Empirical amide I vibrational frequency map: application to 2D-IR line shapes for isotope-edited membrane peptide bundles.
The amide I vibrational mode, primarily associated with peptide-bond carbonyl stretches, has long been used to probe the structures and dynamics of peptides and proteins by infrared (IR) spectroscopy. A number of ab initio-based amide I vibrational frequency maps have been developed for calculating IR line shapes. In this paper, a new empirical amide I vibrational frequency map is developed. To evaluate its performance, we applied this map to a system of isotope-edited CD3-zeta membrane peptide bundles in aqueous solution. The calculated 2D-IR diagonal line widths vary from residue to residue and show an asymmetric pattern as a function of position in the membrane. The theoretical results are in fair agreement with experiments on the same system. Through analysis of the computed frequency time-correlation functions, it is found that the 2D-IR diagonal widths are dominated by contributions from the inhomogeneous frequency distributions, from which it follows that these widths are a good probe of the extent of local structural fluctuations. Thus, the asymmetric pattern of line widths follows from the asymmetric structure of the bundle in the membrane.


Ethnomedicinal uses of plants for various diseases in the remote areas of Changa Manga Forest, Pakistan.
This study aims at reporting the indigenous knowledge of the medicinal flora from the inhabitants of surroundings of the World's largest artificial planted forest "Changa Manga", Pakistan. Data were collected by direct interviews and group meetings from 81 inhabitants including 32 local healers having information regarding the use of indigenous medicinal plants over a period of one year. Different statistical tools were applied to analyze the data including Frequency citation (FC), Relative frequency citation (RFC), Use Value, Factor of informants consensus and fidelity level. This study reported 73 plant species belonging to 37 plant families and 46 genera. The majority of plant species belong to compositae family. The most commonly used medicinal plants were P. hysterophorus L., P. dactylifera L., S. indicum L, P. harmala L., P. emblica L., and A. indica A.Juss. The greatest number of species was used to cure gastrointestinal disorders. The highest fidelity level (68.18%) was of E. helioscopia to cure gastrointestinal disorders. Maximum fresh uses (17) were reported by C. dactylon (L.) Pars. While the highest number of species reporting fresh uses in similar number was 13. In this study, five novel plants are being reported for the first time in Pakistan for their ethnomedicinal worth. Our data reflect unique usage of the medicinal plants in the study area. The statistical tools used in the study proved useful in pointing the most important and disease category specific plants. High use value plant and the new reported medicinal plants might prove an important source of the isolation of pharmacologically active compounds.


An Alzheimer's Disease Genetic Risk Score Predicts Longitudinal Thinning of Hippocampal Complex Subregions in Healthy Older Adults.
Variants at 21 genetic loci have been associated with an increased risk for Alzheimer's disease (AD). An important unresolved question is whether multiple genetic risk factors can be combined to increase the power to detect changes in neuroimaging biomarkers for AD. We acquired high-resolution structural images of the hippocampus in 66 healthy, older human subjects. For 45 of these subjects, longitudinal 2-year follow-up data were also available. We calculated an additive AD genetic risk score for each participant and contrasted this with a weighted risk score (WRS) approach. Each score included APOE (apolipoprotein E), CLU (clusterin), PICALM (phosphatidylinositol binding clathrin assembly protein), and family history of AD. Both unweighted risk score (URS) and WRS correlated strongly with the percentage change in thickness across the whole hippocampal complex (URS: r = -0.40; p = 0.003; WRS: r = -0.25, p = 0.048), driven by a strong relationship to entorhinal cortex thinning (URS: r = -0.35; p = 0.009; WRS: r = -0.35, p = 0.009). By contrast, at baseline the risk scores showed no relationship to thickness in any hippocampal complex subregion. These results provide compelling evidence that polygenic AD risk scores may be especially sensitive to structural change over time in regions affected early in AD, like the hippocampus and adjacent entorhinal cortex. This work also supports the paradigm of studying genetic risk for disease in healthy volunteers. Together, these findings will inform clinical trial design by supporting the idea that genetic prescreening in healthy control subjects can be useful to maximize the ability to detect an effect on a longitudinal neuroimaging endpoint, like hippocampal complex cortical thickness.


Explanation-Driven Deep Learning Model for Prediction of Brain Tumour Status Using MRI Image Data.
Cancer research has seen explosive development exploring deep learning (DL) techniques for analysing magnetic resonance imaging (MRI) images for predicting brain tumours. We have observed a substantial gap in explanation, interpretability, and high accuracy for DL models. Consequently, we propose an explanation-driven DL model by utilising a convolutional neural network (CNN), local interpretable model-agnostic explanation (LIME), and Shapley additive explanation (SHAP) for the prediction of discrete subtypes of brain tumours (meningioma, glioma, and pituitary) using an MRI image dataset. Unlike previous models, our model used a dual-input CNN approach to prevail over the classification challenge with images of inferior quality in terms of noise and metal artifacts by adding Gaussian noise. Our CNN training results reveal 94.64% accuracy as compared to other state-of-the-art methods. We used SHAP to ensure consistency and local accuracy for interpretation as Shapley values examine all future predictions applying all possible combinations of inputs. In contrast, LIME constructs sparse linear models around each prediction to illustrate how the model operates in the immediate area. Our emphasis for this study is interpretability and high accuracy, which is critical for realising disparities in predictive performance, helpful in developing trust, and essential in integration into clinical practice. The proposed method has a vast clinical application that could potentially be used for mass screening in resource-constraint countries.


Brotizolam as a pre-operative hypnotic.
Efficacy of and tolerance to brotizolam when used as a preoperative hypnotic were studied in two double-blind, randomised parallel group studies. Brotizolam (0.25 and 0.50 mg) was superior to placebo. Efficacy was assessed as good-to-satisfactory in 73.0% of patients with 0.25 mg and in 88.0% with 0.5 mg. A similar assessment was reached in 40.0% of patients with placebo. Brotizolam 0.5 mg was superior to 0.25 mg, and with the higher dose subjective assessments of anxiety were reduced. Efficacy of tolerance to 0.5 mg brotizolam and 2.0 mg flunitrazepam were compared, and both drugs were found to be effective and well tolerated. Brotizolam maintained sleep throughout the night more effectively than flunitrazepam. A dose range of 0.25-0.5 mg brotizolam is recommended as a pre-operative hypnotic.


The origin of the tetrapod limb: from expeditions to enhancers.
More than three centuries ago natural philosophers, and later anatomists, recognized a fundamental organization to the skeleton of tetrapod limbs. Composed of three segments, stylopod, zeugopod, and autopod, this pattern has served as the basis for a remarkably broad adaptive radiation from wings and flippers to hands and digging organs. A central area of inquiry has been tracing the origins of the elements of this Bauplan in the fins of diverse fish. Can equivalents of the three segments, and the developmental processes that pattern them, be seen in fish fins? In addition, if so, how do these data inform theories of the transformation of fins into limbs? Answers to these questions come from linking discoveries in paleontology with those of developmental biology and genetics. Burgeoning discoveries in the regulatory biology of developmental genes and in the genomics of diverse species offer novel data to investigate these classical questions.


Detection of skeletal muscle metastasis: torso FDG PET-CT versus contrast-enhanced chest or abdomen CT.
Skeletal muscle metastasis (SMM) in cancer patients has not been sufficiently evaluated regarding prevalence and proper method of detection. To determine the prevalence of SMM and compare the diagnostic competencies for SMM of torso F-18 fluorodeoxyglucose (FDG) positron emission tomography-computed tomography (PET-CT) and contrast-enhanced chest or abdomen CT. We investigated 18,225 PET-CT studies of 6359 cancer patients performed from 2005 to 2012. The PET-CT studies describing potential SMM were retrieved and the corresponding medical records were reviewed. The gold standard for SMM was histopathologically-proven SMM or imaging study-based disease progression. The detectability of SMM was compared between PET-CT and contrast-enhanced CT. Twenty-six patients had 84 SMM lesions, representing a SMM prevalence of 0.41%. Lung cancer was the most common SMM-associated malignancy (54%) and the gluteal/pelvic girdle muscle was the most frequently involved SMM site (37%). All 84 SMM lesions were visualized on PET-CT (100%). Of these PET-CT positive 84 SMM lesions, 51 lesions were in the CT field of view (FOV) (61%), whereas 33 lesions were out of the CT FOV (39%). Among these 51 lesions, 17 lesions showed rim-enhancing nodules/masses (33%), eight lesions showed homogeneously enhancing nodules (16%), three lesions showed heterogeneously enhancing nodules (6%), and 23 SMM lesions (45%) were non-diagnostic by CT. All 51 SMM lesions within CT FOV were detected on PET-CT (100%), whereas only 28 were visualized on CT (54.9%), resulting in a significant difference (P < 0.005). On average, 2.6 more organs with concomitant metastases were found when SMM was revealed by PET-CT. The prevalence of SMM was as low as 0.41% in the current large cohort of cancer patients. Torso PET-CT was a more competent modality than contrast-enhanced CT in the detection of SMM.


Identification of two distinct elements in the long terminal repeat of HTLV-I responsible for maximum gene expression.
Human T-cell leukemia virus type I has a unique sequence, pX, between env and the 3' long terminal repeat (LTR). One of its products, p40, activates gene expression directed by the LTR in a trans-acting manner. We have analysed the mechanism of this trans-activation mediated by p40 in human T cells co-transfected with a plasmid expressing p40 using the transient CAT gene expression. We identified two distinct elements in the LTR which are involved in maximum gene expression. The first was present in a 230-bp fragment upstream from TATA box in the U3 region and behaved as a classical enhancer. This region was also shown to be responsible for trans-activation by p40. This element alone together with functional p40 could direct the gene expression at only approximately 10% of the level achieved by the complete LTR and p40. The second element was present within a 300-bp fragment downstream from the RNA start site and profoundly enhanced the gene expression in a way independent from trans-activation mechanism. This enhancement was observed only when the element was located immediately downstream from the RNA start site without orientation preference. These two elements participate independently in the enhancement of gene expression.


Prevalence, Genotype Richness, and Coinfection Patterns of Hemotropic Mycoplasmas in Raccoons (Procyon lotor) on Environmentally Protected and Urbanized Barrier Islands.
Raccoons (Procyon lotor) are successful urban adapters and hosts to a number of zoonotic and nonzoonotic pathogens, yet little is known about their hemoplasma infections and how prevalence varies across habitat types. This study identifies hemotropic Mycoplasma species infection in raccoons from urban and undisturbed habitats and compares hemoplasma infection in sympatric urban cats (Felis catus) from the same geographic region. We collected blood from raccoons (n = 95) on an urban coastal island (n = 37) and an undisturbed coastal island (n = 58) and from sympatric urban cats (n = 39) in Georgia, USA. Based on 16S rRNA gene amplification, 62.1% (59/95) of raccoons and 17.9% (7/39) of feral cats were positive for hemoplasma. There was a greater percentage of hemoplasma-infected raccoons on the undisturbed island (79.3% [46/58]) than on the urban island (35.1% [13/37]; χ2 = 16.9, df = 1, P = 0.00004). Sequencing of the full-length 16S rRNA gene amplicons revealed six hemoplasma genotypes in raccoons, including five novel genotypes that were distinct from three known hemoplasma species identified in the sympatric cats. In addition, the hemoplasma genotypes detected in raccoons were not identified in sympatric cats or vice versa. Although all six hemoplasma genotypes were found in raccoons from urban and undisturbed islands, coinfection patterns differed between sites and among individuals, with the proportion of coinfected raccoons being greater in the undisturbed site. This study shows that raccoons are hosts for several novel hemoplasmas and that habitat type influences infection patterns.IMPORTANCE This study provides information about novel hemoplasmas identified in raccoons (Procyon lotor), which can be used for assessments of the prevalence of these hemoplasmas in raccoon populations and for future studies on the potential pathogenic impacts of these hemoplasmas on raccoon health. Raccoons from the undisturbed habitat had a higher prevalence of hemoplasma infection than urban raccoons. There does not appear to be cross-species transmission of hemotropic mycoplasmas between urban raccoons and feral cats. Raccoons appear to be hosts for several novel hemoplasmas, and habitat type influences infection patterns.


Mutations of TGFbeta signaling molecules in human disease.
The transforming growth factor beta (TGFbeta) signaling pathway regulates several biological processes including cellular proliferation, differentiation, apoptosis, migration, and extracellular matrix deposition. Ligand and receptor family members signal through two main Smad signaling branches, TGFbeta/activin to Smad2/3 (Sma and MAD-related proteins) and bone morphogenetic protein (BMP) to Smad1/5. At the molecular level, TGFbeta acts by modifying cytoskeletal organization and ultimately regulating expression of specific target genes. Germline disruption of TGFbeta signaling leads to several types of hereditary congenital malformation or dysfunction of the skeletal, muscular and/or cardiovascular systems, and to cancer predisposition syndromes. In this review, the molecular etiology of TGFbeta-associated disorders is examined, together with a discussion of clinical overlap between syndromes and possible biological explanations underlying the variable penetrance and expressivity of clinical characteristics. Increasing our understanding of the molecular etiology underlying genotype-phenotype correlations will ultimately provide a molecular-based approach that should result in better prognostic tools, smart therapeutics and individualized disease management, not only for these rare syndromes, but for more generalized disorders of the cardiovascular and musculoskeletal systems and cancer. The clinical consequence of TGFbeta signaling mutations appears to depend on environmental factors and on the basal levels of ongoing signaling transduction networks specific to each individual. In this respect, genetic background might be a central factor in determining disease outcome and treatment strategy for TGFbeta-associated diseases.


Synthesis of Aldehydes by Organocatalytic Formylation Reactions of Boronic Acids with Glyoxylic Acid.
Reported herein is a conceptually novel organocatalytic strategy for the formylation of boronic acids. New reactivity is engineered into the α-amino-acid-forming Petasis reaction occurring between aryl boronic acids, amines, and glyoxylic acids to prepare aldehydes. The operational simplicity of the process and its ability to generate structurally diverse and valued aryl, heteroaryl, and α,β-unsaturated aldehydes containing a wide array of functional groups, demonstrates the practical utility of the new synthetic strategy.


Perceived discrimination and the adjustment of African American youths: a five-year longitudinal analysis with contextual moderation effects.
Longitudinal links between perceived racial discrimination and later conduct problems and depressive symptoms were examined among 714 African American adolescents who were 10-12 years old at recruitment. Data were gathered 3 times over a 5-year period. Hypotheses were tested via latent curve modeling and multiple-group latent growth modeling. Increases in perceived discrimination were associated with increased conduct problems and depressive symptoms. This association was weaker when youths received nurturant-involved parenting, affiliated with prosocial friends, and performed well academically. For conduct problems, the association was stronger for boys than for girls; for depressive symptoms, no gender differences emerged. The findings thus identify contextual variables that moderate the contribution of perceived discrimination to African American youths' adjustment.


Cotton photosynthetic productivity enhancement through uniform row-spacing with optimal plant density in Xinjiang, China.
Xinjiang is currently the most dominant cotton (Gossypium hirsutum L.)-growing region in China and possesses abundant radiation resource. The cultivation techniques such as wide and narrow row-spacing and high density are widely adopted to obtain high cotton yield in the region. However, the region is facing some problems including poor light transmittance in the field and low exploitation for light resources under the current planting pattern which impedes further growth in cotton yields. Therefore, it is essential to develop some cultivation practices to increase radiation use efficiency (RUE) and cotton yields in Xinjiang. Here we conducted a field experiment to quantify the effects of row spacing pattern and plant density on RUE, intercepted photosynthetically active radiation from May to August (IRAR5-8), and lint yield during 2017 and 2018. In this study, we designed two row-spacing configurations (R1, wide and narrow configuration, 66 cm + 10 cm; R2, uniform row-spacing configuration, 76 cm) and six plant densities (4.5, 9.0, 13.5, 18.0, 22.5, and 27.0 plants m-2). The RUE, lint yield, and number of bolls were higher in R2 than R1 by 4.1-5.9, 2.5-4.8, and 9.1-14.2%, respectively. The RUE significantly increased with plant density, but lint yield stabilized at 18.0 plants m-2. Moreover, RUE had more significant positive effects on boll number and lint yield. Overall, we found that R2 combined with optimal plant densities (13.5-18.0 plants m-2) would be an effective strategy to achieve higher RUE and yields in the Xinjiang cotton system.


Prevalence of serum IgG antibodies against SARS-CoV-2 among clinic staff.
The SARS-CoV-2 pandemic threatens health care providers and society. For planning of treatment capacities, it is of major importance to obtain reliable information on infection and fatality rates of the novel coronavirus. A German community study, the so-called Heinsberg study, found a 5-fold higher infection rate (and thus a remarkably lower fatality rate) than the officially reported cases suggest. We were interested to examine the SARS-CoV-2-IgG antibody status among clinic staff of a large neurological center in Northern Germany. Blood samples and questionnaires (demographic data, medical history) were collected pseudonymously. In total, 406 out of 525 (77.3%) of our employees participated in the study. The infection rate among the staff was as high as 2.7%. Including drop-outs (missing questionnaire but test result available), the infection rate was even higher (2.9%). Only 36% of the positively tested employees did suffer from flu-like symptoms in 2020. None of the nurses-having closest and longest contact to patients-were found to be positive. Despite the fact that the infection rate among clinic staff may not be directly compared to the situation in the surrounding county (due to different testing procedures), one might hypothesize that the infection rate could be more than 30-fold higher than the number of officially reported cases for the county of Hameln-Pyrmont. The high rate of IgG-positive, asymptomatic healthcare workers might help to overcome fears in daily work.


Real-time fluorescence imaging of abdominal, pleural, and lymphatic metastases.
Virally-directed fluorescence imaging has the potential to revolutionize intra-operative oncologic staging and tumor resection. Many viruses genetically engineered to specifically infect tumor cells as cancer therapy can be further modified to have a visible marker gene for cancer staging. In this chapter, we describe such a herpes simplex virus (HSV) modified to be detected by fluorescence. Other viruses so designed can be similarly used in cancer detection and staging. Replication-competent, tumor-specific HSV NV1066 expresses green fluorescent protein (GFP) in infected cancer cells. One single dose of NV1066 administered via intratumor, intracavitary, or systemic injection can spread within and across body cavities to target tumor cells while sparing normal tissue cells from infection. Tumors otherwise invisible by conventional laparoscopy appear green with the use of an endoscope equipped with a fluorescent filter. Furthermore, with GFP expression easily visualized by stereomicroscopy, microscopic, and pathologic analysis is significantly enhanced. This chapter addresses NV1066-directed visualization of peritoneal, pleural, and lymphatic metastases. This chapter also provides protocols for the production of tumor models in various body cavities in rodents.


Report of the 13th Annual International Pachyonychia Congenita Consortium Symposium.
The International Pachyonychia Congenita Consortium (IPCC) is a group of physicians and scientists from around the world dedicated to developing therapies for pachyonychia congenita, a rare autosomal dominant skin disorder. The research presented at the 13th Annual Research Symposium of the IPCC, held on 10-11 May 2016, in Scottsdale, AZ, U.S.A., is reported here.


Impact of Laurel Wilt, Caused by Raffaelea lauricola, on Leaf Gas Exchange and Xylem Sap Flow in Avocado, Persea americana.
Laurel wilt, caused by Raffaelea lauricola, is a destructive disease of avocado (Persea americana). The susceptibility of different cultivars and races was examined previously but more information is needed on how this host responds to the disease. In the present study, net CO2 assimilation (A), stomatal conductance of H2O (gs), transpiration (E), water use efficiency (WUE), and xylem sap flow rates were assessed in cultivars that differed in susceptibility. After artificial inoculation with R. lauricola, there was a close relationship between symptom development and reductions in A, gs, E, WUE, and mean daily sap flow in the most susceptible cultivar, 'Russell', and significantly greater disease and lower A, gs, E, WUE, and sap flow rates were usually detected after 15 days compared with the more tolerant 'Brogdon' and 'Marcus Pumpkin'. Significant differences in preinoculation A, gs, E, and WUE were generally not detected among the cultivars but preinoculation sap flow rates were greater in Russell than in Brogdon and Marcus Pumpkin. Preinoculation sap flow rates and symptom severity for individual trees were correlated at the end of an experiment (r=0.46), indicating that a plant's susceptibility to laurel wilt was related to its ability to conduct water. The potential management of this disease with clonal rootstocks that reduce sap flow rates is discussed.


Temporal characteristics of nasalization in children and adult speakers of American English and Korean during production of three vowel contexts.
The purpose of this study was to identify and compare the temporal characteristics of nasalization in relation to (1) languages, (2) vowel contexts, and (3) age groups. Two distinct acoustic energies from the mouth and nose were recorded during speech production (/pamap, pimip, pumup/) using two microphones to obtain the absolute and proportional measurements on the acoustic temporal characteristics of nasalization. Twenty-eight normal adults (14 American English and 14 Korean speakers) and 28 normal children (14 American English and 14 Korean speakers) participated in this study. In both languages, adults showed shorter duration of nasalization than children within all three vowel contexts. The high vowel context revealed longer duration of nasalization than the low vowel context in both languages. There was no significant difference of temporal characteristics of nasalization between American English and Korean. Nasalization showed different timing characteristics between children and adults across vowel contexts. The results are discussed in association with developmental coarticulation and the relationship between acoustic consequences of articulatory events and vowel height.


Solution conformation of enopeptin A, a depsipeptide antibiotic, using 2D NMR and restrained molecular dynamics studies.
Studies on the solution conformation of the cyclic depsipeptide antibiotic enopeptin A have been carried out using 2D NMR and molecular modelling techniques. The proton resonances of the antibiotic in DMSO-d6 have been assigned by the use of TOCSY and ROESY experiments. The interproton distance information obtained from the ROESY experiments have been used as the basis for elucidating the probable structures in solution. The restrained molecular dynamics technique was applied to calculate the structures in solution, and six resultant structures with fewer distance constraint violations were obtained that satisfy the experimental restraints very well. The conformation of the cyclic moiety of the molecules is well defined whereas the aliphatic chain segment is disordered.


Inter- and intra-observer variability of the Los Angeles classification: a reassessment.
Los Angeles classification is widely adopted for reporting endoscopic gastroesophageal reflux disease. We assessed the inter- and intra-observer variability of the Los Angeles classification. Still images (n = 254) of the lower esophagus were presented to 9 gastroenterologists (6 experts and 3 trainees) and they were asked to report the images according to the Los Angeles classification. After 2 weeks the images were reordered and they were asked to report them again. Kappa statistic was calculated for intra- and inter-observer variability. The kappa for intra-observer agreement was 0.54 (attendings: 0.54; trainees; 0.55; P = not significant) and the inter-observer agreement was 0.22 (attendings: 0.20; trainees: 0.31; P = 0.027). The inter- and intra-observer kappa values in differentiating nonerosive from erosive cases were 0.22 (attendings: 0.21; trainees: 0.31, P = not significant) and 0.57 (attendings: 0.58; trainees: 0.55, P = not significant), respectively. According to our data, the Los Angeles classification has acceptable intra-observer variability, both for detecting presence or absence of erosions and for differentiating between different degrees of esophagitis, while its inter-observer performance seems to be less acceptable. It may be reasonable and timely to have another look at the Los Angeles classification to see whether its performance can be improved even further.


Extracellular HSP60 induces inflammation through activating and up-regulating TLRs in cardiomyocytes.
The molecular events leading from cardiomyocyte ischaemia to inflammatory cytokine production are not well understood. We previously found that heat shock protein 60 (HSP60) appeared in extracellular space after cardiomyocyte ischaemia. This study examined the activation and regulation of toll-like receptors (TLRs) by HSP60 in cardiomyocytes. Cytokine production and TLRs regulation mediated by TLRs signalling were examined in response to exogenous HSP60 (exHSP60) and endogenous HSP60 (enHSP60) released extracellularly under ischaemia. The results showed that exHSP60 induced inflammatory cytokine production in adult rat cardiomyocytes and H9c2 cells (a standard cardiac cell line derived from embryonic cells), through a pathway dependent on TLR4, myeloid differentiation factor 88 (MyD88), p38, and nuclear factor-κB (NF-κB). Further study revealed up-regulated expression of both TLR2 and TLR4 by exHSP60, which was dependent on the activation of TLR4, MyD88, c-Jun NH2-terminal kinase (JNK), and NF-κB, but not on p38. In myocytes exposed to ischaemia, enHSP60 was released into the media, and triggered cytokine production and TLR2/4 overexpression, through the same pathways as exHSP60. In rats subjected to LAD ligation, the released enHSP60 contributed to cytokine production and TLR2/4 overexpression in the ischaemic myocardium. Extracellular HSP60 induces cytokine production via TLR4-MyD88-p38/NF-κB pathway, and up-regulates TLR2/4 expression via TLR4-MyD88-JNK/NF-κB pathway. Both pathways contribute to myocardial inflammation induced by ischaemia.


Vitexin and isovitexin from the Leaves of Ficus deltoidea with in-vivo α-glucosidase inhibition.
The leaves of Ficus deltoidea are used as a traditional medicine by diabetes patients in Malaysia. The objective of the study is to identify and evaluate bioactive constituents with in vivo α-glucosidase inhibition. The partitioned extracts, subfractions and pure bioactive constituents were subjected to α-glucosidase inhibition assay. The identified bioactive constituents were administered orally to sucrose loaded normoglycemic mice and induced diabetic rats. The postprandial blood glucose levels were monitored at 30 min interval. Acute toxicity was evaluated in both normoglycemic mice and induced diabetic rats. Bioactivity guided fractionation led to the isolation of both vitexin (1) and isovitexin (2). Oral administration of 1mg/kg of either vitexin (1) or isovitexin (2) significantly (p<0.05) reduced the postprandial blood glucose level in sucrose loaded normoglycemic mice at 30 min. The percentage of postprandial blood glucose reduction was highest in sucrose loaded induced diabetic rats administered orally with 200mg/kg of vitexin (1) or 100mg/kg of isovitexin (2). Both vitexin (1) and isovitexin (2) did not exert any signs of toxicity at the highest dose of 2g/kg administered orally to normoglycemic mice and induced diabetic rats. Both the C-glycosyl bioflavonoids, namely, vitexin (1) and isovitexin (2) exhibited in vivo α-glucosidase inhibition.


Diagnosis and management of asthma: a brief review of two asthma expert panel reports.
During the past decade, asthma morbidity and mortality have increased noticeably. Although the reasons for the increase are not clear, the prevalence of asthma has been reported to be increasing not only in the United States but also in other countries throughout the world, including the United Kingdom, New Zealand, and Australia. Asthma mortality rates and morbidity trends are increasing despite what appears to be major scientific advances in our understanding of asthma and in the face of a multitude of new therapeutic modalities. This article presents a brief overview of the pathogenesis and diagnosis of asthma, and highlights the major management recommendations of two expert panel reports.


The third group of the B7-CD28 immune checkpoint family: HHLA2, TMIGD2, B7x, and B7-H3.
The B7-CD28 family of ligands and receptors play important roles in T-cell co-stimulation and co-inhibition. Phylogenetically they can be divided into three groups. The recent discovery of the new molecules (B7-H3 [CD276], B7x [B7-H4/B7S1], and HHLA2 [B7H7/B7-H5]/TMIGD2 [IGPR-1/CD28H]) of the group III has expanded therapeutic possibilities for the treatment of human diseases. In this review, we describe the discovery, structure, and function of B7-H3, B7x, HHLA2, and TMIGD2 in immune regulation. We also discuss their roles in important pathological states such as cancers, autoimmune diseases, transplantation, and infection. Various immunotherapeutical approaches are emerging including antagonistic monoclonal antibodies and agonistic fusion proteins to inhibit or potentiate these molecules and pathways in cancers and autoimmune diseases.


Comparison of patients undergoing transurethral resection of the prostate in 1935-36 and 1991-92.
This retrospective study compares two patient groups undergoing transurethral prostatectomy in the same department nearly 60 years apart. The first 100 patients (group A) were operated in 1935-36, the last 100 (group B) in 1991-92. In group A the main indication was residual urine of more than 5o ml, and in 33% total retention. In group B the main indications were elevated symptom score and low flow, supplemented with pressure flow measurements in some patients. To our surprise, total acute retention was the indication in 23% of group B, only a minor reduction from 26% of group A. Median age was increased from 66 years (49-83) in group A to 75 years (57-92) in group B. Of the first 100 patients 7% (3-14%) died within one month after the operation, in group B 0% (0-4%). Preoperative infection rate fell from 29% to 11%, and the number of patients with impaired kidney function fell from 30% to 8%. Postoperative results were satisfactory in 54% and 85%, respectively.


Leukoplakia and incidence of oral cancer.
Reports from India showing comparatively lower prevalence rates of oral leukoplakia and lower malignant transformation rates than in western countries appear paradoxical because of high incidence rates of oral cancer in India. On a closer examination of available data however, it becomes clear that there is no inconsistency.


Incidence of late onset neutropenia associated with rituximab use in B cell lymphoma patients undergoing autologous stem cell transplantation.
Reversible late onset neutropenia associated with rituximab has been reported with incidence rates varying from 15 to 70% in B cell lymphoma patients receiving autologous stem cell transplantation. We conducted a retrospective descriptive study at one tertiary care center in adult B cell lymphoma patients treated with rituximab and autologous stem cell transplantation between 1 January 2004 and 30 June 2014. Late onset neutropenia was defined as an absolute neutrophil count <1.0 × 109 cells/L after neutrophil engraftment and less than six months post autologous stem cell transplantation. The primary objective was to determine the incidence of late onset neutropenia. The secondary objectives were to examine whether the use of rituximab with re-induction therapy, mobilization or high dose chemotherapy regimens increased the risk for late onset neutropenia, and to evaluate infectious complications. Of 315 subjects, 92 (29.2%) developed late onset neutropenia. Mobilization regimens containing rituximab (OR 2.90 95% CI: 1.31-6.40), high dose chemotherapy containing rituximab (OR 1.87 95% CI: 1.14-3.05), and exposure to rituximab in either or both regimens (OR 3.05 95% CI: 1.36-6.88) significantly increased the risk of late onset neutropenia. While neutropenic, 17.4% experienced an infection, 7.6% experienced febrile neutropenia, and 5.4% were hospitalized. In conclusion, rituximab with mobilization or high dose chemotherapy may increase the risk of late onset neutropenia post autologous stem cell transplantation.


Gestational and congenital syphilis across the international border in Brazil.
Brazil lacks data from syphilis in its border areas. We aimed to describe the spatial and temporal distribution of acquired syphilis (AS), in pregnancy (SP) and congenital syphilis (CS) in Brazilian municipalities in the arches border contexts. An ecological, cross-sectional study was conducted from 2010 to 2020. The study was based on the cases of syphilis available in the Notifiable Diseases Information System (SINAN), and on the Primary Health Care Information System. The detection rates of AS and SP, and the incidence of CS were estimated, and the time series was analyzed. Data between the border arches were compared. In 2020, data showed 7,603 cases of AS (detection rate 64.8/100,000 inhabitants), 3,960 cases of SP (detection rate of 21.6/1,000 live births) and 836 cases of CS (incidence of 4.6/1,000 live births) in the border region. Between 2010 and 2020, the mean annual increase of detection rate of SP was 53.4% in Brazil, 48.0% in the border region, 59.6% in the North Arch, 28.8% in the Central and 67.2% in the South. Annual variation on the incidence of CS for the same period was 31.0% in Brazil 38.4% at the border, in the North and South Arcs 18.3% and 65.7% respectively. The Central Arch showed an increase only between 2010 and 2018 (62.7%). A total of 427 (72.6%) municipalities has primary health care coverage ≥ 95% of the population. In 2019, 538 (91.8%) municipalities reported using rapid tests for syphilis, which decreased to 492 (84%) in 2020. In 2019, 441 (75.3%) municipalities reported administering penicillin, and 422 (72%) in 2020. Our data show syphilis reman problem at the Brazilian border, rates in pregnant are high. It was observed a reduction in the detection rates, SP and the incidence of CS between 2018 and 2020. Syphilis should be included on the agenda of all management levels, aiming at expanding access and quality care.


Research progress of imaging technologies for ischemic cerebrovascular diseases.
Cerebrovascular diseases mainly affect the blood supply of the brain, which has a high demand for oxygen and glucose for the nerve tissues to perform its nerve functions. Ischemic cerebrovascular disease can not only cause stroke, but is also associated with a high incidence of asymptomatic infarction and minimal bleeding that can lead to cognitive and behavioral changes. These changes ultimately manifest as vascular dementia or cognitive impairment. In clinical settings, ischemic cerebrovascular disease can be classified as a transient ischemic attack, reversible ischemic neurological deficit, progressive stroke, complete stroke, marginal infarction, or lacunar infarction. In this review, the research progress of imaging technologies for ischemic cerebrovascular diseases was reviewed, with an aim to provide evidence for clinical practitioners.


The diagnostic value of alcohol dehydrogenase (ADH) isoenzymes and aldehyde dehydrogenase (ALDH) measurement in the sera of patients with brain tumor.
Alcohol dehydrogenase (ADH) isoenzymes and aldehyde dehydrogenase (ALDH) exist in the brain. Alcohol dehydrogenase and ALDH are also present in brain tumor cells. Moreover, the activity of class I isoenzymes was significantly higher in cancer than healthy brain cells. The activity of these enzymes in tumor tissue is reflected in the serum and could thus be helpful for diagnostics of brain neoplasms. The aim of this study was to investigate the potential role of ADH and ALDH as markers for brain tumors. Serum samples were taken for routine biochemical investigation from 115 patients suffering from brain tumors (65 glioblastomas, 50 meningiomas). For the measurement of the activity of class I and II ADH isoenzymes and ALDH activity, fluorometric methods were used. The total ADH activity and activity of class III and IV isoenzymes were measured by the photometric method. There was a significant increase in the activity of ADH I isoenzyme and ADH total in the sera of brain tumor patients compared to the controls. The diagnostic sensitivity for ADH I was 78%, specificity 85%, and positive and negative predictive values were 86% and 76% respectively. The sensitivity and specificity of ADH I increased with the stage of the carcinoma. Area under receiver-operating characteristic curve for ADH I was 0.71. The results suggest a potential role for ADH I as a marker for brain tumor.


Resveratrol blunts tumor necrosis factor-alpha-induced monocyte adhesion and transmigration.
The leukocyte recruitment and transmigration across the endothelial barrier into the vessel wall are crucial steps in atherosclerosis. Leukocyte trafficking on the endothelium is elicited by induction of endothelial adhesion molecules, and its transmigration is mediated by degradation of basement membrane proteins through enzymatic activity of matrix metalloproteinases (MMP). The current study investigated whether resveratrol, a polyphenol present in grapes and red wine, was capable of inhibiting leukocyte adhesion to tumor necrosis factor (TNF)-alpha-activated endothelium. It was found that resveratrol inhibited the TNF-alpha-activated endothelial expression of vascular cell adhesion molecule-1 in a dose-dependent manner. In addition, resveratrol hampered THP-1 monocyte adhesion to activated endothelial cells. This study further examined whether resveratrol interfered with transendothelial migration of leukocytes. The MMP-2 gelatinolytic activity of endothelial cells was enhanced by TNF-alpha, which was attenuated by an addition of >/=25 microM resveratrol. In addition, 25 microM resveratrol mitigated the MMP-9 activity of THP-1 cells, followed by a marked inhibition of transendothelial migration. These results demonstrated that resveratrol suppressed monocyte adhesion and migration induced by TNF-alpha through modulating expression of adhesion molecules and gelatinolytic activity of MMP. These findings suggest that dietary resveratrol may be therapeutic agent for inhibiting leukocyte recruitment into the subendothelium during inflammatory atherosclerosis.


Interactive Models in Synthetic Biology: Exploring Biological and Cognitive Inter-Identities.
The aim of this article is to investigate the relevance and implications of synthetic models for the study of the interactive dimension of minimal life and cognition, by taking into consideration how the use of artificial systems may contribute to an understanding of the way in which interactions may affect or even contribute to shape biological identities. To do so, this article analyzes experimental work in synthetic biology on different types of interactions between artificial and natural systems, more specifically: between protocells and between biological living cells and protocells. It discusses how concepts such as control, cognition, communication can be used to characterize these interactions from a theoretical point of view, which criteria of relevance and evaluation of synthetic models can be applied to these cases, and what are their limits.


Role of estrogen receptor binding and transcriptional activity in the stimulation of hyperestrogenism and nuclear bodies.
The effects of estradiol and nafoxidine on nuclear estrogen receptor binding, RNA polymerase activities, and uterine ultrastructure were studied. Animals were either injected with estradiol, implanted with estradiol/paraffin pellets, or injected with nafoxidine. Animals treated with nafoxidine or estradiol implants showed sustained long-term nuclear retention of estrogen receptor and increased nuclear RNA polymerase activities for up to 72 hr. A single injection of estradiol caused initial increases in these variables which returned to control levels by 24 hr after hormone treatment. Uterine tissue was examined by light and electron microscopy 72 hr after hormone treatments. Uteri from eith estradiol-implanted or nafoxidine-treated animals showed markedly increased hypertrophy of the luminal epithelial cells. Nuclei in sections of the uteri of these hyperestrogenized animals displayed a large number and wide array of nuclear bodies composed of a filamentous capsule and granular cores. We conclude that hyperestrogenization, a condition that eventually results in abnormal cell growth, is correlated with increased and sustained nuclear binding of the estrogen receptor, increased and sustained RNA polymerase activity, and the appearance of nuclear bodies.


Measurements of Intracellular Ca2+ Content and Phosphatidylserine Exposure in Human Red Blood Cells: Methodological Issues.
The increase of the intracellular Ca2+ content as well as the exposure of phosphatidylserine (PS) on the outer cell membrane surface after activation of red blood cells (RBCs) by lysophosphatidic acid (LPA) has been investigated by a variety of research groups. Carrying out experiments, which we described in several previous publications, we observed some discrepancies when comparing data obtained by different investigators within our research group and also between batches of LPA. In addition, we found differences comparing the results of double and single labelling experiments (for Ca2+ and PS). Furthermore, the results of PS exposure depended on the fluorescent dye used (annexin V-FITC versus annexin V alexa fluor® 647). Therefore, it seems necessary to investigate these methodological approaches in more detail to be able to quantify results and to compare data obtained by different research groups. The intracellular Ca2+ content and the PS exposure of RBCs separated from whole blood have been investigated after treatment with LPA (2.5 µM) obtained from three different companies (Sigma-Aldrich, Cayman Chemical Company, and Santa Cruz Biotechnology Inc.). Fluo-4 and x-rhod-1 have been used to detect intracellular Ca2+ content, annexin V alexa fluor® 647 and annexin V-FITC have been used for PS exposure measurements. Both parameters (Ca2+ content, PS exposure) were studied using flow cytometry and fluorescence microscopy. The percentage of RBCs showing increased intracellular Ca2+ content as well as PS exposure changes significantly between different LPA manufacturers as well as on the condition of mixing of LPA with the RBC suspension. Furthermore, the percentage of RBCs showing PS exposure is reduced in double labelling compared to single labelling experiments and depends also on the fluorescent dye used. Finally, data on Ca2+ content are slightly affected whereas PS exposure data are not affected significantly by the measuring method (flow cytometry, fluorescence microscopy). The LPA batch used and the mixing procedure of LPA and the RBC suspension has to be taken into consideration when comparing results of intracellular Ca2+ content and PS exposure of RBCs after LPA activation. In addition, one should consider that the results of single and double labelling experiments might be different depending on the fluorescent dyes used.


[Abnormal BCL10 nuclear expression and chromosomal translocation in ocular adnexal mucosa-associated lymphoid tissue lymphomas].
To detect the BCL10 expression and several types of chromosomal translocations [including t (11;18)/API2-MALT1; t (1;14)/IgH- BCL10 and t (14;18)/MALT1-IgH] in ocular adnexal mucosa-associated lymphoid tissue lymphomas (OA-MALT lymphomas). Sixty OA-MALT lymphomas were collected from Cancer Hospital of Fudan University. BCL10 was detected by immunohistochemical studies, and API2-MALT1 fusion gene, BCL10, MALT1 and IgH chromosomal abnormalities were detected by fluorescent in situ hybridization (FISH). Fisher's exact test was used to analyze the relation between nuclear BCL10 expression and chromosomal translocation. BCL10 expressed in 85.0% (51/60) OA-MALT lymphomas. Among these positive cases, 25 cases (41.7%) were expressed in the cytoplasm, and 26 cases (43.3%) were expressed in the nucleus. FISH results showed that no chromosomal abnormalities related with BCL10 and IgH genes, except 2 cases with API2-MALT1 fusion gene. Under statistic of Fisher exact test, nuclear BCL10 expression and API2-MALT1 fusion gene were two independent factors (P > 0.05). BCL10 nuclear expression is common in OA-MALT lymphomas and may be used as a potential marker for the diagnosis of MALT lymphomas arising from ocular adnexa. Aberrant chromosomal translocations reported in the other sites MALT lymphomas are rare in OA-MALT lymphomas.


The importance of standardization of creatinine in the implementation of guidelines and recommendations for CKD: implications for CKD management programmes.
In an attempt to reduce late referral and to improve the care of patients with chronic kidney disease (CKD), different organizations have issued guidelines on when to refer patients to the nephrologist. Most suggest referral of patients with a GFR below 60 ml/min/1.73 m2, and demand referral if the GFR is below 30 ml/min/1.73 m2. It is recommended to use the abbreviated MDRD equation to estimate GFR. This formula is, however, sensitive to the creatinine assay methodology. In addition, the impact of the implementation of such guidelines on the nephrology practice has never been evaluated. This study (i) identifies the true burden of CKD in a population and simulates the effects of a 100% implementation of the guidelines on the nephrology work load, and (ii) evaluates the validity of the estimated GFR using the abbreviated MDRD formula when routinely provided. Different laboratories (both hospital and private) in our region were asked to report on all the serum creatinine values performed during the first week of December 2004. If patients had more than one determination, only the lowest serum creatinine value was retained. Patients already known to a nephrology unit were not included. GFR was calculated using the abbreviated MDRD, using the serum creatinine as reported by these laboratories, or after correction to the MDRD-standard using different published equations. 20,108 patients, with a mean age of 53.4+/-16.2 years, 48% females, had at least one serum creatinine determination in the observation period. According to the K/DOQI CKD classification, 20.2, 1.6 and 0.8% of females and 13.3, 1.6 and 0.6% of males were in stage 3, 4 and 5, respectively, when the abbreviated MDRD formula was used with the serum creatinine value as reported by the laboratories. Important differences in classifications were obtained when the different correction formulae for creatinine were applied. According to the current recommendations, this would lead to a mandatory referral of 1650-2400 CKD stage 4 patients per 100 000 inhabitants and a suggested referral of another 4100-15 360 CKD stage 3 patients per 100,000 inhabitants to a nephrology unit. Implementation of the current guidelines for referral of CKD patients to nephrologists would lead to an overload of the nephrology care capacities. Large differences in estimated GFRs with different corrections for serum creatinine are observed, resulting in important CKD classification differences. Standardization of serum creatinine assays is mandatory before guidelines, and especially the routine provision of the estimated GFR by the abbreviated MDRD formula, can be implemented in clinical practice.


Characteristics of Serum Exosomes after Burn Injury and Dermal Fibroblast Regulation by Exosomes In Vitro.
(1) Background: Exosomes (EXOs) have been considered a new target thought to be involved in and treat wound healing. More research is needed to fully understand EXO characteristics and the mechanisms of EXO-mediated wound healing, especially wound healing after burn injury. (2) Methods: All EXOs were isolated from 85 serum samples of 29 burn patients and 13 healthy individuals. We characterized the EXOs for morphology and density, serum concentration, protein level, marker expression, size distribution, and cytokine content. After a confirmation of EXO uptake by dermal fibroblasts, we also explored the functional regulation of primary human normal skin and hypertrophic scar fibroblast cell lines by the EXOs in vitro, including cell proliferation and apoptosis. (3) Results: EXOs dynamically changed their morphology, density, size, and cytokine level during wound healing in burn patients, which were correlated with burn severity and the stages of wound healing. EXOs both from burn patients and healthy individuals stimulated dermal fibroblast proliferation and apoptosis. (4) Conclusions: EXO features may be important signals that influence wound healing after burn injury; however, to understand the mechanisms by which EXOs regulates the fibroblasts in healing wounds, further studies will be required.


Rhodium-catalyzed oxidative 1:1, 1:2, and 1:4 coupling reactions of phenylazoles with internal alkynes through the regioselective cleavages of multiple C-H bonds.
The direct oxidative coupling of phenylazoles with internal alkynes proceeds efficiently in the presence of a rhodium catalyst and a copper oxidant accompanied by double or quadruple C-H bond cleavages. Thus, as a representative example, 4,5-diphenylpyrazolo[1,5-a]quinoline, 1-(1,2,3,4-tetraphenylnaphthalen-5-yl)pyrazole, and 1-(1,2,3,4,5,6,7,8-octaphenylanthracen-9-yl)pyrazole can be obtained selectively through the coupling of 1-phenylpyrazole and diphenylacetylene in 1:1, 1:2, and 1:4 manners, respectively. The reactions preferentially take place at the electron-deficient sites on the aromatic substrates. A comparison of reactivities of variously substituted and deuterated substrates sheds light on the mechanism of C-H bond cleavage steps. The reaction pathway is highly dependent on reaction conditions employed, especially on the nature of solvent. The influence of solvation of a key rhodacycle intermediate has been investigated computationally. In addition, some of the condensed aromatic products have been found to exhibit intense fluorescence in the solid state.


Priming of influenza virus-specific cytotoxic T lymphocytes vivo by short synthetic peptides.
Influenza virus-specific CTL were primed in vivo by immunization with short synthetic peptides representing major CTL epitopes from the nucleoprotein of type A influenza virus. The resultant CTL after in vitro boosting of primed spleen cells recognized both virus-infected and peptide-pulsed target cells. The requirement of CD4+ T cell activation was investigated in several ways. First the addition of helper epitopes to the CTL epitope did not enhance CTL generation, suggesting that helper activity was either not limiting or not required. However, in vivo depletion of CD4+ T cells completely inhibited the generation of CTL by peptide immunization. The inclusion of anti-CD4 in the in vitro restimulation with peptide also prevented the generation of CTL, whereas in vitro reactivation of virus immune spleen cells with peptide was not inhibited by anti-CD4. Thus there appears to be heterogeneity in the requirement of CD4+ T cell proliferation in CTL generation. One possibility is that virus infected cells can stimulate higher affinity T cells that are less helper T cell dependent.


Current status of insecticide resistance in Helicoverpa armigera after 15 years of Bt cotton planting in China.
Insecticide resistance was an important factor responsible for outbreaks of Helicoverpa armigera (Hübner) in China in the early 1990s. Bt cotton has been adopted in China since 1997, and has resulted in a reduction of insecticide use for H. armigera control. After 15 yr of Bt cotton planting, in 2011 we surveyed resistance to fenvalerate, phoxim, and emamectin benzoate in 16 field populations of H. armigera collected from major cotton production areas of China. Fourteen populations from northern China showed very strong resistance to fenvalerate (from 43- to 830-fold) and low levels of resistance to phoxim (3.0- to 8.9-fold) when compared with the susceptible SCD strain of H. armigera, whereas two populations from northwestern China showed low levels of resistance to fenvalerate (3.0- and 10-fold) and no resistance to phoxim (0.7- and 0.9-fold). Synergist bioassays demonstrated that oxidase-based detoxification was involved in fenvalerate resistance and esterase-based detoxification in phoxim resistance in the resistant field populations. In comparison with the resistance in field populations before Bt cotton adoption, we observed a maintenance of high levels of fenvalerate resistance, but a reversion of phoxim resistance from high levels to low levels in the field populations of H. armigera from northern China. All 16 field populations from both northern China and northwestern China were susceptible to emamectin benzoate (with about two-fold variations in LD50s among populations), but the SCD strain has an inherent tolerance of 11-fold compared with the most susceptible field population (Xiajin-1). Emamectin benzoate is not cross resistant to fenvalerate and may provide an alternative option for H. armigera control in China, if the efficacy of Bt cotton is compromised by Bt resistance in the field.


Superficial venous thrombophlebitis as the initial manifestation of hypereosinophilic syndrome: study of the first 3 cases.
Superficial venous thrombophlebitis (SVT), often perceived as benign, can coexist with hypercoagulable states. Predisposing risk factors for SVT are similar to those observed for deep venous thrombosis. Association of eosinophilia with SVT is a rare situation that can reveal neoplasia, malignant blood disorders, or vasculitis, but it has never been described in hypereosinophilic syndrome (HES). We herein describe the clinical and biological features, outcome, and response to therapy of 3 patients with SVT associated with eosinophilia that revealed HES. Superficial venous thrombophlebitis was the initial manifestation of HES in all 3 patients. The mean eosinophil count at diagnosis was 2.4 x 10(3)/muL. All patients received corticosteroids and anticoagulants as the initial treatment, with marked improvement of SVT and return of the eosinophil count to reference limits. All patients experienced relapse and remained dependent on corticosteroid therapy. Two patients received interferon alfa with dramatic regression of SVT, allowing a decrease in the dose of corticosteroids. We report, to our knowledge, the first 3 cases of SVT related to HES. Superficial venous thrombophlebitis was difficult to treat, with dependence on corticosteroid therapy and partial efficacy of anticoagulant and antiplatelet therapy. Interferon alfa was effective in preventing relapse of SVT related to HES. Mechanisms implied in this thrombogenesis are multiple and remain speculative.


Headache Attributed to Masticatory Myofascial Pain: Clinical Features and Management Outcomes.
To describe the characteristics of headaches attributed to temporomandibular disorders (TMD) and assess the effects of two management strategies used for the management of TMD on headache intensity and frequency. The initial sample (n=60) of this randomized controlled trial comprised patients with masticatory myofascial pain according to the Research Diagnostic Criteria for TMD (RDC/TMD), and headache. The patients were divided into two groups: group 1 received only counseling for behavioral changes, and group 2 received counseling and an occlusal appliance. A 5-month follow-up period included three assessments. TMD-related headache characteristics, eg, headache intensity (scored on a visual analog scale [VAS]) and frequency were measured by a questionnaire. Two-way analysis of variance, chi-square, Friedman, and Mann-Whitney tests were used to test for differences considering a 5% significance level. The main clinical features of headache attributed to masticatory myofascial pain were the long duration (≥4 hours), frontotemporal bilateral location, and a pressing/tightening quality. Forty-one subjects (group 1, 17 subjects; group 2, 24 subjects) were included in the final analysis. There was a reduction in headache intensity and frequency, with no significant differences between groups (P>.05). The mean (±SD) baseline VAS was 7.6 (±2.2) for group 1 and 6.5 (±1.6) for group 2; final values were 3.1 (±2.2) (P<.001) and 2.5 (±2.3) (P<.001), respectively. Headache attributed to masticatory myofascial pain was mainly characterized by long duration, frontotemporal bilateral location, and a pressing/tightening quality. Also, counseling and behavioral management of masticatory myofascial pain improved headache, regardless of the use of an occlusal appliance.


[Relationship between epidemiological evidence and fatality rate in patients with severe acute respiratory syndrome].
To investgate the relationship between epidemiological diagnosis and fatality rate in patients with severe acute respiratory syndrome (SARS). Epidemiological evidences and the outcomes of 304 patients with confirmed SARS were retrospectively analyzed. Patients included highly contagious cases, cluster cases, and sporadic cases. The case fatality rate was 8.2%. The fatality rate had a tendency of increase in an order of sporadic cases, cluster cases, and highly contagious cases (3.9%, 11.4%, and 17.2%, respectively, chi2trend=7.561, P < 0.01). Chi square-test also proved that the fatality rate was higher in older people (chi2trend=27.024, P < 0.01) and in male (male vs female: 12.2% vs 5.5%). A logistic regression model showed that the epidemiological evidence, age, and gender were correlated to fatality rate. By observing changes of the odds ratio for epidemic evidence, age, and gender using forward method, we found epidemic evidence was an independent risk factor related to fatality rate in SARS patients. The epidemiological evidence is an independent factor related to fatality rate in SARS patients. Highly contagious case has a worse prognosis and higher fatality rate than sporadic cases and cluster cases.


[Validity of post-mortem computed tomography angiography (PMCTA) in medico-legal diagnostic management of stab and incised wounds].
The report presents an analysis of results of multi-phase post-mortem computed tomography angiography (MPMCTA) scans performed at the Chair of Forensic Medicine, Jagiellonian University Medical College, in cases of stab wounds and incised wounds, as confronted with the results of conventional medico-legal autopsies. The analysis of the material points to a validity of PMCTA scans as supplementing post-mortem medico-legal examinations and being capable of providing significant data that affect final conclusions and adding new quality to recording post-mortem observations.


Ti-Catalyzed and -Mediated Oxidative Amination Reactions.
Titanium is an attractive metal for catalytic reaction development: it is earth-abundant, inexpensive, and generally nontoxic. However-like most early transition metals-catalytic redox reactions with Ti are difficult because of the stability of the high-valent TiIV state. Understanding the fundamental mechanisms behind Ti redox processes is key for making progress toward potential catalytic applications. This Account details recent progress in Ti-catalyzed (and -mediated) oxidative amination reactions that proceed through formally TiII/TiIV catalytic cycles.This class of reactions is built on our initial discovery of Ti-catalyzed [2 + 2 + 1] pyrrole synthesis from alkynes and azobenzene, where detailed mechanistic studies have revealed important factors that allow for catalytic turnover despite the inherent difficulty of Ti redox. Two important conclusions from mechanistic studies are that (1) low-valent Ti intermediates in catalysis can be stabilized through coordination of π-acceptor substrates or products, where they can act as "redox-noninnocent" ligands through metal-to-ligand π back-donation, and (2) reductive elimination processes with Ti proceed through π-type electrocyclic (or pericyclic) reaction mechanisms rather than direct σ-bond coupling.The key reactive species in Ti-catalyzed oxidative amination reactions are Ti imidos (Ti≡NR), which can be generated from either aryl diazenes (RN═NR) or organic azides (RN3). These Ti imidos can then undergo [2 + 2] cycloadditions with alkynes, resulting in intermediates that can be coupled to an array of other unsaturated functional groups, including alkynes, alkenes, nitriles, and nitrosos. This basic reactivity pattern has been extended into a broad range of catalytic and stoichiometric oxidative multicomponent coupling reactions of alkynes and other reactive small molecules, leading to multicomponent syntheses of various heterocycles and aminated building blocks.For example, catalytic oxidative coupling of Ti imidos with two different alkynes leads to pyrroles, while stoichiometric oxidative coupling with alkynes and nitriles leads to pyrazoles. These heterocycle syntheses often yield substitution patterns that are complementary to those of classical condensation routes and provide access to new electron-rich, highly substituted heteroaromatic scaffolds. Furthermore, catalytic oxidative alkyne carboamination reactions can be accomplished via reaction of Ti imidos with alkynes and alkenes, yielding α,β-unsaturated imine or cyclopropylimine building blocks. New catalytic and stoichiometric oxidative amination methods such as alkyne α-diimination, isocyanide imination, and ring-opening oxidative amination of strained alkenes are continuously emerging as a result of better mechanistic understanding of Ti redox catalysis.Ultimately, these Ti-catalyzed and -mediated oxidative amination methods demonstrate the importance of examining often-overlooked elements like the early transition metals through the lens of modern catalysis: rather than a lack of utility, these elements frequently have undiscovered potential for new transformations with orthogonal or complementary selectivity to their late transition metal counterparts.


Backscattered electron imaging and elemental analysis of rapidly frozen plant cells using variable accelerating voltage.
Rapidly frozen rosemary leaves were observed at variable accelerating voltages in a low-vacuum scanning electron microscope equipped with a cryo transfer system. After water was sublimated from the fractured face of the leaf, distinct backscattered electron (BSE) images were obtained depending on the accelerating voltages applied. At 5 kV, surface cell wall structure was observed, whereas at 10 and 15 kV chloroplasts lining the inside of the cell wall and membrane were visualized. With energy dispersive X-ray microanalysis, elemental information corresponding to the BSE images was obtained. Besides visualization of the structures and elemental composition close to the living state, information on layers at different depths from the surface could be detected by varying the accelerating voltage in this system.


M(o)TOR of aging: MTOR as a universal molecular hypothalamus.
A recent ground-breaking publication described hypothalamus-driven programmatic aging. As a Russian proverb goes "everything new is well-forgotten old". In 1958, Dilman proposed that aging and its related diseases are programmed by the hypothalamus. This theory, supported by beautiful experiments, remained unnoticed just to be re-discovered recently. Yet, it does not explain all manifestations of aging. And would organism age without hypothalamus? Do sensing pathways such as MTOR (mechanistic Target of Rapamycin) and IKK-beta play a role of a "molecular hypothalamus" in every cell? Are hypothalamus-driven alterations simply a part of quasi-programmed aging manifested by hyperfunction and secondary signal-resistance? Here are some answers.


Simultaneous Delivery of Multiple Antibacterial Agents from Additively Manufactured Porous Biomaterials to Fully Eradicate Planktonic and Adherent Staphylococcus aureus.
Implant-associated infections are notoriously difficult to treat and may even result in amputation and death. The first few days after surgery are the most critical time to prevent those infections, preferably through full eradication of the micro-organisms entering the body perioperatively. That is particularly important for patients with a compromised immune system such as orthopedic oncology patients, as they are at higher risk for infection and complications. Full eradication of bacteria is, especially in a biofilm, extremely challenging due to the toxicity barrier that prevents delivery of high doses of antibacterial agents. This study aimed to use the potential synergistic effects of multiple antibacterial agents to prevent the use of toxic levels of these agents and achieve full eradication of planktonic and adherent bacteria. Silver ions and vancomycin were therefore simultaneously delivered from additively manufactured highly porous titanium implants with an extremely high surface area incorporating a bactericidal coating made from chitosan and gelatin applied by electrophoretic deposition (EPD). The presence of the chitosan/gelatin (Ch+Gel) coating, Ag, and vancomycin (Vanco) was confirmed by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). The release of vancomycin and silver ions continued for at least 21 days as measured by inductively coupled plasma (ICP) and UV-spectroscopy. Antibacterial behavior against Staphylococcus aureus, both planktonic and in biofilm, was evaluated for up to 21 days. The Ch+Gel coating showed some bactericidal behavior on its own, while the loaded hydrogels (Ch+Gel+Ag and Ch+Gel+Vanco) achieved full eradication of both planktonic and adherent bacteria without causing significant levels of toxicity. Combining silver and vancomycin improved the release profiles of both agents and revealed a synergistic behavior that further increased the bactericidal effects.


The effect of written information and counselling by an advanced practice nurse on resilience in women with vulvar neoplasia six months after surgical treatment and the influence of social support, recurrence, and age: a secondary analysis of a multicenter randomized controlled trial, WOMAN-PRO II.
Women with vulvar neoplasia often complain about physical and psychological distress after surgical treatment. Lack of information and support can influence resilience. Whether an information-related intervention through an advanced practice nurse supports resilience and which other factors affect resilience in women with vulvar neoplasia has never been investigated. The aims of this study were (a) to analyse whether counselling based on the WOMAN-PRO II program causes a significant improvement in the resilience scores of women with vulvar neoplasia compared to written information and (b) to identify the potential predictors of resilience. A randomized controlled trial was conducted in women with vulvar neoplasia (n = 49) 6 months after surgical treatment in four Swiss hospitals and one Austrian hospital. Analyses of resilience and its predictors were performed using a linear mixed model. Thirty-six women (intervention I, n = 8; intervention II, n = 28) completed the randomized controlled trial. In total, 13 women (26.5%) dropped out of the trial. The resilience score did not differ significantly between the two interventions three and six months after randomisation (p = 0.759). Age (b = .04, p = 0.001), social support (b = .28, p = 0.009), counselling time (b = .03, p = 0.018) and local recurrence (b = -.56, p = 0.009) were identified as significant predictors of resilience in the linear mixed model analyses. The results indicate that the WOMAN-PRO II program as single intervention does not cause a significant change in the resilience scores of women with vulvar neoplasia 6 months after surgery. Predictors that promote or minimise resilience have been identified and should be considered when developing resilience programs for women with vulvar neoplasia. A repetition of the study with a larger sample size is recommended. The WOMAN-PRO II program was registered in ClinicalTrials.gov NCT01986725 on 18 November 2013.


Alterations of ultrastructure and of cytoplasmic filaments in remodelling rat jejunal epithelial cells during recovery from deoxycholate.
Structural features associated with reversibility of lesions induced by deoxycholic acid (DOC) were studied by electron microscopy and immunofluorescence techniques. Tied jejunal loops were incubated in vivo with 2.5-20 mmol/l DOC in isotonic solution. Immediately after this treatment, or after a recovery period of 15 or 150 minutes following washout of the bile acid, the loops were excised and processed. DOC produced epithelial lesions whose severity and reversibility were concentration-dependent. Ultrastructural features associated with the reversibility of the lesions were particularly apparent in specimens exposed to 10-20 mmol/l DOC. These features included cell flattening with the formation of thin, veil-like structures into the eroded area by cells at the edges of the erosions. Immunofluorescence studies showed a redistribution of actin and cytokeratin filaments to the margins and leading edges of the flattened cells. It is suggested that cell flattening and migration are responsible for the rapid morphological recovery of the injured epithelium. Actin and cytokeratin appear to be instrumental in the remodelling and migration.


[Expression of survivin and caspase-3 during the development process in rat cochlea].
To explore the role of some apoptosis regulators during the development in rat cochlea. The morphological development process of cochlea was observed in Wistar rat aged between embryo day 13 to postnatal day 14 in this experiment. Survivin and caspase-3 were respectively detected at protein and mRNA levels by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). The expression of survivin and caspase-3 located in the bottom wall of the cochlear duct. Not only they widespread in the cell proliferation, but also they gradually enhanced in the cell differentiation. Both of them had a crest-time, and survivin was prior to caspase-3. In organ of corti during adult time, caspase-3 was not present and survivin was only expressed. During the development of the rat cochlea, both of them had similar location and trend. But they were derangement. This showed that both of them participated in the cochlear morphological development. It was suggested that the interaction between survivin and caspase-3 regulated the apoptosis, promoted the cochlear morphological development.


MICAL2 promotes breast cancer cell migration by maintaining epidermal growth factor receptor (EGFR) stability and EGFR/P38 signalling activation.
MICAL2, a cytoskeleton dynamics regulator, is identified associated with survival and metastasis of several types of cancers recently. This study was designed to investigate the role of MICAL2 in breast cancer cell migration as well as its underlying mechanisms. The relationship between MICAL2 and EGF/EGFR signalling was analysed by gene overexpression and knock-down techniques. Cell migration was measured by wound-healing assays. Activation of EGF/EGFR signalling pathways were evaluated by immunofluorescence, qPCR, Western blotting and zymography techniques. Rac1 activity was assessed by pull-down assay. Correlation of MICAL2 and EGFR in breast cancer specimens was examined by immunohistochemical analysis. Ectopic expression of MICAL2 in MCF-7 cells augmented EGFR protein level, accompanied by the promotion of cell migration. Silencing MICAL2 in MDA-MB-231 cells destabilized EGFR and inhibited cell migration. In mechanism, the maintaining effect of MICAL2 on EGFR protein content was due to a delay in EGFR degradation. Expression of MICAL2 was also shown positively correlated with the activation of P38/HSP27 and P38/MMP9 signallings, which are the main downstream signalling cascades of EGF/EGFR involved in cell migration. Further analysis indicated that Rac1 activation contributed to the maintaining effect of MICAL2 on EGFR stability. In addition, analysis of breast cancer specimens revealed a positive correlation between MICAL2 and EGFR levels and an association between MICAL2 expression and worse prognosis. MICAL2 is a major regulator of breast cancer cell migration, maintaining EGFR stability and subsequent EGFR/P38 signalling activation through inhibiting EGFR degradation in a Rac1-dependent manner.


The efficacy of laparoscopic cholecystectomy without discontinuation in patients on antithrombotic therapy.
Laparoscopic cholecystectomy (LC) is one of the most commonly performed surgeries in the world today. However, there is no consensus regarding whether LC can be performed in patients with acute cholecystitis while on antithrombotic therapy. The objective of our study was to describe postoperative outcomes of patients who underwent emergent LC without interruption to antithrombotic therapy. We performed a retrospective review of patients who underwent LC for acute cholecystitis while on antithrombotic therapy from 2010 to 2015 at Soonchunhyang Universtiy Cheonan Hospital. Patients were divided into 2 groups as underwent emergent LC and elective LC. A total of 67 patients (emergent group, 22; elective group, 45) were included in the analysis. Elective group had significantly longer duration between the admission and operation (8 [7-10] days vs. 2 [1-3] days, P < 0.001) and longer duration of antithrombotic drugs discontinuation (7 days vs. 1 [0-3] days, P < 0.001). Emergent group had significantly more postoperative anemia (6 patients vs. 0 patient, P = 0.001) and 3 of 6 patients received packed RBC transfusion in postoperative period. However, there was no significant difference in length of postoperative stays, length of intensive care unit stays and mortality rates. Emergent LC without interruption to antithrombotic therapy was relatively safe and useful. A well-designed multicenter study is needed to confirm the safety and efficacy of LC without suspension of antithrombotic therapy and to provide a simple guideline.


Correlation of cortical cholinergic and GABA deficits with quantitative neuropathological findings in senile dementia.
The present paper examines the relationship between choline acetyltransferase (ChAT) activity and gamma aminobutyric acid (GABA) concentrations with neuronal counts, senile plaque counts and estimates of neurofibrillary tangles in a series of 25 cases of senile dementia of Alzheimer type (SDAT) with appropriate controls. ChAT activity was significantly positively correlated with neuronal counts in the frontal and temporal region for the whole group but not in the demented or control subgroups. Significant negative correlations were also found between plaque counts and ChAT activity in all areas studied in the total group of subjects and in most of the SDAT groups. Significant negative correlations between ChAT activity and estimates of neurofibrillary change occurred in the frontal area and midtemporal gyrus in the SDAT cases. All the above correlations were seen most prominently in the youngest dementia patients (less than 79 years of age) who exhibited the most severe neuropathological and neurochemical deficits. The neurochemical distinction between the young and elderly age group of SDAT cases was well illustrated by the paradoxical trends of significantly increased GABA and ChAT levels with increasing age. The few significant correlations observed between GABA and cell counts were positive and occurred in the temporal lobe in the younger age group. Plaque counts and neurofibrillary tangle estimates showed little correlation with GABA concentrations.


Nocturnal oximetry: utility in the respiratory management of amyotrophic lateral sclerosis.
Current recommendations are to institute nocturnal nasal ventilation for amyotrophic lateral sclerosis patients with a forced vital capacity (FVC) of <50% of predicted normal. The purpose of this study was to determine whether this is appropriate. A total of 87 nocturnal oximetry evaluations were performed on 78 consecutive amyotrophic lateral sclerosis patients symptomatic for sleep-disordered breathing. Nocturnal oximetry measurements were compared for those with FVC >50% vs. those with FVC of <50% of normal. FVC was measured sitting and supine. A considerable number of these symptomatic patients manifested evidence of nocturnal hypoxemia as measured by oximetry. However, there was no significant difference between patients with sitting percentage-predicted FVC above and below 50% predicted in minimum oxygen saturation, mean oxygen saturation, percentage of time spent with oxygen saturation of <88%, and number of events per hour. There was no significant difference between patients with supine percentage-predicted FVC above and below 50% predicted in minimum oxygen saturation, mean oxygen saturation, percentage of time spent with oxygen saturation of <88%, and number of events per hour. The recommendation that FVC be <50% of normal is inappropriate for justifying introduction of nocturnal nasal ventilation. Many patients are symptomatic at higher FVC and manifest evidence of nocturnal hypoxemia. Nocturnal oximetry adds additional practical information for justifying earlier respiratory intervention for symptomatic patients.


Stachyose Alleviates Corticosterone-Induced Long-Term Potentiation Impairment via the Gut-Brain Axis.
Stress can induce learning and memory impairment; corticosterone is often used to study the effects and mechanisms of stress in animal models. Long-term potentiation (LTP) has been widely used for tackling the mechanisms of memory. Liuwei Dihuang decoction-active fraction combination (LW-AFC) can improve stress-induced LTP and cognition impairment; stachyose is an oligosaccharide in LW-AFC. The effects and mechanisms of stachyose on stress are unknown. In this study, stachyose showed protective effects against LTP impairment by corticosterone in vivo only via intragastric administration for 7 consecutive days, but there was little effect even after direct intracerebroventricular injection; the protective effect of stachyose could be canceled by non-absorbable antibiotics (ATB) which disturbed gut flora. 16S rRNA sequencing, alpha diversity, and principal coordinate analysis (PCoA) revealed that the gut flora in corticosterone-treated mice was disturbed and stachyose could improve corticosterone-induced gut flora disturbance. Bacteroidetes were decreased and Deferribacteres were increased significantly in corticosterone-treated mice, and stachyose restored Bacteroidetes and Deferribacteres to the normal level. D-serine, a coactivator of NMDA receptors, plays an important role in synaptic plasticity and cognition. Here, corticosterone had little effect on the content of D-serine and L-serine (the precursor of D-serine), but it reduced the D-serine release-related proteins, Na+-independent alanine-serine-cysteine transporter-1 (ASC-1), and vesicle-associated membrane protein 2 (VAMP2) significantly in hippocampus; stachyose significantly increased ASC-1 and VAMP2 in corticosterone-treated mice, and ATB blocked stachyose's effects on ASC-1 and VAMP2. NMDA receptors co-agonists L-serine, D-serine, and glycine significantly improved LTP impairment by corticosterone. These results indicated that stachyose might indirectly increase D-serine release through the gut-brain axis to improve LTP impairment by corticosterone in the hippocampus in vivo.


Fluorescence energy transfer measurements of spatial relationships between sulfhydryl groups of thiolase I from porcine heart.
Mitochondrial thiolase I from pig heart has been found to have at least two and possibly three reactive sulfhydryl residues at or near the active site [Izbicka-Dimitrijević, E., & Gilbert, H. F. (1982) Biochemistry 21, 6112-6118; Izbicka-Dimitrijević, E., & Gilbert, H. F. (1984) Biochemistry 23, 4318-4324]. In the native enzyme, fluorescein mercuric acetate reacts with two of the sulfhydryl groups and inactivates the enzyme with a rate constant of 1.6 X 10(-4) M-1 s-1 in 0.1 M Tris-acetate, pH 7.0. The presence of saturating (250 microM) concentrations of acetoacetyl coenzyme A protects against both modification and inactivation. The acetyl enzyme, a normal intermediate in the reaction catalyzed by thiolase, is not inactivated by fluorescein mercuric acetate although one sulfhydryl group out of five per mole of thiolase subunit is still available for reaction with the reagent. Fluorescein mercuric acetate and S-mercurio-N-dansyl-L-cysteine (Dns-Cys-SHg+) have been used to differentially label two of the sulfhydryl groups in thiolase. The distance between Dns-Cys-SHg+ (donor) and fluorescein mercuric acetate (acceptor) determined by the fluorescence energy transfer is less than 14 A. The fluorescent analogue of coenzyme A, 1,N6-etheno coenzyme A, is recognized by thiolase as a substrate (Km = 21 microM); however, substrate inhibition and equilibrium dialysis show that the affinity of the free enzyme for CoA is quite low (Ki = 100 microM). The quantum yield of the fluorescence of the three thiolase tryptophan residues is low (0.024), corresponding to about 12% of the fluorescence expected from equivalent concentrations of tryptophan.


Synthesis of γ-Cyclodextrin-Reduced Fe(III) Nanoparticles with Peroxidase-like Catalytic Activity for Bacteriostasis of Food.
Foodborne pathogens are a primary cause of human foodborne illness, making it imperative to explore novel antibacterial strategies for their control. In this study, Fe-γ-CD was successfully synthesized as a food antibacterial agent for use in milk and orange juice. The Fe-γ-CD consists of 6/11 Fe(II) and 5/11 Fe(III), which catalyze a Fenton-like catalytic reaction with H2O2 to generate •OH. Consequently, Fe-γ-CD exhibits exceptional peroxidase-like activity and broad-spectrum antibacterial efficacy. Fe-γ-CD not only disrupts the wall structure of ESBL-E. coli but also induces protein leakage and genetic destruction, ultimately leading to its death. Furthermore, Fe-γ-CD inhibits biofilm formation by MRSA and eradicates mature biofilms, resulting in MRSA's demise. Importantly, Fe-γ-CD demonstrates negligible cytotoxicity toward normal mammalian cells, making it an ideal candidate for application as an antibacterial agent in foodstuffs. These findings highlight that Fe-γ-CD is an effective tool for combating the spread of foodborne pathogens and food safety.


Risk factors for burnout among caregivers working in nursing homes.
(i) To assess the level of burnout in nursing home caregivers within a unique healthcare network in France and (ii) to evaluate potential risk factors in this population. Burnout syndrome occurs frequently among nursing home caregivers and has strong detrimental effects on the quality of health care for residents. We used an observational survey to study burnout in nursing home caregivers. The survey was used to quantify burnout level (Maslach Burnout Inventory) and potential risk factors and was implemented from October 2013-April 2014. A logistic regression was used to explore the association between burnout and its risk factors. Three hundred and sixty questionnaires were delivered to caregivers in 14 nursing homes within a unique healthcare network. The response rate was 37% (132/360), and 124/132 (94%) surveys were analysed. Caregiver burnout rate was 40% (49/124). Median age was 41 years (range, 20-70) and most caregivers were female. The most common profession (n = 54; 44%) was nurse caregiver and 90% (n = 112) had an antecedent of bullying by a resident. Risk factors identified were as follows: the presence of institutional protocols (death announcement [OR: 3.7] and pain assessment [OR: 2.8]), working in a profit-making establishment (OR: 2.6) and the antecedent of bullying by a resident (OR: 6.2). Factors most negatively associated with burnout included: practising pastimes (OR: 0.4) and working as a nurse (OR: 0.3). The only significant risk factor in the multivariate analysis was the antecedent of bullying by a resident (OR: 5.3). Several specific risk factors for burnout in nursing home caregivers were identified. In high-risk populations of healthcare professionals, screening and management of risk factors is crucial for preventing burnout.


Time-frequency analysis reveals decreased high-frequency oscillations in writer's cramp.
High-frequency oscillations (HFO) have been suggested to reflect the activity of thalamocortical and/or intracortical neurons bursting at high frequencies. These circuits seem to be involved in pathophysiological mechanisms of focal dystonia. In healthy subjects, we characterized the spectrotemporal properties of HFO patterns evoked by dominant-hand median-nerve stimulation, using magnetoencephalography coupled with time-frequency analysis. Then, we investigated HFO in patients with writer's cramp and found that HFO patterns are strongly decreased in power and disorganized in time. This supports the assumption that abnormal HFOs reflect pathophysiological mechanisms occurring in focal dystonia, possibly resulting from a dysfunction of somatosensory processing.


Transcription termination factor Rho contains three noncatalytic nucleotide binding sites.
The active form of transcription termination factor rho from Escherichia coli is a homohexamer, but several studies suggest that the six subunits of the hexamer are not functionally identical. Rho has three tight and three weak ATP binding sites. Based on our findings, we propose that the tight nucleotide binding sites are noncatalytic and the weak sites are catalytic. In the presence of RNA, the rho-catalyzed ATPase rate is fast, close to 30 s-1. However, under these conditions the three tightly bound nucleotides dissociate from the rho hexamer at a slow rate of 0.02 s-1, indicating that the three tight nucleotide binding sites of rho do not participate in the fast ATPase turnover. These slowly exchanging nucleotide binding sites of rho are capable of hydrolyzing ATP, but the resulting products (ADP and Pi) bind tightly and dissociate from rho about 1500 times slower than the fast ATPase turnover. Both RNA and excess ATP in solution are necessary for stabilizing nucleotide binding at these sites. In the absence of RNA, or when solution ATP is hydrolyzed to ADP, a faster dissociation of nucleotides was observed. Based on these results, we propose that the rho hexamer is similar to the F1-ATPase and T7 DNA helicase-containing noncatalytic sites that do not participate in the fast ATPase turnover. We propose that the three tight sites on rho are the noncatalytic sites and the three weak sites are the catalytic sites.


The biopathologic characteristics of DNA content of hepatocellular carcinomas.
A gross morphologic classification of 66 small hepatocellular carcinomas (SHCC, less than or equal to 3 cm in diameter) was made and DNA content of 30 SHCC and 26 large hepatocellular carcinomas (LHCC, greater than 3 cm in diameter) was determined by image analysis technology. The three types of SHCC are as follows: type I, noncapsule type; type II, capsule type; and type III, infiltrative type. Of the SHCC, 73.3% had diploid DNA content, and 84.6% of LHCC had aneuploid DNA content. The postoperative 5-year survival rate of patients with SHCC was 62.1%, which is higher than the rate of 27.7% of patients with LHCC. There was no correlation between tumor size and serum alpha-fetoprotein (AFP) levels. The results suggest that the period when SHCC are 3 cm in size may be important as the changes of DNA stemlines and biological characteristics would occur then. The SHCC of less than 3 cm reflect relatively benign biological behavior of early hepatocellular carcinoma and this period is the best opportunity for the clinician to get the best prognosis. Currently determination of serum AFP is still one of the possible and effective methods to early finding SHCC.


The xeroderma pigmentosum group B protein ERCC3 produced in the baculovirus system exhibits DNA helicase activity.
The XPB/ERCC3 gene corrects the nucleotide excision-repair defect in the human hereditary disease xeroderma pigmentosum group B and encodes the largest subunit of the basal transcription factor BTF2/TFIIH. The primary sequence of the XPB/ERCC3 protein features the hallmarks of seven helicase motifs found in many known and putative helicases or helicase-related proteins. Recently, the multiprotein BTF2/TFIIH complex has been found to be associated with DNA helicase activity. To explore the properties and functions of XPB/ERCC3, we have used the baculovirus/insect-cell expression system to produce recombinant protein. We report here the construction and analysis of recombinant baculovirus expressing XPB/ERCC3. The XPB/ERCC3 protein is synthesized at a relatively high level in baculovirus-infected insect cells. While the majority of XPB/ERCC3 end up in the insoluble fraction of insect cell lysates, a minor fraction of recombinant protein is present in soluble form which can be purified under native conditions. We have found that a DNA helicase activity is associated with the purified XPB/ERCC3 protein, suggesting that XPB/ERCC3 may function as a DNA helicase in local unwinding of DNA template both in the context of transcription and nucleotide excision repair.


Protective effect of Cox-2 allelic variants on risk of colorectal adenoma development in African Americans.
Recent evidence indicates that single nucleotide polymorphisms (SNPs) in the Cox-2 gene may modulate the risk of colorectal adenoma development. We explored possible associations between Cox-2 polymorphisms and risk of adenoma development in an African American case-control study comprising 72 cases of advanced adenomas and 146 polyp-free controls. An exhaustive approach of genotyping 13 haplotype-tagging SNPs (ht SNPs) distributed over the entire COX-2 gene was used. Statistically significant inverse associations were observed between the heterozygous genotypes at the 5229 G>T polymorphism in intron 5 [odds ratio (OR)=0.42; confidence interval (CI)=0.19-0.92; p=0.03] and at the 10935 A>G polymorphism in the 3' flanking region downstream from the poly A signals (OR=0.39; CI=0.18-0.83;p=0.01) and the risk for colorectal adenoma development. The data from our pilot study suggest that allelic variants of the COX-2 gene significantly influence the risk of adenoma development in the African American population.


Increasing the metabolizable protein supply enhanced growth performance and led to variable results on innate and humoral immune response of preconditioning beef steers.
We evaluated the effects of MP supply on growth performance before and after preconditioning and measurements of innate and humoral immune response of beef steers following vaccination. Angus steers ( = 36; BW = 231 ± 21 kg; age = 184 ± 18 d) were weaned on d -6, stratified by BW and age on d 0, and randomly assigned to 1 of 18 drylot pens (2 steers/pen). Treatments were assigned to pens (6 pens/treatment) and consisted of corn silage-based diets formulated to provide 85%, 100%, or 115% of the daily MP requirements of a beef steer gaining 1.1 kg/d from d 0 to 42. Steers were vaccinated against infectious bovine rhinotracheitis virus, bovine viral diarrhea (BVDV) types 1 and 2 viruses, and clostridium on d 14 and 28. Blood samples were collected on d 0, 14, 15, 17, 21, 28, 29, 30, 35, and 42. Body weight did not differ ( ≥ 0.17) among treatments from d 0 to 28. On d 42, 115% MP steers were heaviest, 100% MP steers were intermediate, and 85% MP steers were lightest ( = 0.05; 297, 290, and 278 ± 7 kg, respectively). Overall, ADG and G:F did not differ ( ≥ 0.13) between 100% and 115% MP steers and were least ( < 0.01) for 85% MP steers (1.2, 1.4, and 0.8 ± 0.07 kg/d and 0.23, 0.24, and 0.19 ± 0.008, respectively). Plasma haptoglobin (Hp) concentrations did not differ among treatments ( ≥ 0.46), whereas plasma ceruloplasmin (Cp) concentrations were greatest ( ≤ 0.04) for 85% MP steers, intermediate for 100% MP steers, and least for 115% MP steers on d 30, 35, and 42. Plasma cortisol concentrations were greater ( ≤ 0.03) for 85% vs. 100% and 115% MP steers on d 14 and 28. Liver mRNA expression of Cp and Hp and muscle mRNA expression of m-calpain, mammalian target of rapamycin, and ubiquitin did not differ among treatments ( ≥ 0.17). Serum neutralization titers to BVDV-1b titers were greater ( ≤ 0.02) for 115% vs. 85% and 100% MP steers on d 42 (5.8, 3.0, and 3.7 ± 0.60 log, respectively), whereas mean serum leukotoxin titers were greater for 85% vs. 100% and 115% MP steers (3.1, 2.4, and 2.5 ± 0.21 log, respectively). Preconditioning MP supply did not affect ( ≥ 0.26) ubsequent finishing growth performance and carcass characteristics. Thus, increasing MP supply from 85% to 115% of daily requirement of preconditioning beef steers had variable results on innate and humoral immune response and enhanced growth performance during a 42-d preconditioning period without affecting carcass characteristics at slaughter.


[Polymorphisms of RAD51(G135C) and XRCC3(C241T) genes and correlations thereof with prognosis and clinical outcomes of acute myeloid leukemia].
To investigate the impacts of RAD51G(135C) and XRCC3(C241T) genotypes on the response, adverse effects, and prognosis of acute myelocytic leukemia (AML). RAD51(G135C), XRCC3(C241T), GSTT1, and GSTM1 genotypes were analyzed in 372 patients with AML, 226 males and 146 females, by PCR-RFLP or PCR. The Complete remission (CR) rate, adverse effects, overall survival (OS), and relapse-free survival (RFS) were compared among the groups with different genotypes. (1) During the induction chemotherapy, XRCC3(C241T) polymorphic allele was significantly associated with the shorter survival of the AML patients with t (15; 17)/PML-RARalpha (OR = 8.750, P = 0.046). Among the non-M3 patients, the complete remission (CR) rate of those with double RAD51(G135C) and GSTT1 wild genotypes was 71.6%, significantly higher than that of those not with double RAD51(G135C) and GSTT1 wild genotypes (54.4%, P = 0.028). (2) The OS of the non-M3 AML patients with double RAD51(G135C) and GSTT1 wild genotypes was (39.1 +/- 7.1) months, significantly longer than those with double variant types [(22.4 +/- 3.2) months, P = 0.042]. The relapse-free survival (RFS) of the M4EO and M2 patients with double XRCC3(C241T) and GSTT1 wild type genotype were 48.3 months and 56.5 months, both significantly longer than those of the patients with double variant genotypes (28.8 months and 10.0 months respectively, both P < 0.05). The OS of the M2 patients with triple RAD51(G135C), GSTT1, and GSTM1 variant genotypes was (22.4 +/- 3.2) months, significantly shorter than those with triple RAD51(G135C), GSTT1, and GSTM1 wild genotypes [(39.1 +/- 7.1) months, P = 0.042]. The RFS of the M2 patients with triple RAD51(G135C), GSTT1, and GSTM1 variant genotypes was 10.0 months, significantly shorter than that of the patients with triple RAD51(G135C), GSTT1, and GSTM1 wild genotypes (64.2 months, P = 0.015). (3) The risk levels of neutropenia, nausea and vomiting, and alopecia of the patients with variant XRCC3(C241T) genotype were all significantly higher than those of the wild type genotype (all P < 0.05). The risk of hematuria of the patients with variant genotype was significantly higher than that of the patients with wild genotype (P = 0.017). The polymorphisms of XRCC3(C241T) and RAD51(G135C) genes are significantly related to response to therapy, adverse effects, and prognosis of AML. Detection of the XRCC3(C241T) and RAD51(G135C) genotypes may be useful in selecting individual chemotherapy regimens for patients with AML.