Co-ordinate action of bacterial adhesins and human carcinoembryonic antigen receptors in enhanced cellular invasion by capsulate serum resistant Neisseria meningitidis.
Neisseria meningitidis (Nm) is a human specific opportunistic pathogen that occasionally penetrates mucosal barriers via the action of adhesins and invasins and evades host immune mechanisms during further dissemination via capsule expression. From in vitro studies, the primary adhesion of capsulate bacteria is believed to be mediated by polymeric pili, followed by invasion via outer membrane adhesins such as Opa proteins. As the latter requires the surface capsule to be down-modulated, invading bacteria would be serum sensitive and thus avirulent. However, there is recent evidence that capsulate bacteria may interact via Opa proteins when host cells express high levels of carcinoembryonic antigen-related cell adhesion molecules (CEACAMs), their target receptors. Such a situation may arise following increased circulation of inflammatory cytokines that upregulate certain adhesion molecules on host cells. In this study, using a tetracycline controlled expression system, we have developed cell lines with inducible CEACAM expression to mimic post-inflammation state of target tissues and analysed the interplay between the three surface components capsule, pili and Opa proteins in cellular interactions. With two distinct cell lines, not only the level but also the rate of adhesion of capsulate Opa-expressing Nm increased concurrently with CEACAM density. Moreover, when threshold levels of receptor were reached, cellular invasion ensued in an Opa-dependent manner. In studies with cell lines intrinsically expressing pilus receptors, notable synergism in cellular interactions between pili and Opa of several meningococcal strains was observed and was independent of capsule type. A number of internalized bacteria were shown to express capsule and when directly isolated from host cells, these bacteria were as serum resistant as the inoculated phenotype. Furthermore, we observed that agents that block Opa-CEACAM binding substantially reduced cellular invasion, while maintaining a low level of cellular adhesion. These studies highlight some of the factors that may determine increased host susceptibility to infection by serum resistant phenotypes; and demonstrate the potential of selective inhibition of key interactions in preventing target tissue penetration while maintaining a level of colonization.


Comparison of linear and nonlinear implementation of the compartmental tissue uptake model for dynamic contrast-enhanced MRI.
Fitting tracer kinetic models using linear methods is much faster than using their nonlinear counterparts, although this comes often at the expense of reduced accuracy and precision. The aim of this study was to derive and compare the performance of the linear compartmental tissue uptake (CTU) model with its nonlinear version with respect to their percentage error and precision. The linear and nonlinear CTU models were initially compared using simulations with varying noise and temporal sampling. Subsequently, the clinical applicability of the linear model was demonstrated on 14 patients with locally advanced cervical cancer examined with dynamic contrast-enhanced magnetic resonance imaging. Simulations revealed equal percentage error and precision when noise was within clinical achievable ranges (contrast-to-noise ratio >10). The linear method was significantly faster than the nonlinear method, with a minimum speedup of around 230 across all tested sampling rates. Clinical analysis revealed that parameters estimated using the linear and nonlinear CTU model were highly correlated (ρ ≥ 0.95). The linear CTU model is computationally more efficient and more stable against temporal downsampling, whereas the nonlinear method is more robust to variations in noise. The two methods may be used interchangeably within clinical achievable ranges of temporal sampling and noise. Magn Reson Med 77:2414-2423, 2017. © 2016 The Authors Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.


[Diagnosis and treatment for the injury of cervical disc and longitudinal ligament].
To study the diagnosis and treatment for the injury of cervical disc and longitudinal ligament. From 2001 to 2005, the clinical data of sixty-three patients with cervical disc and longitudinal ligament injury were studied. Early treatment was done based on spinal cord injuries and spinal stabilities by X rays and MRI. Early operation was done in fifty-four cases and early non-operation in nine cases. The follow-up time was six to forty-one months in all patients. The neurological recovery was found in two of eight complete SCI post-operation, thirty-one in thirty-nine incomplete SCI. Cervical collar or plaster orthotic were used in nine cases with four to six weeks. Evidence of instability was noted in four patients, who were operated with anterior decompression fusion. Neck chronic pain was found in two patients, anterior decompression and fusion was done in one with cervical spinal cord compression. MRI examination is the most value measure for the diagnosis of cervical disc and longitudinal ligament injury. Early anterior decompression and fusion was an important approach for cervical disc and longitudinal ligament injury.


Characteristics of a metastatic variant to the liver of human rectal adenocarcinoma cell line RCM-1.
The interaction of human rectal adenocarcinoma cell line RCM-1 cells with extracellular matrix components, was studied to elucidate the key steps in the liver metastasis of colorectal carcinomas. Highly metastatic variant L-10 cells selected from the metastatic foci of the liver after intrasplenic implantation in nude mice and its parental L-0 cells were used. L-10 cells showed a greater ability to adhere to laminin, fibronectin, and type I and type IV collagens than did L-0 cells but less haptotactic activity than that of L-0 cells to type I or type IV collagen, possibly due to the formation of cellular aggregates. In vitro invasion activities of both cell lines to basement membrane components (Matrigel) or type I collagen were minimal but enhanced by the addition of 12-O-tetradecanoylphorbol-13-acetate (TPA). L-10 cells showed greater ability to invade Matrigel than did L-0 cells, while L-0 cells exhibited higher activity in the invasion of type I collagen than did L-10 cells. TPA did not increase the production of metalloproteinases by both cells when analyzed by gelatin zymography. Based on the differences between the two cell lines, we postulated the following: (1) the high metastatic potential of L-10 cells was due to a greater capacity to attach to and cross the basement membrane; (2) TPA directly enhanced tumor cell invasiveness, not via the increased secretion of metalloproteinases; and (3) haptotactic migration had no significant correlation with the increased metastatic potential of L-10 cells.


Food impaction due to nutcracker esophagus.
Nutcracker esophagus is an esophageal motility disorder characterized by the presence of hypertensive contraction waves. These waves can have very high amplitudes, but they maintain a peristaltic pattern and therefore, bolus passage is minimally affected. Esophageal food impactions are rare in nutcracker esophagus. Our patient was a previously asymptomatic man who presented with an esophageal meat impaction due to nutcracker esophagus in which high-resolution manometry played a key role in the diagnosis. Although a rare etiology, nutcracker esophagus can result in esophageal food impaction. High-resolution manometry plays a critical role in the diagnosis of specific motility disorders, even in the setting of minimal symptoms.


Male-biased autosomal effect of 16p13.11 copy number variation in neurodevelopmental disorders.
Copy number variants (CNVs) at chromosome 16p13.11 have been associated with a range of neurodevelopmental disorders including autism, ADHD, intellectual disability and schizophrenia. Significant sex differences in prevalence, course and severity have been described for a number of these conditions but the biological and environmental factors underlying such sex-specific features remain unclear. We tested the burden and the possible sex-biased effect of CNVs at 16p13.11 in a sample of 10,397 individuals with a range of neurodevelopmental conditions, clinically referred for array comparative genomic hybridisation (aCGH); cases were compared with 11,277 controls. In order to identify candidate phenotype-associated genes, we performed an interval-based analysis and investigated the presence of ohnologs at 16p13.11; finally, we searched the DECIPHER database for previously identified 16p13.11 copy number variants. In the clinical referral series, we identified 46 cases with CNVs of variable size at 16p13.11, including 28 duplications and 18 deletions. Patients were referred for various phenotypes, including developmental delay, autism, speech delay, learning difficulties, behavioural problems, epilepsy, microcephaly and physical dysmorphisms. CNVs at 16p13.11 were also present in 17 controls. Association analysis revealed an excess of CNVs in cases compared with controls (OR = 2.59; p = 0.0005), and a sex-biased effect, with a significant enrichment of CNVs only in the male subgroup of cases (OR = 5.62; p = 0.0002), but not in females (OR = 1.19, p = 0.673). The same pattern of results was also observed in the DECIPHER sample. Interval-based analysis showed a significant enrichment of case CNVs containing interval II (OR = 2.59; p = 0.0005), located in the 0.83 Mb genomic region between 15.49-16.32 Mb, and encompassing the four ohnologs NDE1, MYH11, ABCC1 and ABCC6. Our data confirm that duplications and deletions at 16p13.11 represent incompletely penetrant pathogenic mutations that predispose to a range of neurodevelopmental disorders, and suggest a sex-limited effect on the penetrance of the pathological phenotypes at the 16p13.11 locus.


Epidemiology of hypertension in Japan: where are we now?
Analyses of data from national surveys of the Japanese population have shown a clear decreasing tendency in mean systolic blood pressure (BP) level over the past 50 years in all age groups of men and women; however, mean diastolic BP level clearly did not decrease in men. Hypertension prevalence is high among older people and may increase in the future, especially in men aged ≥50 years. The treatment and control rates of hypertension are not sufficiently high, although they have been continuously improving. Recent epidemiological studies also showed that the burden of cardiovascular diseases and total mortality because of the adverse BP level of the nation is still the highest among other preventable risk factors. To overcome this epidemic, the first priority should be primary prevention of a lifetime increase in BP through lifestyle improvement. Lowering the distribution of BP in the whole population and maintaining BP at optimal levels contributes to the achievement of this goal.


Cost-effectiveness of dual-mobility components in patients with displaced femoral neck fractures.
Total hip arthroplasty (THA) with dual-mobility components (DM-THA) has been shown to decrease the risk of dislocation in the setting of a displaced neck of femur fracture compared to conventional single-bearing THA (SB-THA). This study assesses if the clinical benefit of a reduced dislocation rate can justify the incremental cost increase of DM-THA compared to SB-THA. Costs and benefits were established for patients aged 75 to 79 years over a five-year time period in the base case from the Canadian Health Payer's perspective. One-way and probabilistic sensitivity analysis assessed the robustness of the base case model conclusions. DM-THA was found to be cost-effective, with an estimated incremental cost-effectiveness ratio (ICER) of CAD $46,556 (£27,074) per quality-adjusted life year (QALY). Sensitivity analysis revealed DM-THA was not cost-effective across all age groups in the first two years. DM-THA becomes cost-effective for those aged under 80 years at time periods from five to 15 years, but was not cost-effective for those aged 80 years and over at any timepoint. To be cost-effective at ten years in the base case, DM-THA must reduce the risk of dislocation compared to SB-THA by at least 62%. Probabilistic sensitivity analysis showed DM-THA was 58% likely to be cost-effective in the base case. Treating patients with a displaced femoral neck fracture using DM-THA components may be cost-effective compared to SB-THA in patients aged under 80 years. However, future research will help determine if the modelled rates of adverse events hold true. Surgeons should continue to use clinical judgement and consider individual patients' physiological age and risk factors for dislocation. Cite this article: Bone Joint J 2021;103-B(12):1783-1790.


Physical activity and academic achievement among Norwegian adolescents: Findings from a longitudinal study.
Regular physical activity (PA) has been associated with academic achievement, but the evidence is mainly based on cross-sectional research and self-reported measures of PA. The purpose of the current study was to explore the longitudinal relationship between objectively measured PA and academic achievement among a cohort of adolescents in Norway between 2016 and 2018. As a secondary aim, an indirect relationship via waist circumference (WC) and sleep duration was assessed. Data from 599 adolescents (54.4% female, mean age at baseline ± SD 13.3 ± 0.3 years) were collected annually during their three years at lower secondary school. PA was measured objectively using accelerometry. Academic achievement was assessed using grade point average (GPA) from school records. Linear regression analysis was performed to explore associations between longitudinal changes in measures of PA (Total PA and moderate-to-vigorous PA [MVPA]) and academic achievement directly or via mediators. Results showed no significant associations between Total PA or MVPA and academic achievement, either in the main analyses or through mediation of WC and sleep duration. The results contribute to a growing evidence base of studies showing no association between objectively measured PA and academic achievement among adolescents.


Glucocorticoid-induced tumor necrosis factor receptor family-related ligand triggering upregulates vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 and promotes leukocyte adhesion.
The interaction of glucocorticoid-induced tumor necrosis factor receptor-family related (GITR) protein with its ligand (GITRL) modulates different functions, including immune/inflammatory response. These effects are consequent to intracellular signals activated by both GITR and GITRL. Previous results have suggested that lack of GITR expression in GITR(-/-) mice decreases the number of leukocytes within inflamed tissues. We performed experiments to analyze whether the GITRL/GITR system modulates leukocyte adhesion and extravasation. For that purpose, we first evaluated the capability of murine splenocytes to adhere to endothelial cells (EC). Our results indicated that adhesion of GITR(-/-) splenocytes to EC was reduced as compared with wild-type cells, suggesting that GITR plays a role in adhesion and that this effect may be due to GITRL-GITR interaction. Moreover, adhesion was increased when EC were pretreated with an agonist GITR-Fc fusion protein, thus indicating that triggering of GITRL plays a role in adhesion by EC regulation. In a human in vitro model, the adhesion to human EC of HL-60 cells differentiated toward the monocytic lineage was increased by EC pretreatment with agonist GITR-Fc. Conversely, antagonistic anti-GITR and anti-GITRL Ab decreased adhesion, thus further indicating that GITRL triggering increases the EC capability to support leukocyte adhesion. EC treatment with GITR-Fc favored extravasation, as demonstrated by a transmigration assay. Notably, GITRL triggering increased intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression and anti-ICAM-1 and anti-VCAM-1 Abs reversed GITR-Fc effects. Our study demonstrates that GITRL triggering in EC increases leukocyte adhesion and transmigration, suggesting new anti-inflammatory therapeutic approaches based on inhibition of GITRL-GITR interaction.


A new elderly support system in rural areas in Shanghai.
"In early 1987, we started an experiment in the rural areas in Shanghai suburbs on how to replace the elderly support system then in effect, which was funded by the community, with one that was to be jointly supported by the State, the community and private individuals. The two-year experiment has yielded a new elderly support system applicable to China's rural areas that have a more developed economy and a fairly large number of collectively owned township enterprises.... This article gives a brief introduction to the economic and social background and characteristics of the system."


High dislocation density-induced large ductility in deformed and partitioned steels.
A wide variety of industrial applications require materials with high strength and ductility. Unfortunately, the strategies for increasing material strength, such as processing to create line defects (dislocations), tend to decrease ductility. We developed a strategy to circumvent this in inexpensive, medium manganese steel. Cold rolling followed by low-temperature tempering developed steel with metastable austenite grains embedded in a highly dislocated martensite matrix. This deformed and partitioned (D and P) process produced dislocation hardening but retained high ductility, both through the glide of intensive mobile dislocations and by allowing us to control martensitic transformation. The D and P strategy should apply to any other alloy with deformation-induced martensitic transformation and provides a pathway for the development of high-strength, high-ductility materials.


Diaphragmatic defect or pulmonary sequestration in a five-year-old boy.
This case report presents a 5-year-old boy with diaphragmatic defect with a tardy clinical manifestation. The findings of chest x-ray and chest CT scan were seen by pediatricians and radiologists as interstitial pneumonia and suspected pulmonary sequestration, pediatric surgeons saw it as a benign teratoma and during surgery a muscular defect of hemidiaphragm right was found. This case emphasize the rarity of diaphragmatic hernia in children of older age, the importance of "accented" clinical suspicion based on the maintenance of signs of mild respiratory failure, the need for additional radiological tests and surgery with the aim to diagnose this abnormality and "successfulness" of prenatal ultrasound examination.


Molecular basis of antiangiogenic thrombospondin-1 type 1 repeat domain interactions with CD36.
Antiangiogenic activity of thrombospondin-1 and related proteins is mediated by interactions between thrombospondin type 1 repeat (TSR) domains and the CD36, LIMP-2, Emp sequence homology (CLESH) domain of the endothelial cell receptor CD36. We sought to characterize key molecular determinants of the interaction between thrombospondin-1 TSR domains and the CD36 CLESH domain. Recombinant thrombospondin-1 TSR2 and TSR(2,3) constructs inhibited microvascular endothelial cell migration, microvascular endothelial cell tube formation, and vessel sprouting in aortic ring assays. Interaction with CD36 CLESH decoy peptides negated these effects. Mutational analyses identified a cluster of residues that confer positive charge to the TSR2 surface and mediate interaction with CD36 CLESH. Antiangiogenic activity was significantly reduced by charge-neutralizing mutations of the Arg-Trp ladder in TSR2, but not TSR3. Additionally, I438 and K464 of TSR2 were shown to be required for CD36 CLESH binding to TSR2. A complementary acidic cluster within CD36 CLESH is also required for antiangiogenic activity. Thrombospondin-1 interacts with CD36 CLESH through electrostatic interactions mediated by a positively charged TSR2 surface and multiple negatively charged CD36 CLESH residues. Two key residues serve as specificity determinants that identify other TSR domains that interact with CD36 CLESH.


The effects of 1-year gluten withdrawal on bone mass, bone metabolism and nutritional status in newly-diagnosed adult coeliac disease patients.
To evaluate the impact of a 1-year gluten-free diet on bone metabolism and nutritional status in coeliac disease. Bone mineral density, serum indices of bone remodelling, clinical and biochemical nutritional assessment were evaluated in 86 consecutive newly-diagnosed, biopsy proven, coeliac disease patients (untreated). A complete reevaluation, including intestinal biopsy, was repeated within 1 year of dietary treatment (treated). Untreated: according to WHO criteria, 34% of patients had a normal bone mineral density, 40% had osteopenia and 26% osteoporosis. Between males and females there were no statistical differences in bone metabolism or in most of the nutritional indices, while, between fertile and postmenopausal women, bone mineral density and several bone metabolism markers were significantly different. Compared to subjects with a normal bone mineral density, osteopenics had higher bone specific alkaline phosphatase (BAP) and Bone-Gla-protein (BGP) values. In patients with a concomitant BAP increase and 25OH vitamin D serum level reduction, bone mineral density and several bone turnover markers were statistically different compared to patients without such a serological pattern. Treated: notwithstanding intestinal biopsy which showed a mucosal recovery in only 57%, gluten-free diet led, even in postmenopausal women, to a significant improvement in bone mineral density, bone metabolism and nutrition, except for folic acid, albumin and pre-albumin serum levels which persisted as abnormal in patients with obdurate mucosal impairment. Coeliac disease patients are at high risk for developing a low bone mineral density and bone turnover impairment. A gluten-free diet can improve this situation even in postmenopausal women and in patients with incomplete mucosal recovery.


An exceptionally stable and widespread hydrated amorphous calcium carbonate precipitated by the dog vomit slime mold Fuligo septica (Myxogastria).
Biogenic amorphous calcium carbonate (ACC) is typically metastable and can rapidly transform through aging, dehydration, and/or heating to crystalline calcium carbonate. Gaining insight into its structure and properties is typically hampered by its tendency to crystallize over short time periods once isolated from the host organism, and also by the small quantities that are usually available for study. Here we describe an exceptionally stable hydrated ACC (HACC) precipitated by the cosmopolitan slime mold Fuligo septica (L.) F.H. Wigg. (1780). A single slime mold can precipitate up to a gram of HACC over the course of one night. Powder x-ray diffraction (XRD) patterns, transmission electron microscopy images, infrared absorption spectra, together with the lack of optical birefringence are consistent with an amorphous material. XRD simulations, supported by thermogravimetric and evolved gas analysis data, are consistent with an intimate association of organic matter with ~ 1-nm-sized ACC units that have monohydrocalcite- and calcite-like nano-structural properties. It is postulated that this association imparts the extreme stability of the slime mold HACC by inhibiting loss of H2O and subsequent crystallization. The composition, structure, and thermal behavior of the HACC precipitated by F. septica collected over 8000 km apart and in markedly different environments, suggests a common structure, as well as similar biochemical and biomineralization mechanisms.


Using Recent BCI Literature to Deepen our Understanding of Clinical Neurofeedback: A Short Review.
In their recent paper, Alkoby et al. (2017) provide the readership with an extensive and very insightful review of the factors influencing NeuroFeedback (NF) performance. These factors are drawn from both the NF literature and the Brain-Computer Interface (BCI) literature. Our short review aims to complement Alkoby et al.'s review by reporting recent additions to the BCI literature. The object of this paper is to highlight this literature and discuss its potential relevance and usefulness to better understand the processes underlying NF and further improve the design of clinical trials assessing NF efficacy. Indeed, we are convinced that while NF and BCI are fundamentally different in many ways, both the BCI and NF communities could reach compelling achievements by building upon one another. By reviewing the recent BCI literature, we identified three types of factors that influence BCI performance: task-specific, cognitive/motivational and technology-acceptance-related factors. Since BCIs and NF share a common goal (i.e., learning to modulate specific neurophysiological patterns), similar cognitive and neurophysiological processes are likely to be involved during the training process. Thus, the literature on BCI training may help (1) to deepen our understanding of neurofeedback training processes and (2) to understand the variables that influence the clinical efficacy of NF. This may help to properly assess and/or control the influence of these variables during randomized controlled trials.


Statistical considerations for noninferiority/equivalence trials in vaccine development.
Noninferioritylequivalence designs are often used in vaccine clinical trials. The goal of these designs is to demonstrate that a new vaccine, or new formulation or regimen of an existing vaccine, is similar in terms of effectiveness to the existing vaccine, while offering such advantages as easier manufacturing, easier administration, lower cost, or improved safety profile. These noninferioritylequivalence designs are particularly useful in four common types of immunogenicity trials: vaccine bridging trials, combination vaccine trials, vaccine concomitant use trials, and vaccine consistency lot trials. In this paper, we give an overview of the key statistical issues and recent developments for noninferioritylequivalence vaccine trials. Specifically, we cover the following topics: (i) selection of study endpoints; (ii) formulation of the null and alternative hypotheses; (iii) determination of the noninferioritylequivalence margin; (iv) selection of efficient statistical methods for the statistical analysis of noninferioritylequivalence vaccine trials, with particular emphasis on adjustment for stratification factors and missing pre-or post-vaccination data; and (v) the calculation of sample size and power.


Dual roles of an essential cysteine residue in activity of a redox-regulated bacterial transcriptional activator.
CprK from Desulfitobacterium dehalogenans is the first characterized transcriptional regulator of anaerobic dehalorespiration and is controlled at two levels: redox and effector binding. In the reduced state and in the presence of chlorinated aromatic compounds, CprK positively regulates expression of the cpr gene cluster. One of the products of the cpr gene cluster is CprA, which catalyzes the reductive dehalogenation of chlorinated aromatic compounds. Redox regulation of CprK occurs through a thiol/disulfide redox switch, which includes two classes of cysteine residues. Under oxidizing conditions, Cys11 and Cys200 form an intermolecular disulfide bond, whereas Cys105 and Cys111 form an intramolecular disulfide. Here, we report that Cys11 is involved in redox inactivation in vivo. Upon replacement of Cys11 with serine, alanine, or aspartate, CprK loses its DNA binding activity. C11A is unstable; however, circular dichroism studies demonstrate that the stability and overall secondary structures of CprK and the C11S and C11D variants are similar. Furthermore, effector binding remains intact in the C11S and C11D variants. However, fluorescence spectroscopic results reveal that the tertiary structures of the C11S and C11D variants differ from that of the wild type protein. Thus, Cys11 plays a dual role as a redox switch and in maintaining the correct tertiary structure that promotes DNA binding.


Early transcriptomic response of Arabidopsis thaliana to polymetallic contamination: implications for the identification of potential biomarkers of metal exposure.
Heavy metal contaminated sites are frequently characterized by the simultaneous presence of several heavy metals. However, many studies report metal-induced plant responses after long-term exposure to just one metal. By contrast, whole genome expression microarrays were employed here to investigate the early (3 h) transcriptional responses of Arabidopsis thaliana plants exposed to polymetallic treatment (Pb, Hg, Cu, Cd, Co, Ni, Zn, and Mn) at low (L) and high (H) concentrations. After 3 h of exposure to polymetallic treatment, a total of 1315 noticeably (≥2-fold) and significantly (P < 0.05) differentially expressed genes were identified: 656 and 351 upregulated and 314 and 200 downregulated genes in L and H treatments, respectively. Functional analysis revealed that many genes involved in oxidative stress and perception/signalling/regulation systems were activated. Genes encoding proteins involved in hormone regulation (jasmonic acid, abscisic acid, ethylene, and auxins), glucosinolate metabolism and sulphur and nitrogen transport were also modulated. RT-qPCR analysis of four downregulated (AOP2, SAUR16, BBX31, and MTPC3) and upregulated genes (ASN1, DIN2, BT2, and EXL5), markedly responsive to both L and H treatments, validated our microarray data and suggested the potential of some of these genes (AOP2, SAUR16, ASN1, and DIN2) as early biomarkers of metal exposure. Relevant changes in gene expression occur as early as 3 h after exposure to polymetallic treatment. Four genes deserve further studies as novel putative biomarkers of early metal exposure and also owing to their potential implications in stress-related mechanisms: sulphur balance (AOP2), phytohormone regulation of plant growth and development (SAUR16), ammonium detoxification (ASN1) and senescence (DIN2).


Palytoxin is a non-12-O-tetradecanoylphorbol-13-acetate type tumor promoter in two-stage mouse skin carcinogenesis.
Palytoxon, which is a toxin with a molecular weight of 2681 daltons isolated from a marine coelenterate, is a potent skin irritant. However, it did not induce ornithine decarboxylase in mouse skin, or adhesion of human promyelocytic leukemia cells (HL-60). Moreover, it did not inhibit the specific binding of [3H]12-O-tetradecanoylphorbol-13-acetate (TPA) to a mouse skin particulate fraction or activate protein kinase C isolated from mouse brain in vitro. Since palytoxin showed strong irritation on mouse ear in one short-term screening test for a promoter, it was examined in a two-stage carcinogenesis experiment. The incidence of tumors in a group of mice treated with 7,12-dimethylbenz[a]anthracene plus palytoxin was 62.5% in week 25. These tumors were identified histologically as seven papillomas and one carcinoma. This paper reports the potent tumor-promoting activity of palytoxin, which is classified as a non-TPA-type tumor promoter.


Prognostic Impact of WT-1 and Survivin Gene Expression in Acute Myeloid Leukemia Patients.
Wilms Tumor 1 (WT1) and Survivin genes are important leukemia-associated antigens (LAAs) in AML with potential prognostic impact. We investigated WT1 and Survivin expression levels by RT-PCR in 61 AML patients in correlation with clinical characteristics and outcomes. WT1 was overexpressed in 45 patients (73.8%), associated with higher BM blasts (p = 0.017), lower incidence of favorable-prognosis cytogenetics (p = 0.035), and higher incidence of Flt3-ITD mutations (p = 0.026). Survivin was overexpressed in 17 patients (27.9%) with higher mean WBC count (p = 0.049). Patients with overexpression of either gene showed inferior complete remission (CR) rates and survival rates, patients with overexpression of both genes showed higher mean WBCs (p = 0.035) and higher BM blasts (p = 0.029) while the double negative group showed higher incidence of favorable cytogenetic events (p = 0.021), better CR rates and survival rates. Our findings support the introduced prognostic impact of WT1 and Survivin genes in AML patients and its potential use in MRD monitoring and immunotherapy.


Linear and Branched PEIs (Polyethylenimines) and Their Property Space.
A chemical property space defines the adaptability of a molecule to changing conditions and its interaction with other molecular systems determining a pharmacological response. Within a congeneric molecular series (compounds with the same derivatization algorithm and thus the same brute formula) the chemical properties vary in a monotonic manner, i.e., congeneric compounds share the same chemical property space. The chemical property space is a key component in molecular design, where some building blocks are functionalized, i.e., derivatized, and eventually self-assembled in more complex systems, such as enzyme-ligand systems, of which (physico-chemical) properties/bioactivity may be predicted by QSPR/QSAR (quantitative structure-property/activity relationship) studies. The system structure is determined by the binding type (temporal/permanent; electrostatic/covalent) and is reflected in its local electronic (and/or magnetic) properties. Such nano-systems play the role of molecular devices, important in nano-medicine. In the present article, the behavior of polyethylenimine (PEI) macromolecules (linear LPEI and branched BPEI, respectively) with respect to the glucose oxidase enzyme GOx is described in terms of their (interacting) energy, geometry and topology, in an attempt to find the best shape and size of PEIs to be useful for a chosen (nanochemistry) purpose.


Percutaneous balloon pericardiotomy: a double-balloon technique.
We describe a double-balloon technique for performing a percutaneous balloon pericardiotomy. This technique was employed when the large, single dilation balloon customarily used for this procedure failed to fully inflate across the parietal pericardium. Two smaller balloons were advanced through the same skin tract and simultaneously inflated, thus producing an adequate pericardial window. This double-balloon technique allowed for the more secure anchoring of the balloons across the pericardium and for the delivery of greater dilation pressures.


Paeonol exerts an anticancer effect on human colorectal cancer cells through inhibition of PGE₂ synthesis and COX-2 expression.
Cyclooxygenase-2 (COX-2) and its metabolite prostaglandin E2 (PGE2) can potentially affect most of the events in cancer development, including promotion of proliferation, resistance to apoptosis, angiogenesis, immune suppression and invasion. However, worldwide attention has predominantly centered on the cardiovascular toxicity of selective COX-2 inhibitors. Paeonol is a major active extract from the root bark of Paeonia suffruticosa Andrews with anti‑inflammatory, anti-oxidant, anti-allergic, anti-oxidation and antitumor effects. In the present study, we investigated the underlying mechanisms of paeonol in inducing apoptosis and aimed to ascertain whether its antitumor effect is associated with a reduction in COX-2 expression and a decrease in the levels of PGE2 in colorectal cancer cells. We observed that paeonol inhibited cell proliferation and induced apoptosis in a dose- and time-dependent manner in colorectal cancer cells, which was associated with a reduction in COX-2 expression and PGE2 synthesis. Treatment with the selective COX-2 inhibitor, celecoxib, or transient transfection of colorectal cancer cells with COX-2 siRNA, also inhibited cell proliferation and induced apoptosis. Western blot analysis showed that paeonol inhibited the activation of NF-κB, an upstream regulator of COX-2, and its translocation to the nucleus. Treatment with increasing doses of paeonol led to increased expression of pro-apoptotic factor Bax and decreased expression of anti-apoptotic factor Bcl-2. Caspase-3 and caspase-9 were activated, and paeonol induced loss of mitochondrial membrane potential, suggesting that the apoptosis induced by paeonol was mediated by mitochondrial pathways. In addition, paeonol significantly suppressed tumor growth in a xenograft tumor mouse model in a dose-dependent manner. Our findings indicate that paeonol exerts an antitumor effect on human colorectal cancer cells by inhibiting PGE2 production and COX-2 expression. We expect that paeonol may replace selective COX-2 inhibitors due to their toxic effects, and may offer a new strategy for the therapy of colorectal cancer.


Are Pain Ratings Irrelevant?
Pain intensity ratings have been the basis of pain diagnosis and a fundamental tool in pain research, but are not always used. Frequent comments by physicians that pain ratings, sometimes called pain scores, are not useful in clinical practice and comments by basic scientists that pain ratings may measure the wrong thing, have been in significant part supported by a short survey conducted among members of American Pain Society (APS). Though limited by small number of respondents, the findings of this survey and additional comments by members of APS raise critical questions about why pain ratings do not serve the clinical communities. These findings send an urgent call to the pain community to reassess the status of currently used pain ratings and to find solutions to this fundamental issue.


Rapid purification of iodinated ligands for cyclic nucleotide radioimmunoassays.
The tyrosine methyl esters of succinyl cyclic AMP and succinyl cyclic GMP were iodinated by the chloramine T method and individually applied to C18 cartridges. A solution of 1-propanol/0.1 M sodium acetate pH 4.75 (17.5:82.5) was then pumped onto each cartridge and the eluate collected. A large peak of radioactivity, containing primarily the monoiodo and diiodo derivatives, was eluted. Radioactivity in peak fractions was greater than or equal to 95% the monoiodo derivative and represented 20 to 25% of the starting radioactivity. Contamination by the native cyclic nucleotide analogs was less than 5%. These peak fractions containing primarily monoiodinated products worked well in cyclic nucleotide radioimmunoassays. This fractionation required less than 30 min.


Mutant barley (1-->3,1-->4)-beta-glucan endohydrolases with enhanced thermostability.
The similar three-dimensional structures of barley (1-->3)-beta-glucan endohydrolases and (1-->3,1-->4)-beta-glucan endohydrolases indicate that the enzymes are closely related in evolutionary terms. However, the (1-->3)-beta-glucanases hydrolyze polysaccharides of the type found in fungal cell walls and are members of the pathogenesis-related PR2 group of proteins, while the (1-->3,1-->4)-beta-glucanases function in plant cell wall metabolism. The (1-->3)-beta-glucanases have evolved to be significantly more stable than the (1-->3,1-->4)-beta-glucanases, probably as a consequence of the hostile environments imposed upon the plant by invading microorganisms. In attempts to define the molecular basis for the differences in stability, eight amino acid substitutions were introduced into a barley (1-->3,1-->4)-beta-glucanase using site-directed mutagenesis of a cDNA that encodes the enzyme. The amino acid substitutions chosen were based on structural comparisons of the barley (1-->3)- and (1-->3,1-->4)-beta-glucanases and of other higher plant (1-->3)-beta-glucanases. Three of the resulting mutant enzymes showed increased thermostability compared with the wild-type (1-->3,1-->4)-beta-glucanase. The largest increase in stability was observed when the histidine at position 300 was changed to a proline (mutant H300P), a mutation that was likely to decrease the entropy of the unfolded state of the enzyme. Furthermore, the three amino acid substitutions which increased the thermostability of barley (1-->3,1-->4)-beta-glucanase isoenzyme EII were all located in the COOH-terminal loop of the enzyme. Thus, this loop represents a particularly unstable region of the enzyme and could be involved in the initiation of unfolding of the (1-->3,1-->4)-beta-glucanase at elevated temperatures.


Effect of specific antibody on adherence of Staphylococcus aureus to bovine mammary epithelial cells.
Staphylococcus aureus is a major pathogen in the bovine mammary gland. The ability of S. aureus to adhere to epithelial cells in the ductules and alveoli of the bovine mammary gland is believed to add greatly to its virulence and may be necessary for colonization. Two in vitro methods were developed for the purposes of quantifying adherence and of determining the effect which specific antibody may have in inhibiting the adherence of this organism. Both methods utilize bovine mammary epithelial primary cells as targets for labeled bacteria. In one assay, the bacteria are labeled with [methyl-3H]thymidine and incubated on the primary epithelial monolayers. The second assay involves labeling the bacteria with biotin. An enzyme-linked immunosorbent assay is then performed with streptavidin conjugated to horseradish peroxidase. Both methods have proven to be reliable and allow for the testing of many criteria in one assay. Cows were immunized with a whole-cell vaccine, and immune serum and milk were collected. The bacteria were then incubated in the presence of serum or milk as a test for antiadherent capability. By using the methods described, distinct antiadherent activity in both serum and milk was demonstrated.


Refractory familial hypokalaemic periodic paralysis leading to cardiovascular compromise.
Familial hypokalaemic periodic paralysis (FHPP) is a rare neuromuscular disorder that is classified under periodic paralysis (PP), which is characterised by episodes of muscle weakness. Common triggers include intense exercise, fasting or consumption of carbohydrate-rich meals. Hypokalaemic PP has an incidence of 1 in 100 000; despite the temporal association, cardiac manifestations are exceedingly rare. We present a case of FHPP, a channelopathy presenting with severe refractory hypokalaemia. The challenges with our patient were maintaining potassium levels within normal ranges and initiating a close follow-up plan. Due to the lack of clinical guidance in our case, many aspects of care, including surveillance, medications and genetic testing, remain unaddressed. Medical management includes aggressive correction with supplements, potassium-sparing diuretics and carbonic anhydrase inhibitors. Severe cases of dysrhythmias, especially ventricular fibrillation, require electrophysiology evaluation and possible implantation of a defibrillator to prevent sudden cardiac death.


Polyethylene wear from retrieved acetabular cups.
Laboratory wear testing of ultra high molecular weight polyethylene from 12 Charnley acetabular cups, removed after periods of up to 17.5 years showed that the large patient-to-patient variations in clinical penetration rate cannot be explained by batch-to-batch variation in the wear resistance of the material. Nor was there any evidence of a time-dependent degradation in wear resistance of the material.


[Natural history and eighteenth-century ideas regarding generation and heredity: Buffon and Bonnet].
The intellectual course of natural history reveals three conceptual approaches. The first was the taxonomic point of view, where naturalists worked to name and classify the living beings created by God. The second approach was provided by the eighteenth century's philosophical doctrine of mechanism, which lent natural history its method of endeavoring to comprehend the workings of organisms, inasmuch as the world "ran". Calling into question the adequacy of prior message, the third approach argued that living things display characteristics quite distinct from those of non-living matter, making it necessary to understand processes rather than simply decompose phenomena to then analyze them. This inadequacy became apparent at the moment when ideas of generation and heredity ascribed a reproductive history to living things, a history where the act of one fellow creature being formed by another plays an important role in coming to understand the workings of life. The paper analyzes these conceptual approaches from the perspective of Buffon's and Bonnet's ideas on reproduction and heredity, which represented opposite schools of thought: epigenesis and preformation.


RAR beta 2-mediated growth inhibition in HeLa cells.
Retinoic acid inhibits the growth of a variety of normal and transformed cells in vitro and in vivo. How retinoic acid inhibits cell growth is poorly understood but involves interactions between the ligand and a series of nuclear and cytoplasmic receptors. The nuclear receptors for retinoic acid are of two types, the RARs and the RXRs. Each can function as a ligand-inducible transcription enhancing factor. In previous studies, we have demonstrated that an isoform of one RAR, RAR beta 2, is transcriptionally up-regulated in senescent human dermal fibroblasts and senescent human mammary epithelial cells. Moreover, we have also shown that RAR beta 2 can inhibit oncogene-induced focus formation, in primary rat embryo fibroblasts, as effectively as the tumor suppressor gene p53. Here, we extend our studies of retinoid-regulated signal transduction pathways that inhibit cell proliferation by demonstrating that HeLa cells expressing an RAR beta 2 construct are growth inhibited by greater than 50% when compared to the parent cell lines. The RAR beta 2-expressing cell lines are inhibited further by the addition of exogenous all-trans-retinoic acid. Finally, soft agar assays show that the RAR beta 2-expressing cell lines also demonstrate an inhibition of growth in soft agar, when compared to the parent growth cell lines, and are inhibited further in the presence of added all-trans-retinoic acid. These data definitively show that RAR beta 2 can inhibit cell proliferation in an established tumor cell line and provide more strength to the notion that this isoform is an effective growth inhibitor in vitro and, most likely, in vivo.


Coffee intake and risk of hypertension: the Johns Hopkins precursors study.
Whether the increase in blood pressure with coffee drinking seen in clinical trials persists over time and translates into an increased incidence of hypertension is not known. We assessed coffee intake in a cohort of 1017 white male former medical students (mean age, 26 years) in graduating classes from 1948 to 1964 up to 11 times over a median follow-up of 33 years. Blood pressure and incidence of hypertension were determined annually by self-report, demonstrated to be accurate in this cohort. Consumption of 1 cup of coffee a day raised systolic blood pressure by 0.19 mm Hg (95% confidence interval, 0.02-0.35) and diastolic pressure by 0.27 mm Hg (95% confidence interval, 0.15-0.39) after adjustment for parental incidence of hypertension and time-dependent body mass index, cigarette smoking, alcohol drinking, and physical activity in analyses using generalized estimating equations. Compared with nondrinkers at baseline, coffee drinkers had a greater incidence of hypertension during follow-up (18.8% vs. 28.3%; P =.03). Relative risk (95% confidence interval) of hypertension associated with drinking 5 or more cups a day was 1.35 (0.87-2.08) for baseline intake and 1.60 (1.06-2.40) for intake over follow-up. After adjustment for the variables listed above, however, these associations were not statistically significant. Over many years of follow-up, coffee drinking is associated with small increases in blood pressure, but appears to play a small role in the development of hypertension.


Extracranial repair of cerebrospinal fluid rhinorrhea.
There has been a recent renewal of interest in the extracranial repair of cerebrospinal fluid rhinorrhea because of the relatively high morbidity associated with the transcranial approach. The authors describe an extracranial approach that involves packing of the sphenoid and ethmoid sinuses on the side of the CSF leak. A case of successful treatment of CSF rhinorrhea by this method is presented. The extracranial approach may be advantageous for the repair of CSF rhinorrhea and the authors advocate an increase in its utilization by neurosurgeons and otolaryngologists working as a team.


Erythema Nodosum (Report of two cases of erythema due to oral contraceptive).
Two cases of erythema nodosum probably due to oral contraceptives containing norgestrol and ethinyl oestradiol are reported. In one case, the skin lesions appeared one month after initiation of drug therapy. This seems to be the shortest latent period so far reported between oral contraceptive drug therapy and appearance of erythema nodosum.


The Association Between Cardiorespiratory Fitness and Cardiometabolic Risk in Children is Mediated by Abdominal Adiposity: The HAPPY Study.
It is unclear whether cardiorespiratory fitness (CRF) is independently linked to cardiometabolic risk in children. This study investigated a) the association between CRF level and presence of cardiometabolic risk disorders using health-related cut points, and b) whether these associations were mediated by abdominal adiposity in children. This was a cross-sectional design study. Anthropometry, biochemical parameters and CRF were assessed in 147 schoolchildren (75 girls) aged 10 to 14 years. CRF was determined using a maximal cycle ergometer test. Children were classified as 'fit' or 'unfit' according to published thresholds. Logistic regression was used to investigate the odds of having individual and clustered cardiometabolic risk factors according to CRF level and whether abdominal adiposity mediated these associations. Children classified as unfit had increased odds of presenting individual and clustered cardiometabolic risk factors (P < .05), but these associations no longer remained after adjusting for abdominal adiposity (P > .05). This study suggests that the association between CRF and cardiometabolic risk is mediated by abdominal adiposity in 10- to 14-year-old children and that abdominal adiposity may be a more important determinant of adverse cardiometabolic health in this age group.


Pharmacotherapy of chronic hospitalized schizophrenics: prescription practices.
Prescription practices were examined as part of a multinational study of chronic hospitalized schizophrenic patients. The study included a total of 768 patients from 8 countries. All patients had a diagnosis (ICD-9) of schizophrenia and met defined criteria for chronic hospitalization. The patients were treated with psychotropic drugs from 6 categories, i.e., neuroleptics, antidepressants, lithium salts, anxiolytics, anticonvulsants, and antiparkinsonian medications, as well as with a variety of nonpsychotropic drugs. The majority of the patients received concurrently more than 1 neuroleptic, medications from 2 or more categories, and neuroleptics combined with other agents. Polypharmacy appeared to be universal in this population.


Metamorphosis of spinal-projecting neurons in the brain of the sea lamprey during transformation of the larva to adult: normal anatomy and response to axotomy.
The spinal projecting system of the sea lamprey (Petromyzon marinus) has been used extensively in studies of axonal regeneration in both larvae and adults. However, little is known about the changes that are undergone by this system during metamorphosis. In order to determine the developmental changes in the size of the descending spinal projection and in the morphology of its neurons, larval, transforming, and adult lamprey brains were labeled by retrograde transport of horseradish peroxidase (HRP) injected into the spinal cord at 25% of body length. Examination of brain wholemount preparations revealed that the total number of labeled neurons doubled during metamorphosis. Most of this increase could be explained by elongation of reticulospinal axons from the rostralmost segments of the spinal cord to locations caudal to the injection site. There were no additions or deletions of either identified reticulospinal neurons or of reticulospinal nuclear groups between the larval and the adult stages. The proportions of Müller and Mauthner cells that were labeled reached a maximum of 93% during the early stages of metamorphosis. Axons of these neurons are known to project almost the entire length of the cord, even in larvae. Therefore, the efficiency of retrograde transport appears to be greater during metamorphosis than during larval or adult stages. While changes in efficiency of retrograde transport could account for some of the apparent increase in reticulospinal neuron numbers between larvae and animals undergoing metamorphosis, this could not contribute to the further increase in the apparent size of the reticulospinal system in the adult, since efficiency of retrograde labeling in these animals was lower than that at earlier stages. With retrograde labeling, a significant increase was seen in the profusion of dendritic arborization of some Müller and Mauthner cells during the early stages of metamorphosis. This correlated with an increase in the incidence of extreme axonal die-back, as indicated by the presence of retraction bulbs within the brainstem. However, intracellular injection of Neurobiotin in untransected animals showed similar degrees of dendritic arborization at all examined stages of development. Therefore, the dendritic profusion did not reflect developmental changes in neuronal morphology but rather reflected an increased sensitivity to axotomy during metamorphosis. We conclude that, during the transformation of the lamprey from the large larval to the adult form, there is little change in either the size or the dendritic morphology of the identified giant reticulospinal neurons. With respect to the smaller reticulospinal neurons, the distance of projection of many of their axons increases during metamorphosis, but there is very little increase in the number of reticulospinal neurons.


Evaluation of the Psychometric Properties of the Portuguese Peabody Developmental Motor Scales-2 Edition: A Study with Children Aged 12 to 48 Months.
The purpose of this study was to examine the psychometric properties of Peabody Developmental Motor Scales II (PDMS-2-Folio and Fewell, 2000) using a Portuguese sample. The validation of the Portuguese version of the PDMS-2 was applied according to the manual, for 392 children, from two institutions, from 12 to 48 months, with an analysis of the internal consistency (α Cronbach), of test-retest reliability (ICC) and construct validity (confirmatory factor analysis). The results of the confirmatory factorial analysis (χ2 = 55.614; df = 4; p = 0.06; χ2/df =13.904; SRMR (Standardized Root Mean Square Residual) = 0.065; CFI (Comparative Fit Index) = 0.99, TLI = 0.99, RMSEA (Root Mean Square Error of Approximation) = 0.068) of two factors (Gross Motor and Fine Motor) as the original version but correlated. Most of the subtests had good internal consistency (α = 0.85) and good test-retest stability (ICC = 0.98 to 0.99). The results indicated that the Portuguese version of the PDMS-2 is adequate and valid for assessing global and fine motor skills in children aged 12 to 48 months, and can be used as a reference tool by health and education professionals to assess motor skills and, thus, allowing to detect maladjustments, deficiencies or precocity, so that children can later receive appropriate intervention.


[Baicalin alleviates LPS-induced acute lung injury in rats by regulating macrophage polarization].
Objective To investigate the effect of baicalin on acute lung injury (ALI) induced by lipopolysaccharide (LPS), and to explore the roles of M1/M2 polarization of pulmonary macrophages and M1/M2 macrophage-mediated inflammation in the effect. Methods Sixty SD rats were divided into six groups: normal control group, LPS group, (10, 50, 100) mg/kg baicalin combined with LPS group, and dexamethasone (DEX) group. ALI models were established by intratracheal instillation of LPS. After 24 hours, bronchoalveolar lavage fluid (BALF) and bilateral lung tissues were collected. The pathological changes of rat lung tissue were observed by HE staining, and the wet/dry mass ratio (W/D) of lung tissue was measured; the contents of IL-1β, IL-6, TNF-α, and IL-10 in BALF were detected by ELISA; the M1 macrophage marker inducible nitric oxide synthase (iNOS) and the M2 macrophage marker CD206 in CD68 positive macrophages were detected by immunofluorescence cytochemical staining; the mRNA expressions of iNOS, IL-1β, Arg1, and CD206 were detected by real-time PCR, and the protein expressions of iNOS and Arg1 were detected by Western blot analysis. Results Baicalin significantly reduced lung lesions and lung water content in ALI rats, and down-regulated the secretion levels of pro-inflammatory cytokines TNF-α, IL-6, and IL-1β, while up-regulated the secretion of anti-inflammatory cytokine IL-10 in BALF. Baicalin significantly inhibited the lung macrophage polarization to M1 phenotype, and promoted the polarization to M2 phenotype. Baicalin significantly decreased the mRNA and protein expression levels of IL-1β and iNOS, while increased the mRNA and protein expression levels of CD206 and Arg1 in lung tissues. Conclusion Baicalin can inhibit the lung macrophage polarization to M1 phenotype, promote the polarization to M2 phenotype, and reduce the M1/M2 ratio, thereby alleviating the LPS-induced pulmonary inflammatory response in ALI rats.


Rapid induction by fungal elicitor of the synthesis of cinnamyl-alcohol dehydrogenase, a specific enzyme of lignin synthesis.
A fivefold increase in the extractable activity of cinnamyl-alcohol dehydrogenase, an enzyme of phenylpropanoid metabolism specific for lignin synthesis, was observed within 10 h of treatment of cell-suspension cultures of bean (Phaseolus vulgaris L.) with a high-molecular-mass elicitor preparation heat-released from mycelial cell walls of the bean pathogen Colletotrichum lindemuthianum. Elicitor caused a rapid, marked but transient increase in the synthesis of cinnamyl-alcohol dehydrogenase with maximum rates 2-3 h after elicitation, concomitant with the phase of rapid increase in enzyme activity. There is a close correspondence between increased polysomal mRNA activity encoding cinnamyl-alcohol dehydrogenase, as measured by incorporation of [35S]methionine into immunoprecipitable enzyme subunits in vitro, and the stimulation of enzyme synthesis in vivo in response to elicitor. This marked increase in polysomal mRNA activity represents an increase as a proportion of total cellular mRNA activity, indicating that elicitor does not stimulate synthesis of this enzyme by selective recruitment from the total pool of cellular mRNA. Elicitor stimulation of cinnamyl-alcohol dehydrogenase activity and enzyme synthesis is more rapid than previously observed for other proteins involved inducible defense mechanisms, such as enzymes of phytoalexin biosynthesis or the apoproteins of cell-wall hydroxyproline-rich glycoproteins.


Identification of Dynamic Modes in an Intrinsically Disordered Protein Using Temperature-Dependent NMR Relaxation.
The dynamic modes and time scales sampled by intrinsically disordered proteins (IDPs) define their function. Nuclear magnetic resonance (NMR) spin relaxation is probably the most powerful tool for investigating these motions delivering site-specific descriptions of conformational fluctuations from throughout the molecule. Despite the abundance of experimental measurement of relaxation in IDPs, the physical origin of the measured relaxation rates remains poorly understood. Here we measure an extensive range of auto- and cross-correlated spin relaxation rates at multiple magnetic field strengths on the C-terminal domain of the nucleoprotein of Sendai virus, over a large range of temperatures (268-298 K), and combine these data to describe the dynamic behavior of this archetypal IDP. An Arrhenius-type relationship is used to simultaneously analyze up to 61 relaxation rates per amino acid over the entire temperature range, allowing the measurement of local activation energies along the chain, and the assignment of physically distinct dynamic modes. Fast (τ ≤ 50 ps) components report on librational motions, a dominant mode occurs on time scales around 1 ns, apparently reporting on backbone sampling within Ramachandran substates, while a slower component (5-25 ns) reports on segmental dynamics dominated by the chain-like nature of the protein. Extending the study to three protein constructs of different lengths (59, 81, and 124 amino acids) substantiates the assignment of these contributions. The analysis is shown to be remarkably robust, accurately predicting a broad range of relaxation data measured at different magnetic field strengths and temperatures. The ability to delineate intrinsic modes and time scales from NMR spin relaxation will improve our understanding of the behavior and function of IDPs, adding a new and essential dimension to the description of this biologically important and ubiquitous class of proteins.


Effects of CHADS2 score, echocardiographic and haematologic parameters on stroke severity and prognosis in patients with stroke due to nonvalvular atrial fibrillation.
The aim of this study is to evaluate utility of CHADS2 score to estimate stroke severity and prognosis in patients with ischemic stroke due to non-valvular atrial fibrillation (AF) in addition to evaluate effects of hematologic and echocardiographic findings on stroke severity and prognosis. This prospective study included 156 ischemic stroke cases due to non-valvular AF in neurology ward of Trakya University Medical School between March 2013-March 2015. National Institute of Health Stroke (NIHS) score was used to evaluate severity of stroke at admission. Carotid and vertebral Doppler ultrasonography findings, brain computed tomography (CT) and magnetic resonance imaging (MRI) of the cases were evaluated. Left atrial diameter and ejection fraction (EF) values were measured. CHADS2 score was calculated. Modified Rankin Scale was used to rate the degree of dependence. Effects of age and sex of the patients, presence of diabetes mellitus (DM), Congestive Heart Failure (CHF), Cerebrovascular Disease (CVD) and C-reactive protein (CRP) levels on CHADS2, NIHS, and mRS were evaluated. In patients with age ≥75, mean NIHS score was 3.3 points and mean mRS score was 1.02 points higher, than in patient below 75 years of age. Compared with the mild risk group, cases in the high risk group had older age, higher serum D-dimer, fibrinogen and CRP levels and lower EF. A positive relation was detected between stroke severity and Hemorrhagic Transformation (HT), previous CVD history, and presence of CHF. A significant association was found between increased stroke severity and Early Neurological Deterioration (END) development. Older age, higher serum fibrinogen, D-dimer, CRP and lower EF values were associated with poor prognosis. History of CVD and presence of CHF were associated with poor prognosis. END development was found to be associated with poor prognosis. In the high-risk group, 30.3% (n = 33) had END. Among those in the high-risk group according to the CHADS2 score, END development rate was found to be significantly higher than in the moderate risk group (p <0.05). There was a strong positive correlation between CHADS2 and NIHS scores. mRS score increased with increasing CHADS2 score and there was a strong correlation between them. Effect of stroke severity on prognosis was assessed and a positive correlation was found between NIHS score and mRS value. Our study demonstrated the importance of CHADS2 score, haemostatic activation and echocardiographic findings to assess stroke severity and prognosis. Knowing factors which affect stroke severity and prognosis in patients with ischemic stroke may be directive to decide primary prevention and stroke management.


Differences in innate immune responses correlate with differences in murine susceptibility to Chlamydia muridarum pulmonary infection.
We investigated the phenotypic basis for genetically determined differences in susceptibility and resistance to Chlamydia muridarum pulmonary infection using BALB/c and C57BL/6 mice. Following C. muridarum intranasal inoculation, the intensity of infection was very different between BALB/c and C57BL/6 beginning as early as 3 days post-infection. Intrapulmonary cytokine patterns also differed at early time-points (days 2 and 4) between these two strains of mice. The early recruitment of neutrophils to lung tissue was greater in BALB/c than in C57BL/6 mice and correlated with a higher number of inclusion forming units (IFU) of C. muridarum. At day 12 post-infection, BALB/c mice continued to demonstrate a greater burden of infection, significantly higher lung cytokine levels for tumour necrosis factor-alpha and interleukin-17 (IL-17) and a significantly lower level for interferon-gamma than did C57BL/6 mice. In vitro, bone-marrow-derived dendritic cells (BMDCs) from BALB/c mice underwent less functional maturation in response to C. muridarum infection than did BMDCs from C57BL/6 mice. The BMDCs of BALB/c mice expressed lower levels of activation markers (CD80, CD86, CD40 and major histocompatibility complex class II) and secreted less IL-12 and more IL-23 than BMDCs from C57BL/6 mice. Overall, the data demonstrate that the differences exhibited by BALB/c and C57BL/6 mice following C. muridarum pulmonary infection are associated with differences in early innate cytokine and cellular responses that are correlated with late differences in T helper type 17 versus type 1 adaptive immune responses.


Covered stents may provide extra protection during carotid artery stenting in high risk patients with an excessive thrombus burden.
Carotid artery disease is an important cause of mortality and morbidity related to atherosclerosis. Recently, percutaneous intervention procedures have been widely used to treat atherosclerotic carotid artery disease. We report the case of a 57-year-old male patient with a history of acute amaurosis fugax. Carotid angiography was performed as blood pressure differed between his left and right arms and there was a pan-systolic murmur on the left common carotid artery. Total occlusion of the proximal right brachiocephalic artery and a thrombus occluding 90-99% of the left internal carotid artery were detected by carotid angiogram. A self-expanding graft-covered stent was successfully implanted and there were no complications. This case shows that graft-covered stents may be a good alternative technique in special situations.


[Effects of slope direction on soil nutrient and its ecological stoichiometry in bamboo forest].
We analyzed the effects of slope direction on soil nutrients and ecological stoichiometry by collecting soil samples from different slope directions (shady slope and sunny slope) of the bamboo forest in Longyou County, Zhejiang Province. The results showed that soil nutrients were affected by slope direction and soil depth. The nutrients level of soils in the sampling area showed the trends of shady slope > sunny slope, and surface soil > bottom soil. Compared to sunny slope, the cation exchange capacity (CEC), the contents of total organic carbon, total nitrogen, alkaline hydrolyzed nitrogen, available phosphorus, total potassium and available potassium of shady soils significantly increased by 43.7%, 103.8%, 92.0%, 75.5%, 22.4%, 89.4% and 240.7%, respectively. There was no significant difference in total phosphorus contents between shady slope and sunny slope. At all soil layers, there was no significant difference of C/N ratio between shady and sunny slopes. The average C/P ratio of shady slope was 180.8%, 42.0% and 54.3% higher than that of sunny slope at 0-20 cm, 20-40 cm and 40-60 cm, respectively. At each soil layer, the average C/K and N/K ratios between shady and sunny slopes had no significant difference. The average C/K and N/K ratios of shady slope and sunny slope were all significantly different among the three soil layers. In the shady slope, the contents of soil organic carbon showed significantly positive correlation with total nitrogen, total phosphorus, total potassium, and soil available nutrients. Overall, soil nutrients and ecological stoichiometry characteristics of shady slope of bamboo forest were superior to those of sunny slope.


Delayed diagnosis of thoracic esophageal rupture due to blunt abdominal trauma without chest trauma: A case report.
Thoracic esophageal rupture due to blunt trauma is very rare. Moreover, there have been no reports of thoracic esophageal rupture due to blunt abdominal trauma without chest trauma. We describe a rare case of esophageal rupture due to blunt abdominal trauma in a young female patient. Operation was delayed due to a misdiagnosis of chylothorax, and esophageal repair was performed six days after trauma. Postoperative esophageal leak developed and was treated with esophageal stent. She was discharged two months after surgery without sequelae. It is important to consider esophageal rupture as a differential diagnosis even in patients with only abdominal trauma, when in doubt.


Ranking procedures for repeated measures designs with missing data: Estimation, testing and asymptotic theory.
We develop purely nonparametric methods for the analysis of repeated measures designs with missing values. Hypotheses are formulated in terms of purely nonparametric treatment effects. In particular, data can have different shapes even under the null hypothesis and therefore, a solution to the nonparametric Behrens-Fisher problem in repeated measures designs will be presented. Moreover, global testing and multiple contrast test procedures as well as simultaneous confidence intervals for the treatment effects of interest will be developed. All methods can be applied for the analysis of metric, discrete, ordinal, and even binary data in a unified way. Extensive simulation studies indicate a satisfactory control of the nominal type-I error rate, even for small sample sizes and a high amount of missing data (up to 30%). We apply the newly developed methodology to a real data set, demonstrating its application and interpretation.


Localization of carboxypeptidase I in germinating barley grain.
Activity measurements and Northern blot hybridizations were used to study the temporal and spatial expression of carboxypeptidase I in germinating grains of barley (Hordeum vulgare L. cv Himalaya). In the resting grain no carboxypeptidase I activity was found in the aleurone layer, scutellum, or starchy endosperm. During germination high levels of enzyme activity appeared in the scutellum and in the starchy endosperm but only low activity was found in the aleurone layer. No mRNA for carboxypeptidase I was observed in the resting grain. By day 1 of germination the mRNA appeared in the scutellum where its level remained high for several days. In contrast, little mRNA was observed in the aleurone layer. These results indicate that the scutellum plays an important role in the production of carboxypeptidase I in germinating barley grain.


Cortical network analysis in patients affected by schizophrenia.
In the present study, we studied the structural changes of the brain functional network in a group of schizophrenic (SCHZ) patients during a 2-back working memory task. Cortical signals were obtained from scalp EEG signals through the high-resolution EEG technique, which relies on realistic head models and linear inverse solutions. Functional networks were estimated by computing the spectral coherence--i.e. a measure of synchronization in the frequency domain--between the time series of all the available cortical sources. To analyze those cortical networks we followed a theoretical graph approach by computing the network density as the total number of links and the node degree as the number of links of each cortical source. The major result suggest that in the Alpha2 frequency band (11-13 Hz) the cortical functional networks of the SCHZ patients present the largest differences when compared with those of a group of control (CTRL) subjects. In particular, the structure of the SCHZ network altered radically during the memory task, as the number of links that were different from the REST condition increased sensibly with respect to the CTRL network. In addition, a compensatory mechanism was found in the SCHZ patients during the correct performance of the memory task where the node degree showed a frontal asymmetry with higher activation of the left frontal lobe--i.e. higher number of connections--in the Alpha2 frequency band.


The influence of dental caries on body growth in prepubertal children.
Dental decay and dental treatment are suggested to be related to body growth in children. The aim of this study was first to assess the relation between dental caries and body proportions cross-sectionally in a Suriname caries child population and secondly to investigate whether dental treatment had a significant influence on body growth of these children in a randomised controlled trial using different treatment strategies. Three hundred eighty 6-year-old children with untreated dental decay participated in the study. Caries prevalence and presence of dentogenic infections were recorded. All children were randomly assigned to four different treatment groups ranging from full dental treatment to no invasive treatment at all. Body growth was evaluated by children's height, weight and body mass index. Participants were evaluated after 6 months and 1, 2 and 3 years. Cross-sectionally, negative correlations were observed between anthropometric measures and the number of untreated carious surfaces and caries experience of the children. Next, no significant differences in growth pattern between the treatment groups were observed. Based on these results, it is suggested that caries activity is a negative predictor for body growth in children and dental intervention does not show significant improvement within 3 years.


Excited-state photodynamics of perylene-porphyrin dyads. 5. Tuning light-harvesting characteristics via perylene substituents, connection motif, and three-dimensional architecture.
Seven perylene-porphyrin dyads were examined with the goal of identifying those most suitable for components of light-harvesting systems. The ideal dyad should exhibit strong absorption by the perylene in the green, undergo rapid and efficient excited-state energy transfer from perylene to porphyrin, and avoid electron-transfer quenching of the porphyrin excited state by the perylene in the medium of interest. Four dyads have different perylenes at the p-position of the meso-aryl group on the zinc porphyrin. The most suitable perylene identified in that set was then incorporated at the m- or o-position of the zinc porphyrin, affording two other dyads. An analogue of the o-substituted architecture was prepared in which the zinc porphyrin was replaced with the free base porphyrin. The perylene in each dyad is a monoimide derivative; the perylenes differ in attachment of the linker (either via a diphenylethyne linker at the N-imide or an ethynylphenyl linker at the C9 position) and the number (0-3) of 4-tert-butylphenoxy groups (which increase solubility and slightly alter the electrochemical potentials). In the p-linked dyad, the monophenoxy perylene with an N-imide diphenylethyne linker is superior in providing rapid and essentially quantitative energy transfer from excited perylene to zinc porphyrin with minimal electron-transfer quenching in both toluene and benzonitrile. The dyads with the same perylene at the m- or o-position exhibited similar results except for one case, the o-linked dyad bearing the zinc porphyrin in benzonitrile, where significant excited-state quenching is observed; this phenomenon is facilitated by close spatial approach of the perylene and porphyrin and the associated thermodynamic/kinetic enhancement of the electron-transfer process. Such quenching does not occur with the free base porphyrin because electron transfer is thermodynamically unfavorable even in the polar medium. The p-linked dyad containing a zinc porphyrin attached to a bis(4-tert-butylphenoxy)perylene via an ethynylphenyl linker at the C9 position exhibits ultrafast and quantitative energy transfer in toluene; the same dyad in benzonitrile exhibits ultrafast (<0.5 ps) perylene-to-porphyrin energy transfer, rapid (∼5 ps) porphyrin-to-perylene electron transfer, and fast (∼25 ps) charge recombination to the ground state. Collectively, this study has identified suitable perylene-porphyrin constructs for use in light-harvesting applications.


Trends and Disparities in Inpatient Rehabilitation of Adolescents: The Effect of Demographics, Injury Characteristics, and Facility Type.
Intensive rehabilitation of adolescents occurs in general and pediatric inpatient rehabilitation facilities (IRFs), but differences in admission and outcomes by facility type have not been thoroughly investigated, particularly among persons with spinal cord injury (SCI). To investigate factors related to admission to pediatric or adult IRFs among adolescents and compare the rehabilitation outcomes between facility types. Using a single-center retrospective cohort study design, demographic information, medical data, and rehabilitation outcomes were obtained by chart review of patients aged 15 to 18 years who received a rehabilitation medicine consultation at an urban level 1 trauma center between 2017 and 2019 (N = 96). Analysis was performed using R statistical software. SCI was the second most common diagnosis (21% of patients) and accounted for 36% of inpatient rehabilitation admissions. SCI patients were more likely to undergo rehabilitation at the level 1 trauma versus pediatric facility (relative risk [RR] = 2.43; 95% confidence interval [CI] = 1.08-5.44) compared to traumatic brain injury patients. Admission to trauma versus pediatric IRF was also associated with Black compared to White race (RR = 2.5; CI = 1.12-5.56), violence compared to other etiologies of injury (RR = 2.0; CI = 1.10-3.77), and Medicaid compared to private insurance (RR = 2.15; CI = 1.01-4.59). Admission to pediatric IRF was associated with longer length of stay than admission to adult IRF when adjusted for diagnosis (30.86 ± 21.82 vs. 24.33 ± 18.17 days; p = .046), but Functional Independence Measures did not differ. Adolescents with SCI and those experiencing systemic disadvantages, including racism and poverty, were more likely to be admitted to trauma compared to pediatric IRF.


Cutaneous plasmacytomas with amyloid in six dogs.
Cutaneous plasmacytomas associated with local deposition of amyloid were diagnosed by light microscopy in a series of six older dogs (mean age 10.7 years) consisting of two Cocker Spaniels, a Poodle, a Weimeraner, and two mixed-breed dogs. The neoplasms occurred on the digits (2 dogs), forelimb (2 dogs), lip (1 dog), and ear (1 dog). In most cases, groups of neoplastic plasma cells were widely separated by large homogeneous islands of amyloid. The neoplastic cells had characteristic plasmacytoid features, but the degree of pleomorphism varied greatly between different neoplasms. In four of the six tumors, the diagnosis of plasmacytoma was confirmed by the demonstration of a monoclonal plasma cell population using immunofluorescent staining for anti-canine immunoglobulins. In these tumors, the neoplastic cells reacted with only one class of immunoglobulins (IgG). The amyloid did not react with any of the reagents used. The suspicion that the amyloid was of immunoglobulin origin (primary amyloid) was supported by its retention of birefringence under polarized light after treatment with potassium permanganate and staining with Congo red.


[Clinical analysis of myasthenia gravis accompanied by invasive thymoma].
Over the last 20 years, 6 patients with myasthenia gravis and invasive thymoma have been seen in our Department. These patients underwent non-total excision cases. This represents 2.5 percent of 242 myasthenia gravis patients in our series. We can see 17 such patients in the literature in Japan including our cases. The age ranged 20 to 77 years and the ratio of male to female was 10:7. Post-operative therapeutic methods for invasive thymoma included irradiation, steroid therapy and combination chemotherapy. Every method showed good therapeutic results, but steroid therapy and chemotherapy showed especially good therapeutic effects in the early stage of the disease. Though these therapeutic effects were better than those obtained in other malignant tumors, thymomas tended to reappear within several years, when tumors showed no response to any therapeutic method. It is well known that steroid therapy reduces the level of antiacetylcholine receptor antibody, whereas chemotherapy also reduces antiacetylcholine receptor antibody. Over all 5-year survival rate was 53%, and the 10-year survival rate was 29%. Three out of 6 cases of death were due to myasthenic crisis (50%) and 2 out of 6 cases were due to invasive tumor itself (38%). These results suggested that total excision of the thymoma, if possible, and for the remaining tumor, high doses of adrenocorticosteroids and combination chemotherapy seem to be treatments of choice.


[Radiotherapy concomitant with first-generation epidermal growth factor receptor tyrosine kinase inhibitors in the treatment of brain metastases from non small cell lung cancer: a meta-analysis].
To evaluate the efficacy of whole brain radiotherapy (WBRT) or/and stereotactic radiotherapy (SRT) concomitant with first-generation epidermal growth factor receptor tyrosine kinase inhibitors (TKI), like erlotinib and gefitinib, in the treatment of brain metastases from non-small cell lung cancer (NSCLC). Comprehensive searches were performed in different databases, including EMBASE, MEDLINE and Cochrane Library, etc. All randomized controlled trials (RCTs) and non-randomized controlled trials (non-RCTs) comparing radiotherapy (RT) plus TKI with RT alone in the treatment of brain metastases from NSCLC were included for meta-analysis with Cochrane Collaboration's RevMan5. 2 software. The primary end-points were objective remission (OR), disease control (DC) and toxicity while the secondary end-point was overall survival (OS). Three RCTs and one non-randomized controlled trial were selected and included for final analysis. The RT + TKI group had significant improvements in OR rate (RR = 1. 80, P <0. 01) and DC rate (RR = 1. 31, P <0. 01). Meanwhile, rash (RR = 14. 36, P = 0. 003) and diarrhea (RR = 3. 49, P = 0. 005) increased in the RT + TKI group. Yet there was no significant difference in nausea or vomiting (P = 0. 20). For NSCLC with brain metastases, RT plus TKI could improve OR, DC and OS, but there was a higher incidence of rash and diarrhea. There fore future high-quality and prospective RCTs are warranted to confirm the clinical efficacy of RT plus TKI in the treatment of NSCLC with brain metastases.


Etching conditions for resin-modified glass ionomer cement for orthodontic brackets.
This study reports the tensile bond strength of orthodontic eyelets (RMO, Inc, Denver, Colo) bonded to human extracted teeth with a resin-modified glass ionomer cement (RMGIC) (Fuji Ortho LC, GC America, Alsip, Ill) and various acid etchants (Etch-37 and All-Etch, Bisco, Schaumburg, Ill; Ultra Etch, 3M Unitek, St Paul, Minn) for enamel preparation before bonding. The enamel etch conditions were as follows: 37% phosphoric acid with silica; 37% phosphoric acid, silica-free; 10% phosphoric acid, silica-free; 10% polyacrylic acid; and unetched enamel. Bond strength was measured by pulling in tension on the eyelet with a 0.018-in steel wire perpendicular to the enamel surface with a testing machine (Instron model 1125, Canton, Mass) at a speed of 2 mm/min. A light-cured resin cement (Transbond XT, 3M Unitek, Monrovia, Calif) applied to enamel etched with 37% phosphoric acid containing silica served as a control. Each group included 30 specimens. The Weibull distribution (m) was used for statistical analysis with a 90% CI. The different etchants used with RMGIC did not affect tensile bond strength. The resin cement group had the highest tensile strength. Significantly lower bond strengths were observed when glass ionomer cement was used to bond orthodontic attachments to nonetched teeth. However, unlike resin cement, RMGIC can bond effectively to etched teeth in a moist environment without an additional bonding agent.


Ultrastructure Organization of Collagen Fibrils and Proteoglycans of Stingray and Shark Corneal Stroma.
We report here the ultrastructural organization of collagen fibrils (CF) and proteoglycans (PGs) of the corneal stroma of both the stingray and the shark. Three corneas from three stingrays and three corneas from three sharks were processed for electron microscopy. Tissues were embedded in TAAB 031 resin. The corneal stroma of both the stingray and shark consisted of parallel running lamellae of CFs which were decorated with PGs. In the stingray, the mean area of PGs in the posterior stroma was significantly larger than the PGs of the anterior and middle stroma, whereas, in the shark, the mean area of PGs was similar throughout the stroma. The mean area of PGs of the stingray was significantly larger compared to the PGs, mean area of the shark corneal stroma. The CF diameter of the stingray was significantly smaller compared to the CF diameter in the shark. The ultrastructural features of the corneal stroma of both the stingray and the shark were similar to each other except for the CFs and PGs. The PGs in the stingray and shark might be composed of chondroitin sulfate (CS)/dermatan sulfate (DS) PGs and these PGs with sutures might contribute to the nonswelling properties of the cornea of the stingray and shark.


[Structural and hormonal alterations in the pancreas after hepatectona in dogs].
Humoral factors in the portal blood have often been suggested to play an important role in hepatic regeneration after hepatectomy. However, the correlation among insulin, glucagon and somatostatin, and islet cells of Langerhans has not been studied. Forty percent hepatectomy was carried out in dogs and intravenous glucose administration (0.5 g/kg) was the function of the pancreas to release one week after and one month after surgery to estimate administered intravenously before surgery, the plasma insulin, glucagon and somatostatin. In addition, islet cells of Langerhans were investigated with PAP technique. The following results were obtained. Alterations of carbohydrate metabolism developed as a result of portal plasma insulin deficiency one week after surgery, but granules in B cells were demonstrated as much as preoperatively. It was suggested that hypersomatostatinemia suppressed insulin release from B cells. Alterations of carbohydrate metabolism was restored to normal and portal plasma insulin response was increased one month after surgery. These indicate that hepatic regeneration after hepatectomy may contribute to these changes. Islets cell of Langerhans were hypertrophic and particularly, D cells were increased after surgery.


The thermal stability of immunoglobulin: unfolding and aggregation of a multi-domain protein.
The denaturation of immunoglobulin G was studied by different calorimetric methods and circular dichroism spectroscopy. The thermogram of the immunoglobulin showed two main transitions that are a superimposition of distinct denaturation steps. It was shown that the two transitions have different sensitivities to changes in temperature and pH. The two peaks represent the F(ab) and F(c) fragments of the IgG molecule. The F(ab) fragment is most sensitive to heat treatment, whereas the F(c) fragment is most sensitive to decreasing pH. The transitions were independent, and the unfolding was immediately followed by an irreversible aggregation step. Below the unfolding temperature, the unfolding is the rate-determining step in the overall denaturation process. At higher temperatures where a relatively high concentration of (partially) unfolded IgG molecules is present, the rate of aggregation is so fast that IgG molecules become locked in aggregates before they are completely denatured. Furthermore, the structure of the aggregates formed depends on the denaturation method. The circular dichroism spectrum of the IgG is also strongly affected by both heat treatment and low pH treatment. It was shown that a strong correlation exists between the denaturation transitions as observed by calorimetry and the changes in secondary structure derived from circular dichroism. After both heat- and low-pH-induced denaturation, a significant fraction of the secondary structure remains.


Hair Transplantation Controversies.
Hair transplant surgery creates consistently natural appearing transplanted hair for men. It is increasingly popular procedure to restore natural growing hair for men with hair loss. To review some current controversies in hair transplant surgery. Review of the English PubMed literature and specialty literature in hair transplant surgery. Some of the controversies in hair transplant surgery include appropriate donor harvesting technique including elliptical donor harvesting versus follicular unit extraction whether manual versus robotic, the role of platelet-rich plasma and low-level light surgery in hair transplant surgery. Hair transplant surgery creates consistently natural appearing hair. As with all techniques, there are controversies regarding the optimal method for performing the procedure. Some of the current controversies in hair transplant surgery include optimal donor harvesting techniques, elliptical donor harvesting versus follicular unit extraction, the role of low-level light therapy and the platelet-rich plasma therapy in the procedure. Future studies will further clarify their role in the procedure.


Kinetics of lead in bone and blood after end of occupational exposure.
In 14 retired lead workers, followed for over 18 years after end of exposure, repeated analyses of lead levels in finger bone by an in vivo X-ray fluorescence method revealed a decrease of lead concentration. The data were analysed using an exponential retention model. For the whole group the biological half-time was 16 (asymptotic 95% confidence interval, CI 12,23) years. The median of the estimated bone lead levels at the end of exposure was 85 micrograms.g-1 above the "background" (3 micrograms.g-1). A simultaneous follow-up of blood lead levels displayed a decrease, which could be described by a tri-exponential retention model with group half-times of 34 (CI 29,41) days, 1.2 (CI 0.9,1.8) years, and 13 (CI 10,18) years, respectively. The median of the estimated blood lead levels at the end of exposure for the three components were 0.49, 0.61, and 1.1 mumol.l-1 above the "background" (0.38-0.56 mumol.l-1), respectively. The well-documented decrease of lead exposure in the general population over the years, urged the use of a decreasing "background" of blood lead during the time of the study. The slowest of the three components represented the skeleton (probably mainly cortical bone), as did mainly probably also the intermediate one (trabecular bone). The data show the rather slow turnover of lead in the skeleton, the usefulnes of in vivo skeletal lead measurements as a long-term exposure index, and the importance of bone as a source of "endogenous" lead exposure.


Biodiversity and ecosystem functioning in evolving food webs.
We use computer simulations in order to study the interplay between biodiversity and ecosystem functioning (BEF) during both the formation and the ongoing evolution of large food webs. A species in our model is characterized by its own body mass, its preferred prey body mass and the width of its potential prey body mass spectrum. On an ecological time scale, population dynamics determines which species are viable and which ones go extinct. On an evolutionary time scale, new species emerge as modifications of existing ones. The network structure thus emerges and evolves in a self-organized manner. We analyse the relation between functional diversity and five community level measures of ecosystem functioning. These are the metabolic loss of the predator community, the total biomasses of the basal and the predator community, and the consumption rates on the basal community and within the predator community. Clear BEF relations are observed during the initial build-up of the networks, or when parameters are varied, causing bottom-up or top-down effects. However, ecosystem functioning measures fluctuate only very little during long-term evolution under constant environmental conditions, despite changes in functional diversity. This result supports the hypothesis that trophic cascades are weaker in more complex food webs.


Super-Mini Percutaneous Nephrolithotomy in the Treatment of Pediatric Nephrolithiasis: Evaluation of the Initial Results.
To evaluate the efficacy and safety of super-mini percutaneous nephrolithotomy (SMP) in the treatment of pediatric kidney stones. We reviewed the records of 111 children with renal stones treated with SMP technique in four different centers between September 2014 and September 2015. The indications for SMP treatment in all these kids were either previously failed shock wave lithotripsy or retrograde intrarenal surgery approaches, according to their parents' preferences. Nephrostomy tracts used in the SMP system ranged from 10F to 14F in size. Lithotripsy was performed using either a Holmium laser or pneumatic lithotripter. Perioperative and postoperative parameters along with operative data were recorded in detail and stone components were analyzed by infrared spectroscopy. This study included 71 boys and 40 girls with a mean age of 3.90 ± 3.53 years (range 0.5-15). The mean stone burden was 1.4 ± 0.6 cm (range 0.8-4.8). Mean operative time was 39.4 ± 26.2 minutes (range 7-105). The mean hemoglobin drop was 10.2 ± 7.1 g/L (range 0-25) and no transfusion was needed. Significant complications were observed in 17 (15.3%) children with 10 and 7 cases in Clavien grade I and grade II, respectively. Complete stone clearance on postoperative day 1 and on 3-month follow-up was 84.7% (94/111) and 90.1% (100/111), respectively. Ninety-five (85.6%) children did not require any type of catheters (total tubeless). The mean hospital stay was 2.7 ± 1.5 days (range 1-7). Our preliminary data demonstrated that SMP was safe and effective. SMP could be a feasible treatment option for pediatric stone disease. Further randomized controlled trials are still needed to prove the efficacy of using the SMP system in children, particularly in those with larger stones.


Polymeric indoximod based prodrug nanoparticles with doxorubicin entrapment for inducing immunogenic cell death and improving the immunotherapy of breast cancer.
Immunotherapy based on host immunity has emerged as a powerful therapeutic strategy for tumor treatment. However, utilizing the immune system against tumors often fails to result in a durable immune response due to insufficient immunogenicity and the immunosuppressive conditions in the tumor microenvironment. Herein, we developed prodrug-based nanoparticles (DOX/IND@NPs) for the codelivery of indoximod (IND), an indoleamine 2,3-dioxygenase (IDO) inhibitor that can block the IDO pathway and generate antitumor immunity, and doxorubicin, a DNA-damaging therapeutic agent that can induce tumor immunogenic cell death (ICD). The nanocarrier was designed for tumor chemoimmunotherapy, synergistically promoting immunogenicity and modulating the immunosuppressive tumor microenvironment (ITME). Our data showed that DOX induced tumor immunogenicity and increased the infiltration of CD8 + T cells into the tumor microenvironment; nevertheless, immunosuppressive immune cell components, such like regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs), and tumor associated macrophages (TAMs), hindered the antitumor efficacy. The introduction of IND reduced the levels of these protumor immune cells within the tumor microenvironment and further enhanced CD8 + T cell infiltration and the CD8 +/Treg cell ratio. Moreover, significant reductions in vascular endothelial growth factor (VEGF), MMP9, and CD31 (a vascular marker) expression levels were observed after DOX-IND nanoparticle treatment. This resulted in obvious tumor regression in a murine breast cancer model compared to reference formulations, indicating that the codelivery of DOX and IND is a potent potential strategy for breast cancer chemoimmunotherapy.


Intense fluorescence of metal-to-ligand charge transfer in [Pt(0)(binap)(2)] [binap = 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl].
[Pt(0)(binap)(2)] (binap = 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl) is found to exhibit a luminescence from metal-to-ligand charge transfer state (MLCT) with a quantum yield of 0.12 and a lifetime of 1.2 micros in toluene at an ambient temperature. Prompt fluorescence with a quantum yield of 1.6 x 10(-)(4) is observed by means of a picosecond time-correlated single photon counting technique. The spectrum of the steady-state luminescence is almost identical to that of the prompt fluorescence, indicating that the intense luminescence is mainly delayed fluorescence from thermally activated (1)MLCT. The analysis of the temperature-dependent emission indicates that the energy difference between the (1)MLCT and (3)MLCT is 1.15 x 10(3) cm(-)(1). The lifetime of the prompt fluorescence is determined to be 3.2 ps from the decay of stimulated emission overlapped on subpicosecond transient absorption spectra. The lifetime of the (1)MLCT is much longer than expected from the large spin-orbit coupling constant of 5d (Pt) electrons (4000 cm(-)(1)). Theoretical analysis based on density functional theory reveals that structural distortion in the MLCT states causes large energy splitting between HOMO and HOMO - 1, which prevents a very fast ISC induced by strong spin-orbit interactions between these orbitals. The relatively slow ISC is therefore induced by weak spin-orbit interactions (ca. 50 cm(-)(1)) between ligand-centered molecular orbitals. Theoretical calculations indicate that the phosphorescence observed at lower temperatures is due to intensity borrowing from 4(1)B(2) --> GS transition. However, the large energy difference between HOMO and HOMO - 2 reduces the extent of mixing between the lowest (3)MLCT and 4(1)B(2) due to spin-orbit interaction, thereby decreasing the radiative rate of the phosphorescence.