Associations of Volume and Thyroidectomy Outcomes: A Nationwide Study with Systematic Review and Meta-Analysis.
This study explored how different hospital volumes and surgeon volumes affect thyroidectomy outcomes in terms of length of stay (LOS), costs, and in-hospital mortality. MEDLINE and EMBASE databases. This study retrospectively analyzed a cohort of 125,037 thyroidectomy patients treated at Taiwan hospitals from 1996 to 2010. Relationships between hospital/surgeon volume and patient outcomes were retrospectively analyzed by propensity score matching. In conjunction with the retrospective study, a systematic review and meta-analysis of the relevant literature also were performed. The mean LOS for all thyroidectomies performed during the study period was 3.3 days, and the mean cost was $1193.5. Both high-volume hospitals and high-volume surgeons were associated with significantly shorter LOS and lower costs compared with their low-volume counterparts (P < .001). Different volume groups had similar in-hospital mortality rates. The meta-analysis results consistently showed that the benefits of high-volume hospitals/surgeons are reduced LOS and costs. However, low in-hospital mortality rates were associated with high-volume surgeons but not with high-volume hospitals. This meta-analysis showed that patients who received thyroidectomies performed by high-volume hospitals and surgeons had shorter LOS and lower costs compared with those treated by low-volume hospitals and surgeons. In addition, in-hospital survival rates were better in patients treated by high-volume surgeons. Further research is needed to define the learning curve for thyroidectomy and to clarify how hospital volume and surgeon volume affect its success rate.


Association between Circulating Osteocalcin and Cardiometabolic Risk Factors following a 4-Week Leafy Green Vitamin K-Rich Diet.
Evidence suggests that lower serum undercarboxylated osteocalcin (ucOC) may be negatively associated with cardiometabolic health. We investigated whether individuals with a suppression of ucOC following an increase in dietary vitamin K1 exhibit a relative worsening of cardiometabolic risk factors. Men (n = 20) and women (n = 10) aged 62 ± 10 years participated in a randomized, controlled, crossover study. The primary analysis involved using data obtained from participants following a high vitamin K1 diet (HK; 4-week intervention of increased leafy green vegetable intake). High and low responders were defined based on the median percent reduction (30%) in ucOC following the HK diet. Blood pressure (resting and 24 h), arterial stiffness, plasma glucose, lipid concentrations, and serum OC forms were assessed. Following the HK diet, ucOC and ucOC/tOC were suppressed more (p < 0.01) in high responders (41 and 29%) versus low responders (12 and 10%). The reduction in ucOC and ucOC/tOC was not associated with changes in blood pressure, arterial stiffness, plasma glucose, or lipid concentrations in the high responders (p > 0.05). Suppression of ucOC via consumption of leafy green vegetables has no negative effects on cardiometabolic health, perhaps, in part, because of cross-talk mechanisms.


The influence of inorganic phosphate and citrate anions on the effect of glycosaminoglycans during collagen fibril-formation.
Some anions like phosphate, sulfate and citrate delay the fibril-formation. The interaction of phosphate and citrate with chondroitin sulfate-A (CSA) in binding to collagen was investigated in different environmental conditions. By changing the concentration of phosphate from 5 to 50 mM/l and that of CSA from 0.5 to 16 mM/l some kind of competition of the anions was discovered. When different equilibration, systems were used the affinity of CSA to collagen was found to be 10 times greater than that of phosphate at pH 7.2 and I=0.15. As phosphate anions bind to collagen at physiological concentration. phosphate anions may be supposed to influence the biological fibril-formation. On the other hand the CSA exchanges phosphate for collagen. Therefore, glycosamino-glycans (GAG) in connective tissue ground substance in fibrillogenesis may regulate the ion binding and through this the tendency of aggregation of the collagen molecules. The binding of citrate to collagen and its effect and possible role in fibril-formation was also evaluated.


Population pharmacokinetics and Bayesian estimation of mycophenolic acid concentrations in stable renal transplant patients.
Therapeutic drug monitoring of mycophenolic acid (MPA) may minimise the risk of acute rejection after transplantation. Area under the curve (AUC) rather than trough concentration-based monitoring is recommended and models for AUC estimation are needed. To develop a population pharmacokinetic model suitable for Bayesian estimation of individual AUC in stable renal transplant patients. The population pharmacokinetics of MPA were studied using nonlinear mixed effects modelling (NONMEM) in 60 patients (index group) receiving MPA on a twice-daily basis. Ten blood samples were collected at fixed timepoints from ten patients and four blood samples were collected at sparse timepoints from 50 patients. Bayesian estimation of individual AUC was made on the basis of three blood concentration measurements and covariates. The predictive performances of the Bayesian procedure were evaluated in an independent group of patients (test group) comprising ten subjects in whom ten blood samples were collected at fixed timepoints. A two-compartment model with zero-order absorption best fitted the data. Covariate analysis showed that bodyweight was positively correlated with oral clearance. However, the weak magnitude of the reduction in variability (from 34.8 to 28.2%) indicates that administration on a per kilogram basis would be of limited value in decreasing interindividual variability in MPA exposure. Bayesian estimation of pharmacokinetic parameters using samples drawn at 20 minutes and 1 and 3 hours enabled estimation of individual AUC with satisfactory accuracy (bias 7.7%, range of prediction errors 0.43-15.1%) and precision (root mean squared error 12.4%) as compared with the reference value obtained using the trapezoidal method. This paper reports for the first time population pharmacokinetic data for MPA in stable renal transplant patients, and shows that Bayesian estimation can allow accurate prediction of AUC with only three samples. This method provides a tool for therapeutic drug monitoring of MPA or for concentration-effect studies. Its application to MPA monitoring in the early period post-transplantation needs to be evaluated.


Cerebral network deficits in post-acute catatonic schizophrenic patients measured by fMRI.
Twelve patients with catatonic schizophrenia and 12 matched healthy controls were examined with functional MRI while performing a motor task. The aim of our study was to identify the intracerebral pathophysiological correlates of motor symptoms in catatonic patients. The motor task included three conditions: a self-initiated (SI), an externally triggered (ET) and a rest condition. Statistical analysis was performed with SPM5. During the self-initiated movements patients showed significantly less activation than healthy controls in the supplementary motor area (SMA), the prefrontal and parietal cortex. Our results suggest a dysfunction of the "medial motor system" in catatonic patients. Self-initiated and externally triggered movements are mediated by different motor loops. The "medial loop" includes the SMA, thalamus and basal ganglia, and is necessary for self-initiated movements. The "lateral loop" includes parts of the cerebellum, lateral premotor cortex, thalamus and parietal association areas. It is involved in the execution of externally triggered movements. Our findings are in agreement with earlier behavioral data, which show deficits in self-initiated movements in catatonic patients but no impairment of externally triggered movements.


Pattern recognition analysis of differential mobility spectra with classification by chemical family.
Differential mobility spectra for alkanes, alcohols, ketones, cycloalkanes, substituted ketones, and substituted benzenes with carbon numbers between 3 and 10 were obtained from gas chromatography-differential mobility spectrometry (GC-DMS) analyses of mixtures in dilute solution. Spectra were produced in a supporting atmosphere of purified air with 0.6-0.8 ppm moisture, gas temperature of 120 degrees C, sample concentrations of approximately 0.2-5 ppm, and ion source of 5 mCi (185 MBq) 63Ni. Multiple spectra were extracted from chromatographic elution profiles for each chemical providing a library of 390 spectra from 39 chemicals. The spectra were analyzed for structural content by chemical family using two different approaches. In the one approach, the wavelet packet transform was used to denoise and deconvolute the DMS data by decomposing each spectrum into its wavelet coefficients, which represent the sample's constituent frequencies. The wavelet coefficients characteristic of the compound's structural class were identified using a genetic algorithm (GA) for pattern recognition analysis. The pattern recognition GA uses both supervised and unsupervised learning to identify coefficients which optimize clustering of the spectra in a plot of the two or three largest principal components of the data. Because principal components maximize variance, the bulk of the information encoded by the selected coefficients is about differences between chemical families in the data set. The principal component analysis routine embedded in the fitness function of the pattern recognition GA acts as an information filter, significantly reducing the size of the search space since it restricts the search to coefficients whose principal component plots show clustering on the basis of chemical family. In a second approach, a back propagation neural network was trained to categorize spectra by chemical families and the network was successfully tested using familiar and unfamiliar chemicals. Performance of the network was associated with a region of the spectrum associated with fragment ions which could be extracted from spectra and were class specific.


Recombinant pseudorabies virus expressing E2 of classical swine fever virus (CSFV) protects against both virulent pseudorabies virus and CSFV.
Both classical swine fever (CSF) and pseudorabies are highly contagious, economically significant diseases of swine in China. Although vaccination with the C-strain against classical swine fever virus (CSFV) is widely carried out and severe outbreaks of CSF seldom occur in China, CSF is sporadic in many pig herds and novel sub-subgenotypes of CSFV endlessly emerge. Thus, new measures are needed to eradicate CSFV from Chinese farms. The emergence of a pseudorabies virus (PRV) variant also posed a new challenge for the control of swine pseudorabies. Here, the recombinant PRV strain JS-2012-ΔgE/gI-E2 expressing E2 protein of CSFV was developed by inserting the E2 expression cassette into the intergenic region between the gG and gD genes of the gE/gI-deletion PRV variant strain JS-2012-ΔgE/gI. The recombinant virus was stable when passaged in vitro. A single vaccination of JS-2012-ΔgE/gI-E2 via intramuscular injection fully protected against lethal challenges of PRV and CSFV. Vaccination of piglets with the recombinant JS-2012-ΔgE/gI-E2 in the presence of high levels of maternally derived antibodies (Abs) to PRV can provide partial protection against lethal challenge of CSFV. Vaccination of the recombinant PRV JS-2012-ΔgE/gI-E2 strain did not induce the production of Abs to the gE protein of PRV or to the CSFV proteins other than E2. Thus, JS-2012-ΔgE/gI-E2 appears to be a promising recombinant marker vaccine candidate against PRV and CSFV for the control and eradication of the PRV variant and CSFV.


Generation of induced pluripotent stem cell line(ZZUi006-A)from a patient with myotonic dystrophy type 1.
Myotonic dystrophy type 1 (DM1) is a dominantly inherited neurodegenerative disease caused by a CTG repeat expansion close to the DMPK gene. The fibroblasts from a patient with DM1 were successfully converted to induced pluripotent stem cells (iPSCs), designated ZZUi006-A, by employing episomal plasmids expressing OCT3/4, SOX2, KLF4, LIN28, L-MYC. The ZZUi006-A cell line may provide a robust platform for further study of DM1 pathogenesis as well as drug testing and gene therapy research.


Targeting citrate as a novel therapeutic strategy in cancer treatment.
An important feature shared by many cancer cells is drastically altered metabolism that is critical for rapid growth and proliferation. The distinctly reprogrammed metabolism in cancer cells makes it possible to manipulate the levels of metabolites for cancer treatment. Citrate is a key metabolite that bridges many important metabolic pathways. Recent studies indicate that manipulating the level of citrate can impact the behaviors of both cancer and immune cells, resulting in induction of cancer cell apoptosis, boosting immune responses, and enhanced cancer immunotherapy. In this review, we discuss the recent developments in this emerging area of targeting citrate in cancer treatment. Specifically, we summarize the molecular basis of altered citrate metabolism in both tumors and immune cells, explore the seemingly conflicted growth promoting and growth inhibiting roles of citrate in various tumors, discuss the use of citrate in the clinic as a novel biomarker for cancer progression and outcomes, and highlight the new development of combining citrate with other therapeutic strategies in cancer therapy. An improved understanding of complex roles of citrate in the suppressive tumor microenvironment should open new avenues for cancer therapy.


C-H functionalization at sterically congested positions by the platinum-catalyzed borylation of arenes.
Despite significant progress in the area of C-H bond functionalization of arenes, no general method has been reported for the functionalization of C-H bonds at the sterically encumbered positions of simple arenes, such as mesitylene. Herein, we report the development of the first platinum-based catalyst for C-H borylation of arenes and heteroarenes. Notably, this method exhibited high tolerance toward steric hindrance and provided rapid access to a series of 2,6-disubstituted phenylboronic esters, valuable building blocks for further elaborations.


Chiral correlation between low-birefringent phases with twist grain boundary-like helix and highly birefringent phases with layer chirality as elucidated from circular dichroism observations.
The bent-shaped molecule with a central naphthalene core, 2,7-naphthalene-bis[4-(4-dodecylphenyliminomethyl)]benzoate, forms the low-birefringent B2 (LB-B2) phase with the twist grain boundary (TGB)-like helical structure and the low-birefringent B4 (LB-B4) phase in order of decreasing temperature. By applying the electric field, the LB-B2 phase is altered to the highly birefringent B2 (HB-B2) phase because of unwinding of the TGB-like helix. The HB-B2 phase is transformed to the HB-B4 phase without a loss of birefringence on cooling. These four phases show characteristic circular dichroism spectra, showing the consistent correlation through the transformation between these phases. The source of the chirality in the achiral system and the correlation in the chirality between these phases are discussed.


Contrasting arsenic biogeochemical cycling in two Moroccan alkaline pit lakes.
Pit lakes resulting from the flooding of abandoned mines represent a valuable freshwater reserve. However, water contamination by toxic elements, including arsenic, compromises their use for freshwater supply. For a better management of these reserves, our aim was to gain insight into arsenic cycling in two Moroccan alkaline pit lakes. We first showed that dimethylarsenic dominated in stratified lake ZA whereas in lake ZL1, As(V) was prevailing. Because microbially mediated processes largely contribute to arsenic cycling, the diversity of arsenic-methylating and -oxidizing bacteria was determined through the sequencing of arsM and aioA genes. Diverse arsM-carrying bacteria were thriving in ZA while a low diversity of aioA genes was detected in ZL1. We also determined the structure of the total bacterial communities by fingerprinting (ARISA). Contrasting arsenic speciation and bacterial communities in the two lakes were associated with differences of conductivity, Total Organic Carbon and temperature. In ZA, dissolved oxygen and redox potential were the main factors driving the total bacterial community structure and the ArsM diversity. In ZL1, stable bacterial communities were associated with limited water physico-chemistry variations. Our study provides new insights into the biogeochemical behavior of arsenic and the role of arsenic transforming bacteria in alkaline pit lakes.


Treatment of nonalcoholic fatty liver disease with an anti-osteoporotic medication: A hypothesis on drug repurposing.
Nonalcoholic fatty liver disease (NAFLD) is a disease of high prevalence without any approved treatment. Nonalcoholic steatohepatitis (NASH) is an advanced phenotype of the disease and the main focus of ongoing clinical trials. Denosumab, a human monoclonal antibody, which binds and inhibits the receptor activator of nuclear factor kappa-B ligand (RANKL), is a licensed medication for postmenopausal, male and glucocorticoid-induced osteoporosis, as well as for metastatic bone disease associated with specific cancers. Hepatic RANKL upregulation has been shown in a transgenic mice model. We hypothesized that hepatic RANKL upregulation may be associated with hepatic steatosis and inflammation, thus playing a role in the pathogenesis of NAFLD. If this hypothesis is verified, denosumab, an established anti-osteoporotic medication, may be considered as a candidate for NASH in drug-repurposing studies.


Main factors dominating the development, formation and dissipation of hypoxia off the Changjiang Estuary (CE) and its adjacent waters, China.
Hypoxia off the Changjiang Estuary (CE) and its adjacent waters is purported to be the most severe in China, attracting considerable concern from both the scientific community and the general public. Currently, continuous observations of dissolved oxygen (DO) levels covering hypoxia from its appearance to disappearance are lacking. In this study, twelve consecutive monthly cruises (from February 2015 to January 2016) were conducted. The consecutive spatiotemporal variations in hypoxia throughout the annual cycle were elucidated in detail, and the responses of annual variations in hypoxia to the different influential factors were explored. Overall, hypoxia experienced a consecutive process of expanding from south to north, then disappearing from north to south. The annual variations in hypoxia were mainly contingent on stratification variations. Among different stages, there was significant heterogeneity in the dominant factors. Specifically, low-DO waters initially appeared from the intrusion of nearshore Kuroshio branch current (NKBC), as NKBC intrusion provided a low-DO background and triggered stratification. Thereafter, stratification was enhanced and gradually expanded northward, which promoted the extension of low-DO areas. The formation of hypoxia was regionally selective, and more intense organic matter decomposition at local regions facilitated the occurrence and discontinuous distribution of hypoxia. Hypoxic zones were observed at the Changjiang bank and Zhejiang coastal region from August (most extensively at 14,800 km2) to October. Thereafter, increased vertical mixing facilitated the dissipation of hypoxia from north to south.


A cross-sectional study of environmental, dog, and human-related risk factors for positive canine leptospirosis PCR test results in the United States, 2009 to 2016.
Canine leptospirosis is a reemerging zoonotic disease concern in North America, and a better understanding of its epidemiology is needed. Wide-scale use and subsequent analyses of polymerase chain reaction (PCR) testing may provide additional insight into leptospirosis. This study aimed to describe temporal trends, to descriptively map, and to identify environmental, dog, and human-level factors associated with positive canine leptospirosis PCR test results in the United States. Data obtained from IDEXX Laboratories, Inc. on 40,118 canine leptospirosis PCR tests run in the United States between 2009 and 2016 were evaluated. Climate and socioeconomic (e.g. urban influence, income) data were obtained from public databases. Choropleth maps were created to identify high test-positive proportion areas and a cross-sectional analysis was completed with generalized (univariable, followed by multivariable) mixed logistic regression models accounting for county within state to identify significant predictors for a positive test. Overall test-positive proportion was 5.4% across the United States, with the regional point estimate highest in the southwest (8.1%). In the final multivariable model, the odds of a positive test were greater for male dogs (Odds Ratio [OR] = 1.28) and dogs 0-4 years of age (ORs ranged from 0.35-0.71 for the other age groups). The odds of a positive test were greater for dogs living in areas with wet environmental conditions (OR = 1.24). Season and temperature, as well as the interaction between them, were significant predictors of a positive test. Dogs had a greater probability of testing positive during cool temperatures (< 4 °C) compared to the other temperature categories in the fall season. These findings based on PCR testing allow for an improved understanding of factors influencing a positive canine leptospirosis PCR test and will assist targeted education and prevention efforts.


Care home nursing: Co-creating curricular content with student nurses.
Meeting the complex care needs of an ageing population is a global issue and long term care settings, such as care homes, play an essential role. However, there is a crisis in the recruitment of registered nurses within care homes. Higher educational institutions have a critical part to play in addressing the crisis in recruitment in care homes and it is argued that student nurses can have a significant role to play in co-creating curricular content responsive to population need. To co-create curricular content on care home nursing with student nurses. Co-creation through collaborative enquiry and a three stage thematic analysis. Undergraduate, preregistration nursing programme in a university in the United Kingdom. Student nurses from Years One to Four undertaking a Bachelor in Nursing with Honours degree. Six focus groups and two one to one interviews. Findings revealed predominantly negative attitudes towards care home nursing. Teaching and practice placements appeared to play a minor role in shaping students' attitudes but rather, gave the unspoken message that for the acquisition of necessary knowledge and skills, care homes were less important than other settings. Most students were initially averse to care home nursing as a career choice. During focus groups/interviews, views shifted from seeing care homes as places where you 'lose clinical skills' to places where there is 'a lot of responsibility', and also a potentially rewarding career choice. From this attitudinal shift, students made suggestions for developing better curricular content and more positive learning opportunities. A co-creative framework can create a space for mutual learning between students and staff about challenges and opportunities for equipping nurses to meet the needs of ageing populations. Student nurses are open to learning about care home nursing as part of their education and keen to have a more positive exposure.


Operative mortality rates after elective infrarenal aortic reconstructions.
This study was designed to test the hypothesis that cardiac complications (myocardial infarction, congestive heart failure, fatal arrhythmias) are no longer the leading cause of death after elective aortic reconstructions. The medical records of all elective infrarenal aortic reconstructions performed between January 1982 and June 1994 were retrospectively reviewed. All perioperative deaths were analyzed to determine the cause of death and were compared with a subset of 266 survivors to identify any associated preoperative or intraoperative factors. Seven hundred twenty-two aortic reconstructions were performed for aneurysmal or occlusive disease, and there were 44 deaths (overall mortality rate of 6.1%). The mortality rate after aortic reconstruction alone was 4.9% and increased with the addition of renal (8.9%, p = 0.16) or lower extremity vascular procedures (15.8%, p = 0.01). Multisystem organ failure (MSOF) was the cause of death in 56.8%, of the patients (3.5% overall mortality rate) followed by cardiac events in 25% (1.5% overall mortality rate). Visceral organ dysfunction was the most common cause of MSOF leading to death in 14 patients (56.0%), and postoperative pneumonia was responsible for the fatal MSOF in nine patients (36.0%). Patient age, history of myocardial infarction/congestive heart failure, ejection fraction less than 50%, duration of operative time, and performance of additional procedures were associated with increased operative mortality rates by multivariate analysis. MSOF, predominantly from visceral organ dysfunction, was the leading cause of death after elective infrarenal aortic reconstruction. The risk of MSOF and operative death increases with the complexity of the procedure and the number of comorbid conditions.


Reliability assessment between clinical attachment loss and alveolar bone level in dental radiographs.
The clinical attachment level (CAL) and radiographically assessed bone levels are used to assess the loss of periodontal tissue support in periodontitis, a chronic, multifactorial inflammatory disease of the periodontium. However, few studies have been done to study the relationship between these two parameters. According to our knowledge, this is the first study investigating the relationship between the two measurements using intraclass correlation analysis. The aim of the study is to investigate the relationship between CAL and radiographically assessed bone level in teeth affected with periodontitis. A retrospective cross-sectional study was conducted by selecting a sample of 880 periodontal sites in 104 periodontitis patients, aged 25-60 years. CAL and peri-apical radiographs of the selected sites were obtained from the computerized patient records. The distance from the cemento-enamel junction (CEJ) to the base of the alveolar bone level (ABL) was measured. The data was analyzed using SPSS. Intraclass correlation analysis (ICC) revealed a moderate degree of reliability between CAL and CEJ to ABL measurements. The average ICC was 0.68 with a 95% confidence interval of 0.53-0.77 (p < .001) indicating moderate to good reliability. Comparing the types of teeth, the central incisors, particularly the lower central incisors showed the highest ICC values (ICC: 0.822, CI: 0.77-0.86) indicating good reliability while the premolar and molars showed poor to moderate agreement (Maxillary premolars ICC: 0.464, CI: -0.18-0.74; maxillary first molar ICC: 0.516, CI: -0.154-0.772; mandibular first premolar ICC: 0.662, CI: 0.269-0.782; mandibular first molar ICC: 0.625, CI: 0.31-0.82). A moderate correlation existed between the radiographic and the clinical assessments (r = 0.5, p < .001). Despite the fact that significant varying levels of reliability has been found between CAL and radiographic bone level, both the clinical and radiographic examinations should be performed for the accuracy of diagnosis.


Co-Creation of a School-Based Motor Competence and Mental Health Intervention: Move Well, Feel Good.
Low motor competence (MC) and inhibited psychosocial development are associated with mental health difficulties. Improving children's MC through school-based physical activity interventions emphasising psychosocial development may therefore be a mechanism for promoting positive mental health. This study describes and provides reflective insights into the co-creation of 'Move Well Feel Good', a primary school physical activity intervention to improve children's MC and mental health. Class teachers, school leaders, physical activity specialists, and children (aged 8-9 years) participated in a series of co-creation workshops. Stakeholders' knowledge and experiences were integrated with existing research evidence using creative methods (e.g., post-it note tasks, worksheets, and drawings) to facilitate discussion. The co-creation process culminated in stakeholder consensus voting for one of three proposed intervention ideas. Children cited physical and mental health benefits, enjoyment with friends, and high perceived competence as motives for being physically active. Opportunities to develop MC across the different segments of the school day were identified by adult stakeholders, who perceived children's lack of resilience, an overloaded curriculum, and poor parental support for physical activity as barriers to intervention implementation. The chosen intervention idea received six out of a possible twelve votes. Co-creation projects are specific to the contexts in which they are implemented. This study reinforces the complex nature of school-based intervention development and highlights the value of engaging with stakeholders in co-creation processes.


Prognostic value of cells with more than 5c DNA content in node-negative breast cancer as determined by image cytometry from tissue sections.
The aim of this investigation was to study the prognostic significance of 5c cells (presence of cancer cells with > 5c DNA content; ie, over 18 pg of DNA per nucleus) in axillary node-negative breast cancer. Tissue sections (3 microns) from 134 tumors were stained for DNA using the Feulgen method and screened for the percentage of 5c cells with the CAS 200 image analysis system (Cell Analysis System, Inc, Lombard, IL). Cancer cells with a DNA content exceeding the 5c level were found in 45% (60 of 134) of the cases, accounting for a median of 0.2% (range, 0.05% to 1.05%) of all cells. The presence of 5c cells was associated with a high histologic grade of the tumor (P = .0001), a large number of mitoses (P < .0001), flow cytometric DNA aneuploidy and high S-phase fraction (P = .0002 and P < .0001, respectively), and c-erbB-2 oncoprotein and p53 tumor suppressor gene product overexpression (P = .0002 and P = .0006, respectively). Patients with 5c cell-positive tumors had a significantly worse 8-year survival rate (P = .003) than those with 5c cell-negative tumors. Subgroup analysis showed that the presence of 5c cells had a prognostic impact in low malignancy tumors, ie, in well-differentiated (grade I or II) and slowly proliferating tumors. Our findings suggest that determination of 5c cells may be a useful additional prognostic factor in axillary node-negative breast cancer. It adds prognostic information, especially in cases that are otherwise thought to have a favorable course.


Dobutamine stress echocardiography for cardiac risk assessment before aortic surgery.
This study evaluates dobutamine stress echocardiography (DSE) for perioperative cardiac risk assessment with elective aortic surgery. Dobutamine stress echocardiography was used to evaluate 81 patients before infrarenal aortic surgery. Patients were placed into three groups. Group I (n = 31) had normal DSEs. Group II (n = 25) had resting wall motion abnormalities without dobutamine-induced changes of ischemia. Group III (n = 25) had evidence of dobutamine-induced ischemia. Patient analysis revealed that of 46 patients with clinical indicators of coronary artery disease (CAD), only 23 had DSEs with inducible ischemia. Two of 35 patients without clinical indicators of CAD had DSEs with inducible ischemia. The 56 patients in group I and II underwent aortic reconstruction without cardiac complications or death. Of the 25 patients in group III, surgery was deferred in five (two patients with claudication and three with aneurysms < or = 5 cm), and four underwent coronary artery bypass grafting. Outcome after coronary artery bypass grafting included one death from stroke, one aneurysm rupture, and two uncomplicated aortic reconstructions. The remaining 16 patients in group III underwent aortic surgery, with three postoperative myocardial infarctions (MI) and no deaths. Using DSE for preoperative assessment of cardiac risk allowed us to operate on 74 of 81 patients being considered for elective aortic reconstruction, with no operative deaths and a 4.1% rate of perioperative MI. Dobutamine stress echocardiography has the ability to identify patients with asymptomatic stress-induced ischemic myocardium and its increased risk for perioperative MI (p < 0.001). Equally important, for patients with clinical indicators of CAD but without DSE-inducible ischemia, no further cardiac evaluation is necessary.


[Obstructive disorders of urine transport in the upper urinary tract--vascular and inflammatory causes].
Vascular and inflammatory causes of ureteral obstruction may be difficult in diagnosis and are recognized often too late. Symptoms, diagnostic approach and therapeutic measures are described in detail.


Fruit availability has a complex relationship with fission-fusion dynamics in spider monkeys.
Understanding the ecological and social factors that influence group size is a major focus of primate behavioural ecology. Studies of species with fission-fusion social organizations have offered an insightful tool for understanding ecological drivers of group size as associations change over short temporal and spatial scales. Here we investigated how the fission-fusion dynamics of spider monkeys (Ateles geoffroyi) at Runaway Creek, Belize were affected by fruit availability. When males and females were analyzed together, we found no association between fruit availability and subgroup size. However, when females were analyzed separately, we found that when fruit availability increased, so did subgroup size. In all analyses, higher fruit availability did not influence subgroup spatial cohesion. Our results point to the complexity of understanding grouping patterns, in that while ecological factors make groups of specific sizes advantageous, social factors also play an important determining role.


Selenium and its' role in the maintenance of genomic stability.
Selenium (Se) is an essential micronutrient for humans, acting as a component of the unusual amino acids, selenocysteine (Se-Cys) and selenomethionine (Se-Met). Where Se levels are low, the cell cannot synthesise selenoproteins, although some selenoproteins and some tissues are prioritised over others. Characterised functions of known selenoproteins, include selenium transport (selenoprotein P), antioxidant/redox properties (glutathione peroxidases (GPxs), thioredoxin reductases and selenoprotein P) and anti-inflammatory properties (selenoprotein S and GPx4). Various forms of Se are consumed as part of a normal diet, or as a dietary supplement. Supplementation of tissue culture media, animal or human diets with moderate levels of certain Se compounds may protect against the formation of DNA adducts, DNA or chromosome breakage, and chromosome gain or loss. Protective effects have also been shown on mitochondrial DNA, and on telomere length and function. Some of the effects of Se compounds on gene expression may relate to modulation of DNA methylation or inhibition of histone deacetylation. Despite a large number of positive effects of selenium and selenoproteins in various model systems, there have now been some human clinical trials that have shown adverse effects of Se supplementation, according to various endpoints. Too much Se is as harmful as too little, with animal models showing a "U"-shaped efficacy curve. Current recommended daily allowances differ among countries, but are generally based on the amount of Se necessary to saturate GPx enzymes. However, increasing evidence suggests that other enzymes may be more important than GPx for Se action, that optimal levels may depend upon the form of Se being ingested, and vary according to genotype. New paradigms, possibly involving nutrigenomic tools, will be necessary to optimise the forms and levels of Se desirable for maximum protection of genomic stability in all humans.


GC-MS Analysis of Phthalates and Di-(2-thylhexyl) Adipate in Canadian Human Milk for Exposure Assessment of Infant Population.
Although more information has become available on the occurrence of phthalates and di(2-ethylhexyl) adipate (DEHA) in foods including cow's milk, information on their presence in human milk, the important and recommended sole diet for infants up to six months of age, is very limited, especially for DEHA. To develop a GC-MS method for simultaneous analysis of DEHA and phthalates in human milk samples and generate occurrence data for exposure assessment. Human milk samples were extracted with acetonitrile followed by dispersive solid-phase extraction and GC-MS analysis. Among the 305 human milk samples collected from the Canadian Maternal-Infant Research on Environmental Chemicals Study, some phthalates (DHxP, BBzP, and DOP) were not detected in any of the samples, while DEHA and the other phthalates (DMP, DEP, DBP, DiBP, and DEHP) were detected at low frequencies with levels from 30.4-237 ng/g in up to 31 of the 305 human milk samples. In general, DEHA and phthalates were detected at low frequencies and low levels in the 305 human milk samples. A GC-MS method based on dispersive solid phase extraction was developed for analysis of DEHA and eight phthalates in 305 human milk samples for exposure assessment.


Pregnant women's alcohol consumption: the predictive utility of intention to drink and prepregnancy drinking behavior.
This study had two aims: (1) to examine pregnant women's alcohol consumption across time from prepregnancy until childbirth and (2) to explore whether prepregnancy drinking and intention to drink predict prenatal alcohol consumption while controlling for relevant demographic variables. At 17-21 weeks, 248 pregnant women completed questions about demographics, intention to drink alcohol during the subsequent pregnancy, and retrospective measures of prepregnancy and early pregnancy consumption. After this time, calendars were sent fortnightly assessing daily alcohol consumption until birth. For women who drank both prepregnancy and postpregnancy confirmation, average fortnight alcohol consumption in the first weeks of pregnancy was lower than during prepregnancy, and consumption continued to decrease between gestational weeks 1 and 8, particularly following pregnancy confirmation, after which it remained relatively stable. When predicting whether women drank in late pregnancy, intention accounted for unique variance after controlling for income and prepregnancy drinking. For women who drank after pregnancy confirmation, prepregnancy drinking quantity significantly predicted intention to drink, which in turn predicted fortnight alcohol consumption in later pregnancy, after controlling for prepregnancy drinking and income. Findings highlight the need to measure alcohol consumption at multiple time points across pregnancy, the need for educating and supporting women to reduce consumption when planning pregnancies, and the usefulness of intention to drink as a predictor of drinking during pregnancy.


Aflatoxin contamination of pearl millet during field and storage conditions with reference to stage of grain maturation and insect damage.
Aflatoxin contamination in five varieties of pearl millet (ICMH-451, ICMP-50I, ICTP-8203, WCC-75 and ICMV-155) was studied from field and storage conditions in three districts of Andhra Pradesh State, India and the inter-relationships between various parameters such as stage of grain maturation in the field and insect pest infestation in storage in relation to aflatoxin production were evaluated. Aflatoxin contamination was more frequent in the seed samples collected from the fields during rainy season than winter season. All major aflatoxins were isolated from one or the other varieties of pearl millet, whereas aflatoxin G2 was not commonly observed in the seed samples collected during winter. Among all the varieties tested, ICMH-451 was vulnerable to aflatoxin contamination whereas ICMV-155 was the least susceptible variety. The higher amount of aflatoxins was observed in the matured seed samples followed by pre-matured and milky stage. Among all the toxins reported in the field, aflatoxin B1 was found in higher concentration (185 (μg/kg) followed by B2 (105 μg/kg). The four major types of aflatoxins with higher levels (35, 40, 140, 190 μg/kg of G1, G2, B2, B1 were reported in the rainy season seed samples after six months of storage, whereas aflatoxin G1 was not observed in any variety of stored seed sample from winter. Statistical analysis revealed that the aflatoxin incidence in relation to different parameters studied was significantly different for each factor. The relationship between aflatoxin contamination and insect damaged-grain clearly indicated that the seed samples with 16-40% of insect damage contained higher amounts of aflatoxins (758 μg/kg).


Competitive inhibition by glucose of myo-inositol incorporation into cultured porcine aortic endothelial cells.
To explore the significance of hyperglycaemia as a causal factor for the appearance of diabetic angiopathies we investigated aspects of myo-inositol metabolism in porcine aortic endothelial cells. myo-Inositol was shown to be a long-living metabolite. Its uptake into the cells was mediated by a high-affinity, Na(+)-dependent uptake system inhibitable by ouabain with an apparent KM of 18.6 mumols/l, which was responsible for more than 80% of total uptake at physiological myo-inositol concentrations. Inhibition of inositol uptake by D-glucose was exclusively competitive with an apparent Ki of 24 mmol/l as shown by Lineweaver-Burk- and Dixon-plot analysis. The specificity of competitive inhibition was studied. L-Glucose which is stereochemically related to myo-inositol in the same way as the D-isomer proved to be an equally potent inhibitor. The hexoses D-galactose, D-mannose and D-fructose inhibited myo-inositol uptake to a minor extent. D-allose and 3-O-methyl-D-glucose had no inhibitory effect indicating that the OH-group of the carbon atom in 3 position is essential for the interaction with the carrier. The acyclic hexitol sorbitol also did not compete. As expected, the aldose reductase blocker sorbinil did not influence the carrier since there is no polyol pathway operating in porcine aortic endothelial cells. In accordance with the results of the uptake experiments, the incorporation of exogenous myo-inositol into membrane phosphatidylinositol was reduced at elevated extracellular glucose levels. The results raise the possibility that hyperglycaemia impairs endothelial inositol supply.


Missing binary data extraction challenges from Cochrane reviews in mental health and Campbell reviews with implications for empirical research.
Tο report challenges encountered during the extraction process from Cochrane reviews in mental health and Campbell reviews and to indicate their implications on the empirical performance of different methods to handle missingness. We used a collection of meta-analyses on binary outcomes collated from a previous work on missing outcome data. To evaluate the accuracy of their extraction, we developed specific criteria pertaining to the reporting of missing outcome data in systematic reviews. Using the most popular methods to handle missing binary outcome data, we investigated the implications of the accuracy of the extracted meta-analysis on the random-effects meta-analysis results. Of 113 meta-analyses from Cochrane reviews, 60 (53%) were judged as "unclearly" extracted (ie, no information on the outcome of completers but available information on how missing participants were handled) and 42 (37%) as "unacceptably" extracted (ie, no information on the outcome of completers as well as no information on how missing participants were handled). For the remaining meta-analyses, it was judged that data were "acceptably" extracted (ie, information on the completers' outcome was provided for all trials). Overall, "unclear" extraction overestimated the magnitude of the summary odds ratio and the between-study variance and additionally inflated the uncertainty of both meta-analytical parameters. The only eligible Campbell review was judged as "unclear." Depending on the extent of missingness, the reporting quality of the systematic reviews can greatly affect the accuracy of the extracted meta-analyses and by extent, the empirical performance of different methods to handle missingness.


Author Correction: Serum vitamin D is associated with improved lung function markers but not with prevalence of asthma, emphysema, and chronic bronchitis.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Two-Year Results of the Prospective Spine Treatment Outcomes Study: Analysis of Postoperative Clinical Outcomes Between Patients with and without a History of Previous Cervical Spine Surgery.
History of previous cervical spine surgery is a frequently cited cause of worse outcomes after cervical spine surgery. The purpose of this study was to determine any differences in clinical outcomes after cervical spine surgery between patients with and without a history of previous cervical spine surgery. A multicenter prospective database was reviewed retrospectively to identify patients with cervical spondylosis undergoing surgery with a minimum 2-year follow-up. Patients were divided into 2 groups: patients with (W) or without (WO) previous history of cervical spine surgery. Statistical analyses of Health-Related Quality of Life scores were analyzed with statistical software to fit linear mixed models for continuous longitudinal outcome. A total of 1286 patients (377 W, 909 WO) met criteria for inclusion. Overall, patients in both groups experienced an improvement in their Health-Related Quality of Life scores. However, patients in the W group had significantly decreased improvement compared with WO patients in the Neck Disability Index score and the following SF-36 domain scores: Role Physical, Bodily Pain, General Health, Vitality, Social Functioning, Health Transition, and Physical Component Summary at all time points (P < 0.05). There was no statistically significant difference between the W and WO groups in operative time, estimated blood loss, length of stay, or complications (P > 0.05). Patients with a history of previous cervical spine surgery had inferior improvement in quality of life outcome scores. Patients with a history of previous surgical intervention who elect to undergo subsequent surgeries should be appropriately counseled about expected results.


What makes it difficult to refine protein models further via molecular dynamics simulations?
Protein structure refinement remains a challenging yet important problem as it has the potential to bring already accurate template-based models to near-native resolution. Refinement based on molecular dynamics simulations has been a highly promising approach and the performance of MD-based refinement in the Feig group during CASP12 is described here. During CASP12, sampling was extended well into the microsecond scale, an improved force field was applied, and new protocol variations were tested. Progress over previous rounds of CASP was found to be limited which is analyzed in terms of the quality of the initial models and dependency on the amount of sampling and refinement protocol variations. As current MD-based refinement protocols appear to be reaching a plateau, detailed analysis is presented to provide new insight into the major challenges towards more extensive structure refinement, focusing in particular on sampling with and without restraints.


Regeneration of the native bacteriorhodopsin structure from two chymotryptic fragments.
Chymotrypsin cleaves bacteriorhodopsin to two fragments: C-1, amino acids 72-248, and C-2, amino acids 1-71. Denaturation and renaturation of these fragments have been studied. Following denaturation in sodium dodecyl sulfate, both C-1 and C-2 regain secondary structure in phospholipid/cholate/sodium dodecyl sulfate mixed micelles. When combined, they form a complex with a secondary structure resembling that of bacterio-opsin. However, on further incubation in phospholipid/cholate/sodium dodecyl sulfate, separate fragments as well as the C-1 and C-2 complex denature again. Retinal binds tightly to the C-1 and C-2 complex (Kb greater than 10(7) M-1) and stabilizes the folded conformation. The formation of the complex of C-1, C-2, and retinal is maximal at pH 6.0. The ternary complex contains two species: one which absorbs similarly to the light-adapted purple membrane and a second with a lambda max between 450 and 500 nm. The formation of the latter species is favored at higher temperatures and is reversible. Vesicles formed from the ternary complex of C-1, C-2, and retinal translocate protons at a level close to that of intact bacteriorhodopsin.


Experimental neuropsychopharmacology, yesterday, today and tomorrow. A conversation with Michel Hamon.
For this issue, Luc Zimmer, professor of pharmacology and chair of the Neuropsychopharmacology Committee of the French Society of Pharmacology and Therapeutics, talked with Michel Hamon, honorary director of research at the French National Institute of Health and Medical Research (INSERM) and honorary professor of neuropharmacology at Paris-Sorbonne (Pierre et Marie Curie) University. Some of the leading names in neuropsychopharmacology research are mentioned, pointing to significant conceptual advances that founded this discipline. The links between psychopharmacology and neuropharmacology are also discussed in the light of past collaborations. Finally, priorities are proposed for the emergence of the psychopharmacology of the future.


Changes of Viral Load, Drug Resistance, and CD4 Cell Count in HIV-1-Infected Patients Receiving Antiretroviral Therapy in Hebei Province, China, 2003-2019.
In this study, a total of 462 samples of whole blood were collected from 36 patients enrolled. During the entire course of antiretroviral therapy (ART) from 2003 to 2019, both the CD4 cell count and viral load (VL) of study patients were examined annually, and an HIV-1 genotypic drug resistance (DR) assay was carried out using an in-house method when the HIV-1 VL was >1,000 copies/mL. The results indicated that 13 (36.1%) experienced treatment failure and 23 (63.9%) experienced treatment success among 36 patients. The proportion of patients with effective treatment after the adjustment of ART regimens was significantly higher than before adjustment (χ2 = 33.796, p < .001). Furthermore, the frequencies of HIV-1 DR mutations before adjustment were higher than those after adjustment (t = 3.345, p = .002). In particular, among 23 patients with effective treatment after adjustment, the means ± standard deviations of the VLs and CD4 cell counts before adjustment were 3.85 ± 0.65 log RNA copies/mL and 226.83 ± 106.06 cells/mm3, respectively, compared with 2.19 ± 0.58 log RNA copies/mL and 367.68 ± 174.62 cells/mm3, respectively, after adjustment. Obviously, there were statistically significant differences in the changes in VL (t = 8.728, p < .001) and CD4 cell count (t = -4.476, p < .001) before and after adjustment. Therefore, patients who received updated ART regimens containing LPV/r and TDF after adjustment achieved better therapeutic effects compared with patients who received initial ART regimens harboring D4T/AZT or NVP. This suggests that future research is needed to initiate the surveillance of DR, VLs, and CD4 cell counts immediately after HIV diagnosis, and dynamic changes in these indicators so as to optimize ART effects.


Sepsis in the newborn.
Systemic infection in the newborn is the commonest cause of neonatal mortality. Data from National Neonatal Perinatal Database 2000 suggest that Klebsiella pneumoniae and Staphylococcus aureus are the commonest causes of neonatal sepsis in India. Two forms of clinical presentations have been identified. Early onset sepsis, probably related to perinatal risk factors, usually presents with respiratory distress and pneumonia whthin 72 hours of age. Late onset sepsis, related to hospital acquired infections, usually presents with septicemia and pneumonia after 72 hours of age. Clinical features of sepsis are non-specific in neonates and a high index of suspicion is required for the timely diagnosis of sepsis. Although blood culture is the gold standard for the diagnosis of sepsis, reports are available after 48-72 hours. A practical septic screen for the diagnosis of sepsis has been described and some suggestions for antibiotic use have been included in the protocols.


Pro-apoptotic effect of a nitric oxide-donating NSAID, NCX 4040, on bladder carcinoma cells.
Nitric oxide-releasing non steroidal anti-inflammatory drugs (NO-NSAIDs) are a promising class of compounds that cause cell cycle perturbations and induce apoptosis in cell lines from different tumors. We investigated the activity of a recently developed NO-NSAID (NCX 4040) in bladder cancer cell lines (HT1376 and MCR). Cells were treated with different drug concentrations for different exposure times. Cytostatic and cytocidal activity was tested by SRB assay and apoptosis was evaluated by TUNEL analysis, ANNEXIN V assay and fluorescence microscopy. To further investigate the cell death-inducing mechanisms of NCX 4040, we analyzed gp-170, caspase expression and mitochondrial membrane potential (Delta Psi) depolarization. NCX 4040 showed a striking cytocidal activity in both cell lines, reaching LC(50) at a 10-microM and 50-microM concentrations in HT1376 and in MCR cells, respectively, after an exposure of only 6 h followed by an 18-h washout. Apoptosis was triggered in up to 90% of cells and was associated with active caspase-3 expression and Delta Psi depolarization in both cell lines after a 6-h exposure. In conclusion, NCX 4040, which probably causes apoptosis via a mitochondrial-dependent mechanism, could prove to be a useful agent for improving bladder cancer treatment.


Effect of proguanil and cycloguanil on human lymphocytes in vitro.
The in vitro effect of proguanil and its active metabolite cycloguanil on proliferating human blood mononuclear cells was studied. Proguanil had no effect on 14C-thymidine incorporation or on the number of cells. Cycloguanil, in concentrations corresponding to the plasma levels found in clinical practice, blocked the endogenous synthesis of thymidine and decreased the number of mitogen- and antigen-stimulated cells. The effect on phytohaemagglutinin-stimulated cells was transient. Higher concentrations of cycloguanil, corresponding to intralymphocytic levels in clinical practice, permanently suppressed the growth of lymphocytes. The effect of cycloguanil could be reversed by low doses of folinic acid and high doses of folic acid.


Crystal structure of human uracil-DNA glycosylase in complex with a protein inhibitor: protein mimicry of DNA.
Uracil-DNA glycosylase inhibitor (Ugi) is a B. subtilis bacteriophage protein that protects the uracil-containing phage DNA by irreversibly inhibiting the key DNA repair enzyme uracil-DNA glycosylase (UDG). The 1.9 A crystal structure of Ugi complexed to human UDG reveals that the Ugi structure, consisting of a twisted five-stranded antiparallel beta sheet and two alpha helices, binds by inserting a beta strand into the conserved DNA-binding groove of the enzyme without contacting the uracil specificity pocket. The resulting interface, which buries over 1200 A2 on Ugi and involves the entire beta sheet and an alpha helix, is polar and contains 22 water molecules. Ugi binds the sequence-conserved DNA-binding groove of UDG via shape and electrostatic complementarity, specific charged hydrogen bonds, and hydrophobic packing enveloping Leu-272 from a protruding UDG loop. The apparent mimicry by Ugi of DNA interactions with UDG provides both a structural mechanism for UDG binding to DNA, including the enzyme-assisted expulsion of uracil from the DNA helix, and a crystallographic basis for the design of inhibitors with scientific and therapeutic applications.


Reducing time between referral and diagnosis in paediatric outpatient neurodevelopmental and behavioural clinics.
This study investigated the impact of a transdisciplinary advanced allied health practitioner (AAHP) in a public hospital general paediatric outpatient department for children referred for concerns with neurodevelopment, learning and/or behaviour. It was hypothesised that time from referral to initial consultation and diagnosis was reduced for children initially assessed by AAHP. It was also hypothesised that children seen by AAHP were less likely to require a medical specialist appointment. De-identifiable data were gathered from hospital records both retrospectively and prospectively for each group. Data were collected from: 75 children who attended a medical consultation only clinic in 2014; 75 children who attended an AAHP clinic and a medical officer in 2014 and 75 children who attended an AAHP clinic and a medical officer 2 years after establishment in 2016/2017. On average, 52% children were managed independently by the AAHP. Wait time from referral to consultation significantly decreased from 169 days to 48 days. Similarly, time from referral to diagnosis significantly reduced by almost a half, from 57.2 to 30.59 weeks. This study provides preliminary evidence that utilisation of transdisciplinary AAHPs in general paediatric outpatient departments may reduce waitlists, reduce amount of medical specialist appointments required and provide earlier diagnosis for children with neurodevelopmental, behavioural and/or learning difficulties.


A new in-vitro drug sensitivity test (collagen-gel droplet embedded-culture drug sensitivity test) in carcinomas of pancreas and biliary tract: possible clinical utility.
The collagen-gel droplet embedded-culture drug sensitivity test (CD-DST), a chemosensitivity test, evaluate the efficacy of anticancer drugs and to was used clinically to thus plan rational postoperative chemotherapy for patients with pancreatic and biliary tract carcinomas. The CD-DST solves some problems inherent in other conventional assays. This method: (1) allows evaluation of four chemotherapeutic agents, using small quantities of cells (1 x 10(5) cells), (2) shows high primary culture success rates, (3) maintains the original growth characteristics of the cultured cells, (4) eliminates the effects of fibroblasts by employing image analysis, and (5) permits evaluation using physiologic drug concentrations. Primary cultures of tumor cell samples from all 25 patients with pancreatic or biliary tract carcinomas studied were successful. Against pancreatic carcinomas, the efficacy rates, assessed by CD-DST, of four anticancer drugs evaluated were: 25.0% for mitomycin (MMC), followed by 23.5% for adriamycin, 18.8% for 5-fluorouracil (5-FU), and 11.8% for cisplatin (CDDP). Against biliary tract carcinomas, the rates were highest for 5-FU and MMC (50.0%) and lowest for CDDP (25.0%). The efficacy rates for all four anticancer drugs evaluated were higher against biliary tract carcinomas than against pancreatic carcinomas. Tumor cultures from 10 of 17 patients with pancreatic cancer and 3 of 8 patients with biliary tract cancer showed no sensitivity to any of the drugs tested. The in-vitro results with CD-DST suggest the risk of administering non-selective postoperative chemotherapy to patients with pancreatic and biliary tract carcinomas, and emphasize the importance of carefully selecting effective chemotherapeutic agents based on adequate chemosensitivity testing.


Orientation and flexibility of the choline head group in phosphatidylcholine bilayers.
The average orientation and flexibility of the phosphorylcholine group are deduced from deuterium and phosphorus-31 nuclear magnetic resonance measurements of unsonicated phosphatidylcholine bilayers in the liquid crystalline state. The experimental data are consistent with a model in which the polar head group exhibits a restricted flexibility characterized by rapid transitions between two enantiomeric conformations. A completely flexible or a completely rigid head group structure can be excluded. The phosphorylcholine residue is found to be bent at the position of the phosphate group, due to a gauche-gauche conformation of the phosphodiester linkage. The choline dipole is aligned parallel to the plane of the membrane, which is in agreement with X-ray and neutron diffraction studies. The average orientation of the phosphorylcholine group is therfore the same as that of the phosphorylethanolamine head group.


[The pathomorphology of chronic alcohol-associated myocardial changes].
Forensic cases of sudden cardiac death were reviewed from the point of view of a history of alcohol consumption. A group of known cases of chronic alcoholism and another group of cases known to have been normal social drinkers without evidence of chronic alcoholism were selected. Cases with evidence of hypertension, ischemic or coronary heart disease, as well as cases with positive toxicological findings, were excluded. Applying the same criteria, cases of sudden death from other causes without a known history of alcohol consumption were selected to form a control group. With particular reference to histological myocardial changes, which are often documented in association with alcohol consumption, no noteworthy differences were found between the two major groups despite the significantly much lower frequency in the control group.


Rhodomyrtus tomentosa (Aiton.): A review of phytochemistry, pharmacology and industrial applications research progress.
Rhodomyrtus tomentosa (Aiton) is a flowering plant native to southern and southeastern Asia. Up to date, 106 chemical constituents have been isolated and identified from R. tomentosa. Among these compounds, triterpenoids, flavonoids, phenols and meroterpenoids are the major constituents. Investigations of pharmacological activities of R. tomentosa revealed that this edible medicinal herb exhibits a wide range of therapeutic potential including antibacterial, antitumor, anti-inflammatory and antioxidant activities both in vivo and in vitro. The purpose of this review is to provide an overview of R. tomentosa studies until 2019. This article also intends to review advances in the botanical, phytochemical, pharmacological studies and industrial applications of R. tomentosa, which will provide a useful bibliography for further investigations and applications of R. tomentosa in medicines and foods.


What we see is what we touch? Sex estimation on the pelvis in virtual anthropology.
Computed tomography (CT) scans are a convenient means to study 3D reconstructions of bones. However, errors associated with the different nature of the observation, e.g. visual and tactile (on dry bone) versus visual only (on a screen) have not been thoroughly investigated. We quantified the errors between modalities for sex estimation protocols of nonmetric (categorical and ordinal) and metric data, using 200 dry pelves of archaeological origin and the CT reconstructions of the same bones. In addition, we 3D surface scanned a subsample of 39 pelves to compare observations with dry bone and CT data. We did not focus on the sex estimation accuracy but solely on the consistency of the scoring, hence, the interchangeability of the modalities. Metric data yielded the most consistent results. Among the nonmetric protocols, ordinal data performed better than categorical data. We applied a slightly modified description for the trait with the highest errors and grouped the traits according to consistency and availability in good, intermediate, and poor. The investigated modalities were interchangeable as long as the trait definition was not arbitrary. Dry bone (gold standard) performed well, and CT and 3D surface scans performed better. We recommend researchers test their affinity for using virtual modalities. Future studies could use our consistency analysis and combine the best traits, validating their accuracy on various modalities.


Bio-adsorption and Bio-transformation of Arsenic by Acidithiobacillus ferrooxidans BY3.
Arsenic, a toxic element in the environment, has seriously threatened the health of hundreds of millions of people in the world. Meanwhile, microorganisms play an important role in the adsorption and bio-transformation of arsenic. Here, we compared the biological characteristics of Acidithiobacillus ferrooxidans BY3 in different media systems, such as arsenic bio-adsorption and bio-transformation capacities. We show that arsenic stress significantly affected the pH and Eh of the culture systems, as well as the oxidation rates of Fe2+ and bacteria numbers. Furthermore, arsenic influenced bacterial structure and composition of the cell membrane, caused volume decreased and changed the vibration conditions of characteristic peaks of surface groups (-CH2, -NH, and -OH) on cell membranes. In addition, At.f-BY3 shows high bio-adsorption abilities and certain bio-transformation abilities for iAsIII. Bio-adsorption and conversion efficiency was also shown to be significantly affected by Fe2+ concentrations in the reaction systems. Statistic analysis revealed 10.07-fold increase of the transformation ability of iAsIII into iAsV in the 9 K growth media containing 1600 mg/L NaAsO2 compared with that in the 1 K growth media. Our findings contribute to understand the applications and microbiological mechanisms of Acidithiobacillus ferrooxidans in arsenic pollution. Graphical Abstract Bio-adsorption and bio-transformation are used as a biological method of heavy metals pollution, such as Cu2+, Ni2+, Pb2+, Cr2+, Zn2+, Cd2+, As3+ and As5+ in acid mine water. The aim of this investigation was to assess the performance of arsenite (iAsIII) to adsorption and transformation by Acidithiobacillus ferrooxidans BY3, and application of Acidithiobacillus ferrooxidans BY3 on the aspect of arsenic pollution has great potential of exploration.


Robotic Urethral Reconstruction Outcomes in Men With Posterior Urethral Stenosis.
To evaluate surgical outcomes stratified by posterior urethral obstruction (PUO) etiology in men undergoing definitive robotic posterior urethral reconstruction. A retrospective, single surgeon, review of men undergoing robotic posterior urethral reconstruction between 2018 and 2020 was performed. Differences in complications, reconstructive success (no further intervention), and urinary continence by PUO etiology were assessed. Robotic posterior urethral reconstruction was performed in 21 men. PUO etiology included benign prostatic hypertrophy treatment in 5 (24%), prostatectomy in 10 (48%), radiation in 5 (24%), and trauma in 1 (5%). Median number of prior endoscopic treatments was 3 (benign prostatic hypertrophy), 3 (prostatectomy), and 2 (radiation) with an average time between obstruction and reconstruction of 9, 12, and 15 months (P = .52). Median length of stay after reconstruction was 2, 1, and 2 days (P = .45). Thirty-day complications occurred in 0%, 20%, 40% (P = .19). Post-reconstruction re-intervention was necessary in 0%, 10%, 80% (P = .004). Ultimately, anatomic success was achieved in 100%, 90%, 80% (P = .63), with functional success rates of 100%, 100%, 60% (P = .035). Median postoperative pad/day usage was 0,0, 10.5 (P <.001), and ultimately 0%, 30%, 80% (P = .013) underwent artificial urinary sphincter placement. Endoscopic treatment of posterior urethral obstruction (PUO) secondary to benign and malignant prostate conditions is associated with a high incidence of treatment failure. Robotic posterior urethral reconstruction is a safe and effective surgical solution for men with PUO in the absence of pelvic radiation. Men with pelvic radiation appear to be at increased risk of complications, PUO recurrence, and clinically significant stress urinary incontinence.


Panniculitis caused by Mycobacterium monacense mimicking erythema induratum: a case in Ecuador.
We report the first case of recently characterized species M. monacense associated with chronic nodular vasculitis, infecting a young woman. This case represents the first isolation of M. monacense from Ecuador. The isolate was identified by conventional and molecular techniques.


Unveiling the tumour-regulatory roles of miR-1275 in cancer.
The rapid development of small RNA and molecular biology research in the past 20 years has enabled scientists to discover many new miRNAs that are proven to play essential roles in regulating the development of different cancer types. Among these miRNAs, miR-1275 is one of the well-studied miRNAs that has been described to act as a tumour-promoting or tumour-suppressing miRNA in various cancer types. Even though miR-1275 has been widely reported in different original research articles on its roles in modulating the progression of different cancer types, however, there is scarce an in-depth review that could constructively summarize the findings from different studies on the regulatory roles of miR-1275 in different cancer types. To fill up this literature gap, therefore, this review was aimed to provide an overview and summary of the roles of miR-1275 in modulating the development of different cancers and to unravel the mechanism of how miR-1275 regulates cancer progression. Based on the findings summarized from various sources, it was found that miR-1275 plays a vital role in regulating various cellular signaling pathways like the PI3K/AKT, ERK/JNK, MAPK, and Wnt signaling pathways, and the dysregulation of this miRNA has been shown to contribute to the development of multiple cancer types such as cancers of the liver, breast, lung, gastrointestinal tract and genitourinary tract. Therefore, miR-1275 has great potential to be employed as a biomarker to diagnose cancer and to predict the prognosis of cancer patients. In addition, by inhibiting the expression of its unique downstream targets that are involved in regulating the mentioned cellular pathways, this miRNA could also be utilized as a novel therapeutic agent to halt cancer development.


Absence of evidence for HER2 amplification in nasopharyngeal carcinoma.
HER2 (c-erbB-2) has been suggested to be a prognostic factor in a variety of human cancers including breast, gastric and ovarian cancers. This study is therefore designed to identify changes of HER2 in nasopharyngeal carcinoma (NPC), an epithelia-derived malignancy with strong racial and geographic distribution. Interphase FISH and immunohistochemical (IHC) staining were used to analyze the gene copy number and protein expression of HER2 in 45 cases of NPC from Guangzhou, Southern China, an area with the highest incidence of NPC in the world. Our results, however, found no significant alterations in gene copy number for HER2, although IHC staining detected expression of HER2 oncoprotein in 33% of the 45 NPC tumors. No correlation was observed between HER2 expression and sex, age and clinical outcome of the patients, T stage, lymph node status, site and histopathological grading of the tumors. These results cast doubt on the value of HER2 as a prognostic factor for NPC.


Metastasis from prostatic carcinoma causing acute appendicitis: Report of a case.
Metastasis induced acute appendicitis is extremely rare. We present a 72-year-old man who presented with typical signs of acute appendicitis. He underwent appendectomy and was successfully discharged without any complication. Histopathological examination diagnosed the metastasis to appendix from prostatic cancer. Cancers of the appendix are rare and usually diagnosed incidentally in approximately 1% of all appendectomies. The rate of perforation was found to be higher in metastasis induced acute appendicitis in comparison with simple acute appendicitis in the literature. Tumors other than primary appendix tumors when present in the appendix are easily recognized as metastatic, requiring immunohistochemical studies. Metastasis induced acute appendicitis should be considered as part of the differential diagnosis when a oncologic patient presents with signs of acute appendicitis.


[Effects of contraceptive pills in the field of otorhinolaryngology (author's transl)].
Besides the wellknown alterations of the Eustachian tube there will be discussed the effects of the contraceptive pills on salivarian glands and on inner ear. The influence of the gestagen and estrogen on the regulation mechanism of the above mentioned organs is demonstrated. Especially the erectile tissue of the submandibular and parotic glands and recidiving sudden deafness are discussed.


Double-filtration plasmapheresis combined with immunosuppressive treatment for severe pemphigus: 10 years' experience of a single center in China.
Pemphigus is a group of rare and severe autoimmune blistering disease mediated by pathogenic autoantibodies against desmogleins. Plasmapheresis can directly remove autoantibodies from circulation, which has been applied to the treatment of pemphigus as an adjuvant therapy. But the results of the researches are controversial. This study aims to evaluate the efficacy and safety of double filtration plasmapheresis (DFPP) combined with immunosuppressive treatment for patients with severe pemphigus in our single center. We retrospectively analyzed 17 patients with severe pemphigus who were unresponsive to high-dose corticosteroid and received DFPP treatment between January 2010 and January 2020. The information on demographic characteristics, clinical and laboratory data, treatment regimens, and clinical outcomes were collected. All the patients were diagnosed as severe pemphigus and had a period of at least 1 week of high-dose prednisone (1-1.5 mg/kg/day), but they were unresponsive to corticosteroid and immunosuppressants treatment. They received DFPP treatment as an adjuvant therapy. After DFPP treatment, the titers of desmogleins antibodies significantly decreased (P < .001), Nikolsky's sign became negative and no new blisters appeared. The dosage of corticosteroid could begin to taper down rapidly in 9 ± 4 days. On discharge, the dosage of prednisone decreased significantly (51 ± 3 mg/day, P < .001). No major adverse events happened that could lead to the termination of DFPP treatment. Double filtration plasmapheresis combined with immunosuppressive treatment is an effective and safe therapeutic regimen for severe pemphigus. DFPP can also contribute to the dosage reduction of steroid to avoid more drug-related side effect.


Normal and altered otoliths of guinea pigs. Scanning electron microscopy observations.
Normal morphology and structural alterations of the otoliths were observed in 20 young albino guinea pigs. Ten animals had been exposed to long-term treatment with an aminoglycoside antibiotic and ten others, which had not been treated, have been analyzed as a control group. The observations were carried out using scanning electron microscopy. All the animals survived the period of stabling and treatment. They were sacrificed and the temporal bones were dissected and fixed with 2% osmium tetroxide buffered at pH 7.4. The inner ear was subsequently dehydrated, microdissected, dried with CO2, and coated with gold. Different kinds of deterioration were observed and valued according to the appearance of the otoliths, which looked as if they had been melted, with many pores and wide and prominent cavitations probably due to chemical etching.


Accuracy of femoral tunnel positioning in medial patellofemoral ligament reconstruction: anatomic insertion leads to better clinical outcome.
The medial patello-femoral ligament (MPFL) is considered the primary restraint against patellar dislocation and its reconstruction is indicated in recurrent patellar instability. An anatomical positioning of MPFL femoral insertion is recommended to achieve satisfactory clinical outcomes and prevent osteoarthritis (OA) due to an altered kinematics. The purpose of the study was first to assess the relationship between correct femoral tunnel position and better clinical outcomes and lower patellofemoral osteoarthritis rate. Second, correlation of outcomes with factors potentially affecting the results, such as the type of graft, patellar height and trochlear dysplasia. Fifty-three patients (58 knees) who underwent MPFL reconstruction between 2009 and 2018 by a senior knee surgeon were retrospectively evaluated. Knee radiographs were performed before surgery, 12 months later and at last follow-up to assess trochlear dysplasia, patellar height and patellofemoral OA. The tibial tuberosity-trochlear groove (TT-TG) value was measured on a CT scan. The accuracy of graft positioning was evaluated on sagittal radiographs according to Schöttle et al. Subjective outcomes were collected before surgery, at 12 months and at last follow-up using several validated scores. Forty-six patients (51 knees) with a mean age of 24.1 ± 7.4 years were included in the study. Mean follow-up was 8.9 ± 2.1 years. A significant improvement in all clinical scores was observed at 12 months and final follow-up. Anatomic Insertion (AI) of reconstructed MPFL was considered optimal in 33 (64.7%) and sub-optimal in 18 (35.3%) patients. Sub-optimal AI resulted in lower Kujala, IKCD and higher VAS score (p < 0.01); moreover, for every 1 mm distance in any direction from the ideal insertion, a decrease of 0.8 [95% CI (-1.4; -0.2)] in Kujala score and 0.8 [95% CI (-1.3; -0.3)] in IKDC was observed. At final follow-up, 8 patients presented patellofemoral OA Iwano grade 3 (15.7%): although that incidence did not correlate to graft positioning, the use the artificial ligament in all these cases appeared to be significant. The present study confirms the importance of an optimal anatomic femoral insertion in MPFL reconstruction, reporting a significant positive correlation between graft positioning and clinical outcome. No significant correlations were found between anatomic insertion and patello-femoral OA development. IV.


Supplementing a Phytogenic Feed Additive Modulates the Risk of Subacute Rumen Acidosis, Rumen Fermentation and Systemic Inflammation in Cattle Fed Acidogenic Diets.
Feeding with high-concentrate diets increases the risk of subacute ruminal acidosis (SARA). This experiment was conducted to evaluate whether supplementing a phytogenic feed additive based on L-menthol, thymol, eugenol, mint oil (Mentha arvensis) and cloves powder (Syzygium aromaticum) (PHY) can amend the ruminal fermentation profile, modulate the risk of SARA and reduce inflammation in cattle. The experiment was designed as a crossover design with nine non-lactating Holstein cows, and was conducted in two experimental runs. In each run, cows were fed a 100% forage diet one week (wk 0), and were then transitioned stepwise over one week (0 to 65% concentrate, wk adapt.) to a high concentrate diet that was fed for 4 weeks. Animals were fed diets either with PHY or without (CON). The PHY group had an increased ruminal pH compared to CON, reduced time to pH < 5.8 in wk 3, which tended to decrease further in wk 4, reduced the ruminal concentration of D-lactate, and tended to decrease total lactate (wk 3). In wk 2, PHY increased acetate, butyrate, isobutyrate, isovalerate, and the acetate to propionate ratio compared to CON. Phytogenic supplementation reduced inflammation compared to CON in wk 3. Overall, PHY had beneficial effects on ruminal fermentation, reduced inflammation, and modulated the risk of SARA starting from wk 3 of supplementation.


Granulocytopoietic precursor cells and regulatory factors in irradiated human bone marrow.
The effects of irradiation on committed granulopoietic progenitor cells (CFU-C) and granulopoietic humoral regulators were studied in 17 X-ray treated patients with carcinoma of the breast or of the prostate. After 45 or 70 Gy, the CFU-C decreased to 14 +/- 10% in irradiated areas. This remaining CFU-C population probably reflects a combination of radioresistance and survival in situ, and migration of CFU-C from protected marrow areas. The frequency of peripheral blood CFU-C did not change after irradiation. Endogenous colony stimulating activity (CSA), i.e., release of CSA within the marrow cell population, in post-irradiation bone marrow persisted on roughly the same level, indicating a fairly low radiosensitivity of the CSA-producing cells in the bone marrow. Colony stimulating activity in peripheral blood cells and serum remained unchanged, but there were great interindividual variations, and some of the patients with hypercellularity and CFU-C increase in nonirradiated marrow areas also had increased CSA. Serum lipoprotein inhibitors were higher in the post-treatment patients than in healthy control patients.


Dysfunction of the cochlea contributing to hearing loss in acoustic neuromas: an underappreciated entity.
Hearing loss is a common symptom in patients with cochleovestibular schwannoma. Clinical and histologic observations have suggested that the hearing loss may be caused by both retrocochlear and cochlear mechanisms. Our goal was to perform a detailed assessment of cochlear pathology in patients with vestibular schwannoma (VS). Retrospective analysis of temporal bone histopathology. Multi-center study. Temporal bones from 32 patients with unilateral, sporadic VS within the internal auditory canal. Sections through the cochleae on the VS side and opposite (control) ear were evaluated for loss of inner and outer hair cells, atrophy of the stria vascularis, loss of cochlear neurons, and presence of endolymphatic hydrops and precipitate within the endolymph or perilymph. Observed pathologies were correlated to nerve of origin, VS volume, and distance of VS from the cochlea. Hearing thresholds also were assessed. VS caused significantly more inner and outer hair cell loss, cochlear neuronal loss, precipitate in endolymph and perilymph, and decreased pure tone average, when compared with the opposite ear. Tumor size, distance from the cochlea, and nerve of origin did not correlate with structural changes in the cochlea or the hearing threshold. There is significant degeneration of cochlear structures in ears with VS. Cochlear dysfunction may be an important contributor to the hearing loss caused by VS and can explain certain clinically observed phenomena in patients with VS.


Loss of polymorphic A and B locus HLA antigens in colon carcinoma.
In the present study we have confirmed that approximately one third of human colorectal carcinomas fail to express the HLA - A,B,C monomorphic determinant reactive with the W6/32 MAb, and 44% express class II HLA antigens as shown by reactivity with NFK-1 MAb. Reduced staining with the W6/32 MAb was not always associated with the loss of beta 2m. In addition, the expression of HLA-A2 and Bw4 class I specific haplotypes on normal colon epithelium and tumour biopsy tissue was assessed. All normal colonic epithelia stained positively with MAb against A2 and Bw4 antigens, but a loss of these determinants was shown on tumour biopsies from patients tissue typed for the respective specificities. Loss of the A2 haplotype was shown in 4 of 15 tumour tissue samples, and loss of Bw4 specificities in 5 out of 7 tissue samples. The failure to detect specific loci determinants was not necessarily associated with loss of reactivity with W6/32 MAb.


Professor Leslie H. Blumgart-Pioneering Hepatopancreaticobiliary Surgeon: a Tribute.
Leslie Harold Blumgart was the surgeon behind making Hepatopancreaticobiliary surgery a speciality and his recent demise will always leave a void in that field. He was the person who classified anatomical variations in the biliary tract and devised a special anastomotic technique to decrease the rate of postoperative pancreatic duct fistula after Whipple's pancreaticoduodenectomy. His contribution in managing hepatobiliary oncology cases with chemotherapy and radiotherapy was also significant. No wonder, he was designated as the "living legend" by the International Hepatopancreaticobiliary Association.


Postnatal development of the adult pattern of motor axon distribution in rat muscle.
Establishment of the adult pattern of connections between motoneurones and their muscles is known to be achieved in stages. There is an initial guidance mechanism which in higher vertebrates directs the axons with considerable accuracy to their appropriate muscles. This is followed by a stage when a large proportion of neurones die. Then, after birth, the final modifications are brought about by elimination of redundant motoneuronal branches. We present here evidence that branch elimination does not occur at random from within a motoneurone's initial intramuscular distribution, but in such a way as to sharpen the segmento-topic relationship within a muscle that motoneuronal death had earlier provided.


PI3K-AKT-mTOR signaling pathway: the intersection of allergic asthma and cataract.
Allergic asthma is a chronic inflammatory disease and involves many cells and cellular components. Cataract is a condition that affects the transparency of the lens, which the opacity of the lens caused by any innate or acquired factor degrades its transparency or changes in color. During the establishment of asthma model of rats with chicken ovalbumin nebulization, it was found that asthmatic rats were more likely to have monocular or binocular cataract symptoms than normal rats. Considering that they are all induced by immune imbalance, inflammation, etc., there may be some correlation in the mechanism, and many clues showed that both diseases are associated with activation of the PI3K-AKT-mTOR signaling pathway. Therefore, we hypothesized that the PI3K-AKT-mTOR signaling pathway produces inflammatory or immune imbalance based on allergy leading to cataract.


Chromosomal analysis in mouse eggs fertilized in vitro with sperm exposed to ultraviolet light (UV) and methyl and ethyl methanesulfonate (MMS and EMS).
Chromosome aberrations were analyzed at the first-cleavage metaphase of mouse eggs fertilized in vitro with sperm exposed to ultraviolet light (UV) as well as to methyl and ethyl methanesulfonate (MMS and EMS). The frequencies of chromosome aberrations markedly increased with dose of UV as well as with concentration of MMS and EMS. In the UV-irradiation group, the frequency of chromosome-type aberrations was much higher than that of chromatid-type aberrations. About 90% of chromosome aberrations observed in the eggs following MMS and EMS treatment to sperm were chromosome type in which the frequency of chromosome fragments was the highest. The effects of UV on the induction of chromosome aberrations were clearly potentiated by post-treatment incubation of fertilized eggs in the presence of Ara-C or caffeine, but the effects of MMS and EMS were not pronounced by post-treatment of Ara-C or caffeine. The results indicate a possibility that UV damage induced in mouse sperm DNA is reparable in the eggs during the period between the entry of sperm into the egg cytoplasm and the first-cleavage metaphase.


Co-induction of cell death and survival pathways by phosphoinositide 3-kinase.
A single stimulus can induce both the cell death and survival pathway, suggesting that these pathways share common upstream signaling components. In order to define these components, human U937 cells grown in 10% serum were exposed to serum-free media. This treatment resulted in apoptosis, which was found to be mediated by SAPK/JNK. It was previously reported that the serum withdrawal (SW)-induced SAPK activation is mediated by a positive mutual interaction between the reactive oxygen species (ROS) and phosphoinositide 3-kinase (PI3K). This study shows that the ROS/PI3K interaction also induces a NF-kappaB-dependent survival pathway. Despite the role of PI3K, Akt was found to be irrelevant to the activation of SAPK and NF-kappaB. Comparative analyses of SAPK and NF-kappaB for their responses to exogenous H(2)O(2) revealed that SAPK activation requires much higher H(2)O(2) concentrations than those required for NF-kappaB activation. Moreover, high lethal concentrations of H(2)O(2) were found to activate NF-kappaB and SAPK in a PI3K-independent manner. These results suggest that ROS induce both the SAPK-dependent apoptotic and NF-kappaB-mediated survival pathways, and these inducer signals are amplified by PI3K in the SW-triggered pathway. Cell death appears to be favored as this amplification proceeds.


Identification and characterization of single-stranded DNA-binding protein from the facultative psychrophilic bacteria Pseudoalteromonas haloplanktis.
Single-stranded DNA-binding protein (SSB) plays an important role in DNA metabolism such as DNA replication, repair, and recombination, and is essential for cell survival. This study reports on the ssb-like gene cloning, gene expression and characterization of a single-stranded DNA-binding protein of Pseudoalteromonas haloplanktis (PhaSSB) and is the first report of such a protein from psychrophilic microorganism. PhaSSB possesses a high sequence similarity to Escherichia coli SSB (48% identity and 57% similarity) and has the longest amino acid sequence (244 amino acid residues) of all the known bacterial SSBs with one OB-fold per monomer. An analysis of purified PhaSSB by means of chemical cross-linking experiments, sedimentation analysis and size exclusion chromatography revealed a stable tetramer in solution. Using EMSA, we characterized the stoichiometry of PhaSSB complexed with a series of ssDNA homopolymers, and the size of the binding site was determined as being approximately 35 nucleotides long. In fluorescence titrations, the occluded site size of PhaSSB on poly(dT) is 34 nucleotides per tetramer under low-salt conditions (2mM NaCl), but increases to 54-64 nucleotides at higher-salt conditions (100-300mM NaCl). This suggests that PhaSSB undergoes a transition between ssDNA binding modes, which is observed for EcoSSB. The binding properties of PhaSSB investigated using SPR technology revealed that the affinity of PhaSSB to ssDNA is typical of SSB proteins. The only difference in the binding mode of PhaSSB to ssDNA is a faster association phase, when compared to EcoSSB, though compensated by faster dissociation rate. When analyzed by differential scanning calorimetry (DSC), the melting temperature (Tm) was determined as 63 °C, which is only a few degrees lower than for EcoSSB.


Definition of the carbohydrate response element of the rat S14 gene. Context of the CACGTG motif determines the specificity of carbohydrate regulation.
Transcription of the S14 gene in primary hepatocytes is stimulated in response to increased carbohydrate metabolism. We have demonstrated previously that a 30-base pair (bp) segment of the S14 gene from -1457 to -1428 is a carbohydrate response element (ChoRE). This element contains a (5')CACGTG motif that is essential for control. DNase I footprinting experiments with liver nuclear extract revealed two factors binding within the S14 ChoRE. In transient transfection experiments, mutation of the upstream site between -1457 and -1450 did not affect the response to elevated glucose, whereas the downstream 21-bp site between -1448 and -1428 was sufficient to mediate the glucose induction. Electrophoretic mobility shift assays indicated that the hepatic factor binding to this site in vitro is closely related or identical to the major late transcription factor (MLTF). However, replacement of the 21-bp S14 ChoRE with the authentic MLTF binding site from the adenovirus major late promoter failed to elicit the glucose response. By systematically exchanging bases between the functional S14 and nonresponsive adenovirus sites, the sequence (5')CACGTGNNNGCC was found to be essential for carbohydrate regulation. A segment containing this specific motif from the rat fatty acid synthase gene, another carbohydrate-responsive gene in hepatocytes, conferred a carbohydrate response when linked to the S14 promoter. Thus, the context of the CACGTG motif provides the specificity for regulation by carbohydrate metabolism.


Isotoxic Intensity Modulated Radiation Therapy in Stage III Non-Small Cell Lung Cancer: A Feasibility Study.
Not all patients with stage III non-small cell lung cancer (NSCLC) are suitable for concurrent chemoradiation therapy (CRT). Local failure rate is high for sequential concurrent CRT. As such, there is a rationale for treatment intensification. Isotoxic intensity modulated radiation therapy (IMRT) is a multicenter feasibility study that combines different intensification strategies including hyperfractionation, acceleration, and dose escalation facilitated by IMRT. Patients with unresectable stage III NSCLC, Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 to 2, and unsuitable for concurrent CRT were recruited. A minimum of 2 cycles of platinum-based chemotherapy was compulsory before starting radiation therapy (RT). Radiation dose was increased until a maximum dose of 79.2 Gy was reached or 1 or more of the organs at risk met predefined constraints. RT was delivered in 1.8-Gy fractions twice daily, and an RT quality assurance program was implemented. The primary objective was the delivery of isotoxic IMRT to a dose >60 Gy equivalent dose in 2-Gy fractions (EQD2 assuming an α/β ratio of 10 Gy for acute reacting tissues). Thirty-seven patients were recruited from 7 UK centers. Median age was 69.9 years (range, 46-86 years). The male-to-female ratio was 17:18. ECOG PS was 0 to 5 in 14.2% of patients; PS was 1 to 27 in 77.1% of patients; PS was 2 to 3 in 8.6% of patients. Stage IIIA:IIIB ratio was 22:13 (62.9%:37.1%). Of 37 patients, 2 (5.4%) failed to achieve EQD2 > 60 Gy. Median prescribed tumor dose was 77.4 Gy (range, 61.2-79.2 Gy). A maximum dose of 79.2Gy was achieved in 14 patients (37.8%). Grade 3 esophagitis was reported in 2 patients, and no patients developed grade 3 to 4 pneumonitis. There were 3 grade 5 events: acute radiation pneumonitis, bronchopulmonary hemorrhage, and acute lung infection. Median follow-up at time of analysis was 25.4 months (range, 8.0-44.2) months for 11 of 35 survivors. The median survival was 18.1 months (95% confidence interval [CI], 13.9-30.6), 2-year overall survival was 33.6% (95% CI, 17.9-50.1), and progression-free survival was 23.9% (95% CI, 11.3-39.1). Isotoxic IMRT is a well-tolerated and feasible approach to treatment intensification.


Analysis of the competition between nucleosome formation and transcription factor binding.
We have studied the competitive binding of histones and the Rous sarcoma virus internal enhancer binding factor (IBF) factor (which recent studies indicate is almost certainly cEBP beta). We find that histones and IBF are incapable of forming a ternary complex with a 159-base pair (bp) fragment of DNA containing a single IBF binding site and that histones and factor are mutually exclusive in binding. We have analyzed the various physical parameters of binding, in an attempt to understand how the factor might establish an exclusive binding in the cell. The stability of the nucleosome and the factor-DNA complex have been determined, and in addition a minimum value for the affinity of the histone octamer has been computed. We find that in simple competition the IBF can successfully compete, only if the substrate DNA is shorter than 140 bp. The relevance of these results is discussed in terms of a kinetic model for successful factor competition during the replication of the factor binding site in the cell.


Alcohol consumption and the risk of heart failure: the Suita Study and meta-analysis of prospective cohort studies.
Alcohol consumption is a modifiable lifestyle, but its role in heart failure (HF) development is controversial. Herein, we investigated the prospective association between alcohol consumption and HF risk. A total of 2,712 participants (1,149 men and 1,563 women) from the Suita Study were followed up every two years. Cox regression was applied to calculate the hazard ratios (HRs) and 95% confidence intervals (95% CIs) of HF risk for heavy drinking (≥46 g/day in men or ≥23 g/day in women) and never drinking compared to light drinking (<23 g/day in men or <11.5 g/day in women). Then, we combined the results of the Suita Study with those from other eligible prospective cohort studies in a meta-analysis using the random-effects model. In the Suita Study, within a median follow-up period of 8 years, 319 HF cases (162 in men and 157 in women) were detected. In men, but not women, never and heavy drinking carried a higher risk of HF than light drinking: HRs (95% CIs) = 1.65 (1.00, 2.73) and 2.14 (1.26, 3.66), respectively. Alike, the meta-analysis showed a higher risk of HF among heavy drinkers: HR (95% CI) = 1.37 (1.15, 1.62) and abstainers: HR (95% CI) = 1.18 (1.02, 1.37). We indicated a J-shaped association between alcohol consumption and HF risk among Japanese men. The results of the meta-analysis came in line with the Suita Study. Heavy-drinking men should be targeted for lifestyle modification interventions.


Coexistence of corticotropin-releasing factor-like immunoreactivity and vasotocin in perikarya of the preoptic nucleus in the eel.
A corticotropin-releasing factor (CRF)-like system has been identified in eels (Anguilla anguilla, A. rostrata, and A. japonica) with immunocytochemical techniques. Immunoreactive perikarya were located in the preoptic area, in magno- and parvocellular regions of the preoptic nucleus. Fine fibers reached the pituitary and terminated in the neurohypophysis close to the corticotropic cells of the rostral pars distalis and caudally close to the melanocorticotropic cells of the intermediate lobe. Double immunostainings revealed the presence of three subpopulations of perikarya: one population containing only CRF, a second containing only arginine vasotocin (VT), and a third one containing both CRF and VT. This coexistence of CRF and VT was also demonstrated by comparing adjacent sections stained with anti-CRF and anti-VT, respectively. Because antisera used in the present work were carefully absorbed, these data strongly suggest that CRF and VT coexist in some perikarya, at least in the animals studied.