl was never detected as an intermediate product. While the spectra revealed formation of monomeric BPh, its accumulation did not occur, since its aggregation is very rapid compared to that in the LH1 complex and in the RC-LH1 ensemble. PAG electrophoresis revealed that pheophytinization of BChl in the LH2 complex was accompanied by disruption of the stable cylindrical structure of this complex with emergence of characteristic fragments consisting of α and β peptides and bearing monomeric BPh, as well as of the α peptide aggregates bearing BPh aggregates. Unlike the LH2 complex, BChl pheophytinization in the LH1 complex did not result in its fragmentation. This is an indication of different types of structural stabilization in the LH1 and LH2 complexes. In the LH2 complex, coordination of bacteriochlorophyll Mg2+ by conservative histidine residues of the α and β polypeptides is the main factor responsible for the maintenance of its cylindrical structure. Stability of the LH1 complex is probably based primarily on the highly specific hydrophobic interactions between the surfaces of individual polypeptide chains, since the presence of hydrogen bonds results in autonomy of each αβ3BChl2 subunit, rather than in stabilization of the LH1 complex as a whole.


Minichromosome maintenance 6 complex component identified by bioinformatics analysis and experimental validation in esophageal squamous cell carcinoma.
Esophageal squamous cell carcinoma (ESCC), the main subtype of esophageal cancer (EC), is a common lethal type of cancer with a high mortality rate. The aim of the present study was to select key relevant genes and identify potential mechanisms involved in the development of ESCC based on bioinformatics analysis. Minichromosome maintenance 6 complex component (MCM6) has been identified to be upregulated in multiple malignancies; however, its contributions to ESCC remain unclear. For the purposes of the present study, four datasets were downloaded from the Gene Expression Omnibus (GSE63941, GSE26886, GSE17351 and GSE77861), and the intersection of the differentially expressed genes was obtained using a Venn diagram. The protein‑protein interaction was then constructed, and the modules were verified by Cytoscape, in which the key genes have a high connectivity degree with other genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway were subsequently filtered out to analyze the development of ESCC. MCM6, an upregulated gene, was selected and connected with most of the other genes, for further research validation. The expression levels of MCM6 were then assessed using the Oncomine, GEPIA and UALCAN databases and validated in both ESCC tissues samples and cell lines by immunohistochemistry and RT‑qPCR. Cell counting kit‑8 (CCK‑8), flow cytometry, wound healing and Transwell assays were used to determine the proliferation, apoptosis, cell cycle, migration and invasion of ESCC cells. A total of 24 genes were identified by a series of bioinformatics analyses and the results revealed that the genes were associated with DNA replication and cell cycle. Experimental validation revealed that MCM6 expression was significantly elevated in both ESCC tissues and cell lines. The results were consistent with those of bioinformatics analysis. Furthermore, the knockdown of MCM6 inhibited cell proliferation, migration and invasion and promoted cell apoptosis, and made cells arrested in S stage. In summary, the findings of bioinformatics analysis provided a novel hypothesis for ESCC progression. In particular, the aberrantly elevated expression of MCM6 is a potential biomarker for ESCC diagnosis and treatment.


Complete Genome Sequences of Six South African Rabies Viruses.
South African rabies viruses (RABVs) from dogs and jackals (canid viruses) are highly related and most likely originated from a single progenitor. RABV is the cause of most global human rabies cases. The complete genome sequences of 3 RABVs from South Africa and Zimbabwe are reported here.


New ent-Clerodane and Abietane Diterpenoids from the Roots of Kenyan Croton megalocarpoides Friis & M. G. Gilbert.
The roots of the endangered medicinal plant Croton megalocarpoides collected in Kenya were investigated and twenty-two compounds isolated. Among them were twelve new ent-clerodane (1-12) and a new abietane (13) diterpenoids, alongside the known crotocorylifuran (4 a), two known abietane and four known ent-trachylobane diterpenoids, and the triterpenoids, lupeol and acetyl aleurotolic acid. The structures of the compounds were determined using NMR, HRMS and ECD. The isolated compounds were evaluated against a series of microorganisms (fungal and bacteria) and also against Plasmodium falciparum, however no activity was observed.


Anomeric bond-character in the pyranose sugars.
Evidence from determinations of crystal structure relating to the shortening of carbon-oxygen bonds in the anomeric position in pyranose sugars is presented. There is a high degree of probability that the C(1)-O(1) bond is shortened by about 0.04 A relative to the other C-O single-bond lengths, except in the case of an axially oriented glycosidic group where there is evidence of a distinction between the two ring C-O bonds.


Identification of a characteristic vascular belt zone in human colorectal cancer.
Intra-tumoral blood vessels are of supreme importance for tumor growth, metastasis and therapy. Yet, little is known about spatial distribution patterns of these vessels. Most experimental or theoretical tumor models implicitly assume that blood vessels are equally abundant in different parts of the tumor, which has far-reaching implications for chemotherapy and tumor metabolism. In contrast, based on histological observations, we hypothesized that blood vessels follow specific spatial distribution patterns in colorectal cancer tissue. We developed and applied a novel computational approach to identify spatial patterns of angiogenesis in histological whole-slide images of human colorectal cancer. In 33 of 34 (97%) colorectal cancer primary tumors blood vessels were significantly aggregated in a sharply limited belt-like zone at the interface of tumor tissue to the intestinal lumen. In contrast, in 11 of 11 (100%) colorectal cancer liver metastases, a similar hypervascularized zone could be found at the boundary to surrounding liver tissue. Also, in an independent validation cohort, we found this vascular belt zone: 22 of 23 (96%) samples of primary tumors and 15 of 16 (94%) samples of liver metastases exhibited the above-mentioned spatial distribution. We report consistent spatial patterns of tumor vascularization that may have far-reaching implications for models of drug distribution, tumor metabolism and tumor growth: luminal hypervascularization in colorectal cancer primary tumors is a previously overlooked feature of cancer tissue. In colorectal cancer liver metastases, we describe a corresponding pattern at the invasive margin. These findings add another puzzle piece to the complex concept of tumor heterogeneity.


Three-Step Synthesis of the Antiepileptic Drug Candidate Pynegabine.
Pynegabine, an antiepileptic drug candidate in phase I clinical trials, is a structural analog of the marketed drug retigabine with improved chemical stability, strong efficacy, and a better safety margin. The reported shortest synthetic route for pynegabine contains six steps and involves the manipulation of highly toxic methyl chloroformate and dangerous hydrogen gas. To improve the feasibility of drug production, we developed a concise, three-step process using unconventional methoxycarbonylation and highly efficient Buchwald-Hartwig cross coupling. The new synthetic route generated pynegabine at the decagram scale without column chromatographic purification and avoided the dangerous manipulation of hazardous reagents.


Animals, science, and ethics -- Section II. Animals in science: the justification issue.
Section II, addressing the ethical justification of the uses of animals in science, does not offer "decisive" arguments for or against particular uses. Rather, conflicting values, goods, and responsibilities pertaining to both humans and animals are identified. Particular uses of animals demand the careful consideration of various ethical claims, which will always require the developed art and wisdom of ethical judgment and will never be reduced to a systematic science. Competing ultimate goods or values and the concrete complexity of the real world defy such rational, abstract, or logical harmony.


Spinal infections: clinical and imaging features.
Spinal infections represent a group of rare conditions affecting vertebral bodies, intervertebral discs, paraspinal soft tissues, epidural space, meninges, and spinal cord. The causal factors, clinical presentations, and imaging features are a challenge because the difficulty to differentiate them from other conditions, such as degenerative and inflammatory disorders and spinal neoplasm. They require early recognition because delay diagnosis, imaging, and intervention may have devastating consequences especially in children and the elderly. This article reviews the most common spinal infections, their pathophysiologic, clinical manifestation, and their imaging findings.


Evaluative conditioning in social psychology: Facts and speculations.
The aim of the present paper is to examine the contribution of evaluative conditioning (EC) to attitude formation theory in social psychology. This aim is pursued on two fronts. First, evaluative conditioning is analysed for its relevance to social psychological research. We show that conditioned attitudes can be acquired through simple co-occurrences of a neutral and a valenced stimulus. Moreover, we argue that conditioned attitudes are not confined to direct contact with a valenced stimulus, but can be formed and dynamically reformed indirectly, through association chains. Second, social research is examined in an effort to identify evaluative learning mechanisms. We suggest that several important phenomena in social psychology (e.g., ingroup favouritism, prejudice, name letter effect) are at least partly due to simple mechanisms of evaluative learning. The implications for attitude formation theory and for applied settings are discussed.


Deep Learning-Based Segmentation of Locally Advanced Breast Cancer on MRI in Relation to Residual Cancer Burden: A Multi-Institutional Cohort Study.
While several methods have been proposed for automated assessment of breast-cancer response to neoadjuvant chemotherapy on breast MRI, limited information is available about their performance across multiple institutions. To assess the value and robustness of deep learning-derived volumes of locally advanced breast cancer (LABC) on MRI to infer the presence of residual disease after neoadjuvant chemotherapy. Retrospective. Training cohort: 102 consecutive female patients with LABC scheduled for neoadjuvant chemotherapy (NAC) from a single institution (age: 25-73 years). Independent testing cohort: 55 consecutive female patients with LABC from four institutions (age: 25-72 years). Training cohort: single vendor 1.5 T or 3.0 T. Testing cohort: multivendor 3.0 T. Gradient echo dynamic contrast-enhanced sequences. A convolutional neural network (nnU-Net) was trained to segment LABC. Based on resulting tumor volumes, an extremely randomized tree model was trained to assess residual cancer burden (RCB)-0/I vs. RCB-II/III. An independent model was developed using functional tumor volume (FTV). Models were tested on an independent testing cohort and response assessment performance and robustness across multiple institutions were assessed. The receiver operating characteristic (ROC) was used to calculate the area under the ROC curve (AUC). DeLong's method was used to compare AUCs. Correlations were calculated using Pearson's method. P values <0.05 were considered significant. Automated segmentation resulted in a median (interquartile range [IQR]) Dice score of 0.87 (0.62-0.93), with similar volumetric measurements (R = 0.95, P < 0.05). Automated volumetric measurements were significantly correlated with FTV (R = 0.80). Tumor volume-derived from deep learning of DCE-MRI was associated with RCB, yielding an AUC of 0.76 to discriminate between RCB-0/I and RCB-II/III, performing similar to the FTV-based model (AUC = 0.77, P = 0.66). Performance was comparable across institutions (IQR AUC: 0.71-0.84). Deep learning-based segmentation estimates changes in tumor load on DCE-MRI that are associated with RCB after NAC and is robust against variations between institutions. 2. Stage 4.


An indication for spatial integration in a non-Pacinian mechanoreceptor system?
It is generally accepted that threshold sensitivity for cutaneous vibrational stimuli has a Pacinian component and a non-Pacinian component. While the Pacinian receptor system exhibits spatial integration, such a property has, until now, not been found in any of the non-Pacinian systems. The present measurements of vibration sensitivity for stimuli with spatial contrast in six subjects revealed that at low frequencies (20-80 Hz) the threshold for a matrix of tiny pins is 13 dB lower than the threshold for a single pin, thus suggesting spatial integration in at least one class of non-Pacinian receptors. It is remarkable that the low threshold for matrix stimuli was found under the condition that adjacent rows of pins vibrated with opposite phase while the threshold for a matrix of pins all vibrating with equal phase was similar to the threshold for a single vibrating pin.


[Circulatory effects of stellate ganglion block and high thoracic epidural block].
A comparative study of the circulatory effects of stellate ganglion block (SGB) and high thoracic epidural block (TEB) was conducted in 11 patients. Although blood pressure and heart rate showed no significant changes even after SGB, they both decreased significantly following TEB. The blood flow in the common carotid artery (CCA) increased markedly at 5 minutes after performing SGB, and at 20 minutes it reached its peak value of 163.5 +/- 7.8%, and the increase remained significant up to 75 minutes. On the other hand, it increased significantly from 5 minutes after performing TEB and reached its peak of 122.2 +/- 7.6% at 15 minutes, and this increase remained significant up to 60 minutes. In comparing these two nerve blocks, the blood flow in the CCA showed a more marked increase following SGB than after TEB from 5 to 60 minutes after performing. The blood flow rate in the CCA increased markedly from 5 minutes after performing SGB, reached its peak of 139.1 +/- 11.3% at 20 minutes, and this increase remained significant up to 60 minutes thereafter. After performing TEB no significant changes were observed. In comparing SGB and TEB, from 5 to 60 minutes after performing, the rate became markedly faster after SGB than after TEB. The vascular diameter of the CCA from 5 minutes after performing SGB showed a slight but significant enlargement, and at 20 minutes it reached its maximum of 111.7 +/- 3.0%, and this significant enlargement persisted up to 60 minutes. From 10 minutes after performing TEB, it showed a slight but significant enlargement.(ABSTRACT TRUNCATED AT 250 WORDS)


Sexual abuse and suicidal behavior: a model constructed from a large community sample of adolescents.
To investigate relationships between self-reported sexual abuse, depression, hopelessness, and suicidality in a community sample of adolescents. In 1995, students (mean age 13 years) from 27 high schools in Australia (n = 2,603) completed a questionnaire including measures of depressive symptoms (Center for Epidemiologic Studies Depression Scale), hopelessness, sexual abuse, and suicidality. Data analysis included logistic regression. Sexual abuse is associated with suicidality, both directly and indirectly through hopelessness and depressive symptoms in the model developed. High suicide risk (behavior exceeding thoughts such as plans and threats, or deliberate self-injury) is strongly correlated with suicide attempts (odds ratio 28.8, 95% CI 16-52, p <.001). Hopelessness is associated with high suicide risk only, whereas depressive symptoms are associated with high suicide risk and attempts. Hopelessness is more strongly associated with sexual abuse in boys than girls. Depressive symptoms are more strongly associated with high suicide risk in girls than boys. Controlling for other variables, sexual abuse is independently associated with suicide attempts in girls but not boys. Clinical assessment should consider gender differences in relationships between sexual abuse, depressive symptoms, hopelessness, and suicidality. Sexually abused girls may be at increased risk of attempting suicide, independent of other psychopathology.


Nematode isoenzyme studies and cytogenetics.
The current information on isoenzyme studies of nematode parasites was reviewed. The genetic heterogeneity as reviewed by these studies was highlighted. Application of isoenzyme studies and the role of biotechnological techniques in isoenzyme studies was discussed, and the status of cytogenetic studies on nematode parasites was presented.


ScreenBEAM: a novel meta-analysis algorithm for functional genomics screens via Bayesian hierarchical modeling.
Functional genomics (FG) screens, using RNAi or CRISPR technology, have become a standard tool for systematic, genome-wide loss-of-function studies for therapeutic target discovery. As in many large-scale assays, however, off-target effects, variable reagents' potency and experimental noise must be accounted for appropriately control for false positives. Indeed, rigorous statistical analysis of high-throughput FG screening data remains challenging, particularly when integrative analyses are used to combine multiple sh/sgRNAs targeting the same gene in the library. We use large RNAi and CRISPR repositories that are publicly available to evaluate a novel meta-analysis approach for FG screens via Bayesian hierarchical modeling, Screening Bayesian Evaluation and Analysis Method (ScreenBEAM). Results from our analysis show that the proposed strategy, which seamlessly combines all available data, robustly outperforms classical algorithms developed for microarray data sets as well as recent approaches designed for next generation sequencing technologies. Remarkably, the ScreenBEAM algorithm works well even when the quality of FG screens is relatively low, which accounts for about 80-95% of the public datasets. R package and source code are available at: https://github.com/jyyu/ScreenBEAM. email@example.com, email@example.com, email@example.com Supplementary data are available at Bioinformatics online.


[165 bilateral atherosclerotic carotid occlusive lesions. Surgical and secondary risk].
Operative treatment was applied in 139 patients with hemodynamically significant bilateral stenosis of the carotid arteries due to atherosclerosis. Three groups could be distinguished: group I--stenosis greater than 75% on one side and between 50 and 75% on the other side; group II--stenosis greater than 75% on both sides; group III--obstruction on one side and stenosis greater than 50% on the other side. The risk of clamping did not appear abnormally high, while the peroperative neurologic risk was slightly enhanced due to the possibility of accidents in the region which lies contralateral to that of the operated side, or in the region of the brain stem. The risk in the group III patients did not appear to be increased. Immediate and secondary neurologic prognosis is mainly a function of the presence of multiple peculiar lesions, and also of the presence or absence of parenchymatous ischemic lesions. Preventive surgery on narrowed, stenotic, progressive lesions, detected by ultrasound imaging systematically repeated every 6 months, diminishes the incidence of onset of neurologic complications.


Osteopontin and adiponectin: how far are they related in the complexity of psoriasis?
Increasing attention has been drawn towards the involvement of both osteopontin (OPN) and adiponectin in psoriasis. The relationship between them has been studied before in the context of essential hypertension. To our knowledge, whether a relation between them exists in cases of psoriasis and the metabolic status in such patients have not been investigated. We aimed to verify their possible roles and relations in psoriasis and its metabolic associations. 35 patients with psoriasis vulgaris and 35 controls were included. Patients were clinically assessed by PASI and investigated for the presence of metabolic syndrome (MetS) and/or its components. Plasma levels of OPN and adiponectin were measured using ELISA. On comparing psoriatics to controls, patients showed significantly elevated levels of OPN (90.474 ± 21.22 vs 34.709 ± 13.95 ng/mL) and significantly depressed levels of adiponectin (4,586 ± 1.187 vs 5,905 ± 1.374 ng/mL), (p < 0.001). Strong negative correlation between plasma OPN and adiponectin was detected in patients (r = -0.912, p < 0.001), but not in controls. OPN elevation was related to diabetes mellitus, insulin resistance, and MetS. Adiponectin depression was related to body mass index, and MetS. This study demonstrates for the first time a significant correlation between OPN and adiponectin in psoriasis, hypothesized to be mostly attributed to the inflammatory milieu of psoriasis and MetS as well as the enhanced renin-angiotensin-aldosterone system previously documented in psoriasis. Adjuvant therapies aiming at modulating levels of OPN and adiponectin are speculated to add benefit in psoriasis treatment and protecting against its metabolic risks.


Pyrroloquinoline quinone attenuates iNOS gene expression in the injured spinal cord.
Pyrroloquinoline quinone (PQQ) is a naturally occurring redox cofactor that acts as an essential nutrient, antioxidant, and redox modulator. PQQ has been demonstrated to oxidize the redox modulatory site of N-methyl-d-aspartic acid (NMDA) receptors. Such agents are known to be neuroprotective in experimental stroke models. Therefore, we examined the possible ameliorating effect of PQQ on spinal cord injury (SCI) in adult rats. Intraperitoneal administration of PQQ effectively promoted the functional recovery of SCI rats after hemi-transection, which was preceded by the attenuation of the expression of inducible nitric oxide (NO) synthase (iNOS) mRNA in the injury site. NO is involved in the secondary detrimental mechanisms and has been implicated in NMDA receptor-mediated neurotoxicity. In fact, administration of PQQ induced significantly decreased lesion size and increased axon density adjoining the lesion area. These observations suggest that PQQ protects against the secondary damage by reducing iNOS expression following primary physical injury to the spinal cord.


Protein electrophoresis and immunoglobulin analysis in HIV-infected patients.
We studied the prevalence and nature of immunoglobulin abnormalities in HIV-1-infected patients in the era of highly active antiretroviral therapy. Protein electrophoreses (PEP) were performed on and quantitative immunoglobulin levels obtained in samples from 320 consecutive HIV-1-infected patients. Samples with possible PEP abnormalities underwent immunofixation. The PEP pattern was normal in 83.8% of samples, 8.1% had subtle oligoclonal banding, and 4.4% had a low-concentration (<5% of total protein) monoclonal band. Hypogammaglobulinemia and polyclonal hypergammaglobulinemia accounted for 1.9% each. In multivariate analysis, younger age (odds ratio [OR], 1.06 with each decreasing year of life; 95% confidence interval [CI], 1.02-1.11; P = .016), female sex (OR, 2.4; 95% CI, 1.13-5.11; P = .02), viral load (OR, 1.50 with each increasing logarithmic viral load of 1.0; 95% CI, 1.14-1.98; P = .004), and CD4 cell count (> or =350 vs <350/microL [0.35 x 10(9)/L]) (OR, 2.71; 95% CI, 1.09-6.75; P = .032) were associated with monoclonal or oligoclonal banding. These results suggest that younger HIV-1-infected patients with a more robust immune system (higher CD4 cell count), which is stimulated by uncontrolled viremia, are most likely to have an augmented B-cell response to HIV infection. One manifestation of this B-cell response is low-concentration monoclonal banding in 4.4% of the patients studied.


[Mg2,Ca2+-ATPase activity of sarcolemmas of intestinal smooth muscle cells in the rabbit].
Preparations of rabbit small intestine smooth muscle cell sarcolemma are capable of hydrolyzing ATP in the presence of millimolar concentrations of Mg2+ and Ca2+ and possess the activity of Mg2+,Ca2+-ATPase having a high affinity for Ca2+ (Km = 5.8 X 10(-6) M). The optimal conditions for the Mg2+,Ca2+-ATPase reaction were established. It was demonstrated that sarcolemmal preparations hydrolyze ATP, GTP, ITP and UTP almost at the same rates. The enzyme contains SH-groups that are unequally exposed to the water phase and are inhibited by 50% by p-chloromercurybenzoate and by 90% by dithionitrobenzoate. The Mg2+,Ca2+-ATPase activity is highly sensitive to oxytocin: at the concentration of 10(-7) MU/ml, the hormone completely inhibits the enzyme without affecting its Mg2+-, Ca2+- and Na+,K+-ATPase activities.


[Clinical application of atlas translaminar screws fixation in treatment of atlatoaxial instability].
To explore the effectiveness of fixation of atlas translaminar screws in the treatment of atlatoaxial instability. A retrospective analysis was made on the clinical data of 32 patients with atlatoaxial instability treated with atlantoaxial trans-pedicle screws between March 2007 and August 2009. Of them, 7 patients underwent atlas translaminar screws combined with axis transpedicle screws fixation because of fracture types, anatomic variation, and intraoperative reason, including 5 males and 2 females with an average age of 48.2 years (range, 35-69 years). A total of 9 translaminar screws were inserted. Injury was caused by traffic accident in 4 cases, falling from height in 2 cases, and crushing in 1 case. Two cases had simple odontoid fracture (Anderson type II), and 5 cases had odontoid fracture combined with other injuries (massa lateralis atlantis fracture in 2, atlantoaxial dislocation in 1, and Hangman fracture in 2). The interval between injury and operation was 4-9 days (mean, 6 days). The preoperative Japanese Orthopaedic Association (JOA) score was 8.29 +/- 1.60. The X-ray films showed good position of the screws. Healing of incision by first intention was obtained, and no patient had injuries of the spinal cord injury, nerve root, and vertebral artery. Seven cases were followed up 9-26 months (mean, 14 months). Good bone fusion was observed at 8 months on average (range, 6-11 months). No loosening, displacement, and breakage of internal fixation, re-dislocation and instability of atlantoaxial joint, or penetrating of pedicle screw into the spinal canal and the spinal cord occurred. The JOA score was significantly improved to 15.29 +/- 1.38 at 6 months after operation (t = 32.078, P = 0.000). Atlas translaminar screws fixation has the advantages of firm fixation, simple operating techniques, and relative safety, so it may be a remedial measure of atlatoaxial instability.


Ara AMP- a highly soluble new antiviral drug.
While vidarabine (Ara A) is in several ways superior to idoxuridine (IDU) it still has the disadvantages of rapid inactivation and poor solubility. The monophosphate ester or vidarabine, Ara AMP, however, is metabolized very slowly in humans and is extremely soluble with good tissue penetrance. The present study indicates that Ara AMP drops are equipotent or superior to vidarabine ointment in treatment of type 1 and type 2 herpes simplex keratitis. While iritis was significantly less in all treated groups, a suspicion of chemical side effect was raised but could not be proven in subsequent toxicity tests.


Habitat occupancy of sloth bear Melursus ursinus in Chitwan National Park, Nepal.
Mammals have experienced a massive decline in their populations and geographic ranges worldwide. The sloth bear, Melursus ursinus (Shaw, 1791), is one of many species facing conservation threats. Despite being endangered in Nepal, decades of inattention to the situation have hindered their conservation and management. We assessed the distribution and patterns of habitat use by sloth bears in Chitwan National Park (CNP), Nepal. We conducted sign surveys from March to June, 2020, in 4 × 4 km grids (n = 45). We collected detection/non-detection data along a 4-km trail that was divided into 20 continuous segments of 200 m each. We obtained environmental, ecological, and anthropogenic covariates to understand determinants of sloth bear habitat occupancy. The data were analyzed using the single-species single-season occupancy method, with a spatially correlated detection. Using repeated observations, these models accounted for the imperfect detectability of the species to provide robust estimates of habitat occupancy. The model-averaged occupancy estimate for the sloth bear was 69% and the detection probability was 0.25. The probability of habitat occupancy by sloth bears increased with the presence of termites and fruits and in rugged, dry, open, undisturbed habitats. Our results indicate that the sloth bear is elusive, functionally unique, and widespread in CNP. Future conservation interventions and action plans aimed at sloth bear management must adequately consider their habitat requirements.


Mass analysis by scanning transmission electron microscopy and electron diffraction validate predictions of stacked beta-solenoid model of HET-s prion fibrils.
Fungal prions are infectious filamentous polymers of proteins that are soluble in uninfected cells. In its prion form, the HET-s protein of Podospora anserina participates in a fungal self/non-self recognition phenomenon called heterokaryon incompatibility. Like other prion proteins, HET-s has a so-called "prion domain" (its C-terminal region, HET-s-(218-289)) that is responsible for induction and propagation of the prion in vivo and for fibril formation in vitro. Prion fibrils are thought to have amyloid backbones of polymerized prion domains. A relatively detailed model has been proposed for prion domain fibrils of HET-s based on a variety of experimental constraints (Ritter, C., Maddelein, M. L., Siemer, A. B., Luhrs, T., Ernst, M., Meier, B. H., Saupe, S. J., and Riek, R. (2005) Nature 435, 844-848). To test specific predictions of this model, which envisages axial stacking of beta-solenoids with two coils per subunit, we examined fibrils by electron microscopy. Electron diffraction gave a prominent meridional reflection at (0.47 nm)(-1), indicative of cross-beta structure, as predicted. STEM (scanning transmission electron microscopy) mass-per-unit-length measurements yielded 1.02 +/- 0.16 subunits per 0.94 nm, in agreement with the model prediction (1 subunit per 0.94 nm). This is half the packing density of approximately 1 subunit per 0.47 nm previously obtained for fibrils of the yeast prion proteins, Ure2p and Sup35p, whence it follows that the respective amyloid architectures are basically different.


Neural stem cell migration toward gliomas in vitro.
Various in vivo studies demonstrated a migration tendency of neural stem cells (NSCs) toward gliomas, making these cells a potential carrier for delivery of therapeutic genes to disseminated glioma cells. We analyzed which factors determine NSC migration and invasion in vitro. Conditioned media prepared from 10 different human glioma cell lines, as well as 13 different tumor-associated growth factors, were analyzed for their chemotactic effects on murine C17.2 NSCs. The growth factor receptor status was analyzed by reverse transcriptase-polymerase chain reaction. Invasion of NSCs into multicellular tumor spheroids generated from 10 glioma cell lines was quantified. NSCs displayed a heterogeneous migration pattern toward glioma spheroids as well as toward glioma-cell-conditioned medium. Chemotactic migration was stimulated up to fivefold by conditioned medium as compared to controls. In coculture assays, NSC invasion varied from single cell invasion into glioma spheroids to complete dissemination of NSCs into glioma spheroids of different cell lines. Among 13 different growth factors, scatter factor/hepatocyte growth factor (SF/HGF) was the most powerful chemoattractant for NSCs, inducing a 2.5-fold migration stimulation. An antibody against SF/HGF inhibited migratory stimulation induced by conditioned media. NSC migration can be stimulated by various growth factors, similar to glioma cell migration. The extent to which NSCs infiltrate three-dimensional glioma cell aggregates appears to depend on additional factors, which are likely to include cell-to-cell contacts and interaction with extracellular matrix proteins.


MicroRNA Biomarkers for Infectious Diseases: From Basic Research to Biosensing.
In the pursuit of improved diagnostic tests for infectious diseases, several classes of molecules have been scrutinized as prospective biomarkers. Small (18-22 nucleotide), non-coding RNA transcripts called microRNAs (miRNAs) have emerged as promising candidates with extensive diagnostic potential, due to their role in numerous diseases, previously established methods for quantitation and their stability within biofluids. Despite efforts to identify, characterize and apply miRNA signatures as diagnostic markers in a range of non-infectious diseases, their application in infectious disease has advanced relatively slowly. Here, we outline the benefits that miRNA biomarkers offer to the diagnosis, management, and treatment of infectious diseases. Investigation of these novel biomarkers could advance the use of personalized medicine in infectious disease treatment, which raises important considerations for validating their use as diagnostic or prognostic markers. Finally, we discuss new and emerging miRNA detection platforms, with a focus on rapid, point-of-care testing, to evaluate the benefits and obstacles of miRNA biomarkers for infectious disease.


Expression and immunohistochemical localization of adrenomedullin in the mouse cochlea.
Adrenomedullin (AM) is a novel hypotensive and vasodilator peptide. It has been shown that AM is biosynthesized in various organs and cells and is suggested to play multiple roles including electrolyte homeostasis and body fluid control. The present study describes for the first time the presence of mRNA for AM and AM-like immunoreactivity in the cochlea. A reverse transcription-polymerase chain reaction (RT-PCR) performed with gene-specific AM primer on total RNA extracted from mouse cochlea revealed a PCR product of the expected size. Immunohistochemical examination showed positive immunostaining in vascular smooth muscle cells of the modiolar arterioles and in the stria vascularis of the cochlea. These results suggest that AM may play a role in the regulation not only of cochlear hemodynamics but also of cochlear fluid dynamics.


Deletion of murine Arv1 results in a lean phenotype with increased energy expenditure.
ACAT-related enzyme 2 required for viability 1 (ARV1) is a putative lipid transporter of the endoplasmic reticulum that is conserved across eukaryotic species. The ARV1 protein contains a conserved N-terminal cytosolic zinc ribbon motif known as the ARV1 homology domain, followed by multiple transmembrane regions anchoring it in the ER. Deletion of ARV1 in yeast results in defective sterol trafficking, aberrant lipid synthesis, ER stress, membrane disorganization and hypersensitivity to fatty acids (FAs). We sought to investigate the role of Arv1 in mammalian lipid metabolism. Homologous recombination was used to disrupt the Arv1 gene in mice. Animals were examined for alterations in lipid and lipoprotein levels, body weight, body composition, glucose tolerance and energy expenditure. Global loss of Arv1 significantly decreased total cholesterol and high-density lipoprotein cholesterol levels in the plasma. Arv1 knockout mice exhibited a dramatic lean phenotype, with major reductions in white adipose tissue (WAT) mass and body weight on a chow diet. This loss of WAT is accompanied by improved glucose tolerance, higher adiponectin levels, increased energy expenditure and greater rates of whole-body FA oxidation. This work identifies Arv1 as an important player in mammalian lipid metabolism and whole-body energy homeostasis.


Familial multiple lipomatosis.
A family genealogy, comprising four cases of familial multiple lipomatosis, making their appearance one after the other in three generations is reported. Two cases with impressive clinical features are presented in detail. This report contributes to the opinion that the disease is transmitted by the autosomal dominant route of inheritance.


[Imaging in low back pain: limits and reflexions].
Lumbar imaging is frequently requested in patients with low back pain for various reasons. However, the prevalence of severe lesions, including neoplastic, is only about 1%. Imaging in the absence of clinical suspicion performs poorly. Similarly, the imagery is no more likely to identify the anatomical structures that are the source of pain for patients with low back pain. Finally, the literature shows that the impact on treatment decisions and the effect on quality of life of patients cannot justify an indiscriminate use of these investigative techniques. Without questioning the value of imaging, the practitioner must keep in mind these limitations in order to optimize the use of this resource.


Development of an improved cloning vector and transformation system in Amycolatopsis mediterranei (Nocardia mediterranei).
A versatile plasmid cloning vector pRL60 carrying kanamycin/neomycin resistance (km/neo), erythromycin resistance (ermE) and alpha-amylase (alpha-amy) marker genes that replicates in various Amycolatopsis mediterranei strains and Escherichia coli has been constructed. This cloning vector has been derived from a hybrid plasmid pRL50, which was developed by cloning ermE from pIJ4026 into a pRL1 derivative pULAM2. While cloning ermE into the BamHI site of pULAM2, only a hybrid plasmid pRL50 with an additional copy of pULAM2 was selected. Thus pRL50 (18.7 kb) contained two copies each of the km/neo, alpha-amy, and one copy of ermE. When pRL50 was transformed into A. mediterranei DSM 40773 through electroporation and selected under erythromycin resistance, the plasmid underwent a spontaneous deletion of 8.5 kb fragment resulting in the formation of plasmid pRL60. pRL60 (10.2 kb) is a shuttle vector between A. mediterranei and E. coli with three marker genes: km/neo, ermE and alpha-amy. ermE is expressed in A. mediterranei thus allowing good selection of transformants. The alpha-amy gene of pRL60 is also expressed in A. mediterranei DSM 40773 and its activity can be easily detected on starch containing medium after iodine staining. Most critical parameters evaluated for electrotransformation using pRL60 in A. mediterranei were growth phase, electrical field strength, pulse length, pretreatment of mycelia with lysozyme and use of salt free water. At optimized parameters, a transformation efficiency of 4.0 x 10(4) transformants/microg DNA was reproducibly achieved for A. mediterranei DSM 40773. pRL60 could also be transformed into A. mediterranei DSM 43304, DSM 46095, MTCC-17 and in mutants F1/24 and T-195, (derived from an industrial strain of A. mediterranei N813). The alpha-amy of pRL60 conferred an amylolytic phenotype to all these strains. With the development of pRL60 and a reproducible transformation protocol, the application of recombinant DNA techniques to these industrial microorganisms has now become feasible.


Rotation of spinal curvatures of patients with structural scoliosis.
Scoliosis has obscure aetiopathogenesis and is underreported in our environment. To examine the pattern, magnitude, direction and extent of rotation of 42 structural curves in scoliotic patients. Plain radiographs of the spine in patients with scoliotic deformities were randomly selected from the film libraries of four Nigerian tertiary institutions.These were then assessed radiologically for the indices above, including the use of Cobbs method and the pedicle technique of Moe. In all, 14 thoracic and 28 thoracolumbar curves were evaluated. A double curve was identified in only one patient. Average age (range) of affected males was 31 (3-78) years and for females (n=25), it was 19.4 (3-40) years. Mean Cobb angle was 23.6 degrees (10 degrees-70 degrees) in males and 40.1 degrees (10 degrees-89 degrees) in females. In 29 (69%) of curves, angle magnitude was d" 30 degrees. Direction of spinal curvature was right convexity in 33 (78.6%) of evaluated curves while most curves 35 (84.4%) exhibited +1 vertebral rotation. Females present earlier than the males with a higher Cobb angles. Progression of curves appears to be a higher challenge in females. There is need for scoliosis surveillance programme for early diagnosis.


Traumatic Isolated Avulsion Rupture of the Distal Semitendinosus Tendon in a Non-athlete.
Isolated distal semitendinosus (ST) injuries remain an uncommon hamstring injury, with avulsion ruptures reported even less frequently. These injuries occur due to eccentric overloading seen in sprinting or jumping injuries. Treatment ranges from conservative management to surgical tenotomy or reattachment to the tibial bone. We present a unique case of a 30-year-old male with an isolated avulsion rupture of the distal ST tendon after a fall. To our knowledge, this is the first case reported in the literature of an isolated distal ST injury in a non-athlete due to trauma.


Efficacy of the Community Re-Entry Module for patients with schizophrenia in Beijing, China: outcome at 2-year follow-up.
Few psychosocial interventions have been developed in China that are suitable for use in the community. To evaluate the effectiveness of the Chinese version of the Community Re-Entry Module (CRM; a module of a standardised, structured social skills training programme devised at the University of California, Los Angeles) for patients with schizophrenia compared with standard group psychoeducation. Patients with schizophrenia (n=103) were randomly allocated to CRM or psychoeducation groups and followed up for 24 months. Outcome measures included social functioning, psychiatric symptoms, insight, re-employment, relapse and re-hospitalisation rates. The CRM group significantly improved in terms of social functioning, insight and psychiatric symptoms compared with the psychoeducation group; the re-employment rate was significantly higher and relapse and rehospitalisation rates were significantly lower in the CRM group. The findings support the feasibility and effectiveness of CRM as a psychosocial intervention for Chinese patients with schizophrenia in the community.


[Dementia and mild cognitive impairment in Parkinson's disease: a review].
INTRODUCTION. The cognitive disorders in Parkinson's disease (PD) have traditionally been associated with the presence of dementia in later stages of the disease. Recent studies, however, consider that cognitive impairment can appear as of early stages. Knowing the cognitive profile of PD furthers our understanding of the clinical phenotype, making it easier to reach a timely diagnosis and favouring intervention on the symptoms from the initial stages. AIM. To present a review of the literature on mild cognitive impairment (MCI) and dementia associated with PD. DEVELOPMENT. Several studies report that patients with PD who have a prolonged time to progression develop dementia. Yet, there have also been reports claiming that, as of the early stages, patients can present subtle cognitive alterations known as MCI. The initial neuropsychological profile is mainly of a non-amnesic type, characterised by executive dysfunction, alterations affecting attention, operative memory deficit and faulty retrieval of information. When patients develop dementia, disorders will arise in the storage of information, in semantic fluency, and in visuospatial and visuoperceptual skills. Currently there are criteria available for diagnosing the MCI and dementia associated with PD, as well as valid reliable instruments for detecting those disorders. CONCLUSIONS. Cognitive symptoms are frequent in PD. From the initial stages of the disease onwards patients may present MCI that is mainly characterised by a fronto-subcortical cognitive profile, whereas dementia usually develops at later stages, when a pattern of posterior cortical cognitive disorder is also observed.


An electrochemiluminescent method for glutamate measurement in small microdialysate samples in asphyxiated young rats.
Glutamate (Glu) quantification has been performed by a combination of intracerebral microdialysis through which the samples are obtained and analyzed by high performance liquid chromatography (HPLC); its measurement requires a large expenditure of time (15-30 min per sample) and special training. Therefore, an alternative method is presented here, based on the electrochemiluminescence produced by the use of an enzymatic reactor, containing glutamate-oxidase, mixed and incubated with microdialysate from dorsal striatum (DS) and prefrontal cortex (PFC) of young rats asphyxiated during the neonatal period, under a global asphyxia model in order to test this method. Using this approach, we found high extracellular Glu concentration in the DS of asphyxiated animals, but only during K+ stimulation, while in the PFC, only a delay in the rise of Glu after K+ stimulation was observed, without any difference in extracellular Glu content when compared with controls. This new method permitted a fast measurement of Glu in brain dialysate samples, it significantly reduces the cost of the analysis per sample, since only a single device and pump are needed without using columns and high pressure inside the system or complex hardware and software to control pumps, detector, fraction collector or any other peripheral used in HPLC.


Calmodulin-mediated activation of Akt regulates survival of c-Myc-overexpressing mouse mammary carcinoma cells.
c-Myc-overexpressing mammary epithelial cells are proapoptotic; their survival is strongly promoted by epidermal growth factor (EGF). We now demonstrate that EGF-induced Akt activation and survival in transgenic mouse mammary tumor virus-c-Myc mouse mammary carcinoma cells are both calcium/calmodulin-dependent. Akt activation is abolished by the phospholipase C-gamma inhibitor U-73122, by the intracellular calcium chelator BAPTA-AM, and by the specific calmodulin antagonist W-7. These results implicate calcium/calmodulin in the activation of Akt in these cells. In addition, Akt activation by serum and insulin is also inhibited by W-7. EGF-induced and calcium/calmodulin-mediated Akt activation occurs in both tumorigenic and non-tumorigenic mouse and human mammary epithelial cells, independent of their overexpression of c-Myc. These results imply that calcium/calmodulin may be a common regulator of Akt activation, irrespective of upstream receptor activator, mammalian species, and transformation status in mammary epithelial cells. However, only c-Myc-overexpressing mouse mammary carcinoma cells (but not normal mouse mammary epithelial cells) undergo apoptosis in the presence of the calmodulin antagonist W-7, indicating the vital selective role of calmodulin for survival of these cells. Calcium/calmodulin-regulated Akt activation is mediated directly by neither calmodulin kinases nor phosphatidylinositol 3-kinase (PI-3 kinase). Pharmacological inhibitors of calmodulin kinase kinase and calmodulin kinases II and III do not inhibit EGF-induced Akt activation, and calmodulin antagonist W-7 does not inhibit phosphotyrosine-associated PI-3 kinase activation. Akt is, however, co-immunoprecipitated with calmodulin in an EGF-dependent manner, which is inhibited by calmodulin antagonist W-7. We conclude that calmodulin may serve a vital regulatory function to direct the localization of Akt to the plasma membrane for its activation by PI-3 kinase.


A new cell model for equation of state of polyethylene melt and n-alkane.
The connectivity of successive carbon atoms in polymer decreases the degrees of freedom, and hence, the external degrees of freedom should correspond only to translational motion. We therefore, introduce a coarse-grained particle of a few successive monomers. A cell model (or the hard core model with attractive potential) for the particles can accordingly be used for the derivation of the equation of state of polymers. Modifying the classical cell model by Lennard-Jones and Devonshire, we construct a new equation of state for polyethylene melt and for liquid n-alkane; the free volume term is modified by using the Sutherland potential instead of the Lennard-Jones potential. The characteristic quantities P*, V*0, and T* in the equation of state are almost independent of temperature; the principle of corresponding state holds well. Since our equation of state contains the external degrees of freedom explicitly, we can evaluate the external degrees of freedom, c, for CH2. The value of c for the coarse-grained particle is equal to 1, and hence the particle is composed of 1/c repeating units. The linear length of the particle evaluated, 4.09 A at 0 K, is consistent with that obtained by neutron and x-ray scatterings.


Characterization of the complete mitochondrial genome of the longhorn beetle, Batocerahorsfieldi (Coleoptera, Cerambycidae) and its phylogenetic analysis with suitable longhorn beetles.
Mitochondrial genome analysis is an important tool for studying insect phylogenetics. The longhorn beetle, Batocerahorsfieldi, is a significant pest in timber, economic and protection forests. This study determined the mitochondrial genome of B.horsfieldi and compared it with the mitochondrial genomes of other Cerambycidae with the aim of exploring the phylogenetic status of the pest and the evolutionary relationships among some Cerambycidae subgroups. The complete mitochondrial genome of B.horsfieldi was sequenced by the Illumina HiSeq platform. The mitochondrial genome was aligned and compared with the existing mitochondrial genomes of Batoceralineolata and B.rubus in GenBank (MF521888, MW629558, OM161963, respectively). The secondary structure of transfer RNA (tRNA) was predicted using tRNAScan-SE server v.1.21 and MITOS WebSever. Thirteen protein-coding genes (PCGs) and two ribosomal RNA gene sequences of 21 longhorn beetles, including B.horsfieldi, plus two outgroups, Dryopsernesti (Dryopidae) and Heterocerusparallelus (Heteroceridae), were analyzed. The phylogenetic tree was constructed using maximum likelihood and Bayesian inference methods. In this study, we successfully obtained the complete mitochondrial genome of B.horsfieldi for the first time, which is 15 425 bp in length. It contains 37 genes and an A + T-rich region, arranged in the same order as the recognized ancestor of longhorn beetles. The genome of B.horsfieldi is composed of 33.12% A bases, 41.64% T bases, 12.08% C bases, and 13.16% G bases. The structure, nucleotide composition, and codon usage of the new mitochondrial genome are not significantly different from other longhorn mitochondrial genomes. Phylogenetic analyses revealed that Cerambycidae formed a highly supported single clade, and Vesperidae was either clustered with Cerambycidae or formed a separate clade. Interestingly, B.horsfieldi, B.rubus and B.lineolata were clustered with Monochamus and Anoplophora species in both analyses, with high node support. Additionally, the VesperidaeSpiniphilusspinicornis and Vesperussanzi and the 19 Cerambycidae species formed a sister clade in the Bayesian analysis. Our results have produced new complete mitogenomic data, which will provide information for future phylogenetic and taxonomic research, and provide a foundation for future relevant research.


BST2 regulated by the transcription factor STAT1 can promote metastasis, invasion and proliferation of oral squamous cell carcinoma via the AKT/ERK1/2 signaling pathway.
Oral squamous cell carcinoma (OSCC) is one of the main types of head and neck squamous cell carcinoma. Although progress has been made in treating OSCC, it remains a threat to human health, and novel therapeutic strategies are needed to extend the lifespan of patients with OSCC. The present study, evaluated whether bone marrow stromal antigen 2 (BST2) and STAT1 were potential therapeutic targets in OSCC. Small interfering RNA (siRNA) or overexpression plasmids were used to regulate BST2 or STAT1 expression. Western blotting and reverse transcription‑quantitative PCR were performed to assess changes in the protein and mRNA expression levels of signaling pathway components. The effects of BST2 and STAT1 expression changes on the migration, invasion and proliferation of OSCC cells were assessed using the scratch test assay, Transwell assay and colony formation assay in vitro, respectively. Cell‑derived xenograft models were used to evaluate the impact of BST2 and STAT1 on the occurrence and development of OSCC in vivo. Finally, it was demonstrated that BST2 expression was significantly upregulated in OSCC. Furthermore, it was demonstrated that high expression of BST2 in OSCC contributed to the metastasis, invasion and proliferation of OSCC cells. Moreover, it was demonstrated that the promoter region of BST2 was regulated by the transcription factor STAT1, and that the STAT1/BST2 axis could affect the behavior of OSCC via the AKT/ERK1/2 signaling pathway. In vivo studies also demonstrated that STAT1 downregulation inhibited OSCC growth by down‑regulating BST2 expression via the AKT/ERK1/2 signaling pathway.


Inhibition of human immunodeficiency virus 1 reverse transcriptase by 3'-azidothymidine triphosphate.
In the presence of oligo(dT).poly(rA) as primer-template, 3'-azidothymidine triphosphate (N3'(3)-ddTTP) is a substrate for human immunodeficiency virus 1 reverse transcriptase with an apparent Km value of 3.0 microM. This compares with an apparent Km for thymidine monophosphate (dTMP) incorporation of 2.5 microM. The apparent Vmax value for 3'-azidothymidine monophosphate (N3'(3)-ddTMP) incorporation is 50 times lower than that of dTMP incorporation. Kinetic analysis of the inhibition of reverse transcriptase by N3'(3)-ddTTP shows competitive inhibition with thymidine triphosphate (dTTP) with a Ki of 41 nM and an uncompetitive pattern of inhibition with template-primer having a Ki of 140 nM. This indicates incorporation of the analogue into the primer and inhibition of the enzyme by formation of a dead-end complex. The 3'-azidothymidine-terminated primer-template [N3'(3)-ddT-(dT)15.poly(rA)] is a potent competitive inhibitor versus primer-template with a Ki of 2.4 nM and shows mixed-type inhibition against dTTP with a Ki of 8 nM. The low inhibition constant for this chain-terminated primer suggests that such oligonucleotides can act as potent inhibitors of enzyme catalysis.


Dolphin Morbillivirus in Eurasian Otters, Italy.
We report biomolecular evidence of dolphin morbillivirus in 4 wild Eurasian otters (Lutra lutra) from southern Italy; 2 animals showed simultaneous immunohistochemical reactivity against morbilliviral antigen. These cases add further concern and support to the progressively expanding host range of dolphin morbillivirus in the western Mediterranean Sea.


Spongy degeneration of the CNS in infancy.
We studied six cases of spongy degeneration of the CNS in infancy among Chinese. The main clinical features were cyclical vomiting and wailing, blindness, megalocephaly, convulsions, lethargy, and coma. Chief findings from light microscopic studies were spongiform change, moderate outfall of neurons, and marked astrocytosis, with formation of Alzheimer's cells type 2. Demyelination was only slight, if present. Electron microscopic studies revealed membrane-bound vacuoles, often containing membranous fragments or blisters and electron-dense granules.


Innateness and the instinct to learn.
Concepts of innateness were at the heart of Darwin's approach to behavior and central to the ethological theorizing of Lorenz and, at least to start with, of Tinbergen. Then Tinbergen did an about face, and for some twenty years the term 'innate' became highly suspect. He attributed the change to Lehrman's famous 1953 critique in which he asserted that classifying behaviors as innate tells us nothing about how they develop. Although Lehrman made many valid points, I will argue that this exchange also led to profound misunderstandings that were ultimately damaging to progress in research on the development of behavior. The concept of 'instincts to learn', receiving renewed support from current theorizing among geneticists about phenotypic plasticity, provides a potential resolution of some of the controversies that Lehrman created. Bioacoustical studies, particularly on song learning in birds, serve both to confirm some of Lehrman's anxieties about the term 'innate', but also to make a case that he threw out the genetic baby with the bathwater. The breathtaking progress in molecular and developmental genetics has prepared the way for a fuller understanding of the complexities underlying even the simplest notions of innate behavior, necessary before we can begin to comprehend the ontogeny of behavior.


Magnetic fields for fluid motion.
Three forces induced by magnetic fields offer unique control of fluid motion and new opportunities in microfluidics. This article describes magnetoconvective phenomena in terms of the theory and controversy, tuning by redox processes at electrodes, early-stage applications in analytical chemistry, mature applications in disciplines far afield, and future directions for micro total analysis systems. (To listen to a podcast about this article, please go to the Analytical Chemistry multimedia page at pubs.acs.org/page/ancham/audio/index.html .).


Slowest kinetic modes revealed by metabasin renormalization.
Understanding the slowest relaxations of complex systems, such as relaxation of glass-forming materials, diffusion in nanoclusters, and folding of biomolecules, is important for physics, chemistry, and biology. For a kinetic system, the relaxation modes are determined by diagonalizing its transition rate matrix. However, for realistic systems of interest, numerical diagonalization, as well as extracting physical understanding from the diagonalization results, is difficult due to the high dimensionality. Here, we develop an alternative and generally applicable method of extracting the long-time scale relaxation dynamics by combining the metabasin analysis of Okushima et al. [Phys. Rev. E 80, 036112 (2009)PLEEE81539-375510.1103/PhysRevE.80.036112] and a Jacobi method. We test the method on an illustrative model of a four-funnel model, for which we obtain a renormalized kinematic equation of much lower dimension sufficient for determining slow relaxation modes precisely. The method is successfully applied to the vacancy transport problem in ionic nanoparticles [Niiyama et al., Chem. Phys. Lett. 654, 52 (2016)CHPLBC0009-261410.1016/j.cplett.2016.04.088], allowing a clear physical interpretation that the final relaxation consists of two successive, characteristic processes.


Stereoselective synthesis of the glycosidase inhibitor australine through a one-pot, double-cyclization strategy.
[reaction: see text] A stereocontrolled, convergent synthesis of the alkaloid australine, a glycosidase inhibitor of the pyrrolizidine class, is described. The chiral starting materials were ketone 3, derived from L-erythrulose, and alpha-alkoxy aldehyde 4, prepared from L-malic acid. A key step of the synthesis was the highly stereoselective aldol reaction between 4 and a Z boron enolate derived from 3. Another key step was the one-pot construction of the bicyclic pyrrolizidine system by means of a three-step sequence of SN2 displacements induced by benzylamine on a trimesylate precursor.


Increased intestinal growth in the streptozotocin-diabetic rat occurs prior to changes in hormone secretion.
Depressed somatic growth in the rat with streptozotocin diabetes is associated with greatly enhanced intestinal growth. In patients with diabetes mellitus, growth hormone is elevated. Therefore, we measured the association between gut growth and serum growth hormone in the streptozotocin-diabetic rat. At 10 days growth hormone did not differ in control, untreated diabetic and insulin-treated diabetic groups. At 20 days of diabetes, growth hormone was elevated in untreated diabetic groups, but depressed to control levels by insulin. Intestinal growth was markedly elevated in all diabetic groups by 10 days, prior to the increases in hormone levels. We conclude that the enhanced intestinal growth in diabetes is probably independent of growth hormone.


Using Robotic Systems to Process and Embed Colonic Murine Samples for Histological Analyses.
The understanding of human diseases has been greatly expanded thanks to the study of animal models. Nonetheless, histopathological evaluation of experimental models needs to be as rigorous as that applied for human samples. Indeed, drawing reliable and accurate conclusions is critically influenced by the quality of tissue section preparation. Here, we describe a protocol for histopathological analysis of murine tissues that implements several automated steps during the procedure, from the initial preparation to the paraffin embedding of the murine samples. The reduction of methodological variables through rigorous protocol standardization from automated procedures contributes to increased overall reliability of murine pathological analysis. Specifically, this protocol describes the utilization of automated processing and embedding robotic systems, routinely used for the tissue processing and paraffin embedding of human samples, to process murine specimens of intestinal inflammation. We conclude that the reliability of histopathological examination of murine tissues is significantly increased upon introduction of standardized and automated techniques.


Stimuli-responsive flexible Lewis pair-modified nanoparticles for fluorescence imaging.
A stimuli-responsive fluorophore, encompassing a Lewis acid-base pair, binds to primary amines on mesoporous silica nanoparticles, which may serve as environment-sensitive drug carriers. The fluorophore switches conformation, exhibiting different emission color and lifetimes, allowing for the detection of the water content of the nanoparticles' surroundings through fluorescence spectroscopy and microscopy.


Interactive image-guided neuroendoscopy: development and early clinical experience.
Technical advances and pioneering surgeons have established neuroendoscopy as an accepted diagnostic and therapeutic tool. The clinical indications for endoscopy, variety of operative techniques and number of endoscopic surgeons continue to increase steadily. However, there are fundamental limits to the scope of freehand endoscopy principally governed by the need for direct vision of anatomical and pathological structures. In addition, whilst the expert neuroendoscopist is only occasionally disorientated by complex distorted anatomy, the rising number of novices are likely to be mislead relatively often. We report the integration of neuroendoscopy with an optical neuronavigation system to provide interactive image-guided neuroendoscopy. This combination both removes the constraining requirement for direct vision and provides accurate localisation to guide the surgeon during surgery. We describe the clinical application of this method to two cases where image-guided endoscopy was essential to the safe completion of the procedure.


Revisiting the Longitudinal 90° Limit in High Intensity Linear Accelerators.
Parametric envelope and sum envelope resonances are analyzed to revisit the validity of an assumed stop band and design limit of high intensity linear accelerators at a longitudinal phase advance of 90° per focusing lattice period. While the 90° limit is unquestioned in the transverse plane, we show here that it can be dropped as longitudinal limit for lattices with two or more rf gaps per focusing period. A new limit arises, however, from a novel transverse-longitudinal parametric sum envelope instability. The resulting sum instability rule allows the phase advance to exceed 90° longitudinally provided that transversely it remains correspondingly under 90°. We suggest that the additional design freedom opens the possibility for larger accelerating gradients and stronger longitudinal focusing with potential length and cost saving in the design of advanced superconducting linear accelerator concepts-as long as technological cavity limits are not reached.


Development of a Clinically Relevant Reporter for Chimeric Antigen Receptor T-cell Expansion, Trafficking, and Toxicity.
Although chimeric antigen receptor T (CART)-cell therapy has been successful in treating certain hematologic malignancies, wider adoption of CART-cell therapy is limited because of minimal activity in solid tumors and development of life-threatening toxicities, including cytokine release syndrome (CRS). There is a lack of a robust, clinically relevant imaging platform to monitor in vivo expansion and trafficking to tumor sites. To address this, we utilized the sodium iodide symporter (NIS) as a platform to image and track CART cells. We engineered CD19-directed and B-cell maturation antigen (BCMA)-directed CART cells to express NIS (NIS+CART19 and NIS+BCMA-CART, respectively) and tested the sensitivity of 18F-TFB-PET to detect trafficking and expansion in systemic and localized tumor models and in a CART-cell toxicity model. NIS+CART19 and NIS+BCMA-CART cells were generated through dual transduction with two vectors and demonstrated exclusive 125I uptake in vitro. 18F-TFB-PET detected NIS+CART cells in vivo to a sensitivity level of 40,000 cells. 18F-TFB-PET confirmed NIS+BCMA-CART-cell trafficking to the tumor sites in localized and systemic tumor models. In a xenograft model for CART-cell toxicity, 18F-TFB-PET revealed significant systemic uptake, correlating with CART-cell in vivo expansion, cytokine production, and development of CRS-associated clinical symptoms. NIS provides a sensitive, clinically applicable platform for CART-cell imaging with PET scan. 18F-TFB-PET detected CART-cell trafficking to tumor sites and in vivo expansion, correlating with the development of clinical and laboratory markers of CRS. These studies demonstrate a noninvasive, clinically relevant method to assess CART-cell functions in vivo.


Post-mammoplasty human adjuvant disease.
Morphoea and keratoconjunctivitis sicca developed in a woman with seronegative rheumatoid arthritis six years after augmentation mammoplasty. Previous reports of post-mammoplasty connective tissue disease have followed the use of silicone gel, whereas the more 'biologically inert' saline-filled silastic implants were used in this case.


Spring temperature drives phenotypic selection on plasticity of flowering time.
In seasonal environments, a high responsiveness of development to increasing temperatures in spring can infer benefits in terms of a longer growing season, but also costs in terms of an increased risk of facing unfavourable weather conditions. Still, we know little about how climatic conditions influence the optimal plastic response. Using 22 years of field observations for the perennial forest herb Lathyrus vernus, we assessed phenotypic selection on among-individual variation in reaction norms of flowering time to spring temperature, and examined if among-year variation in selection on plasticity was associated with spring temperature conditions. We found significant among-individual variation in mean flowering time and flowering time plasticity, and that plants that flowered earlier also had a more plastic flowering time. Selection favoured individuals with an earlier mean flowering time and a lower thermal plasticity of flowering time. Less plastic individuals were more strongly favoured in colder springs, indicating that spring temperature influenced optimal flowering time plasticity. Our results show how selection on plasticity can be linked to climatic conditions, and illustrate how we can understand and predict evolutionary responses of organisms to changing environmental conditions.


[Rehabilitation after operations on the hip-joint (author's transl)].
The increase of hip-replacement is also seen in departments of rehabilitation. Among 270 operations during 1974/75 the main age at operations was 50-59 years. Para-articular ossification was more common in men than women. Function was disturbed more often in women, pain more common after operations other than replacement. Unfitness for work and compensation depend on the degree of limitation of mobility and weightbearing related to occupation. Because of relatively common complications and still unsatisfactory postoperative improvement, indication for replacement ought to be more critical. Conservative measures often give astonishingly good results, delay progressive symptoms. Techniques preserving the joint should be exhausted before the proper age for operation is reached.


[What was the fate of patients with leprosy during the plague pandemia in the middle ages (1348-1350)].
The author thinks that, if lepra has suddenly decreased in Europe from the 14th century, it is because the most severe cases, i.e. the most contagious ones, disappeared during the hecatombs caused between 1348 and 1350 by the "Black Death", the black plague, which took most often the pulmonary form. The author disproves the opinion of those who think that lepers died from plague. He thinks that lepers' death was secondary to that of the monks who, at this time, cared for these outcases, and thanks to their self-sacrifice permitted these lepers' survival. The monks were more exposed to contagion; obliged by their vocation and by pope's command to help the dyings and to give them sacraments, they were obliged to leave lepers to their fate. Like domestic animals, the latter died of hunger probably, any corpse or carcass being considered as plague victims. Supporting this opinion, the author reports his observations at Madagascar, where no leper of the leper-houses of Madascar center, a plague focus still to-day but very active between 1922 and 1936, contracted plague. On the other hand, experiments with "leprous" rats (Stefansky bacillus) showed a significant resistance of these animals to an experimental plague infection.


Chiral M3L2 self-assembled capsules through metal coordination of enantiopure ligating benzocyclotrimers: NMR spectroscopic and ESI mass spectrometric investigation.
The synthesis of enantiopure (+)-benzotricamphor syn-5, an important chiral C3-symmetric rigid building block for supramolecular applications, was studied in detail to reduce the number of steps and to increase the diastereoselectivity and overall yield. The new synthetic procedure allowed larger amounts of syn-5 to be obtained and used for the preparation of new derivatives, such as the corresponding tris-trifluoromethanesulfonate syn-12, which was efficiently transformed into (+)-benzotribornenetrinitrile syn-1 and (+)-benzotribornenetris(ethynyl-4-pyridine) syn-2. The previously reported (+)-benzotricamphortrioxime syn-6 was transformed into tris-nitrile syn-3 by Beckman reaction. Compounds syn-1-3 were employed as multidentate ligands for silver(I) and platinum(II) centres in apolar solvents. The linear coordination geometry of Ag(I) and square-planar geometry of cis-chelated Pt(II) in combination with the chiral tripodal ligands syn-1-3 led to the formation of chiral enantiopure capsules with M3 L2 stoichiometry, as confirmed by 2D NMR NOESY and DOSY experiments as well as ESI mass spectrometry.


A new anastomosis technique for large-calibered vein grafts.
A new anastomosis technique for large-calibered vein graft to repair a smaller-sized arterial defect is presented. At the proximal (upstream) anastomosis, a combination of sleeve anastomosis technique and plication of excess vessel wall is performed; and at the distal (downstream) anastomosis, the "fish-mouth" incision technique is used. The patency rate obtained with this technique is quite satisfactory, which justifies its use in clinical situations.


Correlation of chromatin composition with metabolic changes in nuclei of primitive erythroid cells from chicken embryos.
Changes in levels of biosynthesis of DNA, RNA, and histones were compared with relative proportions of each histone class during primitive erythropoiesis in embryonic chicks. We confirmed that erythrocyte-specific histone 5 (H5) was substantial in the earliest accessible, erythroblast-enriched stage and that it doubled in relative amount between polychromatic and orthochromatic stages to about 1 mol per 2 mol of each nucleosomal histone, still considerable less than in adult definitive erythrocytes. No other histones changed during primitive erythropoiesis, but the molar proportion of histone 1 (H1) always exceeded that of H5 in these cells, unlike definitive erythrocytes. The increase in content of H5 was accompanied by continued decline in synthesis of the other histones and DNA. The accumulation of H5 during development appears to occur in steps corresponding to the maturation of the primitive and definitive erythroid cell lines. Lysine-rich histones were more easily extracted from nuclei of the erythrosynthesis in whole cells and in isolated nuclei.


Pulmonary blood flow distribution during partial liquid ventilation.
Regional pulmonary blood flow was investigated with radiolabeled microspheres in four supine lambs during the transition from conventional mechanical ventilation (CMV) to partial liquid ventilation (PLV) and with incremental dosing of perfluorocarbon liquid to a cumulative dose of 30 ml/kg. Four lambs supported with CMV served as controls. Formalin-fixed, air-dried lungs were sectioned according to a grid; activity was quantitated with a multichannel scintillation counter, corrected for weight, and normalized to mean flow. During CMV, flow in apical and hilar regions favored dependent lung (P < 0.001), with no gradient across transverse planes from apex to diaphragm. During PLV the gradient within transverse planes found during CMV reversed, most notably in the hilar region, favoring nondependent lung (P = 0.03). Also during PLV, flow was profoundly reduced near the diaphragm (P < 0.001), and across transverse planes from apex to diaphragm a dose-augmented flow gradient developed favoring apical lung (P < 0.01). We conclude that regional flow patterns during PLV partially reverse those noted during CMV and vary dramatically within the lung from apex to diaphragm.


An Analysis of the Usage of Retinal Imaging Technology in the Detection of Age-Related Macular Degeneration.
Age-related macular degeneration (AMD) is a disease that worsens the central vision of numerous individuals across the globe. Ensuring that patients are diagnosed accurately and that their symptoms are carefully monitored is essential to ensure that adequate care is delivered. To accomplish this objective, retinal imaging technology is necessary to assess the pathophysiology that is required to give an accurate diagnosis of AMD. The purpose of this review is to assess the ability of various retinal imaging technologies such as optical coherence tomography (OCT), color fundus retinal photography, fluorescein angiography, and fundus photography. The statistical methods that were conducted yielded results that suggested that using OCT in conjunction with other imaging technologies results in a higher detection of symptoms among patients that have AMD. Further investigation should be conducted to ascertain the validity of the conclusions that were stated within the review.


Circulation of Bordetella pertussis in the Caribbean Netherlands: a population-based seroepidemiological study.
Pertussis is a respiratory infectious disease caused by Bordetella pertussis. In the Caribbean Netherlands (CN), comprising the islands Bonaire, St Eustatius, and Saba, registration of cases is mandatory for disease surveillance. However, insufficient laboratory facilities hamper case confirmation, and circulation persists. The aim of this seroepidemiological study was to gain insight into B. pertussis circulation in CN, and to investigate what factors contribute to the risk of infection. Blood samples and questionnaires were collected for 1829 participants aged 0-90 years. Concentrations of B. pertussis toxin-specific IgG antibodies (anti-Pt) were determined using a bead-based immunoassay to indicate infections within the previous 12 months (based on anti-Pt ≥ 50 IU/mL) in participants without detectable vaccine-induced humoral immunity. Risk factors for recent infection were analyzed using logistic regression models. An estimated 8.2% (95% CI 6.6-10.1) of CN residents aged ≥ 9 years were found to have been recently infected by B. pertussis. Risk factors for a recent infection were age 12-29 years (13.8-14.6%) and Dutch Caribbean or Surinamese origin (10.7%). B. pertussis infections occur frequently among CN residents aged ≥ 9 years, although few clinical pertussis cases are reported. Transmission to vulnerable individuals seems likely and should be taken into account in optimizing vaccination programs.


Dps-dependent in vivo mutation enhances long-term host adaptation in Vibrio cholerae.
As one of the most successful pathogenic organisms, Vibrio cholerae (V. cholerae) has evolved sophisticated regulatory mechanisms to overcome host stress. During long-term colonization by V. cholerae in adult mice, many spontaneous nonmotile mutants (approximately 10% at the fifth day post-infection) were identified. These mutations occurred primarily in conserved regions of the flagellar regulator genes flrA, flrC, and rpoN, as shown by Sanger and next-generation sequencing, and significantly increased fitness during colonization in adult mice. Intriguingly, instead of key genes in DNA repair systems (mutS, nfo, xthA, uvrA) or ROS and RNS scavenging systems (katG, prxA, hmpA), which were generally thought to be associated with bacterial mutagenesis, we found that deletion of the cyclin gene dps significantly increased the mutation rate (up to 53% at the fifth day post-infection) in V. cholerae. We further determined that the dpsD65A and dpsF46E point mutants showed a similar mutagenesis profile as the Δdps mutant during long-term colonization in mice, which strongly indicated that the antioxidative function of Dps directly contributes to the development of V. cholerae nonmotile mutants. Methionine metabolism pathway may be one of the mechanism for ΔflrA, ΔflrC and ΔrpoN mutant increased colonization in adult mice. Our results revealed a new phenotype in which increased fitness of V. cholerae in the host gut via spontaneous production nonmotile mutants regulated by cyclin Dps, which may represent a novel adaptation strategy for directed evolution of pathogens in the host.


Optimal adaptive two-stage designs for phase II cancer clinical trials.
In oncology, single-arm two-stage designs with binary endpoint are widely applied in phase II for the development of cytotoxic cancer therapies. Simon's optimal design with prefixed sample sizes in both stages minimizes the expected sample size under the null hypothesis and is one of the most popular designs. The search algorithms that are currently used to identify phase II designs showing prespecified characteristics are computationally intensive. For this reason, most authors impose restrictions on their search procedure. However, it remains unclear to what extent this approach influences the optimality of the resulting designs. This article describes an extension to fixed sample size phase II designs by allowing the sample size of stage two to depend on the number of responses observed in the first stage. Furthermore, we present a more efficient numerical algorithm that allows for an exhaustive search of designs. Comparisons between designs presented in the literature and the proposed optimal adaptive designs show that while the improvements are generally moderate, notable reductions in the average sample size can be achieved for specific parameter constellations when applying the new method and search strategy.


[A case of a young woman with tuberculous peritonitis diagnosed owing to high value of ADA].
A 26-year-old woman visited the first hospital due to ascites in August 2003, She had continual abdominal pain diagnosed as Irritable bowel disease after a gastrointestinal and colon fiberscopy was performed. Chest-abdominal CT scan revealed normal chest, massive ascites and swollen ovary. To rule out malignancy, surgical biopsy was performed, which brought no significant findings. We focused on the high value of Adenosin deaminase (ADA) in ascites and strongly suspected tuberculotic peritonitis. Consequently, pathologist confirmed the existence of bacterial bodies stained by acid-fast stain after our consultation. Compared with the poor diagnostic accuracy of surgical biopsy, the value of ADA in ascites has a very high sensitivity and specificity. Considering the high risk of being infertile, to begin diagnostic medication of tuberculotic peritonitis is an acceptable choice for young women with a high value of ADA in the ascites.


Signal transduction through the cAMP-dependent protein kinase.
Temporal cellular events responsible for hormonal activation of responses mediated by the cAMP-dependent protein kinase (PKA) have been studied in living cells. By selectively perturbing molecular function of Gs, the catalytic subunit of PKA (C), or the nuclear factor CREB, in cells through microinjection of inhibitory agents specific for these molecules or activated forms of these molecules, we have obtained evidence for a requirement for the function of each of these molecules in the hormonal stimulation of cAMP-regulated genes. Moreover, by introducing fluorescently labeled PKA subunits into these cells as molecular tracers, or by immunofluorescence of C subunit, we have observed biological translocation of C subunit from the cytoplasm to the nucleus during transcriptional activation and a quenching of this by the inhibitor molecule, PKI. The implications of these cellular and molecular events in the signal transduction of hormonal responses are discussed.


p16Ink4a suppression of lung adenocarcinoma by Bmi-1 in the presence of p38 activation.
Because evasion of tumor suppression is a critical step in cancer development, cancer cells have developed a variety of mechanisms to circumvent the influence of tumor suppressive pathways. Thus, genes that negatively regulate tumor suppressors could be considered novel types of oncogenes such as Bmi-1 repressing p16Ink4a and inhibiting p53 and were found to be frequently up-regulated in a variety of cancers. p38 mitogen-activated protein kinase (MAPK), which reportedly plays a crucial role as a tumor suppressor, is activated in number of lung adenocarcinomas, which is seemingly at odds with its role as a tumor suppressor. We examined 10 lung adenocarcinomas and corresponding normal tissues and determined the expression levels of a variety of tumor suppressor proteins through real-time polymerase chain reaction and immunohistochemistry and measured p38 MAPK activity by immunoblotting or immunohistochemistry analysis. In the in vitro cellular model, p38 activation by H-Ras and consequent senescence induction was achieved through retro-viral gene transduction. Similarly, the suppression of p16Ink4a by Bmi-1 after the introduction of H-Ras was achieved through transient transfection with cationic liposome. We detected several lung adenocarcinomas that were positive for activated p38 MAPK but evidenced reduced levels of p16Ink4a expression. The suppression of p16Ink4a occurred in parallel with an increase in Bmi-1 and/or p16Ink4a promoter hypermethylation. Consistent with these observations, the H-Ras-stimulated induction of p16Ink4a was suppressed significantly through the coexpression of Bmi-1 in vitro. These results demonstrate that the suppression of p16Ink4a by either the induction of Bmi-1 or the hypermethylation of p16Ink4 may be an important step in avoiding tumor surveillance by p38 MAPK during the development of lung cancer.


A Retrospective Study on the Impact of COVID-19 on Emergency General Surgery.
Background The coronavirus pandemic has caused global disruption to all aspects of life. This disturbance has been most notable in the medical world. Political, societal, medical, and behavioral alterations have forced emergency surgical practices to adapt. This study investigated the impact of coronavirus 2019 (COVID-19) at a busy surgical center. Methodology This is a retrospective observational study. Three study periods were analyzed: pre-COVID, first wave, and second wave. Data were collected on referrals, diagnoses, investigations, management pathways, outcomes, patient behavior, and consultant practice. A one-way analysis of variance (ANOVA test) was used for the analysis of parametric data and the Mann-Whitney U test for non-parametric data. Results Declining numbers of patients presented across the three periods. There was a severe disruption in performing emergency general surgeries during the first wave, propagated by alterations in clinical decision-making, as well as fluctuations in societal and patient behavior. Despite the effects of the second wave being significantly more profound in terms of hospitalization and COVID-related mortality, a paradoxical, gradual return to the norm was noted, which was seen in referral pathways, imaging decisions, and management strategies. Conclusion Our data is suggestive of society, both within and outside the medical sphere, adjusting to life with COVID-19.


Appraisal of distal splenorenal shunt in the treatment of esophageal varices: an analysis of prophylactic, emergency, and elective shunts.
From June, 1969 to February, 1987, distal splenorenal shunt was carried out on 78 patients with esophagogastric varices. The operations were urgent in 9, elective in 40, and prophylactic in 29 patients. There were 52 males and 26 females. Age ranged from 16 to 76 years with an average of 53 years. Thirty-seven patients were alcoholics. Hepatitis B surface antigen was positive in only 15.5%. The causes of portal hypertension were cirrhosis of the liver in 67, chronic hepatitis in 5, idiopathic portal hypertension in 4, primary biliary cirrhosis in 1, and fatty liver in 1 patient. Fifty-two patients were in Child's class A, 18 in class B, and 8 in class C. Emergency shunts were performed only when conservative therapy had failed to stop variceal bleeding. Prophylactic operations were done in patients having Child's class A or class B liver disease and risky varices, in varices larger than 5 mm in diameter and/or varices with red color signs such as cherry red spots. Forty-two patients underwent the original Warren shunt, but the remaining 36 had modified distal splenorenal shunt with expanded polytetrafluoroethylene interposition. The operative mortality rates were 11.1% in the emergency group, 2.5% in the elective group, and 3.4% in the prophylactic group. The overall operative and hospital death rates were 3.8% and 7.7%, respectively. The patency rate was 94.1% and the incidence of rebleeding from esophageal varices was 3.8%. Hepatic encephalopathy, although mild to moderate in degree, was observed in 14.7% of 75 patients excluding 3 operative deaths.(ABSTRACT TRUNCATED AT 250 WORDS)


Conservative operation for infective endocarditis of the mitral valve.
Subvalvular preservation is necessary to maintain left ventricular function, but accidental retention of infected tissue could cause postoperative endocarditis. We examined 71 consecutive patients who underwent operation for mitral endocarditis. Endocarditis was uncontrolled and active in 24 patients, partially treated (unfinished antibiotic course) in 17, and healed in 30. Valves were repaired in 17% versus 59% versus 63% and replaced with subvalvular preservation in 25% versus 6% versus 3% of the uncontrolled active, partially treated, and healed groups, respectively. Thirty-day mortality was 29% versus 0% versus 3.3% (p=0.003), total mortality was 46% versus 18% versus 17% (p=0.035), and complications-related mortality was 38% versus 11% versus 13% (p=0.054), respectively. There was a trend toward lower complications-related mortality with subvalvular preservation than without. Postoperative endocarditis occurred in 3 of 30 patients without and 1 of 41 patients with subvalvular preservation. Postoperative mortality in uncontrolled active mitral endocarditis remains high, but results are good with partially treated or healed endocarditis. Subvalvular preservation improves outcome, does not increase postoperative endocarditis rates, and should be performed whenever feasible.


Bone metastases and skeletal-related events in patients with malignant pheochromocytoma and sympathetic paraganglioma.
Bone metastases (BM) can cause severe pain, spinal cord compression, pathological fractures, and/or hypercalcemia. These skeletal-related events (SREs) may cause immobilization, loss of independence, poor quality of life, and reduced survival. There is limited information on the clinical effects of BM and SREs in patients with malignant pheochromocytoma or sympathetic paraganglioma (PHEO/sPGL). We studied the prevalence and clinical characteristics of BM and SREs in patients with PHEO/sPGL and investigated the risk factors for SRE development. Using a large institutional database, we conducted a retrospective study of 128 patients with malignant PHEO/sPGL at The University of Texas MD Anderson Cancer Center from 1967 through 2011. Of the patients, 91 (71%) had BM, and 57 of these (63%) developed metachronous BM at a median time of 3.4 years (range, 5 months to 23 years) after the primary tumor diagnosis. Metastatic disease was confined exclusively to the skeleton in 26 of 128 (20%) patients. Sufficient information to assess SRE occurrence was available for 67 patients, and 48 of 67 (72%) patients had at least 1 SRE. The median overall survival for the 128 patients was 12 years for patients with only BM, 7.5 years for patients with nonosseous metastases, and 5 years for patients with both BM and nonosseous metastases (log rank test P value = .005). We were unable to identify factors predictive of SRE development, but the occurrence of a first SRE was associated with the development of subsequent SREs in 48% of subjects. In responsive patients, the use of systemic therapy was associated with fewer SREs (P < .0001). BM and SREs are frequent in patients with malignant PHEO/sPGL. SREs often develop shortly after the diagnosis of BM; severe pain is the most frequent SRE. These patients should be followed long-term by a multidisciplinary team to promptly identify the need for medical or surgical intervention.


Comparison of the haemodynamic effects of epoprostenol (prostacyclin) and tolazoline.
The haemodynamic effects of infusion of epoprostenol (prostacyclin) and bolus injection of tolazoline were compared in a crossover study in 11 children with pulmonary hypertension caused by pulmonary vascular disease. The children were studied during cardiac catheterisation, while they were anaesthetised, paralysed, and ventilated with 100% oxygen. The order of drug administration was not randomised because tolazoline has a half life of hours whereas epoprostenol has a half life of a few minutes. Both drugs caused pulmonary and systemic vasodilatation, and there were no significant differences between the two. The 95% confidence intervals suggest that tolazoline did not have a clinically important haemodynamic advantage over epoprostenol. Previous reports suggest that serious side effects are common when tolazoline is used in repeated doses; epoprostenol has only a few minor side effects that are rapidly reversible when the infusion is stopped. Epoprostenol is more expensive than tolazoline but this study suggests that epoprostenol is a more suitable pulmonary vasodilator if more than a single dose is required.


Why Do Immigrant and Swedish Adolescents Engage in Ethnic Victimization? Common and Distinct Underlying Factors.
Youth of immigrant background are at risk of experiencing victimization due to their ethnic or cultural background. However, limited knowledge is available regarding why youth victimize their immigrant peers, and whether the factors associated with engagement in ethnic victimization vary across adolescents of different background. To address this gap in knowledge, the present study aimed to elucidate the common or differential factors associated with engagement in ethnic victimization among immigrant and native youth. The analytical sample included seventh grade students residing in Sweden from 55 classrooms (N = 963, Mage = 13.11, SD = 0.41; 46% girls; 38% youth of immigrant background). The results showed that being morally disengaged and engaging in general victimization are the common denominators of engagement in ethnic victimization for immigrant and Swedish youth. Low levels of positive attitudes toward immigrants provide a foundation for ethnic victimization among Swedish youth, but not youth of immigrant background. Classroom ethnic composition was not significantly related to engagement in ethnic victimization in either group. Predictors of engagement in ethnic victimization seem to have similarities and differences among immigrant and Swedish youth. The factors involved require further attention in developing strategies to combat bias-based hostile behaviors in diverse school settings.


Structural constraints for protein repair in plant photosynthetic membranes.
The thylakoid membrane system inside plants chloroplasts defines the structural framework for photosynthetic conversion of sunlight into metabolic energy forms (ATP, NADPH + H(+)). An architectural hallmark of these thylakoid membranes is the tight stacking of part of the membrane into cylindrical flat grana thylakoids, with a diameter of about 500 nm, that are interconnected by unstacked stroma lamellae forming a complex 3D network of alternating grana piles and stroma lamellae. The structural differentiation in the stacked and unstacked thylakoid regions is the basis for a pronounced spatial separation of multisubunit pigment-protein complexes that catalyze energy transformation. The main part of photosystem II (PSII) associated with light-harvesting complex II (LHCII) is concentrated in the grana thylakoids whereas PSI-LHCI and the ATPase complex are excluded from the stacked grana and accumulate in the unstacked thylakoid regions. The fifth protein complex, the cytochrome b 6f complex, is assumed to be homogenously distributed. It is important to recognize that this structural arrangement is not static but highly dynamic and responsive to environmental factors like light intensity and quality or temperature. Knowledge about the interplay between dynamic structural features of the intricate thylakoid architecture, and the functionality, regulation, repair and biogenesis of the photosynthetic machinery is essential for understanding the plasticity of energy conversion in plants living in a fluctuating multi-factorial environment.