Cardiotoxicity in anthracycline therapy: Prevention strategies.
The increasing use of anthracyclines, together with the longer survival of cancer patients, means the toxic effects of these drugs need to be monitored. In order to detect, prevent or mitigate anthracycline-induced cardiomyopathy, it is essential that all patients undergo a rigorous initial cardiovascular assessment, followed by close monitoring. Several clinical trials have shown the cardioprotective effect of non-pharmacological measures such as exercise, healthy lifestyles, control of risk factors and treatment of comorbidities; a cardioprotective effect has also been observed with pharmacological measures such as beta-blockers, angiotensin-converting enzyme inhibitors, angiotensin receptor antagonists, statins, dexrazoxane and liposomal formulations. However, there are currently no guidelines for managing prevention in these patients. In this review the authors discuss the state of the art of the assessment, monitoring, and, above all, the prevention of anthracycline-induced cardiotoxicity.


Biochemical and Structural Insights into Carbonic Anhydrase XII/Fab6A10 Complex.
6A10 is a CA XII inhibitory monoclonal antibody, which was demonstrated to reduce the growth of cancer cells in vitro and in a xenograft model of lung cancer. It was also shown to enhance chemosensitivity of multiresistant cancer cell lines and to significantly reduce the number of lung metastases in combination with doxorubicin in mice carrying human triple-negative breast cancer xenografts. Starting from these data, we report here on the development of the 6A10 antigen-binding fragment (Fab), termed Fab6A10, and its functional, biochemical, and structural characterization. In vitro binding and inhibition assays demonstrated that Fab6A10 selectively binds and inhibits CA XII, whereas immunohistochemistry experiments highlighted its capability to stain malignant glioma cells in contrast to the surrounding brain tissue. Finally, the crystallographic structure of CA XII/Fab6A10 complex provided insights into the inhibition mechanism of Fab6A10, showing that upon binding, it obstructs the substrate access to the enzyme active site and interacts with CA XII His64 freezing it in its out conformation. Altogether, these data indicate Fab6A10 as a new promising therapeutic tool against cancer.


Two Novel Flavonoids and Cytotoxicity Evaluation from Cryptocarya yunnanensis.
Two new flavonoids, cryunchalcone (1) and cryptoyunnanone I (2), were isolated from the leaves and twigs of Cryptocarya yunnanensis. Their structures were elucidated by the detailed spectroscopic data analysis and electronic circular dichroism (ECD) calculations. Cryunchalcone (1) is a biflavonoid constructed by a dihydrochalcone coupled with a chalcone through an unprecedented C-2''-C-6 linkage. Cryptoyunnanone I (2) is a unique complex flavanone bearing a phenylpropanoid moiety.


High Fructose Drives the Serine Synthesis Pathway in Acute Myeloid Leukemic Cells.
A significant increase in dietary fructose consumption has been implicated as a potential driver of cancer. Metabolic adaptation of cancer cells to utilize fructose confers advantages for their malignant growth, but compelling therapeutic targets have not been identified. Here, we show that fructose metabolism of leukemic cells can be inhibited by targeting the de novo serine synthesis pathway (SSP). Leukemic cells, unlike their normal counterparts, become significantly dependent on the SSP in fructose-rich conditions as compared to glucose-rich conditions. This metabolic program is mediated by the ratio of redox cofactors, NAD+/NADH, and the increased SSP flux is beneficial for generating alpha-ketoglutarate from glutamine, which allows leukemic cells to proliferate even in the absence of glucose. Inhibition of PHGDH, a rate-limiting enzyme in the SSP, dramatically reduces leukemia engraftment in mice in the presence of high fructose, confirming the essential role of the SSP in the metabolic plasticity of leukemic cells.


Phosphorylation of soluble pig epidermal proteins by endogenous calcium-activated, phospholipid-dependent protein kinase.
Endogenous calcium-activated, phospholipid-dependent protein kinase phosphorylates pig epidermal protein. Pig epidermis was homogenized and centrifuged at 30,000 X g for 30 min. The supernatant was incubated with or without calcium and phospholipid. A 97 kD soluble protein from pig epidermis was phosphorylated in the presence of calcium and phosphatidylserine. The phosphorylation reaction occurred immediately. With the use of two-dimensional polyacrylamide gel electrophoresis, it was shown that the 97 kD fragment was a basic protein, and that several small proteins were also phosphorylated. The characterization of these proteins is yet to be undertaken.


The changing face of primary open-angle glaucoma in the United States: demographic and geographic changes from 2011 to 2050.
To examine how demographic and geographic variations in US populations from 2011 to 2050 will contribute to estimated numbers of primary open-angle glaucoma (POAG) cases. Cross-sectional study. Prevalence rates from selected population-based studies were used to estimate the number of persons aged 40 years and older with POAG in the United States. For calculation, the age-, sex-, and race/ethnicity-specific prevalence rates were multiplied by the US Census estimates and projections from 2011 to 2050. Main outcome measures are estimated numbers of persons with POAG in different age, sex, and racial/ethnic groups and total and per capita POAG rates by state. In 2011, 2.71 million persons in the United States have POAG, with the highest estimated number among populations aged 70 to 79 years (31%), women (53%), and non-Hispanic whites (44%). The largest demographic group is non-Hispanic white women. In 2050, an estimated 7.32 million persons will have POAG, with the highest number among populations aged 70 to 79 years (32%), women (50%), and Hispanics (50%). The largest demographic group will shift to Hispanic men. During the next 40 years, the highest per capita POAG rates will double in New Mexico, Texas, and Florida. Despite the high prevalence of POAG in African Americans and Hispanics, the largest group in the United States is currently among older non-Hispanic white women but is expected to shift to Hispanic men over the next few decades. Given this shift, the greatest yield from screening programs is likely to be in those states with high numbers of non-Hispanic white women and Hispanic men.


[Giant cell arteritis and polymyalgia rheumatica: diagnostic criteria].
Giant cell arteritis still results in high morbidity related to systemic complications involving the eyes, the central nervous system, the heart and the aorta. Therefore, reliable diagnostic criteria are required in giant cell arteritis and polymyalgia rheumatica (PMR), to detect and manage at early stages patients with giant cell arteritis and PMR. Indeed, diagnostic criteria of giant cell arteritis and PMR have been validated. Taken together, first we propose to review the diagnostic criteria of giant cell arteritis (ACR, GRACG) and PMR (Bird, Jones-Hazleman, Chuang-Hunder, Dasgupta). Second, more recent diagnostic criteria of GCA and PMR are further reviewed, including biochemical/immunological data (especially anti-ferritin antibodies), temporal artery biopsy and imaging findings (artery ultrasonography, angio-CT-scan, angio-MRI, PET-scan, joint echography, and MRI).


The decline in breast cancer mortality in Europe: an update (to 2009).
We updated trends in breast cancer mortality in Europe up to the late 2000's. In the EU, age-adjusted (world standard population) breast cancer mortality rates declined by 6.9% between 2002 and 2006, from 17.9 to 16.7/100,000. The largest falls were in northern European countries, but more recent declines were also observed in central and eastern Europe. In 2007, all major European countries had overall breast cancer rates between 15 and 19/100,000. In relative terms, the declines in mortality were larger at younger age (-11.6% at age 20-49 years between 2002 and 2007 in the EU), and became smaller with advancing age (-6.6% at age 50-69, -5.0% at age 70-79 years). The present report confirms and further quantifies the persisting steady fall in breast cancer mortality in Europe over the last 25-30 years, which is mainly due to advancements in the therapy.


[Comparison of effect between early and delayed in primary intramedullary nailing combined with locked plate fixation for the treatment of multi-segments tibial fractures of type].
To compare the clinical results of early and delayed intramedullary nailing and locked plating for the treatment of multi-segments tibial fractures of type AO/ASIF-42C2. Between January 2010 and January 2013,45 patients with multi-segments closed tibial fractures of AO/ASIF-42C2 were treated by early primary intramedullary nailing and locked plating in 20 cases as early group and delayed in 25 cases as delayed group. In early group,20 cases included 13 males and 7 females with an average age of (37.9±14.3) years old ranging from 20 to 56 years;according to soft tissue injury Tscherne classification, 8 fractures were frade I,12 were grade II. In delayed group, 25 cases included 17 males and 8 females with an average age of (38.7±17.2) years old ranging from 24 to 55 years,4 fractures were grade I ,19 were grade II ,2 were grade III. The operative time, blood loss, hospital stay,fracture healing time and complications were recorded. At final follow-up, the Johner-Wruhs score were used to evaluate functional efficacy, and the posterior-anterior and lateral X-ray to evaluate fracture reduction and alignment. All the patients were followed up for (12.5±2.5) months in early group and (13.2±2.8) months in delayed group (P>0.05). No wounds infections were happened. At the last follow-up, the mean range of knee joint was 10°-0°-120°. According to Johner-Wruhs scoring,there were 15 cases in excellent,3 in good,fair in 2 in early group; 21 in excellent,2 in good,2 in fair. The average operative time,blood loss had no significant differences between two groups (P>0.05), but hospital stay in early group was significantly shorter than those in delayed group(P<0.05). Average fracture healing time of early group and delayed group were (5.3±2.6) months and (6.0±2.9) months, respectively (P>0.05). For multi-segments tibial fractures of type AO/ASIF-42C2 with preoperative minor soft tissue injuries lighter of Tscherne grade I or II, early primary intramedullary nailing and locked plating does not significantly increase the postoperative incidence of soft tissue complications for patients. The early and delayed primary intramedullary nailing and locked plating for treatment of AO/ASIF-42C2 proximal third tibial fractures can get similar curative effect.


Synergistic antibacterial activity of physical-chemical multi-mechanism by TiO2 nanorod arrays for safe biofilm eradication on implant.
Treatment of implant-associated infection is becoming more challenging, especially when bacterial biofilms form on the surface of the implants. Developing multi-mechanism antibacterial methods to combat bacterial biofilm infections by the synergistic effects are superior to those based on single modality due to avoiding the adverse effects arising from the latter. In this work, TiO2 nanorod arrays in combination with irradiation with 808 near-infrared (NIR) light are proven to eradicate single specie biofilms by combining photothermal therapy, photodynamic therapy, and physical killing of bacteria. The TiO2 nanorod arrays possess efficient photothermal conversion ability and produce a small amount of reactive oxygen species (ROS). Physiologically, the combined actions of hyperthermia, ROS, and puncturing by nanorods give rise to excellent antibacterial properties on titanium requiring irradiation for only 15 min as demonstrated by our experiments conducted in vitro and in vivo. More importantly, bone biofilm infection is successfully treated efficiently by the synergistic antibacterial effects and at the same time, the TiO2 nanorod arrays improve the new bone formation around implants. In this protocol, besides the biocompatible TiO2 nanorod arrays, an extra photosensitizer is not needed and no other ions would be released. Our findings reveal a rapid bacteria-killing method based on the multiple synergetic antibacterial modalities with high biosafety that can be implemented in vivo and obviate the need for a second operation. The concept and antibacterial system described here have large clinical potential in orthopedic and dental applications.


Ocular biometry and determinants of refractive error in rural Myanmar: the Meiktila Eye Study.
To describe the ocular biometry and determinants of refractive error in an adult population in Myanmar. A cross-sectional, population-based survey of the inhabitants 40 years of age and over from villages in the Meiktila District was performed; 2481 eligible participants were identified, and 2076 participated in the study. Biometric components including axial length (AL), anterior chamber depth (ACD), vitreous chamber depth (VCD), lens thickness (LT) and corneal curvature (CC) were measured. Lens opalescence was measured using the Lens Opacity Grading System III. Non-cycloplegic refraction was measured with an autorefractor. Complete biometric, refractive and lenticular data were available on 1498 participants. Men had longer ALs, ACDs, VCDs and steeper CCs than women. There was an increase in LT, nuclear opalescence (NO) and myopic shift with increasing age, with no significant change in AL with age. In the 40-59 year age group, VCD was a significant predictor of refractive error, but LT (p<0.001) and NO (p<0.001) were stronger predictors. In the 60+ age group, NO (p<0.001) was also the dominant predictor of refractive error. This Burmese population, particularly women, has a relatively short AL and ACD. NO is the strongest predictor of refractive error across all age groups in this population.


Risk factors for bloodstream infection caused by extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae: A focus on antimicrobials including cefepime.
Extended-spectrum β-lactamase (ESBL)-producing pathogens represent increasing challenges to physicians because of rising prevalence, high mortality, and challenging treatment. Identifying high risks and early appropriate therapy is critical to favorable outcomes. This is a 5-year retrospective case-case-control study performed at the Detroit Medical Center on adult patients with bloodstream infection (BSI) caused by ESBL-producing and non-ESBL-producing Escherichia coli or Klebsiella pneumoniae, each compared with uninfected controls. Data were collected from December 2004-August 2009. Multivariate analysis was performed using logistic regression. Participants included 103 patients with BSI caused by ESBL-producing pathogens and 79 patients with BSI caused by pathogens that did not produce ESBLs. The mean age of patients in the ESBL group was 67 years; of the patients, 51% were men, 77% were black, and 38% (n = 39) died in hospital. The mean age of patients in the non-ESBL group was 58 years; of the patients, 51% were men, 92% were black, and 22% (n = 17) died in hospital. On multivariate analysis, predictors of BSI caused by ESBL-producing pathogens included central venous catheter (odds ratio [OR], 29.4; 95% confidence interval [CI], 3.0-288.3), prior β-lactam-/β-lactamase-inhibitor therapy (OR, 28.1; 95% CI, 1.99-396.5), and prior cefepime therapy (OR, 22.7; 95% CI, 2.7-192.4). The only risk factor for BSI caused by non-ESBL-producing pathogens was urinary catheter insertion (OR, 18.2; 95% CI, 3.3-100.3). Prior antimicrobial therapy, particularly with β-lactam, was the strongest unique risk factor for BSI caused by ESBL-producing E coli or K pneumoniae.


Cutaneous magnetic stimulation reduces rat chronic pain via activation of the supra-spinal descending pathway.
Recent studies have demonstrated that magnetic stimulation (MS) can induce cellular responses such as Ca(2+) influx into the cultured neurons and glia, leading to increased intracellular phosphorylation. We have demonstrated previously that MS reduces rat neuropathic pain associated with the prevention of neuronal degeneration. Thus, we aimed to elucidate the actions of MS in relation to modulation of spinal neuron-glia and the descending inhibitory system in chronic pain. The male SD rats intrathecally implanted with catheters were subjected to sciatic nerve ligation (CCI). MS is a low power apparatus characterized by two different frequencies, 2 KHz and 83 MHz. Rats were given MS to the skin (injured sciatic nerve) for 10 min from the seventh day after CCI. The paw withdrawal latency (PWL) evoked by thermal stimuli was measured for 14 days after CCI. Immunohistochemistry for Iba-1 or GFAP was performed after 4% paraformaldehyde fixation (microscopic analysis). We employed microdialysis for measuring CSF 5-HIAA as a reflection of 5-HT release by MS stimulation. Following CCI, rats showed a decrease in PWL after CCI, and the decrease continued until the 14th day. With MS treatment, the decrease in PWL was reduced during the 10-14 day after CCI. Injection of JNK-1 inhibitors on the 14th day antagonized the analgesic effect of MS. MS also eliminated the CCI-induced decrease in GFAP immunoreactivity. Moreover, MS evoked spinal 5-HT release reflected by increase in spinal 5-HIAA level. Thus, we demonstrate that a novel magnetic stimulator used cutaneously can ameliorate chronic pain by not only preventing abnormal spinal neuron-glia interaction, but also through the activation of the supra-spinal descending inhibitory system.


Danish translation and validation of the Oslo Sports Trauma Research Centre questionnaires on overuse injuries and health problems.
The Oslo Sports Trauma Research Centre Overuse Injury Questionnaire (OSTRC-O) and the Oslo Sports Trauma Research Centre questionnaire on Health Problems (The OSTRC-H) make it possible to monitor illness and injury at regular intervals capturing prevalence and incidence of acute injury, overuse injury, and illnesses. The aim of this study was to translate, culturally adapt, and establish the face validity of the OSTRC-O and the OSTRC-H into a Danish context (DK) through cognitive interviews and the assessment of test-retest reliability. The OSTRC-O.DK was distributed to 57 heterogenous respondents; response rate was 89%. The OSTRC-H was distributed to 58 heterogenous respondents; response rate was 86%. No major disagreements were observed between the original and translated versions of the questionnaires. The OSTRC-O had high internal consistency (Cronbach's alpha 0.80-0.93). The primary reliability analyses including all participants, showed reliability ICC: 0.62 (95% CI: 0.42-0.77. The secondary reliability analyses that only included subjects who did not change injury region from the test to the retest showed an ICC of 0.86 (95% CI: 0.77-0.92).The questionnaires were found to be valid, reliable, and acceptable for use in a Danish population.


Development and initial validation of the Preschooler Gross Motor Quality Scale.
Motor skills have great impact on children in adapting to an environment and developing interpersonal interaction, cognition, and social behavior. Understanding what children can do and how they perform it is essential. Most motor tests seldom contain quality evaluation in the items or criteria. The purpose of this study was to develop and construct validity of the Preschooler Gross Motor Quality Scale (PGMQ) that included 17 items in three subscales to evaluate the quality of fundamental motor skills for the preschooler. The study designed PGMQ items and subscales according to well-described procedures, and recruited 174 children, aged 3-6 (89 boys and 85 girls) from 17 kindergartens. Two independent evaluators who were unaware of each other's results evaluated all the children separately in their kindergartens using PGMQ standardized setting and procedures and Gross Motor Scales of Peabody Developmental Motor Scales II (PDMS-II). Results of this study found significant differences between different ages and genders (Wilks'Λ=0.221, p<.001 and Wilks'Λ=0.690, p<.001 respectively). Total scores and raw scores in three subscales of PGMQ increased as age increased. Confirmatory factor analysis revealed three factors, including locomotion, manipulative ability and balance to explain 51.06% of the variance. The correlations between PGMQ and PDMS-II subscales were moderate to high (r=0.544-0.868, p<.01). The results showed satisfactory validity of PGMQ. PGMQ is useful to evaluate the development and quality of fundamental motor skills for the preschoolers.


Diagnostic Value of MRI Lamellated Hyperintense Synovitis in Periprosthetic Infection of Hip.
To investigate the correlation between magnetic resonance imaging (MRI) lamellated hyperintense synovitis and periprosthetic infection of hip arthroplasty and estimate its value in the diagnosis of infection after hip replacement. A retrospective analysis of 50 patients who underwent MRI from January 2016 to June 2019 after hip replacement was performed. The MRI scanning was performed with a 1.5T clinical imaging unit using SEMAC protocols. A total of 25 patients (cohort 1) showed infected total hip arthroplasty, and 25 patients had non-infected arthroplasty as controls (cohort 2). Two musculoskeletal radiologists, blinded to the clinical diagnosis, reviewed all the images for the presence of lamellated hyperintense synovitis independently. The cases were rereviewed by each reader after 2 weeks. The sensitivity, specificity, positive predictive value, and negative predictive value were calculated using the first reads. The Kappa statistic was used to assess inter-observer and intra-observer reliability. The incidence of lamellated hyperintense synovitis was 76%-88% in the experimental group and 8%-16% in the control group. The sensitivity of lamellated hyperintense synovitis for infection was 0.80-0.88 (95% confidence interval [CI]:0.59 - 0.97), the specificity was 0.84~0.92 (95% CI: 0.64 -0.99), the positive predictive value 0.83-0.92 (95% CI: 0.67 - 0.98), the negative predictive value 0.81 - 0.88 (95% CI: 0.65 - 0.96). The agreement between two readers was substantial (Kappa = 0.76, 95% CI: 0.58 - 0.94, P < 0.05). There were moderate inter-observer agreements for both readers, reader 1 (Kappa = 0.48, 95%CI: 0.23 - 0.72, P < 0.05) and reader 2 (Kappa = 0.44,95% CI: 0.19 - 0.69, P < 0.05). In this cohort, the presence of lamellated hyperintense synovitis in the MRI of hip arthroplasty showed high sensitivity and specificity for infection. This sign had substantial intra-observer reliability and moderate inter-observer reliability in the classification of the synovial pattern.


Prognostic value of a T-cell-based, interferon-gamma biomarker in children with tuberculosis contact.
Enzyme-linked immunospot (ELISpot) assay is an increasingly widely used, T-cell-based, interferon-gamma-release assay for diagnosing tuberculosis infection, but whether positive results are prognostic of active tuberculosis is not known. To determine whether ELISpot results predict the development of active tuberculosis among persons with recent tuberculosis exposure. Longitudinal cohort study of children and adolescents with tuberculosis contact recruited from October 2002 to April 2004. Community-based contact investigations in Turkey. 908 children and adolescents with recent household tuberculosis exposure. Enzyme-linked immunospot assay, incorporating early secretory antigenic target-6 and culture filtrate protein-10, and tuberculin skin test were done at baseline. Incidence rates ratios of progression to active tuberculosis for contacts with positive tuberculin skin test and ELISpot results, and relative incidence rates comparing contacts with positive and negative test results. Isoniazid preventive therapy was given to 688 (76%) contacts according to local guidelines. Fifteen contacts developed active tuberculosis over 1201 person-years of follow-up. Of 381 contacts with positive ELISpot results, 11 developed active tuberculosis over 536 person-years of follow-up (incidence rate, 21 per 1000 person-years [95% CI, 10.2 to 36.7 per 1000 person-years]), a statistically significant 3- to 4-fold increased risk for progression relative to ELISpot-negative contacts. Of 550 contacts with positive tuberculin skin test results, 12 developed active tuberculosis over 722 person-years of follow-up (incidence rate, 17 per 1000 person-years [CI, 8.6 to 29.0 per 1000 person-years]). Only 3 of the 15 incident cases were confirmed by culture. Positive ELISpot results predict subsequent development of active tuberculosis in recent tuberculosis contacts. Although tuberculosis contacts with positive ELISpot results have an incidence rate of tuberculosis similar to that of contacts with positive tuberculin skin test results, ELISpot testing could allow more focused targeting of preventive therapy to fewer contacts.


Dorsal Raphe Serotonergic Neurons Control Intertemporal Choice under Trade-off.
Appropriate choice about delayed reward is fundamental to the survival of animals. Although animals tend to prefer immediate reward, delaying gratification is often advantageous. The dorsal raphe (DR) serotonergic neurons have long been implicated in the processing of delayed reward, but it has been unclear whether or when their activity causally directs choice. Here, we transiently augmented or reduced the activity of DR serotonergic neurons, while mice decided between differently delayed rewards as they performed a novel odor-guided intertemporal choice task. We found that these manipulations, precisely targeted at the decision point, were sufficient to bidirectionally influence impulsive choice. The manipulation specifically affected choices with more difficult trade-off. Similar effects were observed when we manipulated the serotonergic projections to the nucleus accumbens (NAc). We propose that DR serotonergic neurons preempt reward delays at the decision point and play a critical role in suppressing impulsive choice by regulating decision trade-off.


Factors associated with irritable bowel syndrome among medical students at Ain Shams University.
Irritable bowel syndrome (IBS) is one of the most common and potentially disabling gastrointestinal disorders. The pathogenesis of this disorder remains obscure. However, many etiological explanations point toward bacterial etiology. Also, several studies have documented that psychological and social factors may play a role. Medical education is among the most challenging and stressful education, and this may predispose to high rates of IBS. The aims of this study are to estimate the frequency of IBS in a selected sample of students of Faculty of Medicine in Ain Shams University and to find out the determinants associated with this disorder. A cross-sectional study was carried out among medical students from October 2017 to February 2018 at Faculty of Medicine-Ain Shams University. All participants were asked to complete a confidential self-administered questionnaire. An interview questionnaire was used for diagnosis of IBS according to Rome III criteria, while morbid anxiety and depression were diagnosed by using the Arabic version of Hospital Anxiety and Depression Scale (HADS). Three hundred eighty-two students completed the questionnaire. The frequency of IBS was 31.7% with higher proportion among females and among students with positive family history of IBS. However, IBS was significantly less prevalent among students practicing regular exercise. Lastly, based on (HADS), there was a statistical significant relationship between IBS and anxiety (p < 0.05), but not depression. The study revealed that around 31% of the studied group was suffering from IBS. Female gender, suffering from anxiety, and positive family history of IBS were the main associated factors for IBS. Screening of all medical students in the faculty for IBS is suggested. Providing psychological and emotional support along with stress management is highly recommended.


The effect of aluminum hydroxide dissolution on the bleeding of aluminum lake dyes.
The effect of pH on the bleeding of FD&C yellow No. 5 aluminum lake and FD&C red No. 40 aluminum lake was investigated. The pH-bleeding profiles corresponded to the pH-solubility profile of aluminum hydroxide. The similarity of the bleeding profiles of both lake dyes and the pH-solubility profile of aluminum hydroxide indicates that pH related bleeding, other than that occurring by competition with anions, is a result of dissolution of the aluminum hydroxide substrate. This dissolution is related to the properties of the substrate rather than to the structure of adsorbed dye.


Genome-wide characterization on MT family and their expression in response to environmental cues in upland cotton (Gossypium hirsutum L.).
Metallothioneins (MTs) are believed as key metal chelators and scavengers of reactive oxygen species (ROS), which are involved in tolerance and de-toxicity to multiple environmental stresses in plants. The MT gene family was characterized from upland cotton (Gossypium hirsutum L.), compared with its putative genome donors G. arboretum and raimondii. Subsequently, gene functions were predicted by promoter analysis. Moreover, gene expressions subjecting to exogenous stimuli, as well as in terms of developments, were studied. The main findings were shown as follows: 1) 19 GhMTs were identified from G. hirsutum, and the family completely included all four sub-types, namely p1, p2, p3, and pec. Sub-type p2 GhMTs were most conservative in protein motif compositions, gene structures, phylogenic relationships, and group numbers, while p3 GhMTs demonstrated much more diversiform and distant genetic relationships. 2) The GhMT family experienced apparent gene expansion, and the members from the D sub-genome were subjected to stronger environmental selection. 3) GhMTs played differential and overlapped roles in response to environmental cues. 4) GhMT6, GhMT8, and GhMT14 were involved in both vegetative and reproductive developments. These findings must provide valuable insights into understanding the plant MT gene family and novel gene resources for cotton breeding for environmental stresses, phytoremediation, and beyond.


Overexpression of OgPAE1 from wild rice confers fungal resistance against Botrytis cinerea.
A full-length cDNA of the OgPAE1 gene encoding the alpha5 subunit of the 20S proteasome was isolated from wild rice (Oryza grandiglumis) treated by wounding or with a fungal elicitor. The deduced amino acid sequence of OgPAE1 comprises 237 amino acids (25.99 kDa), and shows 94.5% homology with Arabidopsis thaliana AtPAE1. Expression of OgPAE1 is regulated by defense-related signaling chemicals such as cantharidin, endothall and jasmonic acid. Overexpression of OgPAE1 in A. thaliana leads to resistance to the fungal pathogen Botrytis cinerea by lowering disease rate and size of necrotic lesions, and by less penetration and colonization of fungal hyphae. The results indicate that the 20S proteasome from wild rice is involved in the B. cinerea defense pathway via an as yet undetermined mechanism.


Expression and significance of angiostatin, vascular endothelial growth factor and matrix metalloproteinase-9 in brain tissue of diabetic rats with ischemia reperfusion.
To discuss the expression and significance of angiostatin, vascular endothelial growth factor and matrix metalloproteinase-9 in the brain tissue of diabetic rats with ischemia reperfusion. A total of 60 male Wistar rats were randomly divided into the normal group, sham group, diabetic cerebral infarction group and single cerebral infarction group according to the random number table, with 15 rats in each group. The high sucrose diet and intraperitoneal injection of streptozotocin were performed for the modeling of diabetic rats, while the thread-occlusion method was employed to build the model of cerebral ischemia reperfusion. The immunohistochemical staining was performed to detect the expression of angiostatin, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in the brain tissue. The expression of angiostatin after the reperfusion in the brain tissue of rats in the single cerebral infarction group and diabetic cerebral infarction group was increased 6 h after the reperfusion, reached to the peak on 1 d and then decreased gradually. The expression of angiostatin in the diabetic cerebral infarction group 6 h, 1 d, 3 d and 7 d after the reperfusion was significantly higher than that in the single cerebral infarction group (P < 0.05). VEGF began to be increased 1 h after the reperfusion in the single cerebral infarction group and diabetic cerebral infarction group, reached to the peak at 6 h and then decreased gradually. The expression of VEGF in the diabetic cerebral infarction group at each time point after the reperfusion was significantly lower than that in the single cerebral infarction group (P < 0.05). MMP-9 began to be increased 1 h after the reperfusion in the single cerebral infarction group and diabetic cerebral infarction group, reached to the peak on 1 d and then decreased gradually. The expression of MMP-9 in the diabetic cerebral infarction group at each time point after the reperfusion was significantly higher than that in the single cerebral infarction group (P < 0.05). The high glucose environment in which the diabetic cerebral infarction is occurred is to induce the formation of MMP-9 at first and then activate and increase the expression of angiostatin. Afterwards, the expression of VEGF is inhibited, resulting in the poor angiogenesis after cerebral infarction, which thus makes the injury of brain tissue after cerebral infarction even worse than the non-diabetes mellitus.


Primary meningeal malignant fibrous histiocytoma with cerebrospinal dissemination and pulmonary metastasis.
Primary intracranial malignant fibrous histiocytoma (MFH) is very rare, and not much is known about its clinical features. The authors report a case of left temporal leptomeningeal MFH, with consequent cerebrospinal fluid (CSF) dissemination and pulmonary metastasis. The clinical features of this case and the therapeutic prognosis of 17 cases reported previously in the literature were reviewed.


Nano-engineering nanomedicines with customized functions for tumor treatment applications.
Nano-engineering with unique "custom function" capability has shown great potential in solving technical difficulties of nanomaterials in tumor treatment. Through tuning the size and surface properties controllablly, nanoparticles can be endoewd with tailored structure, and then the characteristic functions to improve the therapeutic effect of nanomedicines. Based on nano-engineering, many have been carried out to advance nano-engineering nanomedicine. In this review, the main research related to cancer therapy attached to the development of nanoengineering nanomedicines has been presented as follows. Firstly, therapeutic agents that target to tumor area can exert the therapeutic effect effectively. Secondly, drug resistance of tumor cells can be overcome to enhance the efficacy. Thirdly, remodeling the immunosuppressive microenvironment makes the therapeutic agents work with the autoimmune system to eliminate the primary tumor and then prevent tumor recurrence and metastasis. Finally, the development prospects of nano-engineering nanomedicine are also outlined.


Determining ecoregions for environmental and GMO monitoring networks.
A representative environmental monitoring network at the regional scale cannot use raster-based or random sampling designs, but requires a stratified sampling procedure integrating different information layers, and it has to occur in ecologically differing homogeneous regions (ecoregions). These we have determined using a set of spatial strata with ecological variables which we analysed with classification and regression trees (CART). We present a framework for environmental monitoring, that covers different scales, and we transfer the framework to a potential GMO (genetically modified organisms) monitoring network. We use ecoregion and other environmental strata together with existing environmental monitoring networks to determine GMO monitoring sites more precisely.


A honeybee storage protein gene, hex 70a, expressed in developing gonads and nutritionally regulated in adult fat body.
In preparing for metamorphosis, insect larvae store a huge amount of proteins in hemolymph, mainly hexamerins. Out of the four hexamerins present in the honeybee larvae, one, HEX 70a, exhibited a distinct developmental pattern, especially since it is also present in adults. Here, we report sequence data and experimental evidence suggesting alternative functions for HEX 70a, besides its well-known role as an amino acid resource during metamorphosis. The hex 70a gene consists of 6 exons and encodes a 684 amino acid chain containing the conserved hemocyanin N, M, and C domains. HEX 70a classifies as an arylphorin since it contains more than 15% of aromatic amino acids. In the fat body of adult workers, hex 70a expression turned out to be a nutrient-limited process. However, the fat body is not the only site for hex 70a expression. Both, transcript and protein subunits were also detected in developing gonads from workers, queens and drones, suggesting a role in ovary differentiation and testes maturation and functioning. In its putative reproductive role, HEX 70a however differs from the yolk protein, vitellogenin, since it was not detected in eggs or embryos.


Role of aspartic acid residues D87 and D89 in APS kinase domain of human 3'-phosphoadenosine 5'-phosphosulfate synthase 1 and 2b: A commonality with phosphatases/kinases.
3'-phosphoadenosine 5'-phosphosulfate (PAPS) is synthesized in two steps by PAPS synthase (PAPSS). PAPSS is comprised of ATP sulfurylase (ATPS) and APS kinase (APSK) domain activities. ATPS combines inorganic sulfate with α-phosphoryl of ATP to form adenosine 5'-phosphosulfate (APS) and PPi. In the second step APS is phosphorylated at 3'-OH using another mole of ATP to form PAPS and ADP catalyzed by APSK. The transfer of gamma-phosphoryl from ATP onto 3'-OH requires Mg2 + and purported to involve residues D87GD89N. We report that mutation of either aspartic residue to alanine completely abolishes APSK activity in PAPS formation. PAPSS is an, unique enzyme that binds to four different nucleotides: ATP and APS on both ATPS and APSK domains and ADP and PAPS exclusively on the APSK domain. The thermodynamic binding and the catalytic interplay must be very tightly controlled to form the end-product PAPS in the forward direction. Though APS binds to ATPS and APSK, in ATPS domain, the APS is a product and for APSK it is a substrate. DGDN motif is absent in ATPS and present in APSK. Mutation of D87 and D89 did not hamper ATPS activity however abolished APSK activity severely. Thus, D87GD89N region is required for stabilization of Mg2+-ATP, in the process of splitting the γ-phosphoryl from ATP and transfer of γ-phosphoryl onto 3'-OH of APS to form PAPS a process that cannot be achieved by ATPS domain. In addition, gamma32P-ATP, trapped phosphoryl enzyme intermediate more with PAPSS2 than with PAPSS1. This suggests inherent active site residues could control novel catalytic differences. Molecular docking studies of hPAPSS1with ATP + Mg2+ and APS of wild type and mutants supports the experimental results.


Use of neuromuscular blocking drugs in strabismus surgery: systematic review and ophthalmology survey study.
Nondepolarizing neuromuscular blocking drugs have been advocated for accurate forced duction testing (FDT) during strabismus surgery. The purpose of this study was to evaluate the literature on use of muscle relaxants (MRs) in strabismus surgery and to evaluate the current practice of pediatric ophthalmologists. Systematic review and survey study. Seventy-seven pediatric ophthalmologists. MEDLINE, EMBASE, CENTRAL, Web of Science, and OpenGrey were searched to August 2019. Reporting followed the Preferred Reporting Items for Systematic Review and Meta-analysis (PRISMA) guidelines. Primary studies on depolarizing or nondepolarizing MRs in patients undergoing strabismus surgery were included. Primary outcomes included indications and prevalence of use. A 23-question survey was emailed to pediatric ophthalmologists worldwide to collect practice pattern information involving MRs in strabismus surgery. Three studies were included in the systematic review. MRs were used in 34%-45% of patients undergoing strabismus surgery in one 1999 study for FDT. No evidence exists to support the use of MRs for facilitating FDT. Seven of 77 (9.1%) pediatric ophthalmologists requested MRs for FDT. Those who use MRs were more likely to be practicing outside the United States (p < 0.05) and in academic hospitals or private centres rather than in community settings. Drawbacks appeared to outweigh benefits, with the biggest drawback identified as increased turnover time between surgical cases (44%). Inadequate evidence exists in the literature to support the use of nondepolarizing MRs to maintain paralysis of extraocular muscles during strabismus surgery, and this is an uncommon practice among pediatric ophthalmologists.


Analogous inhibitors of elastase do not always bind analogously.
It has been assumed that the structure of a single inhibitor complex is sufficient to define the available subsites of an enzyme that has a unique binding site and a uniquely defined mode for ligand binding--the specificity for these subsites can thus be probed by kinetic experiments. Elastase is an enzyme for which these traditional assumptions, which underlie such structural and kinetic studies, do not hold. Three new crystal structures of elastase complexed to chemically similar inhibitors with similar binding affinities reveal a diversity of binding modes as well as two new subsites on elastase. The existence of multiple binding sites and different binding modes for such similar inhibitors indicates that researchers must proceed with caution when using kinetics to map out protein subsites.


Dataset of concurrent EEG, ECG, and behavior with multiple doses of transcranial electrical stimulation.
We present a dataset combining human-participant high-density electroencephalography (EEG) with physiological and continuous behavioral metrics during transcranial electrical stimulation (tES). Data include within participant application of nine High-Definition tES (HD-tES) types, targeting three cortical regions (frontal, motor, parietal) with three stimulation waveforms (DC, 5 Hz, 30 Hz); more than 783 total stimulation trials over 62 sessions with EEG, physiological (ECG, EOG), and continuous behavioral vigilance/alertness metrics. Experiment 1 and 2 consisted of participants performing a continuous vigilance/alertness task over three 70-minute and two 70.5-minute sessions, respectively. Demographic data were collected, as well as self-reported wellness questionnaires before and after each session. Participants received all 9 stimulation types in Experiment 1, with each session including three stimulation types, with 4 trials per type. Participants received two stimulation types in Experiment 2, with 20 trials of a given stimulation type per session. Within-participant reliability was tested by repeating select sessions. This unique dataset supports a range of hypothesis testing including interactions of tDCS/tACS location and frequency, brain-state, physiology, fatigue, and cognitive performance.


Psychologic correlates of serum indicators of stress in man: a longitudinal study.
Correlations between serum uric acid, cortisol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol and psychometric indices of stress--namely, anxiety, hostility, and depression--were investigated in 20 students over a 2 1/2 month academic quarter. There were three peak periods of stress, with two occurring during midterm and final exams. The observed changes in serum cortisol were highly correlated with changes in anxiety and depression. Serum uric acid was significantly depressed during the first half of the quarter, but returned to control levels after midterm exams. Serum cholesterol and the LDL cholesterol subfraction were significantly elevated above control levels and remained elevated throughout the quarter after the first day of classes. Absolute levels of HDL cholesterol varied little throughout the quarter, however, the ratio of HDL cholesterol/total cholesterol was significantly depressed twice in the quarter, each time following approximately 10 days after the peak period of stress.


[Physical exercise as a functional test of hgh secretion (author's transl)].
The efficiency of L-dopa and exercise tests have been compared to diagnose the lack of human growth hormone. Both tests gave similar results. The exercise test is recommended as a screening method in large populations to discard growth inhibition due to HGH deficiency, because of its easy, quick and painless application to patients.


Misfit-induced changes of lattice parameters in two-phase systems: coherent/incoherent precipitates in a matrix.
Elastic accommodation of precipitation-induced or thermally induced misfit leads to lattice-parameter changes in crystalline multi-phase systems. Formulae for calculation of such misfit-induced lattice-parameter changes are presented for the aggregate (matrix + second-phase particles) and for the individual matrix and second phase, recognizing the occurrence of either coherent or incoherent diffraction by the matrix and second-phase particles. An overview and an (re)interpretation on the above basis is presented of published lattice-parameter data, obtained by X-ray diffraction analyses of aggregates of matrix plus second-phase particles. Examples for three types of systems consisting of a matrix with misfitting second-phase particles are dealt with, which differ in the origin of the misfit (precipitation or thermally induced) and in the type of diffraction (coherent or incoherent diffraction of matrix plus second-phase particles). The experimental data are shown to be in good to very good agreement with predictions according to the current treatment.


Bacterial molecular syringe for drug delivery.
Different protein-delivery tools are being actively developed for efficient drug delivery into host cells. A recent study by Kreitz et al. in Nature reported a syringe-like protein delivery vector engineered from natural endosymbiotic bacteria for potential human use.


Sugar yields from sunflower stalks treated by hydrothermolysis and subsequent enzymatic hydrolysis.
To produce fermentable sugar with fewer microbial inhibitors via few processes, batch-type hydrothermal treatments of sunflower stalks were performed, followed by enzymatic hydrolysis of pretreatment slurries with Cellic CTec2 and Cellic HTec2 (9:1, v/v, 0.1 ml/g dry biomass, 8.1FPU). The yields of hemicellulosic sugars were maximized at the pretreatment condition of 180°C for 30 min, while the furfural and 5-hydroxymethyl-2-furaldehyde (HMF) concentrations remained low. The glucose yield, however, was only 67.0% of the theoretical glucose yield (TGY). Either the treatment of raw biomass prior to pretreatment or the post-treatment of pretreated residue prior to enzymatic hydrolysis increased the glucose yield as follows: washing the pretreated solid with solvents (90% TGY)>partial removal of liquid fraction from the pretreatment slurry (PS, 83%)>removal of hot-water extractives from the biomass prior to pretreatment (77%)>prewetting of the biomass (70%).


Angiotensin-(1-7) inhibits autophagy in the brain of spontaneously hypertensive rats.
Autophagy is an important cellular process that mediates lysosomal degradation of damaged organelles, which is activated in response to a variety of stress-related diseases, including hypertension. The basal level of autophagy plays an important role in the maintenance of cellular homeostasis, whereas excessive autophagic activity leads to cell death and is considered as a contributing factor to several disorders. Recent works have demonstrated that Angiotensin-(1-7) [Ang-(1-7)] exerted its neuroprotective effects by modulating classic components of renin-angiotensin system associated with reducing oxidative stress and apoptosis in brains of spontaneously hypertensive rats (SHRs). However, the effect of Ang-(1-7) on autophagic activity in brain of hypertensive individual remains unclear. In this study, Wistar-Kyoto rats received intracerebroventricular (I.C.V.) infusion of artificial cerebrospinal fluid (aCSF) while SHRs received I.C.V. infusion of aCSF, Ang-(1-7), Mas receptor antagonist A-779, or angiotensin II type 2 receptor antagonist PD123319 for 4 weeks. Brain tissues were collected and analyzed by western blotting analysis, immunofluorescence assay, and transmission electron microscopic examination. Our study showed that infusion of Ang-(1-7) for 4 weeks inhibited the increase of microtubule-associated protein 1 light chain 3 (LC3)-II and Beclin-1 levels, as well as the autophagosome formation in SHR brain. Meanwhile, the reduction of p62 expression in SHR brain was also reversed by Ang-(1-7). Of note, the anti-autophagic effects of Ang-(1-7) were independent of blood pressure reduction and can be inhibited by A-779 and PD123319. These findings suggest that treatment with Ang-(1-7) may be useful to prevent hypertension-induced excessive autophagic activation in brain.


Serosurvey for Newcastle disease and avian influenza A virus antibodies in great cormorants from France.
Inland great cormorants (Phalacrocorax carbo) culled in France were examined in the winter of 1997-98 and 1998-99 for antibodies to Newcastle disease (ND) and influenza A strains H5 and H7 by the hemagglutination inhibition test. Antibodies to influenza A group antigen were tested by agar gel precipitin test. Ten of 53 adult individuals were seropositive for ND virus. All sera were negative for influenza A antibodies. It is speculated that ND occurred in the sampled population.


Epidemiological study of food allergy in a population of Argentine children.
Food allergies affect mostly children in their first years of life. Epidemiological data obtained in Argentina are scarce. To determine offending foods and describe the prevalence and characteristics of patients with food allergy in an Argentine pediatric population. Observational, retrospective, crosssectional study on patients younger than 18 years. A total of 321 patients were included; food allergy was confirmed in 64 % (207) of cases. An immunoglobulinE-mediated mechanism was observed in 53% (109); cow's milk protein allergy, in 68 % (140); egg allergy, in 20 % (41); and anaphylaxis as clinical manifestation, in 12 % (24). The overall prevalence of food allergy was 0.87 % (95 % confidence interval: 0.7-0.9). The overall prevalence of food allergy was 0.87%. Cow's milk was the main allergen, even among adolescents.


Are there autoantibodies reacting against citrullinated peptides derived from type I and type II collagens in patients with rheumatoid arthritis?
To assess the possible presence in patients with rheumatoid arthritis (RA) of autoantibodies recognising citrullinated peptides derived from type I and II collagens. Firstly, the binding of four pairs of synthetic peptides (arginine-containing and artificially citrullinated forms) related to different regions of human type II collagen were tested with sera from 120 patients with RA and 81 controls. Secondly, two similar pairs of peptides related to the carboxy terminal telopeptides of the alpha1 and alpha2 chains of human type I collagen were tested. 42-53% of the RA sera showed increased binding of arginine peptides related to type II collagen. However, 12 RA sera bound the citrullinated form of the alpha1(II) telopeptide more strongly than the corresponding arginine peptide. 20 RA sera bound the citrullinated carboxytelopeptide from the alpha1 chain of type I collagen (alpha1(I) telopeptide) more strongly than the respective arginine peptide. The correlation between the autoantibodies to type I and II collagen telopeptides was r(s) = 0.576, p < 0.001. Anti-cyclic citrullinated peptide (anti-CCP) assay was positive in 71/120 (59%) patients with RA. An anti-CCP assay detects a different subgroup of antibodies than anti-telopeptide assays. However, both anti-telopeptide and anti-CCP antibodies were increased in patients with RA. Some patients with RA were identified whose sera contained antibodies that specifically bound citrullinated peptides related to the carboxy terminal telopeptides of the alpha1 and alpha2 chains of type I collagen and the alpha1 chains of type II collagen (sequences YYXA, FYXA, and YMXA, where X stands for citrulline).


Three-dimensional echocardiographic pictures of isolated double-orifice mitral valve.
We present a case of a 12-year-old boy with a rare form of cardiac anomaly, isolated double-orifice mitral valve. He was referred to our hospital at 1.5 years old because of heart murmur. Two-dimensional echocardiography showed double-orifice mitral valve without any associated cardiac anomalies. He has been followed carefully without any medication for 11 years. He has had no symptoms and an excellent natural course thus far.


Investigation of Hybrid Nanoparticle-Acid Fluids (HNAFs): Influence of Wettability and Interfacial Tension Mechanisms in Harsh Carbonate Reservoirs for Improved Oil Recovery.
Over the past few years, there has been significant interest in the potential of hybrid nanoparticle-acid fluid (HNAFs) for improved oil recovery. This comprehensive study investigates the effects of nanoparticles and acid on interfacial tension (IFT) to establish a relationship between brine properties and the oil/brine IFT. This investigation is one of the first regional studies conducted utilizing candidate field data from the Middle East. Based on the literature review and screening studies conducted, a seawater (SW)-based HNAF was formulated with nanoparticles (SiO2, Al2O3, and ZnO) and HCl to measure their effect on IFT. A total of 48 formulations of HNAFs, nanofluids with and without acid, were analyzed with crude oil from a candidate field. IFT measurements were subsequently conducted using the pendant drop method under ambient conditions and in a high-pressure, high-temperature reservoir environment. Results showcased that IFT reduction was observed by increasing the acid concentration with SiO2 and Al2O3, although a reverse trend was observed with ZnO. Moreover, it was observed that IFT varied with increasing concentrations of nanoparticles, and at certain acid concentrations, IFT reduced significantly with higher nanoparticle concentrations. From the Amott studies, a clear signature was achieved, with ZnO exhibiting a total of 31.4% oil recovery, followed by SiO2 (27.3%) and Al2O3 (23.7%). The results of this study may assist in defining a screening criterion for future displacement (oil recovery) studies involving the presented nanoparticles. The results also reveal further the mechanisms involved in IFT reduction by hybrid nano-acid fluids and their potential for significant applications in the Middle East.


Quantitative and Qualitative Differences in Endothelial Cell Loss Between Endothelium-In Versus Endothelium-Out Loading in Descemet Membrane Endothelial Keratoplasty.
This study compares endothelial cell loss (ECL) between donor grafts loaded in the Geuder Glass Cannula and the Coronet EndoGlide made specifically for Descemet membrane endothelial keratoplasty (DMEK). Prestripped tissues using the submerged cornea using backgrounds away (SCUBA) technique were obtained from an eye bank. Donor grafts were loaded with the Geuder Glass Cannula or Coronet DMEK EndoGlide. Grafts were then ejected directly onto a glass slide, stained, and imaged with confocal fluorescent microscopy. Trainable Fiji (ImageJ) segmentation software was used to quantify ECL. Twenty-three total tissue samples were tested. Eleven grafts were loaded using the Geuder, and 12 were loaded with the Coronet. Preloading graft endothelial cell density was comparable between Geuder (2436 ± 581 cells/mm) and Coronet (2577 ± 483 cells/mm) groups with P = 0.56. Other baseline characteristics including donor age and time from death to loading were comparable (P = 0.73 and P = 0.66, respectively). Average ECL was 11.3% ± 3.0% in the Geuder group versus 7.2% ± 7.0% in the Coronet group and not significantly different (P = 0.07). When observing patterns of cell loss, grafts loaded in the Coronet DMEK EndoGlide resulted in more focal forceps grabs and trifold patterns. Grafts loaded with the Geuder Glass Cannula yielded more diffuse stippled stress lines. Our results suggest comparable ECL between Geuder (endothelium-out) and Coronet (endothelium-in) DMEK EndoGlide tissue injection techniques. Patterns of ECL in the Coronet group correlated with forceps grabs, but this study suggests that additional manipulation of tissue does not result in higher ECL.


The bone biologic effects of zoledronate in healthy dogs and dogs with malignant osteolysis.
Malignant osteolysis is a process whereby cancer cells in concert with osteoclasts erode bone matrix. Aminobisphosphonates (NBPs) such as zoledronate induce osteoclast apoptosis and thereby decrease malignant skeletal destruction, severity of bone pain, and frequency of pathologic fracture. IV-administered zoledronate will reduce homeostatic bone turnover in healthy dogs and pathologic bone resorption in dogs diagnosed with primary and secondary bone tumors. Six healthy dogs and 20 dogs with naturally occurring primary or metastatic bone tumors were administered zoledronate IV. Prospective study: In all dogs, healthy (n = 6) and with malignant osteolysis (n = 20), the bone biologic effects of zoledronate were evaluated by quantifying changes in serum C-telopeptide (CTx) or urine N-telopeptide (NTx) concentrations or both. In dogs with osteosarcoma (OSA) (n = 10), serial changes in tumor relative bone mineral density (rBMD) assessed by dual-energy x-ray absorptiometry were used to characterize zoledronate's antiresorptive effects within the immediate tumor microenvironment. Additionally, the biochemical tolerability of zoledronate was assessed in 9 dogs receiving multiple (> or =2) consecutive treatments. All dogs had significant reductions in serum CTx or urine NTx concentrations or both after zoledronate administration. In a subset of dogs with appendicular OSA, reduced urine NTx concentrations and increased primary tumor rBMD coincided with improved limb usage as reported by pet owners in dogs treated with zoledronate and concurrent oral analgesics. Multiple zoledronate infusions were not associated with biochemical evidence of toxicosis. In dogs with skeletal neoplasms, IV-administered zoledronate exerts bone biologic effects, appears safe, and can provide pain relief.


The effects of active upper-limb versus passive lower-limb exercise on quality of life among individuals with motor-complete spinal cord injury.
Multi-centre randomized clinical trial. (1) compare the effects of arm-cycle ergometry (ACET) and body weight supported treadmill training (BWSTT) on quality of life (QOL) and intermediary variables in individuals with spinal cord injury (SCI); (2) examine correlations between baseline measures, and changes in physical activity, QOL, and intermediaries. Hospital-based research institutes (Vancouver, Toronto) and University-based exercise program (Hamilton). 35 participants with motor-complete SCI above T6 completed baseline assessments of physical activity, life satisfaction, independence, autonomy, positive and negative affect, and pain. Twenty-eight participants were randomized to 72 sessions of ACET (n = 14) or BWSTT (n = 14) with measures repeated following 36 sessions, 72 sessions, and 6-months post-intervention. Neither intervention significantly impacted QOL. Pain was reduced in ACET compared to BWSTT (interaction effect p = 0.022) and was significantly less at 72 sessions vs. baseline in the ACET group (p = 0.009). At baseline, QOL was positively correlated with independence, autonomy, and positive affect and negatively correlated with negative affect (all p < 0.05). Following BWSTT, changes in moderate-vigorous physical activity correlated with changes in QOL (r = 0.87, p = 0.010). Following ACET, changes in autonomy and independence were positively correlated with changes in QOL (both r > 0.64, p < 0.048). Contrary to previous studies, there was no benefit of either intervention on measures of QOL. The social context of exercise may be important for improving QOL. However, individuals may benefit more from active (ACET) than passive (BWSTT) exercise modalities through reduced pain. Exercise interventions that improve autonomy and independence may lead to improvements in QOL.


De novo copy number variants associated with intellectual disability have a paternal origin and age bias.
De novo mutations and structural rearrangements are a common cause of intellectual disability (ID) and other disorders with reduced or null reproductive fitness. Insight into the genomic and environmental factors predisposing to the generation of these de novo events is therefore of significant clinical importance. This study used information from single nucleotide polymorphism microarrays to determine the parent-of-origin of 118 rare de novo copy number variations (CNVs) detected in a cohort of 3443 patients with ID. The large majority of these CNVs (76%, p=1.14×10(-8)) originated on the paternal allele. This paternal bias was independent of CNV length and CNV type. Interestingly, the paternal bias was less pronounced for CNVs flanked by segmental duplications (64%), suggesting that molecular mechanisms involved in the formation of rare de novo CNVs may be dependent on the parent-of-origin. In addition, a significantly increased paternal age was only observed for those CNVs which were not flanked by segmental duplications (p=0.02). This indicates that rare de novo CNVs are increasingly being generated with advanced paternal age by replication based mechanisms during spermatogenesis.


Perturbation of chromatin structure in the region of the adult beta-globin gene in chicken erythrocyte chromatin.
An EcoRI chromatin fragment containing the adult beta-globin gene and flanking sequences, isolated from chicken erythrocyte nuclei, sediments at a reduced rate relative to bulk chromatin fragments of the same size. We show that the specific retardation cannot be reversed by adding extra linker histones to native chromatin. When the chromatin fragments are unfolded either by removing linker histones or lowering the ionic strength, the difference between globin and bulk chromatin fragments is no longer seen. The refolded chromatin obtained by restoring the linker histones to the depleted chromatin, however, exhibits the original sedimentation difference. This difference is therefore due to a special property of the histone octamers on the active gene that determines the extent of its folding into higher-order structure. That it is not due to the differential binding of linker histones in vitro is shown by measurements of the protein to DNA ratios using CsCl density-gradients. Both before and after selective removal of the linker histones, the globin gene fragment and bulk chromatin fragments exhibit only a marginal difference in buoyant density. In addition, we show that cleavage of the EcoRI fragment by digestion at the 5' and 3' nuclease hypersensitive sites flanking the globin gene liberates a fragment from between these sites that sediments normally. We conclude that the hypersensitive sites per se are responsible for the reduction in sedimentation rate. The non-nucleosomal DNA segments appear to be too long to be incorporated into the chromatin solenoid and thus create spacers between separate solenoidal elements in the chromatin, which can account for its hydrodynamic behaviour.


Physiological properties and afferent connections of the locus coeruleus and adjacent tegmental neurons involved in the generation of paradoxical sleep in the cat.
Results reported here confirm and extend those of early retrograde transport studies of the brainstem in the rat and cat. This study demonstrates substantial and multiple afferent projections to the cat locus coeruleus arising from neurons containing acetylcholine, serotonin, norepinephrine, epinephrine, dopamine, histamine, and neuropeptides such as methionine, enkephaline and substance P. Further, our studies reveal notable differences in afferent projection to the noradrenergic and cholinergic regions of the locus coeruleus.


Clinical characteristics of Epstein-Barr virus infection in the pediatric nervous system.
To investigate the clinical characteristics of Epstein-Barr virus (EBV) infection in the pediatric nervous system (NS). We retrospectively analyzed the clinical data and follow-up results of 89 children with neurological damage caused by EBV who were hospitalized in the children's hospital of Chongqing Medical University from January 2008 to April 2019. EBV infection of the NS can occur at any time of the year. The highest incidence was seen in the age group of 0-4 years. Fever is the main clinical feature (74/89, 83.1%). The main clinical types were encephalitis/meningoencephalitis (64/89, 71.9%), acute myelitis (2/89, 2.2%), acute disseminated encephalomyelitis (ADEM) (3/89, 3.4%), Guillain-Barré Syndrome (GBS) (15/89, 16.9%), neurological damage caused by EBV-hemophagocytic lymphohistiocytosis (EBV-HLH) (4/89, 4.5%), and NS-post-transplant lymphoproliferative disorder (NS-PTLD) (1/89, 1.1%). Anti-N-methyl-D-aspartate receptor encephalitis was found during the convalescence of EBV encephalitis. EBV encephalitis/meningitis showed no symptoms of tonsillitis, lymph node enlargement, skin rash, hepatosplenomegaly. Acute motor axonal neuropathy is the chief complication in GBS caused by EBV. There were significant differences in neurological complications caused by EBV. The prognosis of EBV infection in the NS is generally good. These illnesses are often self-limiting. A few cases may show residual sequelae.


Indicators of malnutrition in leukaemic children.
In 24 children with acute leukaemia a low serum albumin concentration (31 g/l or less) and a median weight:height ratio of less than 0.95 on admission were indicators of severe weight loss.


An outbreak of anaphylactic transfusion reactions to group B plasma and platelets and its possible relationship to Alpha-Gal syndrome.
Alpha-gal syndrome (AGS) is a potentially life-threatening acquired meat allergy associated with tick bites. The allergen Galactose-α-1,3-Galactose is antigenically similar to the B blood group antigen. B blood group status offers some protection against development of the allergy. Although not preferred practice, transfusion of plasma and platelets from group B donors is believed to be safe for group O recipients. Anaphylactic transfusion reactions were reported for three patients in two Washington DC hospitals from Nov 2022 to February 2023. A chart review was performed for all three patients. Patients or family members were interviewed, and IgE levels to alpha-gal were measured in two of the three patients. One reaction was acutely fatal. All reactions were to blood group B Plasma or Platelets in blood group O recipients. One patient had two prior anaphylactic reactions to group O RBCs and group B Plasma in a previous admission. All patients came from southern Maryland where AGS is an emerging problem. Two patients had histories of tick bites, previously unexplained gastrointestinal complaints, and abnormal elevated levels of IgE to alpha gal. Two patients had cat allergies. AGS is an emerging problem which may have implications for blood transfusion practice. Avoidance of blood group B antigen containing components may be prudent in non-blood group B patients with established AGS. Investigation for AGS should be considered in the evaluation of anaphylactic transfusion reactions.


Orteronel (TAK-700), a novel non-steroidal 17,20-lyase inhibitor: effects on steroid synthesis in human and monkey adrenal cells and serum steroid levels in cynomolgus monkeys.
Surgical or pharmacologic methods to control gonadal androgen biosynthesis are effective approaches in the treatment of a variety of non-neoplastic and neoplastic diseases. For example, androgen ablation and its consequent reduction in circulating levels of testosterone is an effective therapy for advanced prostate cancers. Unfortunately, the therapeutic effectiveness of this approach is often temporary because of disease progression to the 'castration resistant' (CRPC) state, a situation for which there are limited treatment options. One mechanism thought to be responsible for the development of CRPC is extra-gonadal androgen synthesis and the resulting impact of these residual extra-gonadal androgens on prostate tumor cell proliferation. An important enzyme responsible for the synthesis of extra-gonadal androgens is CYP17A1 which possesses both 17,20-lyase and 17-hydroxylase catalytic activities with the 17,20-lyase activity being key in the androgen biosynthetic process. Orteronel (TAK-700), a novel, selective, and potent inhibitor of 17,20-lyase is under development as a drug to inhibit androgen synthesis. In this study, we quantified the inhibitory activity and specificity of orteronel for testicular and adrenal androgen production by evaluating its effects on CYP17A1 enzymatic activity, steroid production in monkey adrenal cells and human adrenal tumor cells, and serum levels of dehydroepiandrosterone (DHEA), cortisol, and testosterone after oral dosing in castrated and intact male cynomolgus monkeys. We report that orteronel potently suppresses androgen production in monkey adrenal cells but only weakly suppresses corticosterone and aldosterone production; the IC(50) value of orteronel for cortisol was ~3-fold higher than that for DHEA. After single oral dosing, serum levels of DHEA, cortisol, and testosterone were rapidly suppressed in intact cynomolgus monkeys. In castrated monkeys treated twice daily with orteronel, suppression of DHEA and testosterone persisted throughout the treatment period. In both in vivo models and in agreement with our in vitro data, suppression of serum cortisol levels following oral dosing was less than that seen for DHEA. In terms of human CYP17A1 and human adrenal tumor cells, orteronel inhibited 17,20-lyase activity 5.4 times more potently than 17-hydroxylase activity in cell-free enzyme assays and DHEA production 27 times more potently than cortisol production in human adrenal tumor cells, suggesting greater specificity of inhibition between 17,20-lyase and 17-hydroxylase activities in humans vs monkeys. In summary, orteronel potently inhibited the 17,20-lyase activity of monkey and human CYP17A1 and reduced serum androgen levels in vivo in monkeys. These findings suggest that orteronel may be an effective therapeutic option for diseases where androgen suppression is critical, such as androgen sensitive and CRPC.


[Analysis on adaptive area of Curcuma wenyujin based on GIS].
This stdudy was to determine the suitable area for Curcuma wenyujin in Wenzhou, and expand its growing region, based the growth characteristic of Curcuma wenyujin. The 13 main factors including climate, soil and other environmental conditions were selected. The best genuine production area was regarded as the goal area in similar calculation. The grid data was calculated from the station and sampling sites by Inverse Distance Interpolation on GIS. After normolizing to the grid data, the dimension effect was removed. on this basis, calculating the absolute distance of every cell to the the goal cell. Wenzhou region was divided into best suitable area, suitable area, second suitable area and unsuitable area by clustering analyzing. Most of area in Wenzhou are suit for Curcuma wenyujin, best suitable area, suitable area, second suitable area and unsuitable area reach 98% area of Wenzhou. This result give important reference value of Curcuma wenyujin breeding in Wenzhou.


Autologous fat grafting for enhancement of breast reconstruction with a transverse myocutaneous gracilis flap: A cohort study.
The transverse myocutaneous gracilis (TMG) flap is a viable option for the reconstruction of small- to moderate-sized breasts. The purpose of this series was to examine the outcomes of fat grafting to breast and donor site secondary to breast reconstruction with a TMG flap. A cohort study was conducted to identify all patients who underwent autologous fat grafting after breast reconstruction with a TMG flap at two institutions between January 2009 and October 2013. Data were collected regarding patient demographics, body mass index, and medical history as well as surgical details regarding TMG flap surgery and fat grafting. A total of 145 fat graftings were performed in 83 patients following TMG flap breast reconstruction to improve the contour, shape, and volume of breasts (98%, n = 142), donor site (9%, n = 13) or both (7%, n = 10), mainly after breast cancer surgery (83%). On average, 1.8 fat grafting sessions were performed with an average of 30cc fat per breast and 56.4cc fat per thigh. In 72% of patients, fat injection was performed by other pending reconstructive procedures. Fat was harvested by the tumescent technique, by using a filter syringe system, and injected without any further processing in multiple layers. Autologous fat grafting as a secondary procedure represents a powerful adjunct to enhance the aesthetic outcome of the reconstructed breast and donor site. It can be easily combined with other pending reconstructive procedures. With additional fat grafting, it may be possible to expand indications and offer autologous reconstruction with a TMG flap even in very lean patients.


Plasma repressor element 1-silencing transcription factor levels are decreased in patients with Alzheimer's disease.
Repressor element 1-silencing transcription (REST)/neuron-restrictive silencer factor is considered a new therapeutic target for neurodegenerative disorders such as Alzheimer's disease (AD). However, the relationship between AD and REST remains unclear. This study aimed to 1) examine plasma REST levels and REST gene levels in AD patients and 2) further explore the pathological relationships between REST protein levels and cognitive decline in clinical conditions, including medial temporal lobe atrophy. Participants (n = 252, mean age 68.95 ± 8.78 years) were recruited in Beijing, China, and then divided into a normal cognition (NC) group (n = 89), an amnestic mild cognitive impairment (aMCI) group (n = 79), and an AD dementia group (n = 84) according to diagnostic criteria. All participants underwent neuropsychological assessments, laboratory tests, and neuroimaging scans (magnetic resonance imaging) at baseline. Plasma REST protein levels and the distribution of REST single nucleotide polymorphisms (SNPs) were compared among the three groups. Correlations between cognitive function, neuro-imaging results, and REST levels were determined by a multivariate linear regression analysis. The plasma REST levels in both the NC group (430.30 ± 303.43)pg/ml and aMCI group (414.27 ± 263.39)pg/ml were significantly higher than that in the AD dementia group (NC vs AD dementia group, p = 0.034; aMCI vs AD dementia group, p = 0.033). There was no significant difference between the NC and aMCI groups (p = 0.948). No significant difference was found among the three groups regarding the genotype distribution (rs2227902 and rs3976529 SNPs) of the REST gene. The REST level was correlated with the left medial temporal lobe atrophy index (r = 0.306, p = 0.023). After 6 months of follow-up, the REST level in the NC group was positively correlated with the change in the Mini-Mental State Examination score (r = 0.289, p = 0.02). The plasma REST protein level is decreased in AD dementia patients, which is associated with memory impairment and left temporal lobe atrophy and may have potential value for clinical diagnosis of AD dementia.


New osseous soft markers for trisomy 13, 18 and 21.
For ultrasonographic diagnosis of a fetal trisomy so-called "soft markers" (=ultrasonographically detectable morphological variants) are used. Detection of a certain number of them increases the diagnostic certainty of a fetal trisomy. Up to now there are very few diagnostically accepted osseous soft markers for trisomy. Hence potential osseous soft markers applicable for first and second trimester ultrasound screening for trisomy 21, 18 or 13 were studied. Postmortal fetal X-rays (ap, lateral) of 358 fetuses (trisomy 21: n = 109, trisomy 18: n = 46; trisomy 13: n = 38, control group: n = 165). Not yet described but with trisomy 21 statistically associated soft markers were un-timely os sternale ossification, delayed os sacrum ossification, shortened os maxillare, reduced os maxillare-jaw-corner distance, augmented orbita height, premature os calcaneus ossification, bell-shaped thorax, coronal clefts, trend to wider binocular as well as wider intraocular distances; for trisomy 18: elevated clavicula slope, reduced number of ribs, bell-shaped thorax, coronal clefts, reduced os maxillare-jaw-corner distance, shortened ramus mandibulare, shortened os metacarpale IV and V, augmented ratio between biparietal diameter and (osseus and soft-tissue) shoulder width; for trisomy 13: longer os nasale, elevated clavicula slope, premature sternum, delayed os sacrum ossification, delayed/premature cranium ossification, reduced number of ribs, coronal clefts, reduced os maxillare-jaw-corner distance, shortened ramus mandibulare, augmented orbita height, shortened os metacarpale V and a tendency for a shortened os metacarpale IV. We found several not yet published osseous soft markers statistically associated with trisomy 21, 18 and 13, which can help to ensure sonographically these aneuploidy diagnoses.


Effectiveness of home-based peer counselling to promote breastfeeding in the northeast of Brazil: a randomized clinical trial.
To evaluate the effectiveness of home-based peer counselling to increase breastfeeding rates for unfavourably low birthweight babies. Randomized clinical trial carried out in maternity hospitals and households in Fortaleza, one of the regions in Brazil with very low income; 1003 mothers and their newborns were selected in eight maternity hospitals. Newborns needed were healthy and weighed less than 3000 g. Breastfeeding counselling, conducted by lay counsellors from the community, during home visits carried out on days 5, 15, 30, 60, 90 and 120 after birth. Feeding methods in the fourth month of life. The intervention increased exclusive breastfeeding (24.7% vs 19.4%; p=0.044), delayed the introduction of formula and increased the time infants substituted breastfeeding to bottle milk (bottle milk 33.4% in the control group and 20.1% in the intervention group; p=0.00002). When comparing the frequency of artificial breastfeeding versus all other forms of breastfeeding (exclusive+predominant+partial), the intervention increased breastfeeding rates in 39% (RR=0.61; CI 95%: 0.50-0.75); 15% of children were free from artificial feeding (absolute risk reduction). The number of families to be visited to avoid one child receiving artificial feeding (NNT) was 7 (CI 95%: 5-13). Breastfeeding counselling, promoted by lay counsellors, can impact favourably on exclusive breastfeeding rates and contribute to delaying the utilization of milk formula and weaning. The intervention has great application potential because most cities in the northeast of Brazil count on community health workers that could do the counselling.


The role of language barriers on efficacy of rapid response teams.
Rapid response (RR) systems are associated with decreased hospital mortality. Systemic biases and inequities can negatively impact RR outcomes. Language barriers between patients and providers are associated with worse outcomes, but it is unknown if language barriers are associated with RR outcomes. We analyzed all adult hospitalized patients who experienced a RR over one year (January 2020 to December 2020) at a tertiary care academic medical center. We used an objective scoring system to establish disease severity at the time of the event. We then compared disease severity and outcomes for patients who are primary language Spanish (PLS) and primary language English (PLE) using both univariable and multivariable analyses. Of 1133 patients, 42 identified as PLS and 1091 as PLE. In multivariable analyses, PLS patients had significantly higher disease severity scores, as measured by deterioration index score (8.2, p = 0.021) at the time of their rapid responses. PLS patients also had 18.5% increase in length of stay (LOS) after RRs and this disparity was not mitigated when controlling for disease severity at the time of RRs. PLS was not a significant predictor for hospital mortality after RRs. Our study found that PLS patients had increased disease severity at the time of RRs and increased LOS after RRs. However, the disparity in LOS was not mitigated when controlling for disease severity at the time of RRs. These findings suggest that language barriers may cause both delays in activation of RR systems, as well as the care provided during and after RRs.


Effect of systemic administration of adenosine on brain adenosine levels in pentylenetetrazol-induced seizure threshold in mice.
Adenosine is one of the inhibitory neuromodulators in the brain. The present study was carried out to elucidate the effect of adenosine on the pentylenetetrazol (PTZ)-induced seizure threshold in mice. Further, the study also correlated the brain adenosine levels in PTZ-induced seizure threshold. PTZ (0.5%, w/v) was infused through lateral tail vein of mouse at a constant rate of 0.3 ml/min until various stages of convulsions were observed. Minimal dose of PTZ (mg/kg) needed to induce different phases (myoclonic jerks, generalized clonus and tonic extensor) of PTZ convulsions were noted as an index of seizure threshold. Intravenous infusion of PTZ resulted in a significant decrease in brain adenosine levels. Systemic administration of adenosine (100 and 200 mg/kg, i.p.), 30 min before PTZ infusion, produced a dose-dependent elevation of PTZ-seizure threshold and also enhanced brain adenosine levels as compared to vehicle treated group. The behavioral and neurochemical observations demonstrated a relationship between adenosine levels in the brain and the PTZ seizure threshold in mice.


Physiologic programming of the fetus.
In contrast to conventional theories, a series of provocative epidemiologic findings reported over the last decade suggest that environmental factors in early life are of substantial importance to disease risk in later years. To explain these findings, the idea of early life physiologic programming or imprinting has been advanced. Such programming has been documented in a variety of systems and reflects the action of a factor during a sensitive period or window of development to exert organizational effects that persist throughout life. Programming agents might include growth factors, hormones, and nutrients. These factors may produce adaptations that permanently alter adult metabolism and responses in a direction optimizing survival under continued conditions of malnutrition, stress, or other deprivation, but such responses might be detrimental when the later environment is unexpectedly less challenging.


Synthesis of glycyrrhizin analogues as HMGB1 inhibitors and their activity against sepsis in acute kidney injury.
Glycyrrhizin (GL) is one of the antagonists of highly conserved nuclear protein (HMGB1). The researches have shown that the glycosyl of GL is an important pharmacophore for GL binding to HMGB1, and it is the determinant factor for mechanism of action. To get the HMGB1 inhibitors with higher activity and good pharmacokinetic properties, two classes of GL analogues containing C-N glycoside bond were synthesized, and their anti-inflammatory, anti-oxidative stress and anti-septic kidney injury were evaluated. The results are as follows. First, in the anti-inflammatory assay, all the compounds inhibited NO release in some degree; among them, compound 6 displayed the strongest NO inhibitory effect with IC50 value of 15.9 μM, and compound 15 with IC50 of 20.2 μM. The two compounds not only decreased IL-1β and TNF-α levels in RAW264.7 cells and HK-2 cells, but also downregulated the levels of NLRP3, P-NF-κB p65 and HMGB1 in activated HK-2 cells in a dose-dependent manner. Second, in the renal protection assay with H2O2-stimulated HK-2 cell line, they reduced MDA level and increased SOD in HK-2 cells; additionally, they also inhibited the HK-2 cell apoptosis and downregulated the Caspase-1 p20 level. Third, in the in vivo activity tests of the septic mouse, they also showed good activities just like in vitro, decreasing the IL-1β, TNF-α, MDA, blood creatinine (Scr) and urea nitrogen (BUN) in serum, and increasing SOD levels in a dose-dependent manner. The immunoblotting results showed the two compounds downregulated the levels of HMGB1, P-NF-κB p65, NLRP3 and Caspase-1 p20 protein. All in all, the two compounds improved the renal injury of septic mice, and alleviated the tube wall structure damage and renal tubular dilation in kidney, which further proved by H&E staining. This suggests the two compounds have septic acute kidney injury activity, and they will be potential therapeutic drugs for septic acute kidney injury.


ERK/pERK expression and B-raf mutations in colon adenocarcinomas: correlation with clinicopathological characteristics.
Colorectal (CRC) carcinogenesis through various morphological stages has been linked to several genetic and epigenetic changes. The Raf/MEK/ERK (MAPK) signal transduction cascade is an important mediator of a number of cellular fates. In this study, we investigated the presence of B-raf and K-ras mutations in 94 consecutive cases of primary colon adenocarcinoma in correlation with the immunohistochemical expression of total and activated ERK and the expression of mismatch repair proteins (MMR) hMLH1 and hMSH2 as well as their correlations with standard clinicopathological parameters. The immunostaining pattern for total and activated ERK was nuclear and cytoplasmic. hMLH1 and hMSH2 proteins were preserved in 45/63 (71.43%) cases and 35/53 (66.04%) cases respectively. Total ERK nuclear expression, was positively correlated with tumor stage (p = 0.049), whereas nuclear pERK expression was positively correlated with histological grade (p = 0.0113) and tumor stage (p = 0.0952), although the latter relationship was of marginal significance. DNA sequencing showed that 12 samples (12.7%) had a mutation in B-RAF Exon 15 and none in Exon 11, whereas 22 (23.4%) had a K-ras mutation. Disruption of the MAP kinase pathway-either through K-ras or B-raf mutation-was detected in 37% of all the examined cases, although the overexpression of total and activated ERK1/2 was not correlated with the mutational status of K-ras or B-raf genes. Finally, the preservation of hMLH1 or hMSH2 immunoexpression was not correlated with the presence of B-raf and/or K-ras mutations. In this study, we present evidence that ERK activation occurs in a K-ras or B-raf -independent manner in the majority of primary colon cancer cases. Moreover, B-raf mutations are not associated with mismatch-repair deficiency through loss of hMLH1 or hMSH2 expression. Activated ERK could possibly be implicated in tumor invasiveness as well as in the acquisition of a more aggressive phenotype.


Enterococcus spp. produces slime and survives in rat peritoneal macrophages.
Enterococcal clinical isolates were investigated for the ability to form biofilm on inert surfaces, as a measure of slime production, in an attempt to find new possible virulence factors for these microorganisms. This property was commonly found among Enterococcus faecalis. Also E. faecium isolates were able to form biofilm, although to a lesser extent; for this species, however, biofilm formation seemed more frequently associated with isolates from infection rather than with environmental strains or isolates from healthy individuals. Biofilm formation was strongly affected by the presence of an additional carbohydrate source in the medium, or by iron deprivation, indicating a role of slime for survival in stressful conditions. Slime-producing E. faecalis were able to survive inside peritoneal macrophages for extended periods compared to slime-negative strains or to slime-positive bacteria grown in conditions depressing slime production. In particular, slime-producing and slime-negative cells showed a decrease of 1 and 2 log units, respectively, at 1 h after infection; slime-negative cells were then rapidly killed, with clearance of bacterial cells at 24 h. Slime-producing bacteria persisted up to 48 h, which was the last time point examined, as after that time viability of both infected and non-infected macrophages started to decline. Scanning electron microscopy observations showed the presence of abundant amorphous extracellular material, of possible polysaccharide nature, embedding bacterial cells to form a multilayered biofilm. Even in conditions not supporting biofilm formation, bacterial cells appeared capsulated, suggesting that capsule and slime might represent different structures. Genes belonging to the epa locus or to a putative icaA homolog did not seem to be involved in synthesis and export of slime.


Functional response of fish communities in a multistressed freshwater world.
Freshwater fish communities are impacted by multiple pressures, determining loss of functional diversity and redundancy. Our aim was to disentangle the roles and relevancies of different pressures in shaping fish communities in small streams of the Po plain (North Italy). Long term trend (1998-2018) of functional diversity of 31 fish communities was assessed and modeled in respect to three potential pressures: temperature increase, intensity of exotic fish invasion, and habitat quality degradation. Ecological traits mostly influenced by the pressures were also identified. Reduction of functional richness mostly due to local extinction or contraction of cold adapted predators, such as salmonids, was linked to increasing temperatures. Warming probably also led to a shift of generalist and dominant species, which became more abundant in streams hosting mixed communities of salmonids and cyprinids, and determined the increase of functional dispersion and uniqueness. Reduction of functional redundancy and increasing functional dispersion were both also related to the introduction of new ecological traits brought by expanding exotic species. Low functional overlap was found among native and exotic species, indicating that the invasion process was mainly controlled by competitive interactions and/or resource opportunism. Functional response to habitat quality was not clearly evident. In conclusion, the impact of temperature increase and exotic species on fish functional diversity was effective, idiosyncratic and mediated by the scale of analysis and by the intensity of pressures.


Metal ion-mediated regulation of heme oxygenase induction in cultured avian liver cells.
The induction of heme oxygenase (EC 126.96.36.199) in response to various metal treatments was investigated in monolayer cultures of chick embryo liver cells maintained in a chemically defined serum-free medium. The most potent heme oxygenase-inducing action was exhibited by CO2+, Cd2+, Sb3+, As3+, and Au1+ followed by lesser induction observed with Cu2+, Fe2+, and Fe3+. Mn2+, Ni2+, Se4+, Sn2+, and Zn2+ were without effect. In contrast to the marked inducing effect of Co2+ on heme oxygenase, Co-protoporphyrin IX decreased the enzyme activity in a dose-dependent manner. Addition of Zn2+ (20 microM) to Co2+-treated liver cell cultures revealed a striking ability of Zn2+ to block completely Co2+-induced heme oxygenase. Simultaneous addition of Mn2+ (50 microM) to Co2+-treated cells also blocked Co2+-induced heme oxygenase (approximately 50%). These findings in tissue culture confirm those made earlier in whole animals (Drummond, G. S., and Kappas, A. (1979) Proc. Natl. Acad. Sci. U. S. A. 76, 5331-5335) and indicate that these effects of Zn2+ and Mn2+ are exerted directly in liver cells. Addition of cysteine (400 microM) to the cultures also inhibited heme oxygenase induction by Co2+ substantially. Cycloheximide and actinomycin D blocked the induction of heme oxygenase, indicating that increased heme oxygenase activity by metal treatment is dependent on fresh RNA and protein synthesis. The half-life of the enzyme was calculated to be approximately 15 h after treatment with cycloheximide. These findings provide further evidence that metal ions can regulate heme oxygenase synthesis directly in isolated liver cells and that the metal-metal interactions which lead to blockade of the enzyme induction do not involve extrahepatic tissues.


Auxin's origin: do PILS hold the key?
Auxin is a key regulator of many developmental processes in land plants and plays a strikingly similar role in the phylogenetically distant brown seaweeds. Emerging evidence shows that the PIN and PIN-like (PILS) auxin transporter families have preceded the evolution of the canonical auxin response pathway. A wide conservation of PILS-mediated auxin transport, together with reports of auxin function in unicellular algae, would suggest that auxin function preceded the advent of multicellularity. We find that PIN and PILS transporters form two eukaryotic subfamilies within a larger bacterial family. We argue that future functional characterisation of algal PIN and PILS transporters can shed light on a common origin of an auxin function followed by independent co-option in a multicellular context.


Hospital-associated MRSA and antibiotic resistance-what have we learned from genomics?
In many parts of the world, MRSA are responsible for a high proportion of S. aureus infections in patients in contact with healthcare. Molecular studies have shown this is due to one or more MRSA clones that have become endemic in each hospital or healthcare facility, resulting in hospital- or healthcare-associated MRSA (HA-MRSA). The infection rate and clones responsible for HA-MRSA can vary substantially in different geographical locations. Molecular methods have allowed clones to be categorized, as well as the opportunity to track the evolution and spread of clones in healthcare settings and around the world. The genomes of HA-MRSA isolates belonging to the same clonal group can show dramatic variability particularly in the carriage of mobile genetic elements (MGEs) encoding virulence and resistance genes. HA-MRSA are potentially resistant to all classes of antibiotics, although individual isolates that are fully drug resistant are not reported. The incidence of fluoroquinolone resistance in HA-MRSA is remarkably high, suggesting use of this class of antibiotics as well as the β-lactams contributes to the selection and success of HA-MRSA clones in the hospital setting.


[Mobile seizure monitoring in epilepsy patients].
Wearables are receiving much attention from both epilepsy patients and treating physicians, for monitoring of seizure frequency and warning of seizures. They are also of interest for the detection of seizure-associated risks of patients, for differential diagnosis of rare seizure types and prediction of seizure-prone periods. Accelerometry, electromyography (EMG), heart rate and further autonomic parameters are recorded to capture clinical seizure manifestations. Currently, a clinical use to document nocturnal motor seizures is feasible. In this review the available devices, data on the performance in the documentation of seizures, current options for clinical use and developments in data analysis are presented and critically discussed.


cDNA clone and expression analysis of rodent fast and slow skeletal muscle troponin I mRNAs.
We have characterized the structure and expression of rodent mRNAs encoding the fast and slow skeletal muscle isoforms of the contractile regulatory protein, troponin I (TnIfast and TnIslow). TnIfast and TnIslow cDNA clones were isolated from mouse and rat muscle cDNA clone libraries and were used as isoform-specific probes in Northern blot and in situ hybridization studies. These studies showed that the TnIfast and TnIslow mRNAs are expressed in skeletal muscle, but not cardiac muscle or other tissues, and that they are differentially expressed in individual muscle fibers. Fiber typing on the basis of in situ hybridization analysis of TnI isoform mRNA content showed an excellent correlation with fiber type as assessed by myosin ATPase histochemistry. These results directly demonstrate that the differential expression of skeletal muscle TnI isoforms in the various classes of vertebrate striated muscle cells is based on gene regulatory mechanisms which control the abundances of specific TnI mRNAs in individual muscle cells. Both TnIfast and TnIslow mRNAs are expressed, at comparable levels, in differentiated cultures of rat L6 and mouse C2 muscle cell lines. Thus, although neuronal input has been shown to be an important factor in determining fast versus slow isoform-specific expression in skeletal muscle, both TnIfast and TnIslow genes can be expressed in muscle cells in the absence of nerve. Comparison of the deduced rodent TnI amino acid sequences with previously determined rabbit protein sequences showed that residues with potential fast/slow isoform-specific function are present in several discrete clusters, two of which are located near previously identified actin and troponin C binding sites.


Deletion of the κ genes from mouse hybridomas established with NS-1 myelomas.
Monoclonal antibodies (mAbs) of the IgG class produced by mouse hybridomas raised with NS-1 myelomas have been shown to contain two types of immunoglobulin light (κ) chains derived from the myelomas and antigen-stimulated spleen lymphocytes, and the hybridomas produce three mAb species with light chain heterogeneity (Abe and Inouye, 1993). In the present study, 9 hybridoma lines secreting homogeneous mAbs have been isolated from 63 lines cloned from an established hybridoma line producing three mAbs. They secrete homogeneous mAbs containing light chains derived from either myeloma or spleen cells. They contain either κ gene derived from the respective cells, and the other gene was deleted during the cultivation. The deletion frequency of the κ gene of myelomas is 3 times higher than that of spleen cells, although 80-85% of hybridomas reach the stable state containing both κ genes.