Adaptation to novel spatially-structured environments is driven by the capsule and alters virulence-associated traits.
The extracellular capsule is a major virulence factor, but its ubiquity in free-living bacteria with large environmental breadths suggests that it shapes adaptation to novel niches. Yet, how it does so, remains unexplored. Here, we evolve three Klebsiella strains and their capsule mutants in parallel. Their comparison reveals different phenotypic and genotypic evolutionary changes that alter virulence-associated traits. Non-capsulated populations accumulate mutations that reduce exopolysaccharide production and increase biofilm formation and yield, whereas most capsulated populations become hypermucoviscous, a signature of hypervirulence. Hence, adaptation to novel environments primarily occurs by fine-tuning expression of the capsular locus. The same evolutionary conditions selecting for mutations in the capsular gene wzc leading to hypermucoviscosity also result in increased susceptibility to antibiotics by mutations in the ramA regulon. This implies that general adaptive processes outside the host can affect capsule evolution and its role in virulence and infection outcomes may be a by-product of such adaptation.


Independent component analysis of Gabor features for face recognition.
We present an independent Gabor features (IGFs) method and its application to face recognition. The novelty of the IGF method comes from 1) the derivation of independent Gabor features in the feature extraction stage and 2) the development of an IGF features-based probabilistic reasoning model (PRM) classification method in the pattern recognition stage. In particular, the IGF method first derives a Gabor feature vector from a set of downsampled Gabor wavelet representations of face images, then reduces the dimensionality of the vector by means of principal component analysis, and finally defines the independent Gabor features based on the independent component analysis (ICA). The independence property of these Gabor features facilitates the application of the PRM method for classification. The rationale behind integrating the Gabor wavelets and the ICA is twofold. On the one hand, the Gabor transformed face images exhibit strong characteristics of spatial locality, scale, and orientation selectivity. These images can, thus, produce salient local features that are most suitable for face recognition. On the other hand, ICA would further reduce redundancy and represent independent features explicitly. These independent features are most useful for subsequent pattern discrimination and associative recall. Experiments on face recognition using the FacE REcognition Technology (FERET) and the ORL datasets, where the images vary in illumination, expression, pose, and scale, show the feasibility of the IGF method. In particular, the IGF method achieves 98.5% correct face recognition accuracy when using 180 features for the FERET dataset, and 100% accuracy for the ORL dataset using 88 features.


Sensitivity of osteosarcoma cells to HDAC inhibitor AR-42 mediated apoptosis.
Osteosarcoma (OS) is the most common primary bone tumor in both humans and dogs and is the second leading cause of cancer related deaths in children and young adults. Limb sparing surgery along with chemotherapy has been the mainstay of treatment for OS. Many patients are not cured with current therapies, presenting a real need for developing new treatments. Histone deacetylase (HDAC) inhibitors are a promising new class of anticancer agents. In this study, we investigated the activity of the novel HDAC inhibitor AR-42 in a panel of human and canine OS cell lines. The effect of AR-42 and suberoylanilide hydroxamic acid (SAHA) alone or in combination with doxorubicin on OS cell viability was assessed. Induction of histone acetylation after HDAC inhibitor treatment was confirmed by Western blotting. Drug-induced apoptosis was analyzed by FACS. Apoptosis was assessed further by measuring caspase 3/7 enzymatic activity, nucleosome fragmentation, and caspase cleavage. Effects on Akt signaling were demonstrated by assessing phosphorylation of Akt and downstream signaling molecules. AR-42 was a potent inhibitor of cell viability and induced a greater apoptotic response compared to SAHA when used at the same concentrations. Normal osteoblasts were much less sensitive. The combination of AR-42 with doxorubicin resulted in a potent inhibition of cell viability and apparent synergistic effect. Furthermore, we showed that AR-42 and SAHA induced cell death via the activation of the intrinsic mitochondrial pathway through activation of caspase 3/7. This potent apoptotic activity was associated with the greater ability of AR-42 to downregulate survival signaling through Akt. These results confirm that AR-42 is a potent inhibitor of HDAC activity and demonstrates its ability to significantly inhibit cell survival through its pleiotropic effects in both canine and human OS cells and suggests that spontaneous OS in pet dogs may be a useful large animal model for preclinical evaluation of HDAC inhibitors. HDAC inhibition in combination with standard doxorubicin treatment offers promising potential for chemotherapeutic intervention in both canine and human OS.


Seasonal variations and chemical characteristics of PM(2.5) in Wuhan, central China.
PM2.5 samples were collected at an urban site (WD) and a suburban site (TH) in Wuhan from August 2012 to July 2013. The mass concentrations of water-soluble inorganic ions, carbonaceous species and elements of PM2.5 were measured. The annual mean concentrations of PM2.5 were 106.5 μg/m(3) and 114.9 μg/m(3) at WD and TH, respectively. The chemical compositions of PM2.5 at WD were similar to those at TH and the fractions of the major components of PM2.5 in Wuhan were in the following order of trace elements<chloride<EC (elemental carbon)<ammonium<nitrate<soil dust<sulfate<OM (organic matter). As the secondary ionic aerosols (SIA) and dominant ions, SO4(2-), NO3(-) and NH4(+) all exhibited strong seasonal distributions, consistently with the lowest values in summer and the highest in winter. OM was the most abundant component in PM2.5, the lowest concentrations of which were observed in summer at both sites, while the highest concentrations of OC (organic carbon) appeared in winter at WD and autumn at TH, respectively. The highest OC concentration observed in autumn was tightly related to the biomass burning near the suburban site. The crustal elements (Mg, K, Ca and Fe) dominated the 20 detected elements in PM2.5, with the highest concentrations in spring in Wuhan, which might be due to frequent sandstorm from north carrying abundant soil dusts in spring in China. Ten trace elements (Cu, Ga, Ag, Tl, Ca, As, Zn, Pb, Se and Cd) were enriched in PM2.5 and the higher EF for Ag, Pb, Se and Cd in PM2.5 indicated that the air pollution from vehicle exhaust emission and coal burning in Wuhan was serious and noteworthy.


Prevalence and risk of violence against people with and without disabilities: findings from an Australian population-based study.
There are no population-based estimates of the prevalence of interpersonal violence among people with disabilities in Australia. The project aimed to: 1) estimate the prevalence of violence for men and women according to disability status; 2) compare the risk of violence among women and men with disabilities to their same-sex non-disabled counterparts and; 3) compare the risk of violence between women and men with disabilities. We analysed the 2012 Australian Bureau of Statistics Survey on Personal Safety of more than 17,000 adults and estimated the population-weighted prevalence of violence (physical, sexual and intimate partner violence and stalking/harassment) in the past 12 months and since the age of 15. Population-weighted, age-adjusted, logistic regression was used to estimate the odds of violence by disability status and gender. People with disabilities were significantly more likely to experience all types of violence, both in the past 12 months and since the age of 15. Women with disabilities were more likely to experience sexual and partner violence and men were more likely to experience physical violence. These results underscore the need to understand risk factors for violence, raise awareness about violence and to target policies and services to reduce violence against people with disabilities in Australia.


Acute myeloid leukaemia disrupts endogenous myelo-erythropoiesis by compromising the adipocyte bone marrow niche.
Acute myeloid leukaemia (AML) is distinguished by the generation of dysfunctional leukaemic blasts, and patients characteristically suffer from fatal infections and anaemia due to insufficient normal myelo-erythropoiesis. Direct physical crowding of bone marrow (BM) by accumulating leukaemic cells does not fully account for this haematopoietic failure. Here, analyses from AML patients were applied to both in vitro co-culture platforms and in vivo xenograft modelling, revealing that human AML disease specifically disrupts the adipocytic niche in BM. Leukaemic suppression of BM adipocytes led to imbalanced regulation of endogenous haematopoietic stem and progenitor cells, resulting in impaired myelo-erythroid maturation. In vivo administration of PPARγ agonists induced BM adipogenesis, which rescued healthy haematopoietic maturation while repressing leukaemic growth. Our study identifies a previously unappreciated axis between BM adipogenesis and normal myelo-erythroid maturation that is therapeutically accessible to improve symptoms of BM failure in AML via non-cell autonomous targeting of the niche.


Evaluation of user satisfaction and service responsiveness in municipalities enrolled in the Mais Médicos (More Doctors) Program.
The Mais Médicos (More Doctors) Program (PMM) addresses shortages of doctors, one of whose objectives is to reduce regional inequalities in access to health care. This descriptive cross-sectional study evaluated service responsiveness and user satisfaction with PMM doctors among 263 users of the public health system in 32 municipalities with 20% or more of extreme poverty. Questionnaires of open and closed questions were used. Users were satisfied with patient-doctor relationships, information about diseases and treatment and the clarity and comprehensibility of medical indications. The doctors' technical and humanised conduct helped ensure satisfaction among users, who stressed the importance of the programme's continuance. As regards responsiveness, most users were pleased with the non-medical aspects of care: prompt scheduling of appointments, waiting times of less than one hour and privacy. Users suggested improving infrastructure and making more medicines and more doctors available, which should be considered by SUS managers to help ensure access to the right to health guaranteed in Brazil's constitution.


Retrospective study of the incidence of HFMD and seroepidemiology of antibodies against EV71 and CoxA16 in prenatal women and their infants.
Hand, foot, and mouth disease (HFMD) has been emerging as an important public problem over the past few decades, especially in Asian and Pacific regions. A national program on EV71 vaccine development against HFMD was initiated in China, in 2008, which called for a need for seroepidemiological study for the target population. This was a retrospective study conducted in Jiangsu Province, in October, 2010. We measured the neutralizing antibodies against EV71 and CoxA16 in a cohort of infants aged of 2, 7, 12, and 27-38 months and their mothers just before delivery. Series sera samples from 975 infants and 555 mothers were collected and analyzed. Questionnaires on the history of HFMD were completed in the survey. A total of 143 HFMD cases were collected, but only 11.2% were reported to the National Infectious Disease Information Management System. The level of maternal antibody titers decreased dramatically during the first 7 month and remained at a relatively low level thereafter. But it increased significantly from month 12 to months 27-38. The accumulate incidence density of HFMD demonstrated a significant increase after 14 months of age, resulting in a accumulate incidence density of 50.8/1000 person-years in survey period. Seropositivity of EV71 antibody in infants at the age of 2 months seems to demonstrate a protective effect against HFMD. High seropositive rate of EV71 and CoxA16 antibody was found in prenatal women in mainland China, and there is a need to enhance the HFMD case management and the current surveillance system. We suggest that infants aged between 6 to 14 months should have the first priority to receive EV71 vaccine.


Prevalence of neurodevelopmental differences and autism in Scottish primary schools 2018-2022.
This study investigated the prevalence of neurodevelopmental needs among children in primary schools in Scotland. Two groups were identified: autistic learners and a larger group of learners who had neurodevelopmental differences. These differences encompassed any need for additional support in various domains, including communication, interaction, emotional regulation, coordination, movement, and cognition. A two-phase process was employed, drawing on data from a cross-sectional study followed by a secondary analysis of a population census. In the first phase, a random sample of 688 children with additional support needs from 22 schools participated. Demographics, support characteristics, and neurodevelopmental needs were identified. Results revealed that 76.89% of children with additional support needs exhibited a need type consistent with a neurodevelopmental difference. In the second phase, data from the Scottish Government Annual Pupil Census, covering all state-provided primary school children between 2018 and 2022, were analyzed. Modeling was conducted using data from the first phase to estimate prevalence of neurodevelopmental differences. Data on autism were directly extracted from the census. Analysis revealed an increase in the prevalence of neurodevelopmental differences and autism. The prevalence of autism rose by 31.98%, with 2.60% of primary school children identified as autistic in 2022. Similarly, the prevalence of neurodevelopmental differences increased by 10.57%, with 16.22% of primary school children exhibiting such differences in 2022. Across 32 localities, regional variations in prevalence were observed. These findings show the substantial number of neurodivergent children within Scottish primary schools and emphasize the need for a neurodevelopmentally informed approach to inclusive education.


Co-dependency of PKCδ and K-Ras: inverse association with cytotoxic drug sensitivity in KRAS mutant lung cancer.
Recent studies suggest that the presence of a KRAS mutation may be insufficient for defining a clinically homogenous molecular group, as many KRAS mutant tumors lose reliance on K-Ras for survival. Identifying pathways that support K-Ras dependency may define clinically relevant KRAS subgroups and lead to the identification of new drug targets. We have analyzed a panel of 17 KRAS mutant lung cancer cell lines classified as K-Ras-dependent or -independent for co-dependency on protein kinase C δ (PKCδ). We show that functional dependency on K-Ras and PKCδ co-segregate, and that dependency correlates with a more epithelial-like phenotype. Furthermore, we show that the pro-apoptotic and pro-tumorigenic functions of PKCδ also segregate based on K-Ras dependency, as K-Ras-independent cells are more sensitive to topoisomerase inhibitors, and depletion of PKCδ in this subgroup suppresses apoptosis through increased activation of extracellular signal-regulated kinase (ERK). In contrast, K-Ras-dependent lung cancer cells are largely insensitive to topoisomerase inhibitors, and depletion of PKCδ can increase apoptosis and decrease activation of ERK in this subgroup. We have previously shown that nuclear translocation of PKCδ is necessary and sufficient for pro-apoptotic signaling. Our current studies show that K-Ras-dependent cells are refractive to PKCδ-driven apoptosis. Analysis of this subgroup showed increased PKCδ expression and an increase in the nuclear:cytoplasmic ratio of PKCδ. In addition, targeting PKCδ to the nucleus induces apoptosis in K-Ras-independent, but not K-Ras-dependent non-small-cell lung cancer (NSCLC) cells. Our studies provide tools for identification of the subset of patients with KRAS mutant tumors most amenable to targeting of the K-Ras pathway, and identify PKCδ as a potential target in this tumor population. These subgroups are likely to be of clinical relevance, as high PKCδ expression correlates with increased overall survival and a more epithelial tumor phenotype in patients with KRAS mutant lung adenocarcinomas.


Intrauterine growth restriction is associated with cardiac ultrastructural and gene expression changes related to the energetic metabolism in a rabbit model.
Intrauterine growth restriction (IUGR) affects 7-10% of pregnancies and is associated with cardiovascular remodeling and dysfunction, which persists into adulthood. The underlying subcellular remodeling and cardiovascular programming events are still poorly documented. Cardiac muscle is central in the fetal adaptive mechanism to IUGR given its high energetic demands. The energetic homeostasis depends on the correct interaction of several molecular pathways and the adequate arrangement of intracellular energetic units (ICEUs), where mitochondria interact with the contractile machinery and the main cardiac ATPases to enable a quick and efficient energy transfer. We studied subcellular cardiac adaptations to IUGR in an experimental rabbit model. We evaluated the ultrastructure of ICEUs with transmission electron microscopy and observed an altered spatial arrangement in IUGR, with significant increases in cytosolic space between mitochondria and myofilaments. A global decrease of mitochondrial density was also observed. In addition, we conducted a global gene expression profile by advanced bioinformatics tools to assess the expression of genes involved in the cardiomyocyte energetic metabolism and identified four gene modules with a coordinated over-representation in IUGR: oxygen homeostasis (GO: 0032364), mitochondrial respiratory chain complex I (GO:0005747), oxidative phosphorylation (GO: 0006119), and NADH dehydrogenase activity (GO:0003954). These findings might contribute to changes in energetic homeostasis in IUGR. The potential persistence and role of these changes in long-term cardiovascular programming deserves further investigation.


Preclinical evaluation of microneedle technology for intradermal delivery of influenza vaccines.
Recent clinical studies have suggested that, for certain strains of influenza virus, intradermal (i.d.) delivery may enable protective immune responses using a lower dose of vaccine than required by intramuscular (i.m.) injection. Here, we describe the first preclinical use of microneedle technology for i.d. administration of three different types of influenza vaccines: (i) a whole inactivated influenza virus, (ii) a trivalent split-virion human vaccine, and (iii) a plasmid DNA encoding the influenza virus hemagglutinin. In a rat model, i.d. delivery of the whole inactivated virus provided up to 100-fold dose sparing compared to i.m. injection. In addition, i.d. delivery of the trivalent human vaccine enabled at least 10-fold dose sparing for the H1N1 strain and elicited levels of response across the dose range similar to those of i.m. injection for the H3N2 and B strains. Furthermore, at least fivefold dose sparing from i.d. delivery was evident in animals treated with multiple doses of DNA plasmid vaccine, although such effects were not apparent after the first immunization. Altogether, the results demonstrate that microneedle-based i.d. delivery elicits antibody responses that are at least as strong as via i.m. injection and that, in many cases, dose sparing can be achieved by this new immunization method.


N-terminal specificity of PEGylation of human bone morphogenetic protein-2 at acidic pH.
Site-specific PEGylation offers the possibility to modify a therapeutic protein without interfering with its biological activity. Previously, a preferential N-terminal PEGylation has been reported for several proteins when the reaction was performed at acidic pH. In the present study it was explored if acidic pH favors N-terminal PEGylation of bone morphogenetic protein-2 (BMP-2). PEGylation by poly(ethylene glycol) aldehyde (PEG-AL) or poly(ethylene glycol) carboxymethyl succinimidyl ester (PEG-NHS) was performed at moderate acidic pH of 4. Comparing with PEG-NHS, PEG-AL converted more BMP-2 mainly to mono- or di-PEGylated derivatives at much less molar excess and shorter duration. Analysis of Tryptic fragments of the PEGylated derivatives indicated a partial N-terminal PEGylation specificity. PEG-AL exhibited higher specificity than PEG-NHS. UV spectrometry proved that PEGylation improved the solubility of BMP-2 in PBS. Surface plasmon resonance showed that PEGylation decreased the binding of BMP-2 proteins to a type II receptor. Remarkably, mono-PEGylated BMP-2 with PEG-AL showed higher cellular bioactivity than unmodified protein. Higher N-terminal PEGylation specificity correlates with higher receptor binding affinity and cellular activity. In summary, PEGylation of BMP-2 by PEG-AL and PEG-NHS at acidic pH exhibits a partial N-terminal specificity which however might be sufficient for an efficient site-specific PEGylation process.


Dexamethasone decreases postoperative opioid and antiemetic use in shoulder arthroplasty patients: a prospective, randomized controlled trial.
Multimodal pain control can be beneficial in relieving postoperative pain and limiting narcotic use following orthopedic procedures. Additionally, with increasing interest in outpatient arthroplasty procedures, providers have interest in adequate early postoperative pain control and complications. The purpose of this study was to investigate the effect of dexamethasone on pain, postoperative nausea and vomiting, and length of stay following total shoulder arthroplasty (TSA) and reverse total shoulder arthroplasty (RTSA). One hundred twelve patients undergoing TSA or RTSA by a single surgeon were assessed for inclusion in this investigation. We performed a prospective randomized controlled trial to investigate the effect of 10 mg of dexamethasone administered within 90 minutes of surgery. Primary outcome assessed was the average morphine equivalent use over the first 24 hours postsurgery. Secondary outcomes included postoperative visual analog scale (VAS) scores, antiemetic use, postoperative nausea and vomiting, and complications. A total of 75 patients were included in the final analysis, with 32 patients (42.7%) randomized to the control group and 43 (57.3%) randomized to the dexamethasone group. Body mass index was significantly greater in the control group (33.8 vs. 30.3, P = .014); otherwise, there were no significant demographic differences between groups. Average ondansetron use was significantly lower in the dexamethasone group compared with controls for the 0- to 4-hour interval (0.1 vs. 0.9 mg, respectively, P = .006) and was lower overall for the first 24 hours (0.3 vs. 1.0 mg, P = .025). Differences in VAS scores were significantly lower in the dexamethasone group at all time points (P < .05 for all). The average VAS score over the 24-hour period for the dexamethasone group was also significantly lower than the controls (3 vs. 6, P < .001). Morphine equivalent use was significantly lower in the dexamethasone group compared with controls at 12-16 hours (1.7 vs. 4.0 mg, respectively, P = .004) and at 16-20 hours (1.7 vs. 3.4 mg, respectively, P = .006). When averaged over the first 24 hours, morphine equivalent was also significantly lower in the dexamethasone group (16.1 vs. 25.4 mg, P = .007). There was no significant difference in glucose control or complications between groups. Dexamethasone decreases opioid requirements in the first 24 hours following surgery, provides improved pain control, and decreases antiemetic use following shoulder arthroplasty. Dexamethasone is an important multimodal adjunct for controlling pain and postoperative nausea and vomiting following primary TSA.


NICE atrial fibrillation guideline snubs wearable technology: a missed opportunity?
Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia and a growing public health epidemic. In the UK, over 1.3 million people have a diagnosis of AF and an estimated 400,000 remain undiagnosed. AF-related strokes account for a quarter of all strokes and, as AF episodes are often asymptomatic, are still often the first manifestation of AF. Early diagnosis and initiation of oral anticoagulation, where appropriate, may prevent some of these thromboembolic strokes. Public Health England is committed to decrease the incidence of AF-related strokes and has sponsored initiatives aimed at improving AF detection by promoting the uptake of wearable technologies. However, the National Institute for Health and Care Excellence (NICE) has not recommended wearable technology in their recent AF diagnosis and management guidelines (NG196). Diagnostic accuracy of single-lead electrocardiography (ECG) generated by the latest iteration of wearable devices is excellent and, in many cases, superior to general practitioner interpretation of the 12-lead ECG. High-quality ECG from wearable devices that unequivocally shows AF can expedite AF detection. Otherwise, there is a real risk of delaying AF diagnosis with the potential of devastating consequences for patients and their families.


Birth, growth and death as structuring operators in bacterial population dynamics.
A new model is presented that describes microbial population dynamics that emerge from complex interactions among birth, growth and death as oriented, discrete events. Specifically, birth and death act as structuring operators for individual organisms within the population, which become synchronised as age clusters (called cell generations that are structured in age classes) that are born at the same time and die in concert; a pattern very consistent with recent experimental data that show bacterial group death correlates with temporal population dynamics in chemostats operating at carrying capacity. Although the model only assumes "natural death" (i.e., no death from predation or antimicrobial exposure), it indicates that short-term non-linear dynamic behaviour can exist in a bacterial population growing under longer term pseudo-steady-state conditions (a confined dynamic equilibrium). After summarizing traditional assumptions about bacterial aging, simulations of batch, continuous-flow, and bioreactors with recycle are used to show how population dynamics vary as function of hydraulic retention time, microbial kinetics, substrate level, and other factors that cause differential changes in the distribution of living and dead cells within the system. In summary, we show that population structures induced by birth and death (as discrete and delayed events) intrinsically create a non-linear dynamic system, implying that a true steady state can never exist in growing bacterial populations. This conclusion is discussed within the context of process stability in biotechnology.


Nuclear Medicine in India: A Historical Journey.
Evidence-based historical accounts of critical events, which shaped nuclear medicine in India today, are presented in this article. There was parallel activity happening in the northern and western region of India in the early 60s. Radiation Medicine Center (RMC) at Mumbai inaugurated in September 1963 by Dr. Bhabha; and Institute of Nuclear Medicine and Allied Sciences, Delhi dedicated to the nation in February 1964. The isotope division of Bhabha Atomic Research Center endured as the backbone in the supply of indigenously produced medical radioisotopes in research reactors APSARA (1958) and CIRUS (1960). Design and dispatch of economical generators (loaded with low specific activity 99Mo) with indigenously designed solvent extraction (Methyl Ethyl Ketone) technique had led to rapid growth of nuclear medicine facilities in the country. As per recently released list (July 2018) of the Atomic Energy Regulatory Board, there are 293 nuclear medicine departments in the country. Of which 14% are in the government sector, and the remaining 86% are under private ownership. There are currently 233 functioning gamma cameras (Single-photon emission computed tomography [SPECT]/SPECT-computed tomography [CT]) units in India since 1969 when the first gamma camera was commissioned at RMC. The first medical cyclotron (2002) and first positron emission tomography (PET) (2002) and first PET-CT (2004) in Mumbai had triggered revolution of molecular imaging in India. There are 222 PET-CT, 3 PET-magnetic resonance imaging scanners, and 19 cyclotrons operating currently. India has witnessed relatively slower headways in terms of high dose radionuclide therapy facilities. After first indoor facility at RMC in 1964, only 92 radionuclide therapy (isolation) wards have come up with no more than 200 beds for the entire country in the last 54 years. India started Delhi university approved structured postgraduate diploma in nuclear medicine in 1963 at the Institute of Nuclear Medicine and Allied Sciences (INMAS), first of its kind course in the world at that time. RMC started Mumbai University recognized diploma courses for physicians (Diploma in Radiation Medicine) and technologists (Diploma in Medical Radioisotope Techniques) in 1973. National Board of Examination (Government of India) recognized nuclear medicine as a broad specialty in 1982 and accredited RMC for training for Diplomate of National Board. Doctor of Medicine (MD) started first time in India and Asia at Sanjay Gandhi Postgraduate Institute, Lucknow in 1990. Doctorate of Therapeutic Nuclear Medicine commenced at All India Institute of Medical Sciences Delhi in 2015. There are 18 teaching hospitals currently imparting MD/DNB nuclear medicine residency for physicians with annual intake of 50. Eighteen institutions are offering bachelors and masters programs for nuclear medicine technology with an average annual intake of 110-120 students. Society of Nuclear Medicine, India (SNMI) is the oldest and largest professional body with total life membership of 1425 nuclear medicine professionals. SNMI was established in 1967 and hosted the first Annual Conference at RMC, Mumbai in 1968. Since then, SNMI is organizing its Annual Conferences in various parts of the country with the objective of scientific exchange and popularizing the modality amongst clinicians. Postgraduate Institute of Medical Education Research is hosting the 50th Annual Conference of SNMI (SNMICON-18) as mark of golden jubilee celebration.


Microplate phosphocellulose binding assay for aminoglycoside-modifying enzymes.
We modified the phosphocellulose binding assay for aminoglycoside-modifying enzymes (AMEs) by use of microdilution plates and a multichannel micropipette. Batteries of aminoglycoside substrates for screening organisms for the presence of AMEs as well as for subclassifying enzymes were prepared and stored in microdilution plates. When tested in parallel with the conventional tube reaction assay, the microplate assay yielded comparable radioactive counts and therefore equally correct identifications of AMEs in 32 isolates representing nine bacterial species. Other modifications, such as multichannel dispensing of crude enzyme preparations and radioisotopic precursors, provided a more rapid, convenient, and less expensive means of examining large collections of organisms for AMEs.


[Preoperative identification of ischemic heart disease in patients with surgical diseases of the abdominal cavity].
The authors present the results of preoperative identification of coronary diseases in patients with abdominal surgical diseases. Latent coronary diseases in surgical patients were detected by transesophageal electric stimulation (TEES) of the left atrium. Thirty-eight patients aged 46 to 82 were examined. Manifest symptoms of myocardial ischemia (changed ST segment, T wave inversion) were revealed in 47.3% patients, which is an evidence of the high resolving power of the method. Hence, noninvasive unsophisticated TEES technique is a reliable tool for identifying coronary diseases in surgical patients. The results of such examinations help adequately and purposefully prepare the patients to surgery and select the method of total anesthesia involving the minimal risk.


Violence in mental health care: the experiences of mental health nurses and psychiatrists.
Violence against mental health service personnel is a serious workplace problem and one that appears to be increasing. This study aimed to ascertain the extent and nature of violence against mental health nurses and psychiatrists, and to identify what support, if any, they received following exposure to violence. Mental health staff working within five West Midlands Trusts in the United Kingdom were surveyed using a postal questionnaire to investigate the extent and nature of violence they encountered in their daily work. There was an overall response rate of 47%, which included a response rate for psychiatrists of 60% (n=74) and for mental health nurses of 45% (n=301). Though both groups experienced violence at work, nurses were found: to have been exposed to violence significantly more during their career; to have been a victim of violence within the previous 12 months of the survey; and to have suffered a violent incident involving physical contact. Whilst a higher proportion of nurses than psychiatrists received some support following a violent incident, a large proportion of both groups did not receive any, although most felt in need of it. The implications of this study for training and management are discussed.


Expression of pax8 as a useful marker in distinguishing ovarian carcinomas from mammary carcinomas.
The ovary is a common site of involvement for metastasis and the breast is one of the most common sources. Metastatic breast carcinoma can mimic a primary ovarian carcinoma. Pax8 is a crucial transcription factor for organogenesis of the thyroid gland, kidney, and Müllerian system, and it also regulates Wilms tumor suppressor gene (WT1) expression. A total of 124 cases of ovarian carcinomas (84 serous papillary, 18 endometrioid, 12 mucinous, 10 clear cell) and 243 cases of invasive breast carcinomas (178 ductal, 65 lobular) were immunostained with Pax8 and WT1 by tissue microarrays to see the differential expression. Pax8 reaction was found in 108 of 124 ovarian carcinomas (87.1%) generally in diffuse staining, including 81 of 84 serous papillary carcinomas (96.4%), 16 of 18 endometrioid carcinomas (88.9%), 10 of 10 clear cell carcinomas (100%), and 1 of 12 mucinous carcinomas (8.3%), whereas WT1 expression was seen in 78 of 124 ovarian carcinomas (62.9%), including 73 of 84 serous papillary carcinomas (86.9%), and 5 of 18 endometrioid carcinomas (27.8%), with no expression in all 10 clear cell carcinomas and 12 mucinous carcinomas. All the mammary carcinomas were completely negative for Pax8, but WT1 expression was seen in 5 of 243 cases (2.1%). Pax8 is a useful marker in the differential diagnosis of ovarian and breast carcinomas, and it seems to be superior to WT1 for the diagnosis of all types of nonmucinous ovarian carcinomas, notably clear cell and endometrioid types where WT1 expression is generally negative or only focal.


Sequence composition and context effects on the generation and repair of frameshift intermediates in mononucleotide runs in Saccharomyces cerevisiae.
DNA polymerase slippage occurs frequently in tracts of a tandemly repeated nucleotide, and such slippage events can be genetically detected as frameshift mutations. In long mononucleotide runs, most frameshift intermediates are repaired by the postreplicative mismatch repair (MMR) machinery, rather than by the exonucleolytic proofreading activity of DNA polymerase. Although mononucleotide runs are hotspots for polymerase slippage events, it is not known whether the composition of a run and the surrounding context affect the frequency of slippage or the efficiency of MMR. To address these issues, 10-nucleotide (10N) runs were inserted into the yeast LYS2 gene to create +1 frameshift alleles. Slippage events within these runs were detected as Lys(+) revertants. 10G or 10C runs were found to be more unstable than 10A or 10T runs, but neither the frequency of polymerase slippage nor the overall efficiency of MMR was greatly influenced by sequence context. Although complete elimination of MMR activity (msh2 mutants) affected all runs similarly, analyses of reversion rates in msh3 and msh6 mutants revealed distinct specificities of the yeast Msh2p-Msh3p and Msh2p-Msh6p mismatch binding complexes in the repair of frameshift intermediates in different sequence contexts.


Noble metal nanoparticles applications in cancer.
Nanotechnology has prompted new and improved materials for biomedical applications with particular emphasis in therapy and diagnostics. Special interest has been directed at providing enhanced molecular therapeutics for cancer, where conventional approaches do not effectively differentiate between cancerous and normal cells; that is, they lack specificity. This normally causes systemic toxicity and severe and adverse side effects with concomitant loss of quality of life. Because of their small size, nanoparticles can readily interact with biomolecules both at surface and inside cells, yielding better signals and target specificity for diagnostics and therapeutics. This way, a variety of nanoparticles with the possibility of diversified modification with biomolecules have been investigated for biomedical applications including their use in highly sensitive imaging assays, thermal ablation, and radiotherapy enhancement as well as drug and gene delivery and silencing. Here, we review the available noble metal nanoparticles for cancer therapy, with particular focus on those already being translated into clinical settings.


Knowledge and practice regarding menstrual hygiene in rural adolescent girls of Nepal.
This study was done in the adolescent girls to evaluate the knowledge and practice on different aspects of menstrual hygiene. One hundred and fifty adolescent girls of age 13-15 years from 3 schools of Shivanagar and Patihani village development committees of Chitwan district were involved in this study. Altogether 27 questions were asked to each of them. During our study, we found that they were not properly maintaining the menstrual hygiene. Only 6.0% of girls knew that menstruation is a physiologic process, 36.7% knew that it is caused by hormones. Ninety-four percentages of them use the pads during the period but only 11.3% dispose it. Overall knowledge and practice were 40.6% and 12.9% respectively. Although knowledge was better than practice, both were not satisfactory. So, the girls should be educated about the process and significance of menstruation, use of proper pads or absorbents and its proper disposal. This can be achieved by giving them proper training and health education (by teachers, family members, health educators, and media) so that there won't be any misconception to the adolescent girls regarding menstrual hygiene.


Light-scattering investigation of the dissociation behavior of Lunatia heros and Littorina littorea hemocyanins.
The subunit structure and dissociation of the hemocyanins of two marine snails, Lunatia heros and Littorina littorea, were investigated by light-scattering molecular weight methods. The hemocyanins of both species of snails are readily dissociated to fragments of one-tenth and one-twentieth of the parent proteins of close to 9 X 10(6) daltons by either increasing the pH or using dissociating reagents of the hydrophobic urea series or some of the Hofmeister salts. The lower members of the latter group of reagents, NaCl, and to some extent also NaBr were found to have only marginal effects on the observed molecular weight transitions, suggesting that the two hemocyanins investigated possess beta-type subunits, which are known to be resistant to NaCl dissociation. The molecular weight profiles obtained with the various dissociating reagents were single inverted sigmoidal-shaped curves for both Lunatia and Littorina hemocyanins, suggesting overlapping transitions. The ultracentrifugation patterns and the species-distribution plots based on the urea dissociation data of Littorina hemocyanin suggest the presence of whole, half, and one-tenth molecular weight species in the dissociation transition region. Fitting of the urea dissociation data of Littorina hemocyanin obtained at both pH 5.7 and pH 8.0, assuming a sequential two-step dissociation scheme used in our previous studies [Herskovits, T. T., & Russell, M. W. (1984) Biochemistry 23, 2812-2819], was found to be consistent with a model of a few hydrophobic binding sites at the contact areas of the half-molecules and a much larger apparent number of binding sites (Napp) at the side to side contacts of the one-tenth molecules.(ABSTRACT TRUNCATED AT 250 WORDS)


Early nasal continuous positive airway pressure treatment reduces the need for intubation in very low birth weight infants.
Nasal continuous positive airway pressure (CPAP) applied shortly after birth is said to be an effective treatment of respiratory distress in very low birth weight infants (VLBW). We tested the hypothesis that the use of early nasal CPAP (applied as soon as signs of respiratory distress occurred, usually within 15 min after birth) reduces the need for intubation, the duration of intermittent mandatory ventilation and the incidence of bronchopulmonary dysplasia. All liveborn VLBW infants (birth weight < 1500 g) admitted to our tertiary neonatal intensive care unit in 1990 (historical controls) and in 1993 (early nasal CPAP group) entered the study. The intubation rate was significantly lower after introduction of nasal CPAP (30% vs 53%, P = 0.016). Median duration of intubation was 4.5 days (interquartile range 3-7 days) before versus 6.0 days (2.8-9 days) after nasal CPAP was introduced (P = 0.73). The incidence of bronchopulmonary dysplasia was not reduced significantly (32% vs 30%, P = 0.94). Survival until discharge was 89.5%, before versus 92.9% after introduction of nasal CPAP (P = 0.54). Early nasal CPAP is an effective treatment of respiratory distress in VLBW infants, significantly reducing the need for intubation and intermittent mandatory ventilation, without worsening other standard measures of neonatal outcome. We found no significant decrease in the incidence of bronchopulmonary dysplasia.


Protective effect of alpha-linolenic acid on gentamicin-induced ototoxicity in mice.
Alpha-linolenic acid is one of the fatty acids known as omega 3. Previous studies have shown the antioxidant and anti-inflammatory effects of alpha-linolenic acid, which prevented cell damage by inhibiting apoptotic pathway. Also, it is known that gentamicin activates apoptotic mediators and causes necrosis in the kidney. Due to this reason, we planned a study to evaluate the protective effects of alpha-linolenic acid on gentamicin induced ototoxicity by evaluating inflammation and apoptotic mediators. For this purpose, 100 mg/kg gentamicin (i.p; intraperitoneally) and 200 mg/kg alpha-linolenic acid (gavage) are administered to mice for 9 days. On 9th and 10th days, rotarod performance was assessed to test the effect of gentamicin and alpha-linolenic acid treatment on the motor coordination of mice. Gentamicin treatment decreased fall latency of mice and gentamicin treatment together with alpha-linolenic acid increased fall latency of mice. Gentamicin treatment also increased expression of phospholipase A2(plA2), cyclooxygenase-2(COX-2) and inducible nitric oxide syntheses (iNOS). Furthermore, it increased Bax and caspase-3, which are proapoptotic proteins and decreased bcl-2 that is an antiapoptotic protein. Gentamicin treatment together alpha-linolenic acid recovered the change of expression of these enzymes. In conclusion, this study showed that alpha-linolenic acid will be useful to prevent gentamicin-induced ototoxicity by inhibiting apoptosis and inflammation.


Clinical Efficacy and Tumor Microenvironment Influence in a Dose-Escalation Study of Anti-CD19 Chimeric Antigen Receptor T Cells in Refractory B-Cell Non-Hodgkin's Lymphoma.
Anti-CD19 chimeric antigen receptor (CAR) T cells represent a novel immunotherapy and are highly effective in treating relapsed/refractory B-cell non-Hodgkin's lymphoma (B-NHL). How tumor microenvironment influences clinical response to CAR T therapy remains of great interest. A phase I, first-in-human, dose-escalation study of anti-CD19 JWCAR029 was conducted in refractory B-NHL (NCT03355859) and 10 patients received CAR T cells at an escalating dose of 2.5 × 107(n = 3), 5 × 107(n = 4), and 1 × 108(n = 3) cells. Core needle biopsy was performed on tumor samples collected from diffuse large B-cell lymphoma patients on Day -6 (1 day before lymphodepletion) and on Day 11 after CAR T-cell infusion when adequate CAR T-cell expansion was detected. The overall response rate was 100%, with 6 of 9 (66.7%) evaluable patients achieving complete remission. The most common adverse events of grade 3 or higher were neutropenia (10/10, 100%), anemia (3/10, 30%), thrombocytopenia (3/10, 30%), and hypofibrinogenemia (2/10, 20%). Grade 1 cytokine release syndrome occurred in all patients and grade 3 neurotoxicity in 1 patient. The average peak levels of peripheral blood CAR T cells and cytokines were similar in 3 different dose levels, but CAR T cells were significantly higher in patients achieved complete remission on Day 29. Meanwhile, RNA sequencing identified gene expression signatures differentially enriched in complete and partial remission patients. Increased tumor-associated macrophage infiltration was negatively associated with remission status. JWCAR029 was effective and safe in treating refractory B-NHL. The composition of the tumor microenvironment has a potential impact in CAR T therapy response.


FoxA transcription factor Fork head maintains the intestinal stem/progenitor cell identities in Drosophila.
Understanding how somatic stem cells respond to tissue needs is important, since aberrant somatic stem cell behaviors may lead to tissue degeneration or tumorigenesis. Here, from an in vivo RNAi screen targeting transcription factors that regulate intestinal regeneration, we uncovered a requirement for the Drosophila FoxA transcription factor Fork head (Fkh) in the maintenance of intestinal stem/progenitor cell identities. FoxA/Fkh maintains the expressions of stem/progenitor cell markers and is required for stem cell proliferation during intestinal homeostasis and regeneration. Furthermore, FoxA/Fkh prevents the intestinal stem/progenitor cells from precocious differentiation into the Enterocyte lineage, likely in cooperation with the transcription factor bHLH/Daughterless (Da). In addition, loss of FoxA/Fkh suppresses the intestinal tumorigenesis caused by Notch pathway inactivation. To reveal the gene program underlying stem/progenitor cell identities, we profiled the genome-wide chromatin binding sites of transcription factors Fkh and Da, and interestingly, around half of Fkh binding regions are shared by Da, further suggesting their collaborative roles. Finally, we identified the genes associated with their shared binding regions. This comprehensive gene list may contain stem/progenitor maintenance factors functioning downstream of Fkh and Da, and would be helpful for future gene discoveries in the Drosophila intestinal stem cell lineage.


De novo transcriptome of the Hemimetabolous German cockroach (Blattella germanica).
The German cockroach, Blattella germanica, is an important insect pest that transmits various pathogens mechanically and causes severe allergic diseases. This insect has long served as a model system for studies of insect biology, physiology and ecology. However, the lack of genome or transcriptome information heavily hinder our further understanding about the German cockroach in every aspect at a molecular level and on a genome-wide scale. To explore the transcriptome and identify unique sequences of interest, we subjected the B. germanica transcriptome to massively parallel pyrosequencing and generated the first reference transcriptome for B. germanica. A total of 1,365,609 raw reads with an average length of 529 bp were generated via pyrosequencing the mixed cDNA library from different life stages of German cockroach including maturing oothecae, nymphs, adult females and males. The raw reads were de novo assembled to 48,800 contigs and 3,961 singletons with high-quality unique sequences. These sequences were annotated and classified functionally in terms of BLAST, GO and KEGG, and the genes putatively coding detoxification enzyme systems, insecticide targets, key components in systematic RNA interference, immunity and chemoreception pathways were identified. A total of 3,601 SSRs (Simple Sequence Repeats) loci were also predicted. The whole transcriptome pyrosequencing data from this study provides a usable genetic resource for future identification of potential functional genes involved in various biological processes.


Vertebroplasty versus conservative treatment in acute osteoporotic vertebral compression fractures (Vertos II): an open-label randomised trial.
Percutaneous vertebroplasty is increasingly used for treatment of pain in patients with osteoporotic vertebral compression fractures, but the efficacy, cost-effectiveness, and safety of the procedure remain uncertain. We aimed to clarify whether vertebroplasty has additional value compared with optimum pain treatment in patients with acute vertebral fractures. Patients were recruited to this open-label prospective randomised trial from the radiology departments of six hospitals in the Netherlands and Belgium. Patients were aged 50 years or older, had vertebral compression fractures on spine radiograph (minimum 15% height loss; level of fracture at Th5 or lower; bone oedema on MRI), with back pain for 6 weeks or less, and a visual analogue scale (VAS) score of 5 or more. Patients were randomly allocated to percutaneous vertebroplasty or conservative treatment by computer-generated randomisation codes with a block size of six. Masking was not possible for participants, physicians, and outcome assessors. The primary outcome was pain relief at 1 month and 1 year as measured by VAS score. Analysis was by intention to treat. This study is registered at ClinicalTrials.gov, number NCT00232466. Between Oct 1, 2005, and June 30, 2008, we identified 431 patients who were eligible for randomisation. 229 (53%) patients had spontaneous pain relief during assessment, and 202 patients with persistent pain were randomly allocated to treatment (101 vertebroplasty, 101 conservative treatment). Vertebroplasty resulted in greater pain relief than did conservative treatment; difference in mean VAS score between baseline and 1 month was -5·2 (95% CI -5·88 to -4·72) after vertebroplasty and -2·7 (-3·22 to -1·98) after conservative treatment, and between baseline and 1 year was -5·7 (-6·22 to -4·98) after vertebroplasty and -3·7 (-4·35 to -3·05) after conservative treatment. The difference between groups in reduction of mean VAS score from baseline was 2·6 (95% CI 1·74-3·37, p<0·0001) at 1 month and 2·0 (1·13-2·80, p<0·0001) at 1 year. No serious complications or adverse events were reported. In a subgroup of patients with acute osteoporotic vertebral compression fractures and persistent pain, percutaneous vertebroplasty is effective and safe. Pain relief after vertebroplasty is immediate, is sustained for at least a year, and is significantly greater than that achieved with conservative treatment, at an acceptable cost. ZonMw; COOK Medical.


Pattern of tobacco smoking in north Indian adults.
An exploratory study was conducted among 200 apparently healthy current smokers aged 15-45 years to determine their attitudes and behaviour regarding tobacco smoking by using a precoded questionnaire specifically designed for the purpose. Females constituted 10% of the study group and the 73 participants who smoked cigarettes exclusively were from urban backgrounds and were noted to inhale the smoke more frequently than bidi or hukka smokers. Parental and peer group influence, as well as curiosity in late teenage were the major reasons for starting smoking which was however continued mainly to obtain the stimulatory and or relaxing effects of nicotine. Health hazards of smoking, particularly lung cancer and heart disease, were widely known and fear of these constituted the most important reason for smokers wishing to quit the habit. One-half of the subjects attempted to stop but could not succeed due to withdrawal symptoms and lack of a suitable substitute. The divergence between attitude and behaviour of smokers is highlighted by this study since smokers continued to smoke despite being averse to smoking and disapproval of their habit by their family members. There were important differences in the pattern of smoking and perceptions of various groups of smokers regarding the societal permissiveness, awareness of health hazards, and measures to control smoking.


The impact of COVID-19 on the physical activity and sedentary behaviour levels of pregnant women with gestational diabetes.
The aim of this study was to understand how physical activity and sedentary behaviour levels of pregnant women with gestational diabetes in the UK have been affected by COVID-19. An online survey exploring physical activity and sedentary behaviour levels of pregnant women with gestational diabetes during COVID-19 was distributed through social media platforms. Women who had been pregnant during the COVID-19 outbreak and had gestational diabetes, were resident in the UK, were 18 years old or over and could understand written English were invited to take part. A total of 724 women accessed the survey, 553 of these met the eligibility criteria and took part in the survey. Sedentary time increased for 79% of the women during the pandemic. Almost half of the women (47%) were meeting the physical activity guidelines pre COVID-19 during their pregnancy, this dropped to 23% during the COVID-19 pandemic. Fear of leaving the house due to COVID-19 was the most commonly reported reason for the decline. Significant associations were found between meeting the physical activity guidelines during COVID-19 and educational attainment, fitness equipment ownership and knowledge of how to exercise safely in pregnancy. These results show the impact of COVID-19 on physical activity and sedentary behaviour levels and highlight the need for targeted public health initiatives as the pandemic continues and for future lockdowns. Women with gestational diabetes need to know how it is safe and beneficial to them to engage in physical activity and ways to do this from their homes if fear of leaving the house due to COVID-19 is a barrier for them. Online physical activity classes provided by certified trainers in physical activity for pregnant women may help them remain active when face-to-face appointments are reduced and limited additional resources are available.


The beneficial effects of total cavopulmonary conversion and arrhythmia surgery for the failed Fontan.
Postoperative Fontan patients can develop hemodynamic abnormalities and refractory atrial arrhythmias resulting in significant morbidity and mortality. We present our experience with total cavopulmonary artery conversion and arrhythmia surgery. Between 1994 and 2001, 41 patients underwent total cavopulmonary artery conversion and arrhythmia surgery. Significant hemodynamic lesions were repaired concomitantly: aortic aneurysm (n=1), atrioventricular valve insufficiency (n=8), and pulmonary artery stenosis (n=9). Thirty-five patients were in New York Heart Association class III or IV. Mean age at original Fontan was 7.5+/-6.5 years, at Fontan conversion, 18.7+/-9.0 years. Arrhythmia surgery for atrial re-entry tachycardia evolved from isthmus cryoablation (n=10) to right-sided maze (n=17). Maze-Cox III was used for 14 patients with atrial fibrillation. Atrial (n=34) and dual chamber (n=5) pacemakers were placed. Mortality and reoperation for bleeding rates are 0%. Chest tubes were removed on postoperative day 9.0+/-6.0. Mean hospital stay was 11.8+/-6.6 days. Three patients required cardiac transplantation at 8 days, 9 months, and 33 months postoperatively. There was one long-term death from acute myocardial infarction 2 years postoperatively. For the entire series, arrhythmia recurrence is 12.2% (5/41). Only 9.8% of patients (4/41) receive chronic antiarrhythmic medications; these patients were among the first eight in the series. Most patients are in New York Heart Association I or II. Bruce protocol in 12 patients showed increased tolerance (P<.05) Total cavopulmonary artery conversion with concomitant arrhythmia surgery is excellent therapy for patients with failed Fontan. It is safe, improves New York Heart Association class, improves exercise tolerance, and the incidence of recurrent arrhythmias is low.


In Vitro Bioaccessibility of Carotenoids and Chlorophylls in a Diverse Collection of Spinach Accessions and Commercial Cultivars.
Spinach, a nutrient-dense, green-leafy vegetable, is a rich source of carotenoid and chlorophyll bioactives. While the content of bioactives is known to vary with the genotype, variation in bioaccessibility is unknown. Bioaccessibility was explored in 71 greenhouse-grown spinach genotypes in fall and spring 2018/2019. Spinach was phenotyped for its greenness, leaf texture, leaf shape, and SPAD chlorophyll content. Postharvest, spinach was washed, blanched, and homogenized prior to assessment of bioactive bioaccessibility using a novel high-throughput in vitro digestion model followed by high-performance liquid chromatography with a photodiode array detector analysis. There was a significant variation in the bioaccessible content for all bioactives (p < 0.05), except for chlorophyll b (p = 0.063) in spring-grown spinach. The correlation coefficients of bioaccessible contents between seasons reveal that lutein (r = 0.52) and β-carotene (r = 0.55) were correlated to a greater extent than chlorophyll a (r = 0.38) and chlorophyll b (r = 0.19). The results suggest that carotenoid and chlorophyll bioaccessible contents may vary based on spinach genotypes and may be stable across seasons.


Matter-Wave Tractor Beams.
Optical and acoustic tractor beams are currently the focus of intense research due to their counterintuitive property of exerting a pulling force on small scattering objects. In this Letter we propose a matter-wave tractor beam and utilize the de Broglie waves of nonrelativistic matter particles in analogy to "classical" tractor beams. We reveal the presence of the quantum-mechanical pulling force for the variety of quantum mechanical potentials observing the resonant enhancement of the pulling effect under the conditions of the suppressed scattering known as the Ramsauer-Townsend effect. We also derive the sufficient conditions on the scattering potential for the emergence of the pulling force and show that, in particular, a Coulomb scatterer is always shoved, while a Yukawa (screened Coulomb) scatterer can be drawn. Pulling forces in optics, acoustics, quantum mechanics, and classical mechanics are compared, and the matter-wave pulling force is found to have exclusive properties of dragging slow particles in short-range potentials. We envisage that the use of tractor beams could lead to the unprecedented precision in manipulation with atomic-scale quantum objects.


Mapping change in human pressure globally on land and within protected areas.
It is widely accepted that the main driver of the observed decline in biological diversity is increasing human pressure on Earth's ecosystems. However, the spatial patterns of change in human pressure and their relation to conservation efforts are less well known. We developed a spatially and temporally explicit map of global change in human pressure over 2 decades between 1990 and 2010 at a resolution of 10 km(2) . We evaluated 22 spatial data sets representing different components of human pressure and used them to compile a temporal human pressure index (THPI) based on 3 data sets: human population density, land transformation, and electrical power infrastructure. We investigated how the THPI within protected areas was correlated to International Union for Conservation of Nature (IUCN) management categories and the human development index (HDI) and how the THPI was correlated to cumulative pressure on the basis of the original human footprint index. Since the early 1990s, human pressure increased 64% of the terrestrial areas; the largest increases were in Southeast Asia. Protected areas also exhibited overall increases in human pressure, the degree of which varied with location and IUCN management category. Only wilderness areas and natural monuments (management categories Ib and III) exhibited decreases in pressure. Protected areas not assigned any category exhibited the greatest increases. High HDI values correlated with greater reductions in pressure across protected areas, while increasing age of the protected area correlated with increases in pressure. Our analysis is an initial step toward mapping changes in human pressure on the natural world over time. That only 3 data sets could be included in our spatio-temporal global pressure map highlights the challenge to measuring pressure changes over time.


Defining and categorizing leukoencephalopathies of unknown origin: MR imaging approach.
To categorize leukoencephalopathies of unknown origin into a few major groups by using magnetic resonance (MR) imaging criteria to facilitate further studies, and to assess the possibility of defining "new" (i.e., until now unknown) disease entities within these major groups. MR images of 92 patients (55 male, 37 female; mean age, 9.3 years) with a leukoencephalopathy were examined by using a scoring list of 68 items. Seven major categories were defined according to the predominant location of the white matter abnormalities. Statistical analysis was used to assess the validity of these seven categories. Statistical analysis results showed that the seven categories could be well distinguished by either using the defining variables initially accepted as inclusion criteria or selecting a few other variables found to have discriminating value. The additional variables confirmed that the categories are essentially distinct and vary systematically with regard to items other than the inclusion criteria. The existence of two recently defined leukoencephalopathies was confirmed, but no consistent evidence of other new disease entities could be provided. Establishing these seven categories helps in the interpretation of individual studies by demonstrating features that the patient has in common with other patients, and it may facilitate further research on homogeneous subgroups of patients and allow pooling of data across multiple centers.


Regulation by glucocorticoids of angiotensinogen gene expression and secretion in adipose cells.
Adipose cells are an important source of angiotensinogen (AT). Its activation product, angiotensin II, stimulates in vitro and in vivo the production and release of prostacyclin which acts as a potent adipogenic signal in promoting the terminal differentiation of preadipocytes to adipocytes. Since glucocorticoids are known to promote adipose cell differentiation in vitro as well as in vivo, their role in the regulation of AT gene expression and secretion has been investigated in cultured Ob1771 mouse adipose cells. In contrast with liver cells, which are the major source of AT and the target of several hormones for the regulation of its expression, adipose cells are only responsive to glucocorticoids, which are able to up-regulate AT gene expression and AT secretion rapidly and dose-dependently. On exposure to glucocorticoids, accumulation of AT mRNA appears primarily to be due to transcriptional activation of the gene and is parallelled by secretion of the protein. Similar results on AT mRNA expression and AT secretion were obtained using explants of rat adipose tissue ex vivo demonstrating a major if not exclusive mechanism of regulation of AT production by glucocorticoids in mature adipose cells. Together these results provide a potential link between glucocorticoids, AT, the growth of adipose tissue and increased blood pressure.


Systemic mastocytosis: current concepts and treatment advances.
Systemic mastocytosis (SM), as opposed to cutaneous-only mastocytosis, implies the presence of neoplastic mast cell infiltration in extracutaneous tissue. Mast cell disease in adults is often systemic and often involves the bone marrow. Typical clinical and laboratory features of SM include urticaria pigmentosa, mast cell mediator symptoms (eg, headache, flushing, lightheadedness, urticaria and pruritus, nausea, diarrhea, abdominal pain, and vasodilatory shock), bone pain (eg, osteoporosis, lytic bone lesions, and fractures), hepatosplenomegaly, cytopenia, eosinophilia, elevated serum tryptase and histamine, and bone marrow fibrosis and angiogenesis. SM may be indolent (no evidence of organ dysfunction), aggressive (presence of organ dysfunction), associated with another often chronic myeloid hematologic disease (SM-AHD), or present as mast cell leukemia or sarcoma. Mast cell-mediator symptoms are treated with histamine antagonists and cromolyn sodium. Indolent SM does not require cytoreductive therapy. Aggressive SM and SM-AHD are managed based on their molecular profile. Recent information suggests that FIP1-like-1-platelet-derived growth factor receptor-alpha(+) SM responds well to imatinib mesylate, whereas interferon-alpha should be considered as a first-line treatment in all of the other cases, including patients with Asp816Val(+) SM. Cladribine has been shown to be effective in patients who develop resistance to interferon treatment.


Evaluation of the Uniformity of Protective Coatings on Concrete Structure Surfaces Based on Cluster Analysis.
With the continuous development of urbanization and industrialization in the world, concrete is widely used in various engineering constructions as an engineering material. However, the consequent problem of durability of concrete structures is also becoming increasingly prominent. As an important additional measure, a protective coating can effectively improve the durability of concrete performance. Moreover, the uniformity of the concrete surface coating will directly affect its protective effect. Therefore, we propose a nondestructive inspection and evaluation method of coating uniformity based on infrared imaging and cluster analysis for concrete surface coating uniformity detection and evaluation. Based on the obtained infrared images, a series of processing and analysis of the images were carried out using MATLAB software to obtain the characteristics of the infrared images of the concrete surface. Finally, by extracting the temperature distribution data of the pixel points on the concrete surface, an evaluation method of concrete surface coating uniformity based on a combination of cluster analysis and hierarchical analysis was established. The evaluation results show that the determination results obtained by this method are consistent with the actual situation. This study has a positive contribution to the testing of concrete surface coating uniformity and its evaluation.


[Reference values of erythrocyte ferritin in children and adults].
The erythrocyte ferritin content was measured in 183 healthy subjects in age from 4 to 68 years; 80 were male and 103 were female. In children between the ages of 4 and 12 years there is no significant difference in the mean value between boys and girls. In females the erythrocyte ferritin concentration is independent of age. After 12 years of age the erythrocyte ferritin content is higher in men. Reference intervals were determined by the two quantiles 0.05 and 0.95. The reference interval is 3-24 age by cell in boys between 4 and 12 and females between 4 and 63; the reference interval is 5-38 age by cell in males between 13 and 68 years of age.


[On the assessment of the vertical posture support function based on the results of dynamic posturography].
A group of healthy subjects (n=93) was enrolled in the study designed to obtain the main characteristics of the postural function under effect of polymodal sensory afferentation. The study was carried out with the use of a dynamic stabilometric platform that served to estimate the vertical posture support function by means of an original diagnostic algorithm based on the main principles biological feedback priniciples. The study revealed two types of responses to polymodal afferentation, cortical and subcortical (reflective). The "cortical" type is characterized by slow responses due to the fact that the mechanism of sensory afferentation implies that an impulse has to pass along the entire conducting pathway from the primary receptor cell to the cortical centres that arbitrarily control the posture. The "subcortical" (reflective) mechanism ensures rapid but poorly selective responses to polymodal afferentation. The method for diagnostics of postural problems described in this paper with the use of a dynamic (mobile) stabilometric platform is expected to extend the possibilities for the quantitative evaluation of the vertical posture support function.


Oligoesculin fraction induces anti-tumor effects and promotes immune responses on B16-F10 mice melanoma.
Laccase was used to enzymatically polymerize esculin. Oligoesculin fraction was obtained after ultrafiltration through a 5-kDa membrane. Several studies have been carried out to prove the effectiveness of natural substances such as immunomodulators to promote the anti-cancer activity in situ. The purpose of our report was to explore whether the anti-tumor potential of the oligoesculin fraction in vitro and in vivo is linked to its immunological mechanisms in melanoma-bearing mice. We revealed that oligoesculin fraction reduced B16-F10 proliferation and migration in vitro in a dose-related manner. Moreover, melanin synthesis and tyrosinase activity were inhibited in these melanoma cells in a concentration-dependent way. The anti-tumor potential of oligoesculin fraction was also assessed in vivo. Our results showed that intraperitoneal administration of oligoesculin fraction, at 50 mg/kg body weight (b.w.) for 21 days, reduced tumor size and weight with percentages of inhibition of 94 and 87 %, respectively. Oligoesculin fraction was effective in promoting lysosomal activity and nitric oxide (NO) production by peritoneal macrophages in tumor-implanted mice. In addition, the activities of natural killer (NK), cytotoxic T lymphocytes, and macrophages were significantly enhanced by oligoesculin fraction. These findings suggested that this polymer with its anti-tumor and immunomodulatory properties could be used for the treatment of melanoma.


[A study of nursing systems in transition in developed countries].
The present study was designed to investigate nursing systems in other countries in comparison with the current system in Japan, and thereby assess how nursing personnel should be in order to improve the quality of nursing. The study focused on the actual situation of the nursing system and licensed practical nurses (LPNs) or second-level nurses similar to LPNs in other countries. The survey was conducted in regard to six countries considered to be advanced in the development of nursing activities and likely to provide useful suggestions for the future direction of nursing in Japan: Australia, Canada, France, Sweden, the United Kingdom and the United States. Questionnaires were sent to the heads or staff of a total of 29 organizations, including WHO Collaborating Centers and professional organizations for nursing, in these countries. Nineteen organizations responded (response rate: 65.5%), and an effective response with completed questionnaire forms was obtained from 15 organizations. In addition, an interview survey was conducted on three nursing administration and education experts in the United States in order to acquire clear understanding of the actual state of nursing in health care practice. The results were as follows: 1. Six countries (Japan included) excluding France had LPNs. Educational preparation for LPNs has been discontinued in the United Kingdom. 2. The number of nurses per 1,000 population was lowest in Japan: the total number of registered nurses (RNs) and LPNs in Japan was less than the number of RNs in the United Kingdom. Only in Japan the proportions of LPNs and RNs were similar, while in other countries of number of LPNs was one-third to one-fourth of the number of RNs. 3. In the five other countries having LPNs, the nurse's competency or scope of practice was clearly defined for both LPNs and RNs. In contrast, no clear line was drawn between the two in Japan. 4. The length of education required for LPNs ranged from 11 to 14 years (including the period of compulsory education) and was shortest in Japan (11 years). The educational requirement for admission to LPN school in Japan was 9 years of compulsory education (graduation from junior high school), whereas in other countries it was at the level of senior high school graduation. 5. Four countries had conversion programs for LPNs to become RNs, and the conversion courses were positioned within the framework of higher education comparable to the university level. 6. In the United Kingdom, where the educational preparation for LPNs has been discontinued, nurses are included in a single higher level profession. At the same time there was found to be a need to train and educate auxiliary personnel in order to maintain multi-level care services. Because of the increasing tendency toward advanced medical technology and highly specialized medical care associated with the rapidly increasing care needs in the community, the current educational preparation for LPNs in Japan in unsatisfactory as a training and educational system for nursing manpower to cope with the current situation. The above findings suggest that the education system for LPNs be reviewed with a view to discontinuing it and consolidating nursing education in Japan.


Responses of Manduca sexta larvae to heat waves.
Climate change is increasing the frequency of heat waves and other extreme weather events experienced by organisms. How does the number and developmental timing of heat waves affect survival, growth and development of insects? Do heat waves early in development alter performance later in development? We addressed these questions using experimental heat waves with larvae of the tobacco hornworm, Manduca sexta. The experiments used diurnally fluctuating temperature treatments differing in the number (0-3) and developmental timing (early, middle and/or late in larval development) of heat waves, in which a single heat wave involved three consecutive days with a daily maximum temperature of 42°C. Survival to pupation declined with increasing number of heat waves. Multiple (but not single) heat waves significantly reduced development time and pupal mass; the best models for the data indicated that both the number and developmental timing of heat waves affected performance. In addition, heat waves earlier in development significantly reduced growth and development rates later in larval development. Our results illustrate how the frequency and developmental timing of sublethal heat waves can have important consequences for life history traits in insects.


Role of Mitochondria-ER Contact Sites in Mitophagy.
Mitochondria are often referred to as the "powerhouse" of the cell. However, this organelle has many more functions than simply satisfying the cells' metabolic needs. Mitochondria are involved in calcium homeostasis and lipid metabolism, and they also regulate apoptotic processes. Many of these functions require contact with the ER, which is mediated by several tether proteins located on the respective organellar surfaces, enabling the formation of mitochondria-ER contact sites (MERCS). Upon damage, mitochondria produce reactive oxygen species (ROS) that can harm the surrounding cell. To circumvent toxicity and to maintain a functional pool of healthy organelles, damaged and excess mitochondria can be targeted for degradation via mitophagy, a form of selective autophagy. Defects in mitochondria-ER tethers and the accumulation of damaged mitochondria are found in several neurodegenerative diseases, including Parkinson's disease and amyotrophic lateral sclerosis, which argues that the interplay between the two organelles is vital for neuronal health. This review provides an overview of the different mechanisms of mitochondrial quality control that are implicated with the different mitochondria-ER tether proteins, and also provides a novel perspective on how MERCS are involved in mediating mitophagy upon mitochondrial damage.


The association between daily calcium intake and sarcopenia in older, non-obese Korean adults: the fourth Korea National Health and Nutrition Examination Survey (KNHANES IV) 2009.
Recent data suggest that variations in calcium intake may influence body weight and composition; however, the relationship between daily calcium intake and muscle mass has not been well established. The objective of this study was to assess the relationship between daily calcium intake and sarcopenia. We analyzed data for older adults (over 60 years) from the fourth Korea National Health and Nutrition Examination Survey (KNHANES) conducted in 2009. A total of 1339 Non-Obese (BMI between 18.5 and 25 kg/m²), older adults (592 men and 707 women) were enrolled. Dietary variables were assessed using a nutrition survey that used a 24-hour recall method. Daily calcium intake based on the consumption of each food item was calculated. Sarcopenia was defined as an appendicular skeletal muscle mass divided by body weight less than 2 SD below the sex-specific mean for young adults. We found that daily calcium intake was negatively correlated with total body fat percentage and positively correlated with appendicular skeletal mass (p<0.001). Participants with sarcopenia appear to have significantly lower daily calcium intakes than participants without sarcopenia (p<0.001). The unadjusted prevalence of sarcopenia according to daily calcium intake tertiles were 6.3%, 4.3%, and 2.7% in tertiles 1, 2, and 3, respectively. After adjustment for age, sex, BMI, total energy intake, and lifestyle factors, compared with those in the lowest tertile of daily calcium intake, participants in the highest tertile had an odds ratio for sarcopenia of 0.295 (95% confidence interval, 0.087-0.768; p for trend = 0.014). We found that daily calcium intake, corrected for total energy intake and serum 25(OH)D status, was significantly lower in subjects with sarcopenia than in those without. Our results suggest a strong inverse association between daily calcium intake and sarcopenia in non-obese, older Korean adults.


Genotoxicity of anti-tumor necrosis factor therapy in patients with juvenile idiopathic arthritis.
To assess the possible effects of both inflammation and the anti-tumor necrosis factor agents (anti-TNF) on DNA damage with a specific assay, and their effects on the repair capacity of DNA. From a group of 20 children with juvenile idiopathic arthritis (JIA), 16 patients who completed the study and 16 control subjects were evaluated. DNA damage and repair capacity were analyzed by the comet assay at the level of peripheral lymphocytes before anti-TNF (etanercept) injections and on the 15th, 90th, and 180th days after the first injection. The amount of damage as detected by the aforementioned assay was higher in patients with JIA compared with controls. On the 15th day after the initial anti-TNF injection, there was a decrease in the mean DNA tail length of JIA patients, however on the 90th day an increase was observed; thereafter, an upward trend was observed until the end of the study. JIA patients had a DNA repair capacity that was significantly less than that of controls. The results of the comet technique suggests that JIA patients already have increased basal DNA damage before anti-TNF therapy; they are more sensitive to the DNA damage produced by H(2)O(2), and have a less efficient DNA repair system in comparison with control cells. After an initial improvement at 2 weeks, parameters of genotoxicity worsened, and DNA repair was further impaired 6 months after the addition of an anti-TNF agent to treatment.


A prospective examination of the interpersonal-psychological theory of suicidal behavior among psychiatric adolescent inpatients.
The challenge of identifying suicide risk in adolescents, and particularly among high-risk subgroups such as adolescent inpatients, calls for further study of models of suicidal behavior that could meaningfully aid in the prediction of risk. This study examined how well the Interpersonal-Psychological Theory of Suicidal Behavior (IPTS)--with its constructs of thwarted belongingness (TB), perceived burdensomeness (PB), and an acquired capability (AC) for lethal self-injury--predicts suicide attempts among adolescents (N = 376) 3 and 12 months after hospitalization. The three-way interaction between PB, TB, and AC, defined as a history of multiple suicide attempts, was not significant. However, there were significant 2-way interaction effects, which varied by sex: girls with low AC and increasing TB, and boys with high AC and increasing PB, were more likely to attempt suicide at 3 months. Only high AC predicted 12-month attempts. Results suggest gender-specific associations between theory components and attempts. The time-limited effects of these associations point to TB and PB being dynamic and modifiable in high-risk populations, whereas the effects of AC are more lasting. The study also fills an important gap in existing research by examining IPTS prospectively.


Phase II study of weekly high dose fluorouracil in previously treated patients with metastatic colorectal cancer.
The purpose of this study was to evaluate the efficacy, toxicity, and safety of outpatient chemotherapy with weekly high-dose 5-fluorouracil (HD-5FU) in previously treated patients (pts) with metastatic colorectal cancer. Previously treated and failed pts with histologically confirmed, measurable metastatic colorectal cancer, performance status (WHO) 0-2 and adequate bone marrow function were eligible for treatment. Patients received 5FU (2.6 g/m2) as a 24-hour continuous infusion. Treatment was repeated weekly. A total of 202 cycles were given. Eighteen pts were enrolled. No pts achieved objective response. In 6 pts (33%), after 4 weeks of treatments, CEA level decreased 25% or more, and after 8 weekly treatments it rose again. Mild myelosuppression rarely occurred. Grade I nausea and vomiting occurred in 2 pts and Grade I diarrhea occurred in 2 pts. Mucositis was not observed. In our experience single agent, weekly, high dose 5-FU is well tolerated, but is ineffective as second-line treatment for metastatic colorectal cancer, and has only a marginal effect on CEA level.


Direct quantification of norepinephrine spillover and hormone output from the pancreas of the conscious dog.
To estimate pancreatic neural activity and to assess the potential role of the pancreatic nerves in the regulation of hormone secretion, the methodology necessary to quantify neurotransmitter spillover and hormone output in the conscious dog was developed. A femoral artery and the superior pancreaticoduodenal vein (SPDV) were chronically cannulated, and a flow probe was placed on the SPDV. Hormone output was calculated using the pancreatic arteriovenous concentration difference and the SPDV plasma flow. Basal glucose levels were 103 +/- 1 mg/dl; the pancreatic outputs of insulin, glucagon, and pancreatic polypeptide (PP, an index of parasympathetic neural activity) were 2,900 +/- 700 microU/min, 1,900 +/- 400 pg/min, and 9.3 +/- 4.6 ng/min, respectively. Pancreatic norepinephrine (NE) spillover was calculated similarly; however, pancreatic extraction of epinephrine was used as an index of NE extraction. Basal NE spillover was 3,600 +/- 700 pg/min, greatly exceeding that reported using anesthetized, laparotomized dogs. Intravenous glucose infusion increased plasma glucose to 146 +/- 13 mg/dl, increased insulin output approximately twofold, and suppressed glucagon output by approximately 50%. Hyperglycemia markedly reduced PP output. Hyperglycemia failed to influence pancreatic NE spillover. Insulin-induced hypoglycemia (36 +/- 2 mg/dl) completely suppressed insulin output and stimulated glucagon output (> 10-fold). Hypoglycemia increased NE spillover and PP output to 19,900 +/- 4,600 pg/min and 117 +/- 22 ng/min, respectively. We conclude that pancreatic neurotransmitter spillover in the basal state is much higher than previously appreciated and that neural signaling to the pancreas is responsive to physiological and pathophysiological changes in the metabolic state.


Case report: Treatment of mild traumatic brain injury with hyperbaric oxygen.
Two United States Air Force Airmen were injured in a roadside improvised explosive device (IED) blast in Iraq in January 2008. Both airmen suffered concussive injuries and developed irritability, sleep disturbances, headaches, memory difficulties and cognitive difficulties as symptoms of mild traumatic brain injury (mTBI). Six months after injury, repeat Automated Neuropsychological Assessment Metrics (ANAM) testing showed deterioration, when compared to pre-injury baseline ANAM assessment, in all measured areas (simple reaction time, procedural reaction time, code substitution learning, code substitution delayed, mathematical processing, and matching to sample). The airmen were treated with hyperbaric oxygen in treatments of 100% oxygen for one hour at 1.5 atmospheres absolute, resulting in rapid improvement of headaches and sleep disturbances, improvement in all symptoms and resolution of most symptoms. Repeat ANAM testing after completion of the hyperbaric treatments - nine months after initial injury - showed improvement in all areas, with most measures improving to pre-injury baseline levels. The airmen received no other treatment besides medical monitoring. Repeat neuropsychologic testing confirmed the improvement. We conclude that the improvement in symptoms and ANAM performance is most likely attributable to HBO treatment.


Molecular characterization and functional importance of pectate lyase secreted by the cyst nematode Heterodera schachtii.
SUMMARY To analyse the parasitic behaviour of the plant-parasitic nematode Heterodera schachtii, proteins secreted by this nematode were purified and separated by two-dimensional gel electrophoresis. Mass spectrometric analysis identified one of the spots as a pectate lyase (EC 18.104.22.168). The corresponding gene was cloned from a cDNA library using primers derived from the peptide tag. A second pectate lyase was cloned based on similarity to known pectate lyases of related cyst nematodes. The predicted proteins are only 29% identical. Despite the low homology, the proteins have a similar secondary structure and it is likely that they fold into a similar right-handed beta-helix. Both proteins have a putative signal peptide for secretion, and in situ hybridization revealed that expression of the genes was limited to the subventral secretory glands. RT-PCR showed that both genes were expressed in the migratory preparasitic stage although the level of expression between the two genes was different. Post-transcriptional gene silencing by soaking the nematodes in double-stranded RNA against the gene with the highest expression level affected the infection process of the nematode, which is in agreement with the general idea that pectate lyases are essential during migration of the nematode in the plant root.


Surface engineering of macrophages with nanoparticles to generate a cell-nanoparticle hybrid vehicle for hypoxia-targeted drug delivery.
Tumors frequently contain hypoxic regions that result from a shortage of oxygen due to poorly organized tumor vasculature. Cancer cells in these areas are resistant to radiation- and chemotherapy, limiting the treatment efficacy. Macrophages have inherent hypoxia-targeting ability and hold great advantages for targeted delivery of anticancer therapeutics to cancer cells in hypoxic areas. However, most anticancer drugs cannot be directly loaded into macrophages because of their toxicity. In this work, we designed a novel drug delivery vehicle by hybridizing macrophages with nanoparticles through cell surface modification. Nanoparticles immobilized on the cell surface provide numerous new sites for anticancer drug loading, hence potentially minimizing the toxic effect of anticancer drugs on the viability and hypoxia-targeting ability of the macrophage vehicles. In particular, quantum dots and 5-(aminoacetamido) fluorescein-labeled polyamidoamine dendrimer G4.5, both of which were coated with amine-derivatized polyethylene glycol, were immobilized to the sodium periodate-treated surface of RAW264.7 macrophages through a transient Schiff base linkage. Further, a reducing agent, sodium cyanoborohydride, was applied to reduce Schiff bases to stable secondary amine linkages. The distribution of nanoparticles on the cell surface was confirmed by fluorescence imaging, and it was found to be dependent on the stability of the linkages coupling nanoparticles to the cell surface.


A novel mechanism for anti-EGFR antibody action involves chemokine-mediated leukocyte infiltration.
Overexpression of the epidermal growth factor receptor (EGFR) is a hallmark of squamous cell carcinoma of the head and neck (SCCHN). Monoclonal antibodies (mAbs) against EGFR are currently used for therapy of recurrent or metastatic disease; however, their mode of action is not completely understood. To investigate the immunological effects of anti-EGFR mAb, we generated a three-dimensional spheroid model of EGFR-expressing SCCHN and used this model to study the effect of anti-EGFR mAb on leukocyte migration toward tumors. Pretreatment with the blocking anti-EGFR mAb EMD 72000, its F(ab')2 fragments or an EGFR tyrosine kinase inhibitor led to substantially increased leukocyte infiltration into EGFR overexpressing tumor spheroids, but not into those with low EGFR expression. Nonblocking anti-EGFR mAb or fibroblast-specific mAb did not affect leukocyte infiltration, suggesting that the observed increase in leukocyte infiltration depends on interference with EGFR activation. Using a human cytokine macroarray, we demonstrated that the blockade of EGFR by anti-EGFR mAb in EGFR-overexpressing SCCHN cells leads to differential expression of several cytokines and chemokines, including the chemokine MCP-1/CCL-2. The significant upregulation of MCP-1/CCL2 on exposure to anti-EGFR mAb was confirmed by quantitative PCR and enzyme-linked immunospot analyses. Moreover, blocking anti-MCP-1 antibody inhibited leukocyte migration toward tumor cells induced by anti-EGFR mAb, pointing to an important role of MCP-1/CCL2 in anti-EGFR mAb-induced leukocyte migration. Our findings demonstrate that anti-EGFR mAb induces leukocyte infiltration to tumor spheroids by upregulating chemokine expression. This novel mechanism for anti-EGFR mAb action may contribute to the antitumor effects of anti-EGFR mAb in vivo.


Electroacupuncture alleviates ischaemic brain injury by regulating the miRNA-34/Wnt/autophagy axis.
Electroacupuncture (EA), a modern form of acupuncture therapy, has been widely used for the treatment of ischaemic brain injury. However, the molecular mechanism by which EA improves ischaemic brain injury remains unclear. In the current study, middle cerebral artery occlusion (MCAO) rats were treated with EA. The infarct volumes and apoptosis of neurocytes were assessed to determine the therapeutic effect of EA. The differentially expressed miRNAs between the control, MCAO and MCAO treated with EA groups were detected by high-throughput sequencing. The results indicated that EA treatment decreased neurocyte apoptosis and ischaemic infarct volume. Between the three groups, miR-34, miR-235 and miR-275 were found to be significantly different. Furthermore, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway data suggest that the Wnt pathway may play an important role in ischaemic brain injury and in the treatment of EA. Our data documented that miR-34 was obviously increased in the MCAO group, while EA treatment decreased miR-34 expression. WNT1 was the target of miR-34 and was confirmed by a luciferase reporter assay. A previous study suggested that the Wnt pathway mediates autophagy in EA-pretreated MCAO mice. Our data further confirmed that EA treatment after MCAO also alleviated autophagy in the MCAO group. Our results suggest that EA treatment alleviates ischaemic brain injury by inhibiting autophagy through the miR-34/Wnt pathway.


Rapid and Facile Microwave-Assisted Surface Chemistry for Functionalized Microarray Slides.
We describe a rapid and facile method for surface functionalization and ligand patterning of glass slides based on microwave-assisted synthesis and a microarraying robot. Our optimized reaction enables surface modification 42-times faster than conventional techniques and includes a carboxylated self-assembled monolayer, polyethylene glycol linkers of varying length, and stable amide bonds to small molecule, peptide, or protein ligands to be screened for binding to living cells. We also describe customized slide racks that permit functionalization of 100 slides at a time to produce a cost-efficient, highly reproducible batch process. Ligand spots can be positioned on the glass slides precisely using a microarraying robot, and spot size adjusted for any desired application. Using this system, we demonstrate live cell binding to a variety of ligands and optimize PEG linker length. Taken together, the technology we describe should enable high-throughput screening of disease-specific ligands that bind to living cells.


Genotoxic stress stimulates eDNA release via explosive cell lysis and thereby promotes streamer formation of Burkholderia cenocepacia H111 cultured in a microfluidic device.
DNA is a component of biofilms, but the triggers of DNA release during biofilm formation and how DNA contributes to biofilm development are poorly investigated. One key mechanism involved in DNA release is explosive cell lysis, which is a consequence of prophage induction. In this article, the role of explosive cell lysis in biofilm formation was investigated in the opportunistic human pathogen Burkholderia cenocepacia H111 (H111). Biofilm streamers, flow-suspended biofilm filaments, were used as a biofilm model in this study, as DNA is an essential component of their matrix. H111 contains three prophages on chromosome 1 of its genome, and the involvement of each prophage in causing explosive cell lysis of the host and subsequent DNA and membrane vesicle (MV) release, as well as their contribution to streamer formation, were studied in the presence and absence of genotoxic stress. The results show that two of the three prophages of H111 encode functional lytic prophages that can be induced by genotoxic stress and their activation causes DNA and MVs release by explosive cell lysis. Furthermore, it is shown that the released DNA enables the strain to develop biofilm streamers, and streamer formation can be enhanced by genotoxic stress. Overall, this study demonstrates the involvement of prophages in streamer formation and uncovers an often-overlooked problem with the use of antibiotics that trigger the bacterial SOS response for the treatment of bacterial infections.


Atrial Fibrillation-Associated Ischemic Stroke Patients With Prior Anticoagulation Have Higher Risk for Recurrent Stroke.
Background and Purpose- Ischemic stroke associated with nonvalvular atrial fibrillation (NVAF) despite prior anticoagulation may indicate underlying problems that nullify the stroke-preventing effects of oral anticoagulants. We aimed to evaluate the risk for recurrent stroke in patients with NVAF with prior anticoagulation, compared with that in patients without prior anticoagulation. Methods- This study comprised pooled individual patient data on NVAF-associated acute ischemic stroke or transient ischemic attack from 2011 to 2014 arising from the Clinical Research Collaboration for Stroke in Korea (15 South Korean stroke centers) and the Stroke Acute Management With Urgent Risk-Factor Assessment and Improvement-NVAF registry (18 Japanese stroke centers). Data on 4841 eligible patients from the Clinical Research Collaboration for Stroke in Korea registry were pooled with data on all patients (n=1192) in the Stroke Acute Management with Urgent Risk-factor Assessment and Improvement-NVAF registry. The primary outcome was recurrent ischemic stroke. The secondary outcomes were hemorrhagic stroke and all-cause death. Outcome events were captured up to 1 year after the index event. Results- Among the 6033 patients in the full cohort, 5645 patients were analyzed, of whom 1129 patients (20.0%) had received prior anticoagulation. Median age was 75 years (interquartile range, 69-81 years), and 2649 patients (46.9%) were women. Follow-up data of 4617 patient-years (median follow-up 365 days, interquartile range 335-365 days) were available. The cumulative incidence of recurrent ischemic stroke in patients with prior anticoagulation was 5.3% (60/1129), compared with the 2.9% (130/4516) incidence in patients without prior anticoagulation. The risk for recurrent ischemic stroke was higher in patients with prior anticoagulation than in those without (multivariable Cox shared-frailty model, hazard ratio 1.50 [95% CI, 1.02-2.21]). No significant differences in the risks for hemorrhagic stroke and mortality were seen between the 2 groups. Conclusions- The risk for recurrent ischemic stroke may be higher in NVAF-associated stroke patients with prior anticoagulation than in those without prior anticoagulation. Registration- URL: https://www.clinicaltrials.gov; Unique identifier: NCT01581502.