NBS1 protein from Physcomitrium patens confers protection against oxidative damage by limiting the accumulation of cellular reactive oxygen species.
Nijmegen breakage syndrome 1 (NBS1) protein is a core member of the MRE11-RAD50-NBS1 (MRN) complex that plays a crucial role in DNA damage sensing and repair in plants. Here we report that NBS1 from moss Physcomitrium patens reduces oxidative damage by lowering the cellular ROS in addition to its known role in oxidative DNA damage recovery. Real-time transcript analysis showed up-regulation of the PpNBS1 transcript under different stress conditions. Bacterial cells showed better cell survivability upon over-expressing PpNBS1 protein as compared to untransformed cells. Likewise, overexpression of PpNBS1 in tobacco plants provides improved protection against oxidative damage and exhibited a lesser amount of ROS upon exposure to oxidative stress. Moreover, PpNBS1 contributes to the antioxidant defense mechanism by positively regulating the expression of the antioxidant genes under stress conditions in transgenic tobacco plants. PpNBS1 expressing transgenic tobacco plants resulted in lesser membrane damage, lower lipid peroxidation level, and higher chlorophyll content under stress conditions. Taken together, we conclude in addition to its known role as DNA damage sensor, PpNBS1 also plays a definite role in oxidative stress mitigation by minimizing ROS accumulation in the cell.


Missed opportunities for influenza vaccination in children with chronic medical conditions.
To assess the frequency and characteristics of missed opportunities for influenza immunization in children with chronic medical conditions and, among unimmunized children in that group, to explore parent-reported reasons for not vaccinating their child. Prospective cohort study. Data were obtained from billing and immunization registry databases and telephone interviews of parents. Four pediatric practices in metropolitan Denver, Colo, during the 2002-2003 influenza season. Children aged 6 to 72 months with 1 or more chronic medical conditions. Main Outcome Measure A missed opportunity for influenza immunization, defined as having a billed encounter, being eligible for immunization, and not receiving vaccine. Subjects with asthma were analyzed separately from those with other conditions. We identified 926 children with chronic conditions: 820 (89%) with asthma only and 106 (11%) with other conditions. Missed opportunities occurred at 78% of all vaccine-eligible visits for children with asthma and 74% of all visits for children with other conditions. For children with asthma, 92% of vaccine-eligible visits in December and January resulted in a missed opportunity vs 69% in October and November; for children with other conditions, corresponding frequencies were 86% vs 68%, respectively. For children with asthma, 86% of non-well-child visits resulted in a missed opportunity, compared with 62% of well-child visits; similar frequencies were seen among children with other chronic conditions. Parents of unimmunized children reported lack of a physician recommendation and low perceived susceptibility to influenza as the primary reasons for not immunizing their child. Missed opportunities contribute significantly to low influenza immunization rates among children with chronic medical conditions.


Evaluating different web applications to assess the toxicity of plasticizers.
Plasticizers increase the flexibility of plastics. As environmental leachates they lead to increased water and soil pollution, as well as to serious harm to human health. This study was set out to explore various web applications to predict the toxicological properties of plasticizers. Web-based tools (e.g., BOILED-Egg, LAZAR, PROTOX-II, CarcinoPred-EL) and VEGA were accessed via an 5th-10th generation computer in order to obtain toxicological predictions. Based on the LAZAR mutagenicity assessment was only bisphenol F predicted as mutagenic. The BBP and DBP in RF; DEHP in RF and XGBoost; DNOP in RF and XGBoost models were predicted as carcinogenic in the CarcinoPred-EL web application. From the bee predictive model (KNN/IRFMN) BPF, di-n-propyl phthalate, diallyl phthalate, dibutyl phthalate, and diisohexyl phthalate were predicted as strong bee toxicants. Acute toxicity for fish using the model Sarpy/IRFMN predicted 19 plasticizers as strong toxicants with LC50 values of less than 1 mg/L. This study also considered plasticizer effects on gastrointestinal absorption and other toxicological endpoints.


Alpha tocopherol and blood pressure in PN/n mice.
The tail cuff blood pressure of PN/n mice, previously studied for arteritis and glomerulitis, was found to be raised before histological changes appeared. The pressure of mice bred for two generations on a diet containing a low level of alpha tocopherol (E) fell to that of control CBA mice. Replacement of E in the diet, or by intraperitoneal injection, led to a rise of pressure. The pressure reached a mean of 150 mm/Hg after breeding the mice for a further three generations on a more complete diet with additional E. An inhibitory effect of E on prostaglandin production was postulated. E had no effect on the pressure of CBA/MAC mice.


28-day sepsis mortality prediction model from combined serial interleukin-6, lactate, and procalcitonin measurements: a retrospective cohort study.
Sepsis is a global medical issue owing to its unacceptably high mortality rate. Therefore, an effective approach to predicting patient outcomes is critically needed. We aimed to search for a novel 28-day sepsis mortality prediction model based on serial interleukin-6 (IL-6), lactate (LAC), and procalcitonin (PCT) measurements. We enrolled 367 septic patients based on Sepsis-3 (Third International Consensus Definitions for Sepsis and Septic Shock). Serum IL-6, LAC, and PCT levels were measured serially. Results collected within 24 and 48-72 h of admission were marked as D1 and D3 (e.g., IL-6D1/D3), respectively; the IL-6, LAC, and PCT clearance (IL-6c, LACc, PCTc) at D3 were calculated. Data were split into training and validation cohorts (7:3). Logistic regression analyses were used to select variables to develop models and choose the best one according to the Akaike information criterion (AIC). Receiver operating characteristic curves (ROC), calibration plots, and decision curve analysis (DCA) were used to test model performance. A nomogram was used to validate the model. There were 314 (85.56%) survivors and 53 (14.44%) non-survivors. Logistic regression analyses showed that IL-6D1, IL-6D3, PCTD1, PCTD3, and LACcD3 could be used to develop the best prediction model. The areas under the curves (AUC) of the training (0.849, 95% CI: 0.787-0.911) and validation cohorts (0.828, 95% CI: 0.727-0.929), calibration plot, and the DCA showed that the model performed well. Thus, the predictive value of the risk nomogram was verified. Combining IL-6D1, IL-6D3, PCTD1, PCTD3, and LACcD3 may create an accurate prediction model for 28-day sepsis mortality. Multiple-center research with a larger quantity of data is necessary to determine its clinical utility.


Stable isotope dilution assay for prostaglandin E metabolite: 24-hour urinary output in healthy male subjects.
A stable isotope dilution assay for the major urinary metabolite of prostaglandins E1 and E2 (PGE-M) was developed. After extraction and purification, the compound was converted to the dimethyl ester bis(O-methyloxime)-trimethylsilyl ether derivative and quantified with the 13C-labeled dimethyl ester analog as internal standard. The interassay coefficient of variation was 0.8%, and the intraassay coefficient of variation ranged from 0.9 to 1.7%. The recovery of authentic, underivatized PGE-M added to urine was 99.5 +/- 3.2% (mean +/- SE, N = 9). The equation of the regression curve correlating the amounts added and recovered was Y = 1.05X - 5.84, with R = 0.998. The level of excreted metabolite in healthy male subjects was 13.86 +/- 1.46 micrograms/24 h (N = 24), in good agreement with data from other sources.


School Age Effects of the Newborn Individualized Developmental Care and Assessment Program for Medically Low-Risk Preterm Infants: Preliminary Findings.
By school age, even low risk moderately preterm-born children show more neuro-cognitive deficits, underachievement, behavioral problems, and poor social adaptation than full-term peers. To evaluate the outcomes at school-age for moderately preterm-born children (29-33 weeks gestational age), appropriate in growth for gestational age (AGA) and medically at low-risk, randomized to Newborn Individualized Developmental Care and Assessment Program (NIDCAP) or standard care in the Newborn Intensive Care Unit. At school-age, the experimental (E) group will show better neuropsychological and neuro-electrophysiological function, as well as improved brain structure than the control (C) group. The original sample consisted of 30 moderately preterm-born infants (29 to 33 weeks), 23 (8C and 15E) of them were evaluated at 8 years of age, corrected-for-prematurity with neuropsychological, EEG spectral coherence, and diffusion tensor magnetic resonance imaging (DT MRI) measures. E-performed significantly better than C-group children on the Kaufman Assessment Battery for Children-Second Edition (KABC-II) and trended towards better scores on the Rey-Osterrieth Complex Figure Test. They also showed more mature frontal and parietal brain connectivities, and more mature fiber tracts involving the internal capsule and the cingulum. Neurobehavioral results in the newborn period successfully predicted neuropsychological functioning at 8 years corrected age. Moderately preterm infants cared for with the NIDCAP intervention showed improved neuropsychological and neuro-electrophysiological function as well as improved brain structure at school-age.


Role of adiponectin in sphingosine-1-phosphate induced airway hyperresponsiveness and inflammation.
Epidemiological data suggest that obesity represent an important risk factor for asthma, but the link between excess fat and airway hyperresponsiveness (AHR) and inflammation is not fully understood. Recently, a key role in physiopathologic conditions of lungs has been given to adiponectin (Acrp30). Acrp30 is one of the most expressed adipokines produced and secreted by adipose tissue, showing an intriguing relationship with metabolism of sphingolipids. Sphingosine-1-phosphate (S1P) has been proposed as an important inflammatory mediator implicated in the pathogenesis of airway inflammation and asthma. In the present study we analyze the effects of recombinant Acrp30 administration in an experimental model of S1P-induced AHR and inflammation. The results show that S1P is able to reduce endogenous Acrp30 serum levels and that recombinant Acrp30 treatment significantly reduce S1P-induced AHR and inflammation. Moreover, we observed a reduction of Adiponectin receptors (AdipoR1, AdipoR2 and T-cadherin) expression in S1P treated mice. Treatment with recombinant Acrp30 was able to restore Acrp30 serum levels and adiponectin receptors expression. These results could indicate the ability of S1P to modulate the Acrp30 action, by modulating not only the serum levels of the protein, but also its receptors. Taken together, these data suggest that adiponectin could represent a possible biomarker in obesity-associated asthma.


Occurrence and significance of odontoid lateral mass interspace asymmetry in trauma patients.
The odontoid lateral mass interspace (OLMI) is the space between the lateral aspect of the dens axis and the medial circumference of the massa lateralis atlantis. The position of OLMI asymmetry as a normal variant or pathologic finding is an area of debate and clinical interest in trauma patients. We designed this prospective study to lay a framework for proposing strategies for the appropriate use of OLMI. A total of 301 adult patients admitted for trauma were included. Computed tomography (CT) and magnetic resonance imaging (MRI) of the cervical spine were performed and examined for the presence OLMI asymmetry and bony/ligamentous lesions of the occipitoatlantoaxial complex. Head rotation is linked to the occurrence of OLMI asymmetry. Reliable OLMI asymmetry evaluation is limited by observer agreement under a threshold value of 1.0 mm. In all, 86 patients (28.6 %) were found to have OLMI asymmetry on CT after trauma. Among these patients, 17.4 % had a bony/ligamentous lesion of the occipitoatlantoaxial complex. Among the patients without OLMI asymmetry, 8.8 % were found to have such lesions. OLMI asymmetry should only be investigated by CT scans of the head in optimal position and with the threshold value of 1.0 mm. OLMI asymmetry should not be used alone as a sign of a cervical spine lesion. MRI should be performed if: (1) the physician has a high degree of suspicion of a cervical spine lesion; (2) OLMI asymmetry was demonstrated on a technically adequate CT scan; (3) clinical symptoms persist in patients with OLMI asymmetry when no acute MRI was performed.


Infection of replication-deficient adenoviral vector enhances interleukin-8 production in small airway epithelial cells more than in large airway epithelial cells.
In clinical trials or experiments of gene therapy, airway administration of an adenoviral-based vector (E1A-deleted) elicits a dose-dependent inflammatory response with limitation in the duration of transgene expression. The purpose of this study was to evaluate the possibility that the adenoviral-based vector directly enhances IL-8 production independent of adenoviral E1A in normal human airway epithelial cells and to examine the different responses between primary human bronchial epithelial cells (HBE) and primary human small airway epithelial cells (HSAE) in production of IL-8 following exposure to an adenovirus vector. Interleukin (IL)-8 levels were evaluated in the culture medium from HBE and HSAE treated with increasing doses of E1A-deleted adenoviral vector contained the Escherichia coli LacZ reporter gene (AdCMVLacZ). To clarify the mechanism of enhancing IL-8 production in airway epithelial cells by infection with adenovirus vector, alphavbeta5 agonistic antibody as an analogue of adenoviral capsid and adenoviral capsid vector denatured by exposure to ultraviolet (UV) light were used in the present study. Inoculation of HBE with AdCMVLacZ at a multiplicity of infection (MOI) of between 1 and 200 resulted in a dose-dependent expression of LacZ, and maximal expression was observed at a MOI of 100. In contrast, inoculation of HSAE with AdCMVLacZ resulted in maximum expression of LacZ at a MOI of 10. Interleukin-8 levels in culture media from the same experiments revealed significantly greater production of IL-8 in HSAE inoculated with AdCMVLacZ at a MOI of 50, compared to HBE under the same conditions. The capsid-denatured adenoviral vector did not enhance IL-8 production, and alphavbeta5 agonistic antibody induced IL-8 enhancement. These results suggest that the adenoviral vector directly induces the expression of airway epithelial inflammatory cytokines in the pathogenesis of inflammation and that small airway cells have a greater affinity for adenovirus than other airway epithelial cells.


[The synthesis of chalcone and flavanone semicarbazones from 1,3- and 1,4-dibenzodioxane and 1,5-benzodioxepane analogues of chalcones].
In dependence on endurance, the reaction of chalcones with thiosemicarbazide resulted in chalcone or flavanone thiosemicarbazones, whose structures were proven by 1H NMR.


Tetrachloroethylene (PCE, Perc) levels in residential dry cleaner buildings in diverse communities in New York City.
Fugitive tetrachloroethylene (PCE, perc) emissions from dry cleaners operating in apartment buildings can contaminate residential indoor air. In 1997, New York State and New York City adopted regulations to reduce and contain perc emissions from dry cleaners located in residential and other buildings. As part of a New York State Department of Health (NYSDOH) study, indoor air perc levels were determined in 65 apartments located in 24 buildings in New York City where dry cleaners used perc on site. Sampling occurred during 2001-2003, and sampled buildings were dispersed across minority and nonminority as well as low-income and higher income neighborhoods. For the entire study area, the mean apartment perc level was 34 microg/m3, 10-fold lower than mean apartment levels of 340-360 microg/m3 documented before 1997. The maximum detected perc level was 5,000 microg/m3, 5-fold lower than the maximum of 25,000 microg/m3 documented before 1997. Despite these accomplishments, perc levels in 17 sampled apartments still exceeded the NYSDOH residential air guideline of 100 microg/m3, and perc levels in 4 sampled apartments exceeded 1,000 microg/m3. Moreover, mean indoor air perc levels in minority neighborhoods (75 microg/m3) were four times higher than in nonminority households (19 microg/m3) and were > 10 times higher in low-income neighborhoods (256 microg/m3) than in higher income neighborhoods (23 microg/m3). Logistic regression suitable for clustered data (apartments within buildings) indicated that perc levels on floors 1-4 were significantly more likely to exceed 100 microg/m3 in buildings located in minority neighborhoods (odds ratio = 6.7; 95% confidence interval, 1.5-30.5) than in nonminority neighborhoods. Factors that may be contributing to the elevated perc levels detected, especially in minority and low-income neighborhoods, are being explored.


The aging reproductive neuroendocrine axis.
It is well known that the reproductive system is one of the first biological systems to show age-related decline. While depletion of ovarian follicles clearly relates to the end of reproductive function in females, evidence is accumulating that a hypothalamic defect is critical in the transition from cyclicity to acyclicity. This minireview attempts to present a concise review on aging of the female reproductive neuroendocrine axis and provide thought-provoking analysis and insights into potential future directions for this field. Evidence will be reviewed, which shows that a defect in pulsatile and surge gonadotropin hormone-releasing hormone (GnRH) secretion exists in normal cycling middle-aged female rats, which is thought to explain the significantly attenuated pulsatile and surge luteinizing hormone (LH) secretion at middle-age. Evidence is also presented, which supports the age-related defect in GnRH secretion as being due to a reduced activation of GnRH neurons. Along these lines, stimulation of GnRH secretion by the major excitatory transmitter glutamate is shown to be significantly attenuated in middle-aged proestrous rats. Corresponding age-related defects in other major excitatory regulatory factors, such as catecholamines, neuropeptide Y, and astrocytes, have also been demonstrated. Age-related changes in hypothalamic concentrations of neurotransmitter receptors, steroid receptors, and circulating steroid hormone levels are also reviewed, and discussion is presented on the complex interrelationships of the hypothalamus-pituitary-ovarian (HPO) axis during aging, with attention to how a defect in one level of the axis can induce defects in other levels, and thereby potentiate the dysfunction of the entire HPO axis.


Extremity gossypiboma mimicking sarcoma: case report and review.
A 70-year-old man with a history of bladder and colon cancer presented with an enlarging mass in his right lower extremity. Forty years before presentation, he had injured his right lower extremity in a motor vehicle accident. Imaging findings indicated suspected sarcoma, which led to biopsy. Biopsy and further surgical exploration revealed the presence of a surgical sponge and surrounding local inflammatory reaction. No neoplasm was found, and the sponge and involved tissues were removed. Gossypiboma is exceedingly rare in the extremities. Imaging of retained foreign material can appear suggestive of sarcoma because of strong inflammatory responses and local tissue mass-like derangement resulting in heterogeneous signal changes. Ultimately, biopsy must be performed to ensure that no oncological pathological condition is present.


Application of magnetic chromatography to the isolation of lysosomes from fibroblasts of patients with lysosomal storage disorders.
A method for the purification of lysosomes from fibroblasts has been developed which uses endocytosis of superparamagnetic colloidal iron dextran particles followed by separation of the iron-containing lysosomes in a magnetic field. This permitted isolation of lysosomes from fibroblasts from patients with infantile sialic acid storage disorder and other lysosomal storage diseases in which a shift in lysosomal density induced by the storage material prevents purification by centrifugation in a Percoll gradient. The magnetic lysosomes isolated from these cells are very similar to those from normal cells as judged by lysosomal marker enzyme activity and 2D-PAGE analysis of the enriched proteins.


Mechanisms of Disruptive Action of Dichlorodiphenyltrichloroethane (DDT) on the Function of Thyroid Follicular Epitheliocytes.
The mechanism of disruptive action of dichlorodiphenyltrichloroethane (DDT) on the function of thyroid follicular epitheliocytes was examined in mature male Wistar rats administered with a daily dose of DDT (2 μg/kg) for 6 or 10 weeks. In 6 weeks, the serum level of Na(+)/I(-) symporter responsible for the transport of iodides into follicular thyrocytes decreased, while serum concentration of thyroperoxidase increased. In 10 weeks, both the number of follicles in the thyroid gland and serum level of Na(+)/I(-) symporter increased. In circulating blood, the concentrations of thyroglobulin as well as α and β receptors of the thyroid hormones did not change. DDT down-regulated the iodine-accumulated function of follicular thyrocytes by suppressing Na(+)/I(-) symporter synthesis. Recovery of Na(+)/I (-) symporter production resulted from up-regulation of the proliferative processes in thyroid gland.


The hope theory and specific learning disorders and/or attention deficit disorders (SLD/ADHD): Developmental perspectives.
Research on disabilities has generally used a deficit-oriented approach. However, the recent appreciation of the personal strengths and positive traits of the disabled has raised awareness of them as empowering and activating factors. According to the hope theory, the ability to embrace future perspectives, set meaningful goals, and plan specific paths to reach these goals predict well-being and adjustment. We review the research on the role of hope in the lives of individuals with SLD/ADHD from a developmental perspective. We investigate hope as a resource in predicting these people's ability to deal with personal risks through developmental stages and contextual conditions. We conclude by proposing future research and intervention directions.


Community-acquired pneumonia.
Community-acquired pneumonia (CAP) is a significant cause of morbidity and mortality in all age groups, especially the elderly, which is a patient population that continues to grow. Recently the spectrum and clinical picture of pneumonia has been changing as a reflection of this aging population; this requires a reassessment of and a new approach to the patient with pneumonia. Currently, pneumonia patients are classified as having either community-acquired or hospital-acquired infection rather than typical versus atypical. Patients who have CAP are categorized by age, presence of a coexisting medical illness, and the severity of the pneumonia. The rationale behind categorizing patients is to stratify them in terms of mortality risk to help determine the location of therapy (e.g., outpatient, inpatient, intensive care unit) and focus the choice of initial antimicrobial therapy. Once the decision to hospitalize a patient with pneumonia is made, the next step is to decide on an appropriate diagnostic evaluation and antibiotic therapy. Both decisions have evolved over the last several years since the publication of the American Thoracic Society's CAP guidelines. The current approach to the diagnostic work-up of pneumonia stresses a limited role of diagnostic tests and procedures. The antimicrobial regimen has now evolved into one that is empiric in nature and based on the age of the patient, the presence of coexisting medical disease, and the overall severity of the pneumonia. This process is a dynamic once because bacterial resistance to commonly used antibiotics can further complicate the course of pneumonia therapy, but the impact of resistance on outcome is less clear. Resistance of Streptococcus pneumoniae to penicillin is a prime example of this growing problem, and adjustment to pneumonia therapy may be required. A difficult but not uncommon problem in pneumonia patients is slow recovery and delayed resolution of radiographic infiltrates. Factors that impact negatively on pneumonia resolution include advanced age and the presence of serious comorbid illnesses such as diabetes mellitus, renal disease, or chronic obstructive pulmonary disease. In addition, certain organism factors (e.g., intrinsic virulence) may interact with host factors and advanced age to delay pneumonia resolution. For example, 50% of patients with pneumococcal pneumonia have radiographic clearing at 5 weeks, and the majority clear within 2 to 3 months. Recent data demonstrate that radiographic resolution of CAP is most influenced by the number of lobes involved and the age of the patient. Radiographic clearance of CAP decreases by 20% per decade after age 20, and patients with multilobar infiltrates take longer to clear than those with unilobar disease. In general, when approaching slowly resolving infiltrates after pneumonia, bronchoscopic evaluation and lung biopsy are more likely to yield a specific diagnosis if the patient is a nonsmoker younger than 55 years old with multilobar disease. If the patients has either no identifiable factors associated with prolonged pneumonia resolution or the repeat chest radiograph at 1 month shows no appreciable change, further diagnostic testing is indicated. The route and duration of antibiotic therapy, another detail of the management of CAP patients that has changed recently, is complicated by the fact that the majority of patients with CAP have no pathogen identified. Therefore, in most instances the physician initiates empiric antibiotics on the basis of epidemiologic data. If an etiologic pathogen is identified (either initially or at a later time), then the antibiotic spectrum can be narrowed. When no pathogen is discovered, broad-spectrum empiric antibiotics are continued. (ABSTRACT TRUNCATED)


Socio-economic and cultural vulnerabilities to cervical cancer and challenges faced by patients attending care at Tikur Anbessa Hospital: a cross sectional and qualitative study.
Cervical cancer is a leading cause of death among women in Ethiopia, affecting them at a time of their life when they are critical to social and economic stability. This study was mainly focused on assessing different socioeconomic and cultural related factors that make women vulnerable to cervical cancer and challenges women face in the process of diagnosis and treatment. To achieve the objective of the study both qualitative and quantitative methods were utilized.198 participants were enrolled in a cross- sectional survey and 10 in-depth interviews were conducted with patients in Tikur Anbessa Hospital. A consecutive sampling method was used to select participants in the survey while purposive sampling was employed to select participants in the qualitative design. For the population covered in the study, poverty along with other socio-cultural practices such as early marriage, high parity and to certain extent polygamy were identified as factors that increased the vulnerability of women to cervical cancer. In addition, the study has uncovered several challenges faced by cervical cancer patients in the diagnosis and treatment process. Three types of challenges that include, health care based, psychological and economic were identified. System and practitioner delay were found as the main hurdles within the variable of health care related challenges. What's more, the psychological challenges identified included, fear of recurrence, negative social attitude and distress associated with the side effects from treatments such as fecal & urinary leakage. Furthermore, inability of bearing costs related to treatment and accommodation were cited as the main economic challenges. The study showed various socio-economic and cultural vulnerabilities that expose women to cervical cancer and the challenges encountered by cervical cancer patients after their diagnosis. Addressing this issue largely lies in strengthening primary and secondary preventive mechanisms, providing public education about safer sex practices, family planning and eliminating cultural practices such as early marriage and polygamy in connection to cervical cancer. Furthermore, improving the diagnostic and treatment procedures and facilities is also a crucial component that needs due emphasis in the fight against cervical cancer.


Endoscopic management of biliary tract stones and stenosis.
The role of endoscopy in managing biliary tract stones and stenosis is surveyed. Rapid advances in endoscopic videoimaging with small diameter maneuverable or large instrument channel endoscopes allows diagnosis and therapy under visual or radiographic control at almost any site in the biliary tree. Techniques for cytology, biopsy, sphincterotomy, mechanical or laser lithotripsy, dilatation with balloon or bougie, large (15 F) or small stent insertion, "polypectomy" or excision with laser or electrosurgery, and brachytherapy provide a therapeutic armamentarium that must be considered as painless, low risk, and cost-effective alternatives to surgical or percutaneous therapy.


The markers of pig heart mitochondrial sub-fractions. II. - On the association of malate dehydrogenase with inner membrane.
Malate dehydrogenase, reputed to be a soluble matricial enzyme, is shown to be also strongly associated with the inner membrane, in pig heart mitochondria. Repeated sonications, water washes, freezing-thawing cycles are not very effective to remove malate dehydrogenase activity from inner membranes, which whatever the treatment, remains important. This activity is only partly solubilized by the substrates, malate or oxaloacetate. High ionic strength treatments by either NaCl-carbonate or 3M KCl have a strong effect, but they also remove cytochrome c oxidase and rotenone-sensitive NADH-cytochrome c reductase, reputed inner membrane intrinsic enzymes, thus strongly damaging the inner membrane. After the action of phospholipase A from Naja Naja Venom, the residual activity is about twenty per cent and only phosphatidyl choline and phosphatidyl ethanolamine decreased significantly, the other phospholipids being unchanged. It is suggested that the enzyme is deeply buried in the membrane and mainly interacts with phosphatidyl choline.


Structural basis for cofactor and substrate selection by cyanobacterium succinic semialdehyde dehydrogenase.
Aldehyde dehydrogenase (ALDH) catalyzes the oxidation of aldehydes to carboxylic acids. Cyanobacterium Synechococcus contains one ALDH enzyme (Sp2771), together with a novel 2-oxoglutarate decarboxylase, to complete a non-canonical tricarboxylic acid cycle. However, the molecular mechanisms for substrate selection and cofactor preference by Sp2771 are largely unknown. Here, we report crystal structures of wild type Sp2771, Sp2771 S419A mutant and ternary structure of Sp2771 C262A mutant in complex with NADP(+) and SSA, as well as binary structure of Gluconobacter oxydans aldehyde dehydrogenase (Gox0499) in complex with PEG. Structural comparison of Sp2771 with Gox0499, coupled with mutational analysis, demonstrates that Ser157 residue in Sp2771 and corresponding Pro159 residue in Gox0499 play critical structural roles in determining NADP(+) and NAD(+) preference for Sp2771 and Gox0499, respectively, whereas size and distribution of hydrophobic residues along the substrate binding funnel determine substrate selection. Hence, our work has provided insightful structural information into cofactor and substrate selection by ALDH.


A metal-organic framework based PCR-free biosensor for the detection of gastric cancer associated microRNAs.
We report herein five sensing platforms for the detection of five gastric cancer associated microRNAs (miRNAs). The sensing platforms are hybrids formed from a water-stable metal organic framework (MOF) {[Cu(dcbb)2(H2O)2]·10H2O}n (1, H2dcbbBr=1-(3,5-dicarboxybenzyl)-4,4'-bipyridinium bromide), respectively with five carboxyfluorescein (FAM) labeled probe single-stranded DNA (probe ss-DNA, denoted as P-DNA). Within the hybrid, MOF 1 tightly interacts with the P-DNA through electrostatic and/or π-stacking interactions and results in fluorescence quenching of FAM via a photo-induced electron transfer (PET) process. In the presence of the complementary target miRNAs miR-185, miR-20a, miR-92b, miR-25 and miR-210, which are expressed abnormally in the plasma of gastric carcinoma patients, P-DNA is released from the surface of MOF 1 ascribed to the stronger base pair matching, leading to the FAM fluorescence recovery. Each P-DNA@1 system is effective and reliable for the detection of its complementary target miRNA with the detection limits from 91 to 559pM, and is not interfered by other four miRNA sequences.


Radiocaesium (137Cs) in marine mammals from Svalbard, the Barents Sea and the North Greenland Sea.
Specific activities of the anthropogenic radionuclide, 137Cs, were determined in marine mammals from Svalbard and the Barents and North Greenland Seas. Muscle samples were collected from 12 polar bears, 15 ringed seals, 10 hooded seals, 7 bearded seals, 14 harp seals, one walrus, one white whale and one blue whale in the period 2000-2003. The mean concentrations (+/-SD) of 137Cs were: 0.72+/-0.62 Bq/kg wet weight (w.w.) for polar bears; 0.49+/-0.07 Bq/kg w.w. for ringed seals; 0.25+/-0.10 Bq/kg w.w. for hooded seals; 0.22+/-0.11 Bq/kg w.w. for bearded seals; 0.36+/-0.13 Bq/kg w.w. for harp seals; 0.67 Bq/kg w.w. for the white whale sample; 0.24 Bq/kg w.w. for the blue whale; and below detection limit for the walrus. Significant differences in 137Cs specific activities between some of the species were found. Ringed seals had higher specific activities than the other seal species in the study. Bearded seals and hooded seals had similar values, which were both significantly lower than the harp seal values. The results in the present study are consistent with previous reported results, indicating low specific activities of 137Cs in Arctic marine mammals in the Barents Sea and Greenland Sea region during the last 20 years. The species specific differences found may be explained by varying diet or movement and distribution patterns between species. No age related patterns were found in specific activities for the two species (polar bears and hooded seals) for which sufficient data was available. Concentration factors (CF) of 137Cs from seawater were determined for polar bears, ringed, bearded, harp and hooded seals. Mean CF values ranged from 79+/-32 (SD) for bearded seals sampled in 2002 to 244+/-36 (SD) for ringed seals sampled in 2003 these CF values are higher than those reported for fish and benthic organisms in the literature, suggesting bioaccumulation of 137Cs in the marine ecosystem.


Reductive chain separation of botulinum A toxin--a prerequisite to its inhibitory action on exocytosis in chromaffin cells.
Cleavage of the disulfide bond linking the heavy and the light chains of tetanus toxin is necessary for its inhibitory action on exocytotic release of catecholamines from permeabilized chromaffin cells [(1989) FEBS Lett. 242, 245-248; (1989) J. Neurochem., in press]. The related botulinum A toxin also consists of a heavy and a light chain linked by a disulfide bond. The actions of both neurotoxins on exocytosis were presently compared using streptolysin O-permeabilized bovine adrenal chromaffin cells. Botulinum A toxin inhibited Ca2+-stimulated catecholamine release from these cells. Addition of dithiothreitol lowered the effective doses to values below 5 nM. Under the same conditions, the effective doses of tetanus toxin were decreased by a factor of five. This indicates that the interchain S-S bond of botulinum A toxin must also be split before the neurotoxin can exert its effect on exocytosis.


Transformations, NMR studies and biological testing of some 17β-isoxazolyl steroids and their heterocyclic ring cleavage derivatives.
The synthesis and NMR structure analysis of a group of oxygenated steroids containing isoxazole, dihydrofuran, tetrahydrofuran rings or enamino carbonyl fragment in the side chain have been fulfilled. The prepared compounds were tested toward several enzymes (human cytochrome P450s CYP17, CYP19, CYP51 and CYP51 of pathogenic fungus Candida glabrata) as their potential inhibitors. A number steroids show a high level affinity (micro- and submicromole) for the enzyme-ligand complexes of the tested compounds with human CYP51, CYP19 and CYP51 of C. glabrata.


Biological effect of radiation-degraded alginate on flower plants in tissue culture.
Alginate with a weight-average molecular mass (Mw) of approx. 9.04 x 10(5) Da was irradiated at 10-200 kGy in 4% (w/v) aqueous solution. The degraded alginate product was used to study its effectiveness as a growth promoter for plants in tissue culture. Alginate irradiated at 75 kGy with an Mw of approx. 1.43 x 10(4) Da had the highest positive effect in the growth of flower plants, namely limonium, lisianthus and chrysanthemum. Treatment of plants with irradiated alginate at concentrations of 30-200 mg/l increased the shoot multiplication rate from 17.5 to 40.5% compared with control. In plantlet culture, 100 mg/l irradiated alginate supplementation enhanced shoot height (9.7-23.2%), root length (9.7-39.4%) and fresh biomass (8.1-19.4%) of chrysanthemum, lisianthus and limonium compared with that of the untreated control. The survival ratios of the transferred flower plantlets treated with irradiated alginate were almost the same as the control value under greenhouse conditions. However, better growth was attained for the treated plantlets.


Discrimination of half-siblings when maternal genotypes are known.
Given the DNA profiles of two individuals and one parent (say the mother) of each, we present likelihood ratios (LRs) comparing the hypothesis that they have the same father with the hypothesis of unrelated fathers. If the individuals have the same mother, the problem is to distinguish full- from half-siblings, otherwise we are comparing a half-sibling relationship with unrelated. We simulate STR profiles at up to 60 loci, based on allele proportions observed at 15 loci in three populations, and use them to approximate misclassification rates both for binary classification (e.g. "half-sib" versus "unrelated"), and when a third "cannot say" category is included. We find that reliable inferences in the absence of the mothers' profiles require many more STR loci than the 10-25 loci that are currently routinely available. However, profiling the two mothers conveys more discriminatory power than profiling the same number of additional loci in the individuals themselves. Our likelihood ratio formulas include a theta (or Fst) adjustment to allow for the individuals concerned to have recent shared ancestry (coancestry), relative to the population from which the allele frequency database is drawn. We illustrate that using an appropriate value of theta can reduce the average misclassification rate.


Analysis of clinical trials on biomaterial and therapeutic applications of chitosan: A review.
Chitosan is a modified natural carbohydrate polymer derived from chitin that occurs in many natural sources. It has a diverse range of applications in medical and pharmaceutical sciences. Its primary and permitted use is biomaterial in medical devices. Chitosan and its derivatives also find utility in pharmaceuticals as an excipient, drug carrier, or therapeutic agent. The USFDA has approved chitosan usage as a biomaterial but not for pharmaceutical use, primarily because of the concerns over its source, purity, and immunogenicity. A large number of clinical studies are underway on chitosan-based materials/ products because of their diverse applications. Herein, we analyze clinical studies to understand their clinical usage portfolio. Our analysis shows that >100 clinical studies are underway to investigate the safety/efficacy of chitosan or its biomaterials/ nanoparticles, comprising ~95% interventional and ~ 5% observational studies. The regulatory considerations that limit the use of chitosan in pharmaceuticals are also deliberated. TEASER: Clinical Trials of Chitosan.


A broadly applicable approach to prepare monoclonal anti-cephalosporin antibodies for immunochemical residue determination in milk.
A simple, efficient and rapid method for the synthesis of cephalosporin-protein conjugates was established. These conjugates were used as immunogens to produce monoclonal antibodies (mAbs) and as solid phase antigens in competitive indirect enzyme immunoassays (EIAs). With this generic approach, a novel set of monoclonal antibodies for cephalosporins was prepared, including ceftiofur and cephalexin as well as, reported here for the first time, cefoperazone, cefquinome and cephapirin. All 5 EIAs were highly sensitive, with standard curve IC(50) values of 0.7 (ceftiofur), 1.1 (cefquinome), 5.2 (cephalexin), 13.8 (cefoperazone) and 40.3 ng mL(-1) (cephapirin). Detection limits (IC(30)) ranged from 0.3 (ceftiofur mAb 1D7) to 17.2 ng mL(-1) (cephapirin mAb 2F10). Specificity studies revealed that cephalosporin-antibody binding was strongly determined by the side chain residues of the cephem nucleus. Therefore all mAbs, to some extent, recognized other beta-lactam antibiotics with similar side chain residues. Within the group of cephalosporins approved for use in veterinary medicine, however, the final EIAs were highly selective for their respective antigen, except for the ceftiofur EIA which showed cross-reactions with cefquinome. The applicability of the five assays for drug residue testing in milk was demonstrated. In each EIA the target drug could be determined in milk with high accuracy and precision at concentrations far below the European Union maximum residue limits.


Periosteal thickening as a manifestation of trauma in infancy.
This paper reports the findings in a study of the incidence of periosteal elevation in children and its possible relationship to child abuse. Two separate sets of radiographs of the skeleton of children were taken for a variety of diagnostic purposes. The suspected abuse set consisted of 59 radiographs taken for suspected child abuse. The mean age for all patients examined was 1.22 years whereas, the mean age for patients with cortical thickening was 0.5 years, suggesting that cortical thickening occurs in a relatively young population. Cortical thickening was assessed by reason for examination. A significant difference (p = .05) was detected with 6/8 (75%) of the patients with cortical thickening from the suspected child abuse groups and only 2/8 (25%) of the patients from the seizure-diagnostic category. The two infants who were noted to have periosteal elevation but were not suspected of abuse had experienced unusual circumstances, one was wearing an abduction splint and the other was a severely hypotonic premature. It is our impression that periosteal thickening is not a normal finding in infants and does not represent a consequence of normal infant care practices. In each case in which cortical thickening was detected, there was evidence to suggest that the child had experienced abnormal or rough handling. As a result of these findings, we believe that cortical thickening of the long bones detected on radiograph is an indication of child abuse.


Comparison of five segmentation tools for 18F-fluoro-deoxy-glucose-positron emission tomography-based target volume definition in head and neck cancer.
Target-volume delineation for radiation treatment to the head and neck area traditionally is based on physical examination, computed tomography (CT), and magnetic resonance imaging. Additional molecular imaging with (18)F-fluoro-deoxy-glucose (FDG)-positron emission tomography (PET) may improve definition of the gross tumor volume (GTV). In this study, five methods for tumor delineation on FDG-PET are compared with CT-based delineation. Seventy-eight patients with Stages II-IV squamous cell carcinoma of the head and neck area underwent coregistered CT and FDG-PET. The primary tumor was delineated on CT, and five PET-based GTVs were obtained: visual interpretation, applying an isocontour of a standardized uptake value of 2.5, using a fixed threshold of 40% and 50% of the maximum signal intensity, and applying an adaptive threshold based on the signal-to-background ratio. Absolute GTV volumes were compared, and overlap analyses were performed. The GTV method of applying an isocontour of a standardized uptake value of 2.5 failed to provide successful delineation in 45% of cases. For the other PET delineation methods, volume and shape of the GTV were influenced heavily by the choice of segmentation tool. On average, all threshold-based PET-GTVs were smaller than on CT. Nevertheless, PET frequently detected significant tumor extension outside the GTV delineated on CT (15-34% of PET volume). The choice of segmentation tool for target-volume definition of head and neck cancer based on FDG-PET images is not trivial because it influences both volume and shape of the resulting GTV. With adequate delineation, PET may add significantly to CT- and physical examination-based GTV definition.


Severe corrosion after malpositioning of a metallic head over the Morse taper of a cementless hip arthroplasty. A case report.
Morse tapers are frequently used in total hip replacement to achieve precise adjustment of lengths and femoral offset. Mechanically, they do not raise any specific problems so long as strict positioning requirements are observed and elements from different manufacturers are not mixed together. We report a case in which the implant induced unexplained pain at 2 years, in relation to a defective fit between the metallic head and the Morse taper. Asymmetric partial fit of the head onto the taper was detected on control X-ray and was implicated as causing metallosis due to excessive release of metal debris from the Morse taper. Revision required femoral stem exchange because of the damage to the Morse taper as well as replacing the cup with new metal-metal bearings. Evolution was favorable at 3 years' follow-up. Most hip replacements include a Morse taper; the present clinical case is a reminder that strict positioning rules are to be respected, without which corrosion and wear may lead to mechanical failure.


Changes in the intracellular microenvironment in the aging human brain.
Normal brain aging is associated with changes occurring at all levels. This study investigates age-related differences in the brain intracellular microenvironment by comparing the apparent diffusion coefficients (ADC) and apparent transverse relaxation time constants (T2) of 5 neurochemicals (i.e., total N-acetyl-aspartate, total creatine, total choline, glutamate, and myo-inositol) between young and older adults. Thirty-two young healthy adults (18-22 years) and 26 older healthy adults (70-83 years) were recruited. Three brain regions were studied at 3 T: prefrontal, posterior cingulate and occipital cortices. ADC and T2 were measured using stimulated echo acquisition mode and localization by adiabatic selective refocusing sequences, respectively. This study shows that the diffusivities of several neurochemicals are higher in older than in younger adults. In contrast, shorter apparent T2 values for several metabolites were measured in older adults. Age-related difference in ADC and apparent T2 of metabolites seem to be region-specific. Furthermore, this study shows that it is feasible to observe age-related differences in the cellular microenvironment of neurochemicals in the normal aging brain.


Capturing domain knowledge from multiple sources: the rare bone disorders use case.
Lately, ontologies have become a fundamental building block in the process of formalising and storing complex biomedical information. The community-driven ontology curation process, however, ignores the possibility of multiple communities building, in parallel, conceptualisations of the same domain, and thus providing slightly different perspectives on the same knowledge. The individual nature of this effort leads to the need of a mechanism to enable us to create an overarching and comprehensive overview of the different perspectives on the domain knowledge. We introduce an approach that enables the loose integration of knowledge emerging from diverse sources under a single coherent interoperable resource. To accurately track the original knowledge statements, we record the provenance at very granular levels. We exemplify the approach in the rare bone disorders domain by proposing the Rare Bone Disorders Ontology (RBDO). Using RBDO, researchers are able to answer queries, such as: "What phenotypes describe a particular disorder and are common to all sources?" or to understand similarities between disorders based on divergent groupings (classifications) provided by the underlying sources. RBDO is available at http://purl.org/skeletome/rbdo. In order to support lightweight query and integration, the knowledge captured by RBDO has also been made available as a SPARQL Endpoint at http://bio-lark.org/se_skeldys.html.


Clomiphene citrate stimulated cycles - additional gonadotrophin stimulation increases endometrium thickness without increasing implantation rate.
It is known that Clomiphene citrate (CC) reduces endometrial thickness, but it is unknown if additional gonadotrophin stimulation increases endometrial thickness and if this has an effect on implantation rate in in vitro fertilization (IVF). The retrospective study included 263 minimal stimulation IVF-cycles stimulated with 25 mg CC per day (CC-IVF), and 161 IVF-cycles stimulated with CC plus 75IU hMG (human Menopausal Gonadotrophin) per day (CC/hMG-IVF). Endometrial and oestradiol (E2) measurements were analysed between day -4 and 0 (0 = day of oocyte retrieval) and the association of endometrial thickness and treatment on implantation rates were studied after multiple adjustments. It was shown that on day 0, endometrium was significantly thicker in CC/hMG-IVF versus CC-IVF cycles (9.81 ±2.68 versus 9.06 ±2.54 mm, p = 0.005). However, increased endometrial thickness did not have an effect on implantation and live birth rates. In conclusion, gonadotrophins should not be added to low dose CC treated IVF cycles just to increase endometrial thickness as increased endometrial thickness does not increase implantation rate.


Dose verification in intensity modulation radiation therapy: a fractal dimension characteristics study.
This study describes how to identify the coincidence of desired planning isodose curves with film experimental results by using a mathematical fractal dimension characteristic method to avoid the errors caused by visual inspection in the intensity modulation radiation therapy (IMRT). The isodose curves of the films delivered by linear accelerator according to Plato treatment planning system were acquired using Osiris software to aim directly at a single interested dose curve for fractal characteristic analysis. The results were compared with the corresponding planning desired isodose curves for fractal dimension analysis in order to determine the acceptable confidence level between the planning and the measurement. The film measured isodose curves and computer planning curves were deemed identical in dose distribution if their fractal dimensions are within some criteria which suggested that the fractal dimension is a unique fingerprint of a curve in checking the planning and film measurement results. The dose measured results of the film were presumed to be the same if their fractal dimension was within 1%. This quantitative rather than qualitative comparison done by fractal dimension numerical analysis helps to decrease the quality assurance errors in IMRT dosimetry verification.


Nanoparticle Conjugation of Human Papillomavirus 16 E7-long Peptides Enhances Therapeutic Vaccine Efficacy against Solid Tumors in Mice.
Treatment of patients bearing human papillomavirus (HPV)-related cancers with synthetic long-peptide (SLP) therapeutic vaccines has shown promising results in clinical trials against premalignant lesions, whereas responses against later stage carcinomas have remained elusive. We show that conjugation of a well-documented HPV-E7 SLP to ultra-small polymeric nanoparticles (NP) enhances the antitumor efficacy of therapeutic vaccination in different mouse models of HPV+ cancers. Immunization of TC-1 tumor-bearing mice with a single dose of NP-conjugated E7LP (NP-E7LP) generated a larger pool of E7-specific CD8+ T cells with increased effector functions than unconjugated free E7LP. At the tumor site, NP-E7LP prompted a robust infiltration of CD8+ T cells that was not accompanied by concomitant accumulation of regulatory T cells (Tregs), resulting in a higher CD8+ T-cell to Treg ratio. Consequently, the amplified immune response elicited by the NP-E7LP formulation led to increased regression of large, well-established tumors, resulting in a significant percentage of complete responses that were not achievable by immunizing with the non-NP-conjugated long-peptide. The partial responses were characterized by distinct phases of regression, stable disease, and relapse to progressive growth, establishing a platform to investigate adaptive resistance mechanisms. The efficacy of NP-E7LP could be further improved by therapeutic activation of the costimulatory receptor 4-1BB. This NP-E7LP formulation illustrates a "solid-phase" antigen delivery strategy that is more effective than a conventional free-peptide ("liquid") vaccine, further highlighting the potential of using such formulations for therapeutic vaccination against solid tumors. Cancer Immunol Res; 6(11); 1301-13. ©2018 AACR.


Heavy metal complexation in polluted molluscs II. Oysters (Ostrea edulis and Crassostrea gigas).
Cadmium, zinc and copper, accumulated from polluted habitats by the oysters Ostrea edulis and Crassostrea gagas, were studied. Three distinct low molecular weight (less than 3000 daltons) zinc complexes were separated from highly contaminated C. gigas, one of which may be a complex with the amino acid taurine. Only one of these zinc complexes was present in less contaminated specimens of O. edulis. On gel permeation chromatography of oyster extracts, copper was eluted together with amino acids (principally taurine) and the betaine homarine. No evidence of metallothionein type proteins was found.


Adaptive changes in autophagy after UPS impairment in Parkinson's disease.
Ubiquitin-proteasome system (UPS) and autophagosome-lysosome pathway (ALP) are the most important machineries responsible for protein degradation in Parkinson's disease (PD). The aim of this study is to investigate the adaptive alterations in autophagy upon proteasome inhibition in dopaminergic neurons in vitro and in vivo. Human dopaminergic neuroblastoma SH-SY5Y cells were treated with the proteasome inhibitor lactacystin (5 μmol/L) for 5, 12, or 24 h. The expression of autophagy-related proteins in the cells was detected with immunoblotting. UPS-impaired mouse model of PD was established by microinjection of lactacystin (2 μg) into the left hemisphere of C57BL/6 mice that were sacrificed 2 or 4 weeks later. The midbrain tissues were dissected to assess alterations in autophagy using immunofluorescence, immunoblotting and electron microscopy assays. Both in SH-SY5Y cells and in the midbrain of UPS-impaired mouse model of PD, treatment with lactacystin significantly increased the expression levels of LC3-I/II and Beclin 1, and reduced the levels of p-mTOR, mTOR and p62/SQSTM1. Furthermore, lactacystin treatment in UPS-impaired mouse model of PD caused significant loss of TH-positive neurons in the substantia nigra, and dramatically increased the number of autophagosomes in the left TH-positive neurons. Inhibition of UPS by lactacystin in dopaminergic neurons activates another protein degradation system, the ALP, which includes both the mTOR signaling pathway and Beclin 1-associated pathway.


Guidelines for the performance of fusion procedures for degenerative disease of the lumbar spine. Part 7: intractable low-back pain without stenosis or spondylolisthesis.
Class I medical evidence exists in support of the use of lumbar fusion as a treatment standard for carefully selected patients with low-back pain intractable to the best medical management. There is Class III medical evidence that suggests that a course of intensive cognitive and physical therapy may be an efficacious treatment option for the treatment of patients with chronic disabling low-back pain.


An overview of developments in research on persons with intellectual disabilities.
Intellectual disabilities (ID) are one of the largest, most complex, and frequently studied of the mental health and medical specialty areas. Thousands of papers have been published dating back to the early work on IQ testing more than a century ago. Many reviews have been published on specific topics with the field of ID, but to date no papers have appeared providing an overview of trends in this vast literature. In this paper we looked at studies published in referred journals from 1979 to 2008. Fifty-six thousand, three hundred and twenty studies were identified using the terms mental retardation and ID. The number of papers published per year ranged from 1038 to 2075 with the fewest papers published in 1980 and the greatest number of studies appearing in 2005. Genetics was by far the most frequently studied topic. The implication of these and related trends in the literature are discussed.


CircSAMD4A aggravates H/R-induced cardiomyocyte apoptosis and inflammatory response by sponging miR-138-5p.
Hypoxia/reoxygenation (H/R)-induced myocardial cell injury is the main cause of acute myocardial infarction (AMI). Many proofs show that circular RNA plays an important role in the development of AMI. The purpose of this study was to investigate the role of circSAMD4A in H/R-induced myocardial injury. The levels of circular SAMD4A (circSAMD4A) were detected in the heart tissues of AMI mice and H/R-induced H9C2 cells, and the circSAMD4A was suppressed in AMI mice and H/R-induced H9C2 cells to investigate its' function in AMI. The levels of circSAMD4A and miR-138-5p were detected by real-time quantitative PCR, and MTT assay was used to detect cell viability. TUNEL analysis and Annexin V-FITC were used to determine apoptosis. The expression of Bcl-2 and Bax proteins was detected by Western blot. IL-1β, TNF-α and IL-6 were detected by ELISA kits. The study found that the levels of circSAMD4A were up-regulated after H/R induction and inhibition of circSAMD4A expression would reduce the H/R-induced apoptosis and inflammation. MiR-138-5p was down-regulated in H/R-induced H9C2 cells. circSAMD4A was a targeted regulator of miR-138-5p. CircSAMD4A inhibited the expression of miR-138-5p to promote H/R-induced myocardial cell injury in vitro and vivo. In conclusion, CircSAMD4A can sponge miR-138-5p to promote H/R-induced apoptosis and inflammatory response.


Effect of different nuclear localization sequences on the immune responses induced by a MIDGE vector encoding bovine herpesvirus-1 glycoprotein D.
To optimize the efficiency of a Minimalistic Immunogenically Defined Gene Expression (MIDGE) vector, peptides containing proven (SV40 T-antigen and bovine herpesvirus-1 VP8) or putative (herpes simplex virus-1 VP22) nuclear localization signals (NLSs) were linked to a MIDGE vector encoding a truncated, secreted form of BHV-1 glycoprotein D (tgD) (MIDGE-tgD). Conjugation of an NLS to the MIDGE-tgD vector improved the tgD expression in vitro and the humoral and cellular immune responses induced in mice in vivo. The NLS from BHV-1 VP8 was most efficient at enhancing the tgD production and tgD-specific immune responses when conjugated to the MIDGE-tgD vector.


Nerve growth factor prevents apoptosis of cord blood-derived human cultured mast cells synergistically with stem cell factor.
Stem cell factor (SCF) has been identified as a critical survival factor of human mast cells. Other cytokines which possess survival promotion activity on human mast cells are less known. We examined the survival promotion activity of nerve growth factor (NGF) on cord blood-derived human cultured mast cells. Expression and function of NGF receptors on the mast cells were examined by RT PCR, flowcytometric analysis, immunoprecipitaion and western blotting. The survival promotion activity of NGF to the mast cells was examined. To evaluate the proliferating activity of NGF on the human cultured mast cells, flow cytometric analysis with propidium iodide staining was applied. To confirm whether the human mast cell growth activity of NGF was caused by a suppression of apoptosis, the proportion of the cells containing in situ DNA fragmentation was counted. The human cultured mast cells expressed the high affinity receptor p140trk but not the low affinity receptor p75LNGFR. NGF induced the phosphorylation of p140trk. NGF alone could not support the survival of the mast cells, however, the addition of NGF to the culture medium containing recombinant SCF led to a significant increase of the number of survival mast cells. No significant changes of the cell cycle from G0/G1 phase to the S/G2 + M phases were observed by NGF. In contrast, the addition of NGF to the medium with SCF showed a significant inhibitory effect on the apoptosis of the mast cells. NGF may act as a key factor to promote the survival of human mast cells synergistically with SCF through the prevention of apoptosis.


The effect of immunosuppression on manifestations of sepsis in an animal model of cecal ligation and puncture.
The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified response to infection. Our experiment in minipigs was designed to compare responses to sepsis between experimental groups of septic minipigs with and without immunosuppression. Minipigs with identical baseline parameters were randomized into 3 groups: Sepsis (n = 10); immunosuppression (n = 11), including cyclosporine, methylprednisolone, and mycophenolate mofetil treatment before surgery, and a sham group (n = 6). Sepsis was induced by cecal ligation and puncture (CLP). We recorded selected clinical and laboratory parameters up to 24 hours postoperatively. All CLP animals developed septic shock with a febrile response, tachycardia, and hypotension requiring noradrenaline administration. The hemodynamic responses to sepsis in septic groups with and without immunosuppression were similar. Noradrenaline infusion was started on average later in the immunosuppression than in the group without immunosuppression; however, the difference was not significant. The kinetics of the plasma levels of most selected cytokines and C-reactive protein were similar in both septic groups. At 10 hours after surgery, the immunosuppression group showed significantly lower interleukin (IL)-6 levels compared with the sepsis group. At 19, 22, and 25 hours after surgery immunosuppressed animals displayed significantly greater increases in IL-10 levels compared with the cohort without immunosuppression. CLP is a simple, reproducible model of sepsis in minipigs. All CLP animals developed sepsis within 24 hours on average. Significant differences in IL-6 and IL-10 plasma levels were recorded between septic animals with versus without immunosuppression.


The necessity of education and hepatitis B vaccination for young people: A study of high risk behaviour for blood borne viruses in the United Kingdom.
Blood borne virus (BBV) infection in adults involved in high risk behaviour is well recognized. There are limited reported data on young people involved in high risk behaviour. A descriptive questionnaire was used to ascertain risk behaviour at the Young People's Substance Misuse Service (Birmingham). Data collection included risk behaviour and serological tests for hepatitis B, C and HIV. Sixty-five of one-hundred three (63%) young people participated; 37/65 male; age range 13.9-18.9 (median 17.4 years). Risk behaviour included 6 intravenous drug, 58 cannabis, and 61 had sexual partners, of whom 52 (85%) engaged in unprotected sex. Sixty-five participants were negative for BBV infection: 9 were HBV immune. HB vaccination was not available at the centre (for <18 year), and all refused referral to their general practitioner for vaccination due to fear of disclosure. The main risk for BBV acquisition was unprotected sex with multiple sexual partners and illicit drug use. Most were unaware of the risks related to high risk behaviour. Effective education programmes of relevant risk factors with HBV vaccination should be implemented during preadolescence. We recommend an integrated service via specialized centres, to work together to improve awareness and increase efforts to vaccinate adolescents at risk for HBV infection.


Role of cyclic AMP response element binding protein in human leukemias.
Acute myeloid leukemia (AML) in adults has a 20% 5-year disease-free survival despite treatment with aggressive cytotoxic chemotherapy. Previous work from our laboratory demonstrated that the majority of patients with acute lymphoid and myeloid leukemia overexpress CREB in the bone marrow. CREB overexpression is associated with poor initial outcome of clinical disease in AML patients. CREB is a transcription factor that functions in glucose homeostasis, growth-factor-dependent cell survival, and memory. Signaling by hematopoietic growth factors, such as GM-CSF, results in activation of CREB and up-regulation of CREB target genes. To study its role in hematopoiesis, we overexpressed CREB in leukemia cell lines and in mice. CREB overexpression resulted in increased survival and proliferation of myeloid cells and blast-transformation of bone marrow progenitor cells from transgenic mice expressing CREB in the myeloid lineage. CREB transgenic mice also develop myeloproliferative disease after 1 year. Thus, CREB acts as a protooncogene to regulate hematopoiesis and contributes to the leukemia phenotype. Our results suggest that CREB-dependent pathways may serve as targets for directed therapies in leukemia in the future.


[Role of a stabilized chronic pathology RPN in mental health].
While our health care system is compartmentalized by specialty, the implementation of an advanced practice nurse mentioning stabilized chronic pathologies in a public mental health institution may seem ambitious. However, it is interesting, for patients suffering from a mental pathology, for caregivers practicing in psychiatry and for the institution, to integrate it into the care pathway.


Evidence for induction and repair of potentially lethal damage in plateau-phase V79 cells after exposure to adriamycin. The importance of removal of adriamycin released from the cells during the post-treatment incubation period.
Plateau-phase Chinese hamster V79 cells were exposed to various concentrations of adriamycin (0-21 micrograms ml-1) in conditioned medium from plateau-phase cultures (C-med). Cells were plated for colony formation, either immediately after adriamycin treatment or after a 24 h incubation either in fresh medium (F-med) or C-med. A potentiation of cell killing was observed in cells plated 24 h after treatments which was larger for cells incubated in F-med than for cells incubated in C-med. Trypsinization of the cells and replating for 24 h in the same volume of medium (total amounts of cells) but at lower surface density to reduce intercellular contact, did not modify the killing potentiation observed after delayed plating. Four to 6 changes of medium, carried out at 1 h intervals, starting immediately after treatment, led to an elimination of the killing potentiation otherwise found in cells kept after treatment in F-med and resulted in survival levels similar to those of cells plated immediately after treatment. On the other hand, survival levels higher by a factor 1.5 to 10 than those obtained for cells plated immediately after treatment were observed for cells kept in C-med when four medium changes were carried out during the first 5 h of the 6 or 24 h post-treatment period. Incubation with 150 microM beta-arabinofuranosyladenine (araA) for 6 h (C-med) after exposure to adriamycin (4 changes of medium at 1 h intervals) prevented the increase in survival observed after incubation in C-med but also caused an additional potentiation in killing resulting in survival levels lower than those of cells plated immediately after treatment. These results are interpreted as indicating the induction by adriamycin of potentially lethal damage (PLD), sensitive to araA, similar to that observed after exposure to low LET ionizing radiation. Repair and/or fixation of this form of PLD can only be shown if precautions are taken to circumvent toxicity induced by adriamycin released from the cells during the post-treatment time interval, for example, by frequent changes in medium.


The effects of atypical antipsychotic medications on psychosocial outcomes.
This review examines the question--Do atypical antipsychotic medications improve psychosocial outcomes?--by examining studies that compared the effects of atypicals to conventional antipsychotics or to other atypical medications. The authors reviewed randomized clinical trials of atypical antipsychotic medication that included psychosocial variables as outcomes. Findings from 31 published studies on more than 12,000 individuals generally showed that atypical medications led to significant improvements in negative symptoms compared to conventional antipsychotics. Effects on global assessment of psychosocial functioning and on the quality of life were mixed with only about half the studies reporting significant improvements. Olanzapine yielded the best results on psychosocial functioning; remoxipride was found to yield few significant changes on these variables. This review provides evidence that some atypical antipsychotics may have direct effects on some of the psychosocial disabilities that result from serious mental illness.


Quantitative evaluation of the optic nerve head in patients with unilateral visual field loss from primary open-angle glaucoma.
Measurable structural alterations of the optic nerve head may precede visual field abnormalities in early open-angle glaucoma. The authors studied the optic nerve heads of 10 patients with unilateral visual field loss from primary open-angle glaucoma, and 12 age- and sex-matched normal subjects. Topographic optic nerve head parameters were measured with a system of computerized image analysis (Rodenstock Analyzer, G. Rodenstock Instrumente GMBH, Munich, W. Germany). In patients with asymmetric primary open-angle glaucoma, eyes with normal visual fields had a slightly larger mean (+/- standard error of the mean) disc rim area (0.90 +/- 0.04 mm2) than eyes with glaucomatous visual field defects (0.78 +/- 0.05 mm2). However, both sets of eyes in the asymmetric primary open-angle glaucoma patients had smaller mean disc rim areas (P less than 0.0007) than did the control group (1.27 +/- 0.09 mm2). These findings support the hypothesis that loss of the optic disc rim can be detected before perimetric abnormalities develop in primary open-angle glaucoma.


Apolipoprotein E epsilon4 allele and neurobehavioral status after on-pump coronary artery bypass grafting.
Abstract Background and Aim: The presence of apolipoprotein E epsilon4 allele is being considered as a risk factor for cognitive decline after cardiac surgery. We sought the effect of apolipoprotein E epsilon4 allele on neurobehavioral status after on-pump coronary artery bypass grafting. Prior to the operation, neurologic examination and neurobehavioral cognitive status test (COGNISTAT) were performed. Both procedures were repeated on the day of discharge and 3 months after surgery. Apolipoprotein E epsilon4 allele positive and apolipoprotein E epsilon4 allele negative patients' performance on COGNISTAT were compared. There was no statistically significant demographic and operative data difference between two groups. No neurological impairment was observed on examinations. There was no statistically significant neurocognitive decline difference between two groups' postoperative performances. It seems that apolipoprotein E epsilon4 allele may not affect neurobehavioral status in the intermediate period after on-pump coronary artery bypass grafting.


Isolation and characterization of the active cDNA of the human cell cycle gene (RCC1) involved in the regulation of onset of chromosome condensation.
The human RCC1 gene was cloned after DNA-mediated gene transfer into the tsBN2 cell line, which shows premature chromosome condensation at nonpermissive temperatures (39.5-40 degrees C). This gene codes for a 2.5-kb poly(A)+ RNA that is well conserved in hamsters and humans. We isolated 15 cDNA clones from the Okayama-Berg human cDNA library, and found two that can complement the tsBN2 mutation with an efficiency comparable to that of the genomic DNA clone. The base sequences of these two active cDNA clones differ at the 5' proximal end, yet both have a common open reading frame, encoding a protein of 421 amino acids with a calculated molecular weight of 44,847 and with seven homologous repeated domains of about 60 amino acids. This human RCC1 gene was located to human chromosome 1 using sorted chromosomal fractions.


Relationship between hepatitis C and kidney stone in US females: Results from the National Health and Nutrition Examination Survey in 2007-2018.
The main objective of this study is to explore the effects of hepatitis C (HCV) on the prevalence rate of kidney stones in US women. Dates for HCV infection and kidney stones were collected from National Health and Nutrition Examination Survey (NHANES) database, a cross-sectional study. The analysis samples included adults aged ≥20 years and women from six consecutive cycles of the NHANES 2007-2018. The association between HCV infection and kidney stones was performed by using logistic regression models. Subgroup analyses were conducted to find sensitive crowds. A total of 13,262 participants were enrolled, including 201 infected with HCV. After adjustment for potential confounders, we revealed a positive relationship between HCV and kidney stones (OR = 1.70, 95%CI:1.13-2.56). The crowds' statistically significant difference was characterized by other races (OR = 8.17, 95%CI:1.62-41.22) and BMI within 25-29.9 kg/m2 (OR = 2.45, 95%CI:1.24-4.83). HCV infection may affect the prevalence of urolithiasis in US women, even the causal relationship remains unclear, the relation deserves special attention. We considered such a study an ideal way to begin exploring the effects of HCV on kidney stones.


Measurements of serum procollagen-III peptide and M30 do not improve the diagnostic accuracy of transient elastography for the detection of hepatic fibrosis in patients with nonalcoholic fatty liver disease.
Transient elastography (TE) has recently emerged as an accurate noninvasive imaging method for the diagnosis of liver fibrosis in nonalcoholic fatty liver disease (NAFLD). Here, we tested whether adding serum measures of serum procollagen-III peptide (PIIIP) and caspase-cleaved cytokeratin 18 fragment (M30) to TE could improve its diagnostic accuracy in patients with biopsy-proven NAFLD. TE was performed in 87 patients with NAFLD. Serum PIIIP and M30 levels were determined by enzyme-linked immunosorbent assay. Liver histology was considered the gold standard. The diagnostic accuracies were assessed by measuring the area under the receiver operating curve. At histopathological examination, 34 patients (39.1%) had significant fibrosis (F2-F4) and 19 patients (21.8%) had advanced fibrosis (F3-F4). Both TE and serum M30 levels were independent predictors of fibrosis, whereas no association was found with PIIIP. No significant differences in terms of sensitivity and specificity for both significant and advanced fibrosis were evident for TE, serum M30, or their combination. Moreover, the area under the receiver operating curves of serum M30 combined with TE did not differ significantly from those of either test alone. Both TE and serum M30 levels are accurate for the noninvasive diagnosis of fibrosis in NAFLD. However, their combination did not improve the overall diagnostic accuracy.


Preclinical discovery of ixabepilone, a highly active antineoplastic agent.
The epothilones and their analogs constitute a novel class of antineoplastic agents, produced by the myxobacterium Sorangium cellulosum. These antimicrotubule agents act in a similar manner to taxanes, stabilizing microtubules and resulting in arrested tumor cell division and apoptosis. Unlike taxanes, however, epothilones and their analogs are macrolide antibiotics, with a distinct tubulin binding mode and reduced susceptibility to a range of common tumor resistance mechanisms that limit the effectiveness of taxanes and anthracyclines. While natural epothilones A and B show potent antineoplastic activity in vitro, these effects were not seen in preclinical in vivo models due to their poor metabolic stability and unfavorable pharmacokinetics. A range of epothilone analogs was synthesized, therefore, with the aim of identifying those with more favorable characteristics. Here, we describe the preclinical characterization and selection of ixabepilone, a semi-synthetic epothilone B analog, among many other epothilone analogs. Ixabepilone demonstrated superior preclinical characteristics, including high metabolic stability, low plasma protein binding and low susceptibility to multidrug resistance protein-mediated efflux, all of which were predictive of potent in vivo cell-killing activity. Ixabepilone also demonstrated in vivo antitumor activity in a range of human tumor models, several of which displayed resistance to commonly used agents such as anthracyclines and taxanes. These favorable preclinical characteristics have since translated to the clinic. Ixabepilone has shown promising phase II clinical efficacy and acceptable tolerability in a wide range of cancers, including heavily pretreated and drug-resistant tumors. Based on these results, a randomized phase III trial was conducted in anthracycline-pretreated or resistant and taxane-resistant metastatic breast cancer to evaluate ixabepilone in combination with capecitabine. Ixabepilone combination therapy showed significantly superior progression-free survival and tumor responses over capecitabine alone.


Mutational specificity of N-nitrosodimethylamine: comparison between in vivo and in vitro assays.
We have determined the DNA alterations recovered after treatment with Aroclor 1254-treated rat S9-activated N-nitrosodimethylamine (NDMA) in the N-terminal region of the lacI gene of E. coli. A total of 125 independent LacI-d mutants of E. coli were characterized by DNA sequencing. Consistent with the known methylating ability of this compound, the predominant mutation was the G:C-->A:T transition, which accounted for 90% of all the mutations recovered. Non G:C-->A:T events include 2 G:C-->T:A, 2 G:C-->C:G, 2 A:T-->G:C, 3 A:T-->T:A and 3 frameshifts. Contingency analysis reveals that NDMA-induced mutations recovered after in vitro activation (S9) have a spectrum very similar to that previously obtained after in vivo activation employing a mouse host-mediated assay. In both systems, G:C-->A:T events clearly dominate and their distribution reveals similar site-specificity. Moreover, the proportion and kind of non-G:C-->A:T events are also similar.


Desorption/ionization of biomolecules from aqueous solutions at atmospheric pressure using an infrared laser at 3 microm.
A new atmospheric pressure (AP) infrared (IR) matrix-assisted laser desorption/ionization (MALDI) ion source was developed and interfaced with a Thermo Finnigan LCQ ion trap mass spectrometer. The source utilized a miniature all-solid-state optical parametric oscillator (OPO)-based IR laser system tunable in the lambda = 1.5-4 microm spectral range and a nitrogen ultraviolet (UV) laser (lambda = 337 nm) for use in comparative studies. The system demonstrated comparable performance at 3 microm and 337 nm wavelengths if UV matrices were used. However, AP IR-MALDI using a 3 microm wavelength showed good performance with a much broader choice of matrices including glycerol and liquid water. AP IR-MALDI mass spectra of peptides in the mass range up to 2000 Da were obtained directly from aqueous solutions at atmospheric conditions for the first time. A potential use of the new AP IR-MALDI ion source includes direct MS analysis of biological cells and tissues in a normal atmospheric environment as well as on-line coupling of mass spectrometers with liquid separation techniques.


Molecular monitoring of BCR-ABL transcripts after allogeneic stem cell transplantation for chronic myeloid leukemia.
The monitoring of minimal residual disease (MRD) through low sensitivity real-time (RT) polymerase chain reaction (PCR) analysis of BCR-ABL transcripts allows early detection of chronic myeloid leukemia (CML) relapse after allogeneic hematopoietic stem cell transplantation (HSCT). The introduction of more sensitive techniques, such as RT quantitative (Q)-PCR, may lead to an overestimation of the risk of CML relapse. In this study, we reviewed the results of peripheral blood RT Q-PCR in CML patients who underwent allogeneic HSCT from 1983 to 2007. In our laboratory, RT Q-PCR analysis was routinely performed since 2002. Eighty-seven of 189 patients had available RT Q-PCR data; 63 patients had at least 3 RT Q-PCR analyses assessable. Fifty-two of 63 patients (83%) had, at least once, detectable transcript levels, but with an BCR-ABL/ABL ratio <.1% defined as <MR3 (molecular remission <0,1%), whereas 11 (17%) had persistent undetectable BCR-ABL transcripts. Six of 52 patients with <MR3 relapsed, defined as BCR-ABL transcript numbers >.1% confirmed by the finding of Ph+ cells in bone marrow. No patients with persistent undetectable transcripts relapsed (P = .19). Relapse did not correlate with the number of occurrences of <MR3 or with the time to the first <MR3 result. Finally, of 46 patients with detectable transcripts who did not relapse, 35 had undetectable transcripts at last contact. RT Q-PCR analysis had low specificity (19%) and low positive predictive value (12%) in predicting relapse of CML patients after allogeneic HSCT. Our data suggest that detection of low BCR-ABL transcript levels by RT Q-PCR analysis has a poor accuracy in defining the risk of CML relapse and should not be considered as the unique indication to treatment. Fluctuation of BCR-ABL transcripts levels is common as late as ≥10 years posttransplantation, possibly suggesting the long-term persistence of CML stem cells.


Effect of Cadherin-11 Expression on the Prognosis of a Newly Diagnosed Primary Glioblastoma.
Cadherin-11, a cell-to-cell adhesion molecule, is associated with higher tumor grade and decreased patient survival. The purpose of this study was to investigate the clinical significance of cadherin-11 expression in the progression and prognosis of a newly diagnosed primary glioblastoma (GBL). Between 2007 and 2016, 52 out of 178 patients diagnosed with a GBL and satisfied the following criteria: 1) a new primary GBL, 2) gross-total resection, 3) immunohistochemically-available tissue, and 4) standardized adjuvant treatment. In terms of staining intensity, the low-intensity cadherin-11 group showed longer progression-free survival (PFS) than the high-intensity cadherin-11 group (median PFS, 12.0 months [95% CI, 11.1-12.9] vs. median PFS, 6.0 months [95% CI, 3.7-8.3]; p<0.001). The low-intensity cadherin-11 group revealed longer overall survival (OS) than the high-intensity cadherin-11 group (median OS, 20.0 months [95% CI, 11.8-16.6] vs. median OS, 15.0 months [95% CI, 11.8-18.2]; p=0.003). The staining intensity of cadherin-11 was a statistically significant factor in PFS and OS in terms of univariate and multivariate analyses (univariate analysis: p<0.001 and p=0.005; multivariate analysis: p<0.001 and p=0.005). Our clinical study demonstrates high cadherin-11 expression may be associated with poor PFS and OS for a newly diagnosed primary GBL.


Bone morphogenetic protein-2 enhances bone regeneration mediated by transplantation of osteogenically undifferentiated bone marrow-derived mesenchymal stem cells.
We have hypothesized that human bone marrow-derived mesenchymal stem cells (BMMSCs), that are not osteogenically differentiated prior to implantation, would regenerate bone extensively in vivo once exogenous bone morphogenetic protein-2 (BMP-2) was delivered to the implantation site. BMP-2 released from heparin-conjugated poly(lactic-co-glycolic acid) (HCPLGA) scaffolds stimulates osteogenic differentiation of cultured BMMSCs. Upon implantation, undifferentiated BMMSCs on BMP-2-loaded HCPLGA scaffolds induce far more extensive bone formation than either undifferentiated BMMSCs or osteogenically differentiated BMMSCs on HCPLGA scaffolds. These BMP-2-loaded HCPLGA scaffolds could prove invaluable for in vivo regeneration of bone from undifferentiated human BMMSCs.


Denis Browne's thoracotomy revised.
The three main issues involved in thoracotomy technique for the repair of oesophageal atresia (OA) are: (1) prevention of chest wall deformities; (2) adequate surgical exposure; and (3) the cosmetic appearance of the skin scar. Adequate surgical exposure should be the first priority. The technique described is an endeavour to address all the above concerns, a modification of the technique described by Denis Browne that eliminates the risk of injury to the long thoracic nerve (LTN). A retrospective analysis from case notes of 70 children (1974-1997) with OA from a single surgeon's practice was performed. Data collected included the presence of complications such as wound problems, chest-wall deformities, and evidence of nerve palsies including winged scapula. A uniform technique of a vertical skin incision in the mid-axillary line, preservation of the latissimus dorsi, and division of the serratus anterior muscle anterior to its nerve was followed in all cases. No wound infection, breakdown, or seroma formation was seen in any case. On long-term followup (range 1-22 years, mean 10.5 years) no chest wall deformities or nerve palsies were observed. This modification gives excellent exposure, avoids injury to the LTN, does not lead to chest-wall deformity, and gives a pleasing cosmetic result. The scar is hardly noticeable with the arm by the side of the body.


Zinc supplementation in children and adolescents with acute leukemia.
Zinc is known as an essential micronutrient for human health because of its structural and biochemical functions, influencing growth and affecting multiple aspects of the immune system. Zinc has been extensively studied in neoplastic processes but its role in children with leukemia still remains to be elucidated in several aspects. The aim of this study was to evaluate the effects of oral zinc supplementation on weight gain and infectious episodes in children and adolescents with acute leukemia. This study included 38 patients, and was carried out as a randomized, double-blind, placebo-controlled investigation. The dosage of plasma zinc levels and the evaluation of nutritional status were performed during a period of 60 days. Zinc was supplemented orally, 2 mg/kg/day, in the form of amino acid salt. The results showed that plasma zinc concentrations did not increase significantly with the addition of the micronutrient. However, from a clinical point of view, it has become evident that supplementary zinc exerts a positive effect on nutritional status as positive weight gain. Moreover, the number of infection episodes was significantly reduced, possibly because of the immune stimuli. In conclusion, zinc supplementation can prevent some of the chemotherapy adverse effects in children with leukemia, improving their quality of life.


A Combination Treatment of Raloxifene and Vitamin D Enhances Bone-to-Tendon Healing of the Rotator Cuff in a Rat Model.
Tearing and degeneration of the rotator cuff at the tendon-to-bone junction are common in adults aged ≥50 years. Few studies have reported on the relationship between estrogen and the rotator cuff enthesis. In addition to preventing bone loss, selective estrogen receptor modulators have been shown to improve tendon and muscle quality. To evaluate the effects of raloxifene (RLX) and vitamin D on rotator cuff tendon-to-bone healing in a rat model. Controlled laboratory study. A total of 29 female rats (58 shoulders) were assigned to 4 groups: (1) control group, (2) ovariectomy (OVX)-only group, (3) no RLX group (OVX and rotator cuff repair [RCR]), and (4) RLX group (OVX, RCR, and RLX). Rats that did not undergo rotator cuff tear (RCT) surgery were divided into the control and OVX-only groups according to OVX surgery. Rats that underwent RCT surgery and RCR were divided into the no RLX and RLX groups according to RLX and vitamin D administration. An estrogen-deficient state was induced by OVX at 12 weeks of age. Bone mineral density (BMD) and trabecular bone characteristics were measured by micro-computed tomography, and healing of the tendon-to-bone junction was evaluated by biomechanical testing, histomorphometry, and micro-magnetic resonance imaging (MRI). The mean final body weight (BW; 461.6 ± 47.3 g) of the OVX-only group was significantly higher and BMD (0.25 ± 0.07 g/cm3) was significantly lower (P < .001) than the mean final BW (338.5 ± 35.1 g) and BMD (0.48 ± 0.05 g/cm3) of the control group. In contrast, the RLX group showed that the BW (369.6 ± 35.8 g) and BMD (0.41 ± 0.08 g/cm3) were not significantly different from the control group. The RLX group had a significantly higher histomorphometric total score (8.50 ± 1.05) than the no RLX group (4.83 ± 2.48). On biomechanical testing, the RLX group (29.7 ± 9.1 N) showed a significantly higher load to failure than the no RLX group (19.4 ± 8.8 N). On micro-MRI, the RLX group had a more homogeneous low signal and tendon continuity than the no RLX group. The combination treatment of RLX and vitamin D prevented a decrease in local BMD (greater tuberosity of the proximal humerus) and enhanced tendon-to-bone healing of the rotator cuff in a rat model. This study induced an estrogen-deficient state similar to the human postmenopausal state and used drugs that are actually being prescribed in a clinical situation.


Improved selectivity of molecularly imprinted polymers based on the synergistic action of hydrogen bond and electrostatic interaction.
Based on the synergistic action of hydrogen bond and electrostatic interaction, provided by methacrylic acid and 2-aminoethyl ester hydrochloride (FM2), respectively, novel molecularly imprinted polymers (SA-MIPs) were designed to improve its selective recognition ability. Diclofenac sodium (DFC) was chosen as the template molecule of this study. The interaction and their recognition sites between two functional monomers and templates were confirmed by nuclear magnetic resonance hydrogen spectroscopy. Because of the synergistic action of hydrogen bond and electrostatic interaction, the imprinting factor (IF) of SA-MIPs (IF = 2.26) is superior to the corresponding monofunctional monomer imprinting materials (IF = 1.52, 1.20) and the materials using two functional monomers with an only single type of interaction (IF = 1.54, 1.75). The results of selective adsorption experiments indicate that the selective recognition ability of SA-MIPs is significantly better than that of the other four MIPs, and the difference in selectivity coefficient for methyl orange is the largest between SA-MIPs and the MIPs only using FM2, which is about 70 times. In addition, x-ray photoelectron spectroscopy was used to verify the interaction between SA-MIPs and the template. This work and its explanation of the interaction mechanism at the molecular level will be helpful for the rational design of novel MIPs with higher selectivity. Besides, SA-MIPs have good adsorption performance (37.75 mg/g) for DFC in aqueous solutions, which could be used as potential adsorption materials for the effective removal of DFC in the aquatic environment.


Influence of reverse signaling via membrane TNF-alpha on cytotoxicity of NK92 cells.
Membrane tumor necrosis factor-alpha (mTNF-alpha) serves as a receptor transducing signals into mTNF-alpha-bearing cells. Among human peripheral blood mononuclear cells, natural killer (NK) cells have been reported to be the only cell type constitutively expressing mTNF-alpha, which is involved in the cytotoxicity of resting NK cells. Using an IL-2-dependent human NK cell line, NK92, which constitutively expresses mTNF-alpha, we examined the effect of reverse signaling via mTNF-alpha on cellular activities. When the cells were prestimulated with soluble TNFR1 (sTNFR1) which activated mTNF-alpha-mediated reverse signaling, the cytotoxicity of NK92 cells was significantly increased. Further investigation demonstrated that prestimulation with sTNFR1 augmented exocytosis and mRNA transcription of two cytotoxic molecules, perforin and granzyme B, which could serve as underlying molecular mechanisms by which mTNF-alpha-mediated reverse signaling promoted cytotoxicity of NK cells toward K562 cells. On the other hand, pretreatment of NK92 with sTNFR1 boosted the expression of FasL and TNF-alpha, including both the secretory and membrane forms. These molecules also contribute to the NK-mediated cytotoxicity, although K562 cells are Fas-negative and sTNF-alpha-resistant. Interestingly, the mTNF-alpha reverse signaling was found to act synergistically with IL-2 on NK-mediated cytotoxicity. This synergy markedly promoted the production of secretory as well as membrane cytotoxic molecules which may be responsible for the enhanced NK92-mediated cytotoxicity. Our observations suggest that, via reverse signaling, constitutively expressed mTNF-alpha may sensitize NK cells to activating stimuli, such as IL-2, resulting in increased NK-mediated cytotoxicity through promoting the production of multiple cytotoxic effector molecules.


Formation of osteoclast-like cells on HA and TCP ceramics.
An essential property of bone substitute materials is that they are integrated into the natural bone remodelling process, which involves the resorption by osteoclast cells and the formation by osteoblast cells. If monocyte cells adhere to a calcium phosphate surface (bone or bone substitute material), they can fuse together and form multinucleated osteoclast cells. In this study we show that osteoclast-like cells derived from a human leukoma monocytic lineage responded in a different way to tricalciumphosphate (TCP) than to hydroxyapatite (HA) ceramics. Both ceramics were degraded by resorbing cells; however, HA enhanced the formation of giant cells. The osteoclast-like cells on HA formed a more pronounced actin ring, and larger lacunas could be observed. TCP ceramics are medically used as bone substitute materials because of their high dissolution rate. On the other hand, highly soluble calcium phosphate ceramics like TCP seem to be inappropriate for osteoclast resorption because they produce a high calcium concentration in the osteoclast interface and in the environment.


Mask is required for the activity of the Hippo pathway effector Yki/YAP.
The Drosophila Yorkie (Yki) protein and its mammalian homolog Yes-associated protein (YAP) are potent growth promoters, and YAP overexpression is associated with multiple types of cancer. Yki and YAP are transcriptional coactivators and function as downstream effectors of the Hippo tumor suppressor pathway. The regulation of Yki and YAP by the Hippo signaling pathway has been extensively investigated; however, how they regulate gene expression is poorly understood. To identify additional regulators of Yki activity, we performed a genome-wide RNAi screen in Drosophila S2 cells. In this screen, we identified the conserved protein Mask (Multiple ankyrin repeats single KH domain) as a novel promoter of Yki activity in vitro and validated this function in vivo in Drosophila. We found that Mask is required downstream of the Hippo pathway for Yki to induce target-gene expression and that Mask forms complexes with Yki. The human Mask homolog MASK1 complexes with YAP and is required for the full activity of YAP. Additionally, elevated MASK1 expression is associated with worsened outcomes for breast cancer patients. We conclude that Mask is a novel cofactor for Yki/YAP required for optimal Yki/YAP activity during development and oncogenesis.


Selective expression of the beta-subunit of nucleoid-associated protein HU during cold shock in Escherichia coli.
Expression of Escherichia coli hupA and hupB, the structural genes encoding the most abundant nucleoid-associated proteins HUalpha and HUbeta has been studied during cold shock. This article demonstrates that: (i) transcriptional expression of hupA is blocked following a sudden temperature downshift (from 37 degrees C to 10 degrees C), whereas transcription of hupB from the P2 and P3 promoters is maintained at a constitutive level and is activated de novo from the P4 promoter; (ii) all three hupB mRNAs (transcribed from the three natural promoters P2, P3 and P4) become much more stable than the single hupA transcript; and (iii) the hupB transcripts, unlike that of hupA, are efficiently translated in vivo during cold acclimation and can be actively translated in vitro at low temperature. Taken together, the results indicate that during cold shock the expression of the HUbeta subunit is preferentially stimulated and that of HUalpha repressed, suggesting that an altered HUalpha to HUbeta expression ratio resulting in an increase of HUalpha/HUbeta heterodimers and/or (HUbeta)2 homodimers may play an important role during cold adaptation.