[Music for institutionalized elderly: perceptions of the intervening musicians].
This qualitative study analyzed musicians' perception of their musical activities with institutionalized elderly in Portugal and France using a questionnaire and the Collective Subject Discourse to organize the data. The musicians see their performance as a human experience; they intentionally look for proximity and communication; they recognize the old persons' reality and wish to transform it; they present difficulties in interpersonal communication and in professional development, and adopt musical, personal and intuitive criteria when choosing their repertoire. The results indicate social, human and professional commitment from the musicians analyzed, and may be used as a source for reflection for the health team.


Identification of lipids that accumulate during the routine storage of prestorage leukoreduced red blood cells and cause acute lung injury.
Lipids accumulate during the storage of red blood cells (RBCs), prime neutrophils (PMNs), and have been implicated in transfusion-related acute lung injury (TRALI). These lipids are composed of two classes: nonpolar lipids and lysophosphatidylcholines based on their retention time on separation by high-pressure liquid chromatography. Prestorage leukoreduction significantly decreases white blood cell and platelet contamination of RBCs; therefore, it is hypothesized that prestorage leukoreduction changes the classes of lipids that accumulate during storage, and these lipids prime PMNs and induce acute lung injury (ALI) as the second event in a two-event in vivo model. RBC units were divided: 50% was leukoreduced (LR-RBCs), stored, and sampled on Day 1 and at the end of storage, Day 42. Priming activity was evaluated on isolated PMNs, and the purified lipids from Day 1 or Day 42 were used as the second event in the in vivo model. The plasma and lipids from RBCs and LR-RBCs primed PMNs, and the LR-RBC activity decreased with longer storage. Unlike RBCs, nonpolar lipids comprised the PMN-priming activity from stored LR-RBCs. Mass spectroscopy identified these lipids as arachidonic acid and 5-, 12-, and 15-hydroxyeicsotetranoic acid. At concentrations from Day 42, but not Day 1, three of four of these lipids individually, and the mixture, primed PMNs. The mixture also caused ALI as the second event in a two-event model of TRALI. We conclude that the nonpolar lipids that accumulate during LR-RBC storage may represent the agents responsible for antibody-negative TRALI.


The impact of certificate of need laws on heart attack mortality: Evidence from county borders.
Certificate of need (CON) regulations requires that health care providers obtain state approval before offering a new service or expanding existing facilities. The purported goal of CON regulations is to reduce health care costs by generating regional economies of scale and reducing redundant investments resulting from excessive competition. Critics of CON regulations note that the regulatory environment increases the costs of expansion and may incentivize health care providers to forgo capital investment, which can have a negative effect on health outcomes. To estimate the net effect of CON regulations, I use a border discontinuity design to measure within-regional heart attack mortality spanning 1968 to 1982. I estimate that CON regulations led to an increase in heart attack deaths, by 6%-10%, three years after the policy was enacted.


There are no racial, age, sex, or weight differences in the effect of salt on blood pressure in salt-sensitive hypertensive patients.
This report is part of a larger, multicenter, placebo-controlled study designed to test the effects of low and high salt intake on the antihypertensive action of enalapril maleate or isradipine in salt-sensitive, hypertensive patients. To present our findings with respect to the effects of race, age, sex, and weight on the blood pressure response to low and high salt intake in salt-sensitive hypertensive patients before randomization into the larger study. After 3 week (weeks -9 to -6) of ad lib salt intake (100-200 mmol/d of sodium), 1916 patients whose sitting diastolic blood pressure was between 95 and 115 mm Hg entered a 3-week period (week -6 to -3) of low salt intake (50-80 mmol/d of sodium) and then a 3-week period (week -3 to 0) of high salt intake (200-250 mmol/d of sodium). Of the 1916 patients, 624 were identified as being sensitive to salt by demonstrating an increase in sitting diastolic blood pressure of equal to or more than 5 mm Hg from the low to high salt intake. Of these patients, 367 were white, 156 were black, 92 were Hispanic, 8 were Asian, and 1 was American Indian. Also, 315 were men and 309, women; 351 were 55 years or younger and 273 were older than 55 years; and 195 had a body mass index of 27 or less and 429 had a body mass index higher than 27. The sitting blood pressure decreased with salt restriction and increased with salt load in all groups of patients (P < .001). There were no statistically significant differences in the blood pressure changes to salt changes by race, age, sex, and weight. This large, multicenter study did not demonstrate any statistically significant effect of race, age, sex, and weight on blood pressure response to salt changes in salt-sensitive hypertensive patients.


Surgery versus epilation for the treatment of minor trichiasis in Ethiopia: a randomised controlled noninferiority trial.
Trachomatous trichiasis can cause corneal damage and visual impairment. WHO recommends surgery for all cases. However, in many regions surgical provision is inadequate and patients frequently decline. Self-epilation is common and was associated with comparable outcomes to surgery in nonrandomised studies for minor trichiasis (<six lashes touching eye). This trial investigated whether epilation is noninferior to surgery for managing minor trichiasis. 1,300 individuals with minor trichiasis from Amhara Regional State, Ethiopia were recruited and randomly assigned (1:1) to receive trichiasis surgery or epilation. The epilation group were given new forceps and epilation training. The surgical group received trichiasis surgery. Participants were examined every 6 months for 2 years by clinicians masked to allocation, with 93.5% follow-up at 24 months. The primary outcome measure ("failure") was ≥five lashes touching the eye or receiving trichiasis surgery during 24 months of follow-up, and was assessed for noninferiority with a 10% prespecified noninferiority margin. Secondary outcomes included number of lashes touching, time to failure, and changes in visual acuity and corneal opacity. Cumulative risk of failure over 24 months was 13.2% in the epilation group and 2.2% in the surgical group (risk difference = 11%). The 95% confidence interval (8.1%-13.9%) includes the 10% noninferiority margin. Mean number of lashes touching the eye was greater in the epilation group than the surgery group (at 24 months 0.95 versus 0.09, respectively; p<0.001); there was no difference in change in visual acuity or corneal opacity between the two groups. This trial was inconclusive regarding inferiority of epilation to surgery for the treatment of minor trichiasis, relative to the prespecified margin. Epilation had a comparable effect to surgery on visual acuity and corneal outcomes. We suggest that surgery be performed whenever possible but epilation be used for treatment of minor trichiasis patients without access to or declining surgery. ClinicalTrials.gov NCT00522912.


An amino-terminal fragment of hemokinin-1 has an inhibitory effect on pruritic processing in rats.
Hemokinin-1 (HK-1) is a peptide encoded by the preprotachykinin gene, TAC-4, and shares the hydrophobic carboxyl-terminal (C-terminal) region common to mammalian tachykinin peptides, such as substance P (SP). It is generally believed that C-terminal fragments of SP elicit an excitatory effect, while pretreatment with amino-terminal (N-terminal) fragments of SP inhibits the function of SP; however, there is no available information on HK-1. Therefore, to clarify the characteristics of C-terminal and N-terminal fragments of HK-1, HK-1 was divided into HK-1 (1-5) as the N-terminal fragment and HK-1 (6-11) as the C-terminal fragment based on the similarity of amino acids between HK-1 and SP. Intrathecal administration of HK-1 (6-11) induced scratching behavior similar to HK-1, while HK-1 (1-5) hardly induced scratching. Pretreatment with HK-1 (1-5), however, attenuated scratching induced by HK-1 and SP, whereas pretreatment with SP (1-5) attenuated SP-induced scratching, but not HK-1. Furthermore, intrathecal administration of HK-1 (1-5) and SP (1-5) markedly attenuated the induction of flinching and enhancement of c-Fos expression in the spinal cord following the intradermal administration of formalin, a noxious stimulant, while pretreatment with HK-1 (1-5), but not SP (1-5), markedly attenuated the induction of scratching behavior by subcutaneous administration of pruritic agents, such as serotonin or histamine. Taken together, these findings indicate that HK-1 (1-5) suppresses pruritic and nociceptive processing, while SP (1-5) suppresses nociceptive processing. Therefore, it is suggested that HK-1 (1-5) may be a useful tool for revealing pruritic processing and HK-1 may play a crucial role in pruritic processing.


Bifurcation from stable holes to replicating holes in vibrated dense suspensions.
In vertically vibrated starch suspensions, we observe bifurcations from stable holes to replicating holes. Above a certain acceleration, finite-amplitude deformations of the vibrated surface continue to grow until void penetrates fluid layers, and a hole forms. We studied experimentally and theoretically the parameter dependence of the holes and their stabilities. In suspensions of small dispersed particles, the circular shapes of the holes are stable. However, we find that larger particles or lower surface tension of water destabilize the circular shapes; this indicates the importance of capillary forces acting on the dispersed particles. Around the critical acceleration for bifurcation, holes show intermittent large deformations as a precursor to hole replication. We applied a phenomenological model for deformable domains, which is used in reaction-diffusion systems. The model can explain the basic dynamics of the holes, such as intermittent behavior, probability distribution functions of deformation, and time intervals of replication. Results from the phenomenological model match the linear growth rate below criticality that was estimated from experimental data.


From genes to proteins: in vitro expression of rickettsial proteins.
The availability of the complete genome sequences of several organisms allows the comparative analysis of genomes, a branch of bioinformatics known as genomics. With this approach, much can be learned about the biology of organisms that are difficult to culture, even when few, if any, of their proteins have been isolated and studied directly. We have focused our interest on Rickettsia conorii, an obligate intracellular bacterium responsible for Mediterranean spotted fever, a disease endemic in southern Europe. While bioinformatic annotation of the complete genome of this bacteria has allowed identification of 1,374 genes, a large number of them remain functionally uncharacterized. The final goal of many experiments in molecular biology is to use biological systems to synthesize the protein encoded by the gene being studied. Because three-dimensional structures are more resilient to evolution and change than amino acid sequences, structure determination of some open reading frames should also exhibit structural similarity to previously described protein families. We have thus initiated a systematic expression and structure determination program for the proteins encoded by rickettsial genes of interest. We have cloned different genes of R. conorii by recombinational cloning (GATEWAY), Invitrogen) a method that uses in vitro site-specific recombination to accomplish a directional cloning of PCR products and the subsequent automatic subcloning of the DNA segment into new vector backbones at high efficiency. The constructions in p-Dest17 yielded several clones able to express recombinant proteins with a C-terminal histidine tag. Expression of corresponding proteins was then performed using a cell-free protein expression system (Rapid Translation System, RTS, Roche Diagnostics). The recombinational cloning approach coupled to RTS provides an approach to rapid optimization of protein expression and is very useful to express rickettsial proteins. Moreover, this system is able to overcome some of the limitations encountered with rickettsial proteins highly toxic for E. coli or insect cells.


Prevalence rates of infection in intensive care units of a tertiary teaching hospital.
To determine the prevalence rates of infections among intensive care unit patients, the predominant infecting organisms, and their resistance patterns. To identify the related factors for intensive care unit-acquired infection and mortality rates. A 1-day point-prevalence study. A total of 19 intensive care units at the Hospital das Clínicas-University of São Paulo, School of Medicine (HC-FMUSP), a teaching and tertiary hospital, were eligible to participate in the study. All patients over 16 years old occupying an intensive care unit bed over a 24-hour period. The 19 intensive care unit s provided 126 patient case reports. Rates of infection, antimicrobial use, microbiological isolates resistance patterns, potential related factors for intensive care unit-acquired infection, and death rates. A total of 126 patients were studied. Eighty-seven patients (69%) received antimicrobials on the day of study, 72 (57%) for treatment, and 15 (12%) for prophylaxis. Community-acquired infection occurred in 15 patients (20.8%), non-intensive care unit nosocomial infection in 24 (33.3%), and intensive care unit-acquired infection in 22 patients (30.6%). Eleven patients (15.3%) had no defined type. The most frequently reported infections were respiratory (58.5%). The most frequently isolated bacteria were Enterobacteriaceae (33.8%), Pseudomonas aeruginosa (26.4%), and Staphylococcus aureus (16.9%; [100% resistant to methicillin]). Multivariate regression analysis revealed 3 risk factors for intensive care unit-acquired infection: age > or = 60 years (p = 0.007), use of a nasogastric tube (p = 0.017), and postoperative status (p = 0.017). At the end of 4 weeks, overall mortality was 28.8%. Patients with infection had a mortality rate of 34.7%. There was no difference between mortality rates for infected and noninfected patients (p = 0.088). The rate of nosocomial infection is high in intensive care unit patients, especially for respiratory infections. The predominant bacteria were Enterobacteriaceae, Pseudomonas aeruginosa, and Staphylococcus aureus (resistant organisms). Factors such as nasogastric intubation, postoperative status, and age 60 years were significantly associated with infection. This study documents the clinical impression that prevalence rates of intensive care unit-acquired infections are high and suggests that preventive measures are important for reducing the occurrence of infection in critically ill patients.


What Do They Want from Us? A Survey of EM Program Directors on EM Application Criteria.
Although a relatively young specialty, emergency medicine (EM) is popular among medical students and is one of the most competitive large specialties. Consequently, students increasingly seek more opportunity to differentiate themselves from their colleagues by pursuing more clerkships at the cost of taking out additional loans: this despite the fact that those who match in EM typically do so in their top three choices. We sought to ascertain what factors EM program directors seek in their typical candidate. We recruited EM program directors via the Council of Emergency Medicine Residency Directors email listserv to participate in an anonymous survey regarding the United States Medical Licensing Examination (USMLE), the number of standardized letters of evaluation (SLOE), and the number of EM rotations during the fourth year. 135 respondents completed the anonymous survey: 59% of respondents stated their program did not have a minimum USMLE Step 1 score, but 39% reported a minimum score of 210 or higher; 95% of programs do not require Step 2 to grant an interview, but 46% require it to place the student on the rank list; 80% require only one EM rotation to grant an interview and none require more than two; 95% of programs will accept two SLOEs for both application and rank list placement. For the typical EM applicant, there is likely little benefit to performing more than two rotations and obtaining more than two SLOEs. Students can defer USMLE Step 2 but must complete it by the time rank lists are due. Our study was limited by the anonymity of the survey, and comments by the respondents revealed the questions did not account for some nuances programs apply to their application review process.


Concordance of self-reported and medical chart information on cancer diagnosis and treatment.
Self-reported information is an important tool for collecting clinical information for epidemiologic studies and in clinical settings where electronic medical records are not employed and shared. Using data collected from the Shanghai Breast Cancer Survival Study (SBCSS), a population-based, prospective cohort study of 5,042 women diagnosed with breast cancer in Shanghai, China, we compared the concordance of patient questionnaire responses to a survey administered approximately 6 months after cancer diagnosis with medical chart information obtained from the diagnostic hospitals for several disease and treatment-related variables. Of 5,042 SBCSS participants, medical chart information was available for 4,948 women (98.1%). Concordance between patient self-reported and medical chart information was high for the majority of disease-related variables, including: diagnosing hospital (agreement: 98.7%, kappa: 0.99), type of surgery conducted (94.0%, 0.53), ER/PR status (94.5%, 0.91), and tumor position (98.2%, 0.97), as well as for important calendar dates, such as date of diagnosis, surgery, and first chemotherapy treatment. The 10 most commonly used chemotherapeutic drugs were all reported with agreement rates of at least 82%, with associated kappa values that ranged from 0.41 for calcium folinate to 0.76 for vinorelbine. Our study found high validity for patient self-reported information for a variety of disease and treatment-related variables, suggesting the utility of self-reports as an important source of clinical information for both epidemiological research and patient care.


IL-6 originated from breast cancer tissue-derived mesenchymal stromal cells may contribute to carcinogenesis.
Tumor microenvironment is an important factor, which sustains and promotes the tumors by inflammatory signals. Interleukin-6 (IL-6) is known as a multifunctional cytokine, which is a major activator of the signaling pathway of Janus kinases (JAKs)/signal transducer and activator of transcription 3 (STAT3). In this study, we aimed to investigate the effect of IL-6 in the tumor microenvironment on carcinogenesis. For this purpose, healthy breast tissue-derived stromal cells (HBT-SCs) and malign breast tissue-derived stromal cells (MBT-SCs) were co-cultured with MCF-7 (human breast adenocarcinoma cell line) cells using semipermeable membranes. The cell proliferation was monitored with water-soluble tetrazolium (WST) and carboxyfluorescein succinimidyl ester (CFSE) assays. Protein levels were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot hybridization, while gene expressions were measured by real-time PCR. The results demonstrated that IL-6 protein levels increased significantly in the supernatants of MBT-SCs when they were co-cultured with MCF-7 cells. In accordance with this, the expression of IL-6 was significantly higher in MBT-SCs. Additionally, the expression of STAT3 in MCF-7 cells increased slightly when they were co-cultured with MBT-SCs. Considering together, there is an important interaction between tumor microenvironment and tumor cells mediated by IL-6 signaling. Thereby, the targeting on IL-6 signaling in the treatment of cancer might effectively prevent the tumor progression.


Enhanced Rice Resistance to Sheath Blight through Nitrate Transporter 1.1B Mutation without Yield Loss under NH4+ Fertilization.
Nitrogen fertilization can promote rice yield but decrease resistance to sheath blight (ShB). In this study, the nitrate transporter 1.1b (nrt1.1b) mutant that exhibited less susceptibility to ShB but without compromising yield under NH4+ fertilization was screened. NRT1.1B's regulation of ShB resistance was independent of the total nitrogen concentration in rice under NH4+ conditions. In nrt1.1b mutant plants, the NH4+ application modulated auxin signaling, chlorophyll content, and phosphate signaling to promote ShB resistance. Furthermore, the findings indicated that NRT1.1B negatively regulated ShB resistance by positively modulating the expression of H+-ATPase gene OSA3 and phosphate transport gene PT8. The mutation of OSA3 and PT8 promoted ShB resistance by increasing the apoplastic pH in rice. Our study identified the ShB resistance mutant nrt1.1b, which maintained normal nitrogen use efficiency without compromising yield.


Color constancy for multiple light sources.
Color constancy algorithms are generally based on the simplifying assumption that the spectral distribution of a light source is uniform across scenes. However, in reality, this assumption is often violated due to the presence of multiple light sources. In this paper, we will address more realistic scenarios where the uniform light-source assumption is too restrictive. First, a methodology is proposed to extend existing algorithms by applying color constancy locally to image patches, rather than globally to the entire image. After local (patch-based) illuminant estimation, these estimates are combined into more robust estimations, and a local correction is applied based on a modified diagonal model. Quantitative and qualitative experiments on spectral and real images show that the proposed methodology reduces the influence of two light sources simultaneously present in one scene. If the chromatic difference between these two illuminants is more than 1°, the proposed framework outperforms algorithms based on the uniform light-source assumption (with error-reduction up to approximately 30%). Otherwise, when the chromatic difference is less than 1° and the scene can be considered to contain one (approximately) uniform light source, the performance of the proposed method framework is similar to global color constancy methods.


Predicting Recruitment Feasibility for Acute Spinal Cord Injury Clinical Trials in Canada Using National Registry Data.
Traumatic spinal cord injury (SCI) represents a significant burden of illness, but it is relatively uncommon and heterogeneous, making it challenging to achieve sufficient subject enrollment in clinical trials of therapeutic interventions for acute SCI. The Rick Hansen Spinal Cord Injury Registry (RHSCIR) is a national SCI Registry that enters patients with SCI from acute-care centers across Canada. To predict the feasibility of conducting clinical trials of acute SCI within Canada, we have applied the inclusion/exclusion criteria of six previously conducted SCI trials to the RHSCIR data set and generated estimates of how many Canadian persons would have been eligible theoretically for enrollment in these studies. Data for SCI cases were prospectively collected for RHSCIR at 18 acute and 13 rehabilitation sites across Canada. RHSCIR patients enrolled between 2009-2013 who met the following key criteria were included: non-penetrating traumatic SCI; received acute care at a RHSCIR site; age more than 18, less than 75 years, and had complete admission single neurological level of injury data. Inclusion and exclusion criteria for the Minocycline in Acute Spinal Cord injury (Minocycline), Riluzole, Surgical Timing in Acute Spinal Cord Injury Study (STASCIS), Cethrin, Nogo antibody study (NOGO), and Sygen studies were applied retrospectively to this data set. The numbers of patients eligible for each clinical trial were determined. There were 2166 of the initial 2714 patients (79.8%) who met the key criteria and were included in the data set. Projected annual numbers of eligible patients for each trial were: Minocycline, 117; Riluzole, 62; STASCIS, 109; Cethrin, 101; NOGO, 82; and Sygen, 70. An additional 8.0% of the sample had a major head injury (Glasgow Coma Scale [GCS] score ≤12) and would have been excluded from the trials. RHSCIR provides a comprehensive national data set that may serve as a useful tool in the planning of multicenter clinical SCI trials.


Availability of schizophrenic patients and their families for genetic linkage studies: findings from the Maryland epidemiology sample.
It has been suggested that collections of affected sib pairs, or their nuclear families, may be an efficient method for screening for genetic linkages in schizophrenia. We present the data collected in five years from 15 hospitals in the state of Maryland in an effort to determine if such a collection scheme will be feasible. Probands in our sample were eligible for inclusion in the sample if they were white, were age 16 years or older, and carried a research diagnosis of schizophrenia. Family data are reported for 258 probands. Using the most stringent category of affected (RDC schizophrenia) revealed ten families with two or more affected sibs. The broadest category of affected (any psychotic disorder or psychiatric hospitalization) identified only 36 families with two or more affected sibs. We conclude that, if schizophrenia is a heterogeneous disorder with decreased penetrance, an effort to collect multiplex nuclear families is unlikely to provide enough data to identify genetic linkage. Alternatively, an effort to seek out and collect larger multiplex, multigenerational families rather than a collection of affected sib pairs may be more efficacious.


Undergraduate midwifery students' first experiences with stillbirth and neonatal death.
While midwifery practice predominantly deals with happy experiences, unexpected and unpredictable events occur. This paper reports on a study that explored undergraduate midwifery students' first experiences of stillbirth and neonatal death. It sought to better understand their encounters to ensure curricula were responsive to students' needs. Semi-structured interviews were conducted with eight Bachelor of Midwifery students. Many were found to have been exposed to perinatal death early in their courses and were ill prepared for the confrontations. Furthermore, the existence of support services was varied. Dealing with these experiences appears to occur through a developmental process and resonates with previous research around compassion, fatigue and posttraumatic stress disorder. Overall, students require specific preparation prior to undertaking placements, as well as clearly identified support strategies if such experiences are encountered.


Diverse groups of plant RNA and DNA viruses share related movement proteins that may possess chaperone-like activity.
Amino acid sequences of plant virus proteins mediating cell-to-cell movement were compared to each other and to protein sequences in databases. Two families of movement proteins have been identified, the members of which show statistically significant sequence similarity. The first, larger family (I) encompasses the movement proteins of tobamo-, tobra-, caulimo- and comoviruses, apple chlorotic leaf spot virus (ACLSV) and geminiviruses with bipartite genomes. Thus this family includes viruses which move by two methods, those requiring the coat protein for the cell-to-cell spread (comoviruses) and those not having this requirement (tobamoviruses). The previously unsuspected relationship between the movement proteins of RNA and DNA viruses having no RNA stage in their life cycle (geminiviruses) suggested that their movement mechanisms might be similar. The second, smaller family (II) consists of the movement proteins of tricornaviruses (bromoviruses, cucumoviruses, alfalfa mosaic virus and tobacco streak virus) and dianthoviruses. Alignment of the sequences of family I movement proteins highlighted two motifs, centred at conserved Gly and Asp residues, respectively, which are assumed to be crucial for the movement protein function(s). Screening the amino acid sequence database revealed another conserved motif that is shared by a large subset of family I movement proteins (those of caulimo- and comoviruses, and ACLSV) and the family of cellular 90K heat shock proteins (HSP90). Based on the analogy to HSP90, it is speculated that many plant virus movement proteins may mediate virus transport in a chaperone-like manner.


Cloning and characterization of a 92 kDa soluble phosphatidylinositol 4-kinase.
A phosphatidylinositol (PtdIns) 4-kinase cDNA cloned from a rat brain cDNA library encoded a protein of 816 amino acids with a calculated molecular mass of 91654 Da. This molecule contained a lipid-kinase-unique domain and a presumed lipid/ protein kinase homology domain that are found in other PtdIns 4-kinases and PtdIns 3-kinases. Furthermore, this kinase molecule had 43.3% shared identity with the presumed catalytic domain of yeast PtdIns 4-kinase, PtdInsK1, and the two molecules had a region of similarity that is not conserved in other lipid kinases. By examining PtdIns kinase activity in transfected COS-7 cells using epitope tag immunoprecipitation as well as conventional methods, the product PtdIns phosphate was identified as phosphatidylinositol 4-phosphate (PtdIns4P), but not phosphatidylinositol 3-phosphate (PtdIns3P). The PtdIns 4-kinase activity was recovered predominantly from the soluble fraction and the activity was markedly enhanced in the presence of Triton X-100 and was relatively insensitive to inhibition by adenosine. In addition, the PtdIns 4-kinase activity was completely inhibited in the presence of 10 microM wortmannin. When examined by epitope tag immunocytochemistry, the immunoreactivity for the PtdIns 4-kinase molecule was dominantly aggregated in a cytoplasmic region juxtaposed to the nuclei and was faintly but widely dispersed in the cytoplasm. By in situ hybridization analysis, the mRNA for PtdIns 4-kinase was expressed ubiquitously and was detected in most neurons throughout the grey matter of the brain, with higher expression intensity found in fetal than in adult brain.


Transformation of NIH-3T3 mouse cells by avian retroviral DNAs.
NIH-3T3 cells were found to be transformable by RSV DNA in the absence of progeny virus production. Cells transformed by intact RSV DNAs contained rescuable RSV genomes that were integrated into cellular DNA and were colinear with unintegrated RSV DNA. NIH-3T3 cells were also transformable by subgenomic fragments of RSV DNA, synthesized in vitro or generated by restriction endonuclease digestion of intracellular RSV DNA. These cells did not contain rescuable RSV genomes but did contain RSV DNA fragments that efficiently induced transformation of NIH-3T3 cells in secondary transfection assays. Further analysis of the RSV DNA sequences present in these cells and transformation assays of defined fragments of RSV DNA may contribute to the elucidation of the sequences required for expression of the src gene of RSV. DNAs of the avian acute leukemia viruses MC29 and AEV also induced transformation of NIH-3T3 cells. The use of these cells as recipients may thus provide a system suitable for functional analysis of the transforming genes of avian leukemia viruses as well as sarcoma viruses.


Endocrine Disruptors: Focus on the Adrenal Cortex.
Endocrine-disrupting chemicals (EDCs) are exogenous substances known to interfere with endocrine homeostasis and promote adverse health outcomes. Their impact on the adrenal cortex, corticosteroids and their physiological role in the organism has not yet been sufficiently elucidated. In this review, we collect experimental and epidemiological evidence on adrenal disruption by relevant endocrine disruptors. In vitro data suggest significant alterations of gene expression, cell signalling, steroid production, steroid distribution, and action. Additionally, morphological studies revealed disturbances in tissue organization and development, local inflammation, and zone-specific hyperplasia. Finally, endocrine circuits, such as the hypothalamic-pituitary-adrenal axis, might be affected by EDCs. Many questions regarding the detection of steroidogenesis disruption and the effects of combined toxicity remain unanswered. Not only due to the diverse mode of action of adrenal steroids and their implication in many common diseases, there is no doubt that further research on endocrine disruption of the adrenocortical system is needed.


Expression of ATM in ataxia telangiectasia fibroblasts rescues defects in DNA double-strand break repair in nuclear extracts.
Ataxia telangiectasia (A-T) is a human genetic disorder characterized by progressive cerebellar degeneration, hypersensitivity to ionizing radiation (IR), immunodeficiency, and high cancer risk. At the cellular level, IR sensitivity and increased frequency of spontaneous and IR-induced chromosomal breakage and rearrangements are the hallmarks of A-T. The ATM gene, mutated in this syndrome, has been cloned and codes for a protein sharing homology with DNA-PKcs, a protein kinase involved in DNA double-strand break (DSB) repair and DNA damage responses. The characteristics of the A-T cellular phenotypes and ATM gene suggest that ATM may play a role similar to that of DNA-PKcs in DSB repair and that there is a primary DNA repair defect in A-T cells. In the current study, the function of ATM in DNA DSB repair was evaluated in an in vitro system using two plasmids, carrying either an EcoRI-induced DSB within the lacZalpha gene or various endonuclease-induced DSB in the SupF suppressor tRNA gene. We found that the DSB repair efficiency in A-T nuclear extracts was comparable to, if not higher than, that in normal nuclear extracts. However, the repair fidelity in A-T nuclear extracts was decreased when repairing DSB with short 5' and 3' overhangs (<4 base pairs (bp)) or blunt ends, but not 5' 4-bp overhangs. Sequencing of the mutant plasmids revealed that deletions involving 1-6 nucleotide microhomologies were the major class of mutations in both A-T and normal extracts. However, the size of the deletions in plasmids from A-T nuclear extracts was larger than that from normal nuclear extracts. Expression of the ATM protein in A-T cells corrected the defect in DSB repair in A-T nuclear extracts. These results suggest that ATM plays a role in maintaining genomic stability by preventing the repair of DSB from an error-prone pathway.


Psychological mechanisms in a digital intervention to improve physical activity: A multicentre randomized controlled trial.
The randomized controlled trial examined factors that might be responsible for individual differences in physical activity change among men and women who participated in a lifestyle intervention. The main purpose of the analyses regarded the role of psychological mechanisms involving motivation, planning, self-monitoring, and habit strength. A two-arm digital intervention was conducted in Italy, Spain, and Greece to improve physical activity levels, with follow-ups at 3 and 6 months after baseline assessment. Participants were 1,564 adults at baseline, n = 638 at 6-month follow-up. Linear mixed models examined the intervention effects, and a two-group longitudinal structural equation model explored which psychological constructs (motivation, planning, self-monitoring, habit strength) were associated with changes in physical activity. In addition to an overall increase in self-reported activity, there were interactions between time and sex and between time and experimental groups, and a triple interaction between time, sex, and experimental groups, indicating that men reported an increase in activity independent of groups, whereas women in the active control group did not benefit from the intervention. Planning, self-monitoring, and habit strength mediated sequentially between initial motivation and follow-up physical activity. Although the intervention produced overall improvements in physical activity, the time-by-treatment interaction emerged only for women. The mechanism included a sequence leading from motivation via planning, self-monitoring, and habit strength towards physical activity. Statement of contribution What is already known on this subject? Digital lifestyle interventions can be effective in terms of physical activity performance gains. Men are on average more physically active than women. Long-term adherence rates to digital interventions are usually low. What does this study add? Giving users of an online platform more interactive options did not make a difference. Women gained more than men from adaptive, dynamic online platform content. Individual characteristics (motivation, planning, self-monitoring, habit) were more important than online treatment features.


Production of reactive oxygen species after reperfusion in vitro and in vivo: protective effect of nitric oxide.
Thrombolytic treatments for ischemic stroke can restore circulation, but reperfusion injury, mediated by oxygen free radicals, can limit their utility. The authors hypothesized that, during reperfusion, nitric oxide (NO) provides cytoprotection against oxygen free radical species. Levels of NO and oxygen free radicals were determined in both reoxygenation in vitro and reperfusion in vivo models using an NO electrochemical probe and high-performance liquid chromatography with the 2,3- and 2,5-dihydroxybenzoic acid trapping method, before and after addition of the NO donor diethanolamine nitric oxide (DEA/NO). Reoxygenation after anoxia produced a twofold increase in NO release by human fetal astrocytes and cerebral endothelial cells (p < 0.005). In both cell lines, there was also a two- to threefold increase in oxygen free radical production (p < 0.005). In human fetal astrocytes and cerebral endothelial cells given a single dose of DEA/NO, free radical production dropped fivefold compared with peak ischemic levels (p < 0.001). In a study in which a rat global cerebral ischemia model was used, NO production in a vehicle-treated group increased 48 +/- 16% above baseline levels after reperfusion. After intravenous DEA/NO infusion, NO reached 1.6 times the concentration of the postischemic peak in vehicle-treated animals. In vehicle-treated animals during reperfusion, free radical production increased 4.5-fold over basal levels (p < 0.01). After intravenous DEA/NO infusion, free radical production dropped nearly 10-fold compared with peak levels in vehicle-treated animals (p < 0.006). The infarct volume in the vehicle-treated animals was 111 +/- 16.9 mm3; after DEA/NO infusion it was 64.8 +/- 23.4 mm3 (p < 0.01). The beneficial effect of early restoration of cerebral circulation after cerebral ischemia is limited by reperfusion injury. These results indicate that NO release and oxygen free radical production increase during reperfusion, and suggest a possible early treatment of reperfusion injury using NO donors.


The Minority and Rural Coronavirus Insights Study: Design and Baseline Characteristics of a Minority Cohort.
The Minority and Rural Coronavirus Insights Study (MRCIS) is an ongoing prospective cohort study examining health disparities associated with SARS-CoV-2 infection among medically underserved populations. This report describes procedures implemented to establish the MRCIS cohort and examines the factors associated with the molecular and serological assessment of SARS-CoV-2 infection status at participant enrollment. Participants were recruited from 5 geographically dispersed federally qualified health centers between November 2020 and April 2021. At baseline, participants completed a detailed demographic survey and biological samples were collected for testing. SARS-CoV-2 infection status was determined based on the combined molecular and serological test results. Chi-squared and logistic regression analyses were conducted to examine associations between sociodemographic factors, COVID-19 safety measures, existing comorbidities, and SARS-CoV-2 infection status. The final cohort included 3238 participants. The mean age of participants was 50.2 ± 15.8 years. Most participants identified as female (60.0%), heterosexual or straight (93.0%), White (47.6%), and Hispanic or Latino (49.1%). Approximately 26.1% of participants had at least one positive SARS-CoV-2 test result. The main effect model included age, sex, and race/ethnicity. Compared with adults ≥65 years, participants in all other age groups had ∼2 times increased odds of a positive SARS-CoV-2 test result. In addition, racial/ethnic minorities had ∼2 times increased odds of a positive SARS-CoV-2 infection status compared with non-Hispanic Whites. A unique cohort of a traditionally medically underserved minority population was established. Significant racial and ethnic disparities in SARS-CoV-2 infection status at baseline were discovered.


Keratinization in atypical glandular cell clusters as a cytological clue to endometrioid carcinoma on cervical cytology.
Specific diagnosis of endometrial carcinomas on cervical cytology is difficult with few useful cytomorphological clues reported. This study reviews a cohort of cervical cytology to investigate the presence of keratinization in atypical glandular cells (AGC), an undescribed cytomorphological clue for identifying endometrial endometrioid carcinomas on cervical cytology. Cervical cytology slides from patients with a histologic diagnosis of endometrial endometrioid carcinoma were reviewed for the presence of keratinization associated with AGCs. Corresponding histology slides were reviewed for tumour grading and degree of squamous differentiation. In total, 42 cases of cervical cytology specimens from 41 patients were retrieved, including 7 (16.7%) with keratinization associated with AGCs seen and 35 (83.3%) without. Comparison of histologic grading did not demonstrate an association with the presence of keratinization on cytology (p = 0.565). Corresponding histology slides were available for 37 cases. Cytologic and histologic keratinization were associated statistically (p = 0.002). Frank keratinization was seen on histologic slides of five cases, with four also showing cytologic keratinization. Area of squamous differentiation, including squamous morule formation, did not correlate with keratinization on cytologic preparation (p = 0.185). Histologic and cytologic keratinization are observed in endometrioid endometrial carcinomas. Such is reflected in cervical cytology by the presence of orangeophilic, rigid and acellular fragments within or associated with AGC clusters. Keratinization, when identified with AGCs, should be regarded as a cytologic clue suggestive of an endometroid carcinoma of endometrial origin.


Factors affecting the uptake and retention of Vibrio vulnificus in oysters.
Vibrio vulnificus, a bacterium ubiquitous in oysters and coastal water, is capable of causing ailments ranging from gastroenteritis to grievous wound infections or septicemia. The uptake of these bacteria into oysters is often examined in vitro by placing oysters in seawater amended with V. vulnificus. Multiple teams have obtained similar results in studies where laboratory-grown bacteria were observed to be rapidly taken up by oysters but quickly eliminated. This technique, along with suggested modifications, is reviewed here. In contrast, the natural microflora within oysters is notoriously difficult to eliminate via depuration. The reason for the transiency of exogenous bacteria is that those bacteria are competitively excluded by the oyster's preexisting microflora. Evidence of this phenomenon is shown using in vitro oyster studies and a multiyear in situ case study. Depuration of the endogenous oyster bacteria occurs naturally and can also be artificially induced, but both of these events require extreme conditions, natural or otherwise, as explained here. Finally, the "viable but nonculturable" (VBNC) state of Vibrio is discussed. This bacterial torpor can easily be confused with a reduction in bacterial abundance, as bacteria in this state fail to grow on culture media. Thus, oysters collected from colder months may appear to be relatively free of Vibrio but in reality harbor VBNC cells that respond to exogenous bacteria and prevent colonization of oyster matrices. Bacterial-uptake experiments combined with studies involving cell-free spent media are detailed that demonstrate this occurrence, which could explain why the microbial community in oysters does not always mirror that of the surrounding water.


ATM augments nuclear stabilization of DYRK2 by inhibiting MDM2 in the apoptotic response to DNA damage.
The tumor suppressor p53 is a transcription factor that regulates cell cycle, DNA repair, senescence, and apoptosis in response to DNA damage. Phosphorylation of p53 at Ser-46 is indispensable for the commitment to apoptotic cell death. A previous study has shown that upon exposure to genotoxic stress, DYRK2 translocates into the nucleus and phosphorylates p53 at Ser-46, thereby inducing apoptosis. However, less is known about mechanisms responsible for intracellular control of DYRK2. Here we show the functional nuclear localization signal at N-terminal domain of DYRK2. Under normal conditions, nuclear and not cytoplasmic DYRK2 is ubiquitinated by MDM2, resulting in its constitutive degradation. In the presence of proteasome inhibitors, we detected a stable complex of DYRK2 with MDM2 at the nucleus. Upon exposure to genotoxic stress, ATM phosphorylates DYRK2 at Thr-33 and Ser-369, which enables DYRK2 to escape from degradation by dissociation from MDM2 and to induce the kinase activity toward p53 at Ser-46 in the nucleus. These findings indicate that ATM controls stability and pro-apoptotic function of DYRK2 in response to DNA damage.


Simulation and CRM.
Patients are harmed as a result of incidents. Both poor interdisciplinary communication and teamwork are contributing factors to such events. The principles of crisis resource management are meant to help prevent and manage difficulties and reflect both, the social-team-oriented and cognitive-individual-oriented aspects of human factors. This article explores the importance of human factors training for safe care of patients and the role of simulation. Based on the available literature, the need to integrate this type of training to increase awareness of the importance of human factors and to change attitudes appears obvious. A combination of different training methods appears to be useful. Simulation-based training appears to be favourable, although the number of studies demonstrating the impact of training is limited. It is important to develop training programmes for individual teams, based on the knowledge of challenges and deficiencies, and to monitor behavioural change. Several methods, including patient safety data, interviews, observational studies and simulations, can be used to specify learning objectives. The training should be established for the real team(s). Furthermore, leaders need to implement training in the organisation and establish databases to monitor the impact on patient outcome.


The emotional well-being of young people having a parent with younger onset dementia.
Younger onset dementia (YOD) not only affects the person with the diagnosis but the whole family, which often includes young people. A limited body of research on this group of young people indicates that they experience varying degrees of emotional trauma. We explored the lived experiences of young people having a parent with YOD from the perspective of the social model of disability. Data were available from semi-structured interviews with 12 young people who had a parent with YOD looking at their lived experiences between 8 and 24 years. Thematic analysis identified four main themes: the emotional toll of caring, keeping the family together, grief and loss and psychological distress The social model of disability theory provides a helpful framework for these families who experience significant emotional distress, demonstrating that the disability is often socially constructed by a society, which marginalizes and excludes them. A 'whole family' approach is proposed, where the needs of young people and their parents are respected and responded to age appropriately.


Differences in color vision make passerines less conspicuous in the eyes of their predators.
Sexual selection often favors brighter and exaggerated traits, which also increase the risk of detection by predators. Signals that are preferentially conspicuous to conspecifics would reduce the predation cost of signaling and, therefore, might facilitate the evolution of stronger sexual and social signals. This selective signaling is possible if predators and prey have differently tuned sensory systems. By using a retinal model to compare reflectance from the plumages of Swedish songbirds to the reflectance of their natural backgrounds, we found their color badges to be significantly more conspicuous to other songbirds (which have a UV-tuned visual system) than to raptors and corvids (which have a violet-tuned system) in both coniferous and deciduous forests, consistent with an adaptive private communication system.


Identification of membrane-bound calcium, calmodulin-dependent protein kinase II in canine heart.
Phospholamban, the putative regulatory proteolipid of the Ca2+/Mg2+ ATPase in cardiac sarcoplasmic reticulum, was selectively phosphorylated by a Ca2+/calmodulin (CaM)-dependent protein kinase associated with a cardiac membrane preparation. This kinase also catalyzed the phosphorylation of two exogenous proteins known to be phosphorylated by the multifunctional Ca2+/CaM-dependent protein kinase II (Ca2+/CaM-kinase II), i.e., smooth muscle myosin light chains and glycogen synthase a. The latter protein was phosphorylated at sites previously shown to be phosphorylated by the purified multifunctional Ca2+/CaM-kinase II from liver and brain. The membrane-bound kinase did not phosphorylate phosphorylase b or cardiac myosin light chains, although these proteins were phosphorylated by appropriate, specific calmodulin-dependent protein kinases added exogenously. In addition to phospholamban, several other membrane-associated proteins were phosphorylated in a calmodulin-dependent manner. The principal one exhibited a Mr of approximately 56,000, a value similar to that of the major protein (57,000) in a partially purified preparation of Ca2+/CaM-kinase II from the soluble fraction of canine heart that was autophosphorylated in a calmodulin-dependent manner. These data indicate that the membrane-bound, calmodulin-dependent protein kinase that phosphorylates phospholamban in cardiac membranes is not a specific calmodulin-dependent kinase, but resembles the multifunctional Ca2+/CaM-kinase II. Our data indicate that this kinase may be present in both the particulate and soluble fractions of canine heart.


Diet-Related Risk Factors for Chronic Noncommunicable Diseases in Italian Prisoners: B.A.C.I. (Benessere All'interno delle Carceri Italiane, Well-Being Inside the Italian Prisons) Project by the Italian Society of Penitentiary Medicine and Public Health (S.I.M.S.Pe. Società Italiana di Medicina e Sanità Penitenziaria).
The review aims to present an overview of inmate health, focusing on lifestyle-related diseases, physical activity levels, and nutritional status. It also presents the B.A.C.I. (Benessere All'interno delle Carceri Italiane, well-being inside the Italian prisons) project, which aims to offers an innovative path of prevention, diagnosis, and treatment of noncommunicable diseases (NCDs) related to unhealthy lifestyles in prisons in the Campania region, Italy. The global prison population has risen by 24% since the year 2000, with over 10.77 million people detained worldwide in 2021. In Italy alone, there are currently over 57,000 inmates. Inmates face a higher risk of NCDs such as cardiovascular disease due to unhealthy lifestyles characterized by poor diets and lack of physical activity. Additionally, sleep disorders, particularly insomnia, are prevalent among inmates, further contributing to health disparities. While physical activity has shown positive effects on inmate well-being, there is limited research on nutritional status and interventions in prison populations. Providing quality healthcare to inmates is an international policy norm, but the standards vary globally and are often inadequate. The economic burden of NCDs is rising, and this is exacerbated in prisons, making it challenging for individuals to reintegrate into society after release.


Comparison of Different Phenotypic Approaches To Screen and Detect mecC-Harboring Methicillin-Resistant Staphylococcus aureus.
Similar to mecA, mecC confers resistance against beta-lactams, leading to the phenotype of methicillin-resistant Staphylococcus aureus (MRSA). However, mecC-harboring MRSA strains pose special difficulties in their detection. The aim of this study was to assess and compare different phenotypic systems for screening, identification, and susceptibility testing of mecC-positive MRSA isolates. A well-characterized collection of mecC-positive S. aureus isolates (n = 111) was used for evaluation. Routinely used approaches were studied to determine their suitability to correctly identify mecC-harboring MRSA, including three (semi)automated antimicrobial susceptibility testing (AST) systems and five selective chromogenic agar plates. Additionally, a cefoxitin disk diffusion test and an oxacillin broth microdilution assay were examined. All mecC-harboring MRSA isolates were able to grow on all chromogenic MRSA screening plates tested. Detection of these isolates in AST systems based on cefoxitin and/or oxacillin testing yielded overall positive agreements with the mecC genotype of 97.3% (MicroScan WalkAway; Siemens), 91.9% (Vitek 2; bioMérieux), and 64.9% (Phoenix, BD). The phenotypic resistance pattern most frequently observed by AST devices was "cefoxitin resistance/oxacillin susceptibility," ranging from 54.1% (Phoenix) and 83.8% (Vitek 2) to 92.8% (WalkAway). The cefoxitin disk diffusion and oxacillin broth microdilution assays categorized 100% and 61.3% of isolates to be MRSA, respectively. The chromogenic media tested confirmed their suitability to reliably screen for mecC-harboring MRSA. The AST systems showed false-negative results with varying numbers, misidentifying mecC-harboring MRSA as methicillin-susceptible S. aureus This study underlines cefoxitin's status as the superior surrogate mecC-positive MRSA marker.


Ultrasound guided percutaneous microwave ablation of benign breast lesions.
The benign breast lesions (BBLs) share a high incidence for women and therapy methods with minimal invasion and better cosmetic outcome are thirsted for. In this study, 122 patients with 198 biopsy-proved BBLs were enrolled. Ultrasound (US)-guided microwave ablation (MWA) was performed with local anesthesia from November, 2013 to April, 2016. The mean longest tumor size assessed was 1.6±0.7 cm (ranging 0.7-4.9 cm). MWA was successfully performed in all cases including 85 lesions adjacent to the skin, pectoralis and areola. The mean ablation time was 3.2mins (ranging 0.5-18.3 mins). 99.5% of BBLs showed complete ablation when assessed by magnetic resonance imaging and 100% of them by US. At the median 14-month follow-up, the BBLs were not palpable in 45.9 % of the cases (palpable in 90.2 % of the cases before MWA) and the mean volume reduction ratio was 78.4±33.5% for total lesions and 89.3±20.8%, 84.7±27.6% and 55.9±32.9% for ≤1.0 cm, 1.1-2.0cm and >2.0 cm lesions in 12-month follow-up, respectively. Cosmesis were reported as good or excellent in 100 % by physician and patients. No side effect was found. The MWA of the BBLs proved feasible and effective, while showing meaningful reduction in volume, palpability and cosmetic satisfying outcomes.


Multimodal phenotypic axes of Parkinson's disease.
Individuals with Parkinson's disease present with a complex clinical phenotype, encompassing sleep, motor, cognitive, and affective disturbances. However, characterizations of PD are typically made for the "average" patient, ignoring patient heterogeneity and obscuring important individual differences. Modern large-scale data sharing efforts provide a unique opportunity to precisely investigate individual patient characteristics, but there exists no analytic framework for comprehensively integrating data modalities. Here we apply an unsupervised learning method-similarity network fusion-to objectively integrate MRI morphometry, dopamine active transporter binding, protein assays, and clinical measurements from n = 186 individuals with de novo Parkinson's disease from the Parkinson's Progression Markers Initiative. We show that multimodal fusion captures inter-dependencies among data modalities that would otherwise be overlooked by field standard techniques like data concatenation. We then examine how patient subgroups derived from the fused data map onto clinical phenotypes, and how neuroimaging data is critical to this delineation. Finally, we identify a compact set of phenotypic axes that span the patient population, demonstrating that this continuous, low-dimensional projection of individual patients presents a more parsimonious representation of heterogeneity in the sample compared to discrete biotypes. Altogether, these findings showcase the potential of similarity network fusion for combining multimodal data in heterogeneous patient populations.


Cell Cycle-Regulated Transcription of CENP-A by the MBF Complex Ensures Optimal Level of CENP-A for Centromere Formation.
The centromere plays an essential role in chromosome segregation. In most eukaryotes, centromeres are epigenetically defined by the conserved histone H3 variant CENP-A. Proper centromere assembly is dependent upon the tight regulation of CENP-A level. Cell cycle regulation of CENP-A transcription appears to be a universal feature across eukaryotes, but the molecular mechanism underlying the temporal control of CENP-A transcription and how such regulation contributes to centromere function remains elusive. CENP-A in fission yeast has been shown to be transcribed before S phase. Using various synchronization methods, we confirmed that CENP-A transcription occurs at G1, leading to an almost twofold increase of the protein during S phase. Through a genetic screen, we identified the MBF (MluI box-binding factors) complex as a key regulator of temporal control of CENP-A transcription. The periodic transcription of CENP-A is lost in MBF mutants, resulting in CENP-A mislocalization and chromosome segregation defects. We identified the MCB (MluI cell cycle box) motif in the CENP-A promoter, and further showed that the MBF complex binds to the motif to restrict CENP-A transcription to G1. Mutations of the MCB motif cause constitutive CENP-A expression and deleterious effects on cell survival. Using promoters driving transcription to different cell cycle stages, we found that timing of CENP-A transcription is dispensable for its centromeric localization. Our data instead indicate that cell cycle-regulated CENP-A transcription is a key step to ensure that a proper amount of CENP-A is generated across generations. This study provides mechanistic insights into the regulation of cell cycle-dependent CENP-A transcription, as well as its importance on centromere function.


Cytological screening for cervical cancer and human papillomavirus in general practice.
In a retrospective study of cervical screening in a general practice in Birmingham 156 out of 1913 smears taken over three years showed some abnormality. Smears from 65 women showed severe non-specific inflammation, and 91 women had various grades of cervical intraepithelial neoplasia, of whom 53 were aged under 30 and 13 over 40. Of 35 women with clinical evidence of human papillomavirus, 21 had normal results on cervical testing and 14 abnormal results. The incidence of genital warts among sexually active young people is growing, but the association of human papillomavirus with abnormal cervical smears is not clear. The efficacy of screening in the United Kingdom must be improved by actively encouraging younger patients to attend for regular screening.


Differential expression of alpha2-6 sialylated polylactosamine structures by human B and T cells.
We found that human peripheral B and T cells differed in the surface expression of alpha2-6 sialylated type 2 chain glycans. In contrast to B cells, T cells expressed only sialoglycans with repeated N-acetyllactosamine (Galss1-4GlcNAc) disaccharides. This finding was based on the specificity of the monoclonal antibodies HB6, HB9 (CD24), HD66 (CDw76), FB21, and CRIS4 (CDw76) with the alpha2-6 sialylated model gangliosides IV6NeuAcnLc4Cer (2-6 SPG), VI6NeuAcnLc6Cer (2-6 SnHC), VIII6NeuAcnLc8Cer (2-6 SnOC), and X6NeuAcnLc10Cer (2-6 SnDC). We found that, in addition to their common requirement of an alpha2-6 bound terminal sialic acid for binding, the antibodies displayed preferences for the length of the carbohydrate backbones. Some of them bound mainly to 2-6 SPG with one N-acetyllactosamine (LacNAc) unit (HB9, HD66); others preferentially to 2-6 SnHC and 2-6 SnOC, with two and three LacNAc units, respectively (HB6 and FB21); and one of them exclusively to very polar alpha2-6 sialylated type 2 chain antigens (CRIS4) such as to 2-6 SnOC and even more polar gangliosides with three and more LacNAc units. These specificities could be correlated with the cellular binding of the antibodies as follows: whereas all antibodies bound to human CD 19 positive peripheral B cells, their reactivity with CD3 positive T cells was either nearly lacking (HD66, HB9), intermediate (about 65%: HB6, FB21) or strongly positive (CRIS4, 95%). Thus, the binding of the antibodies to 2-6 sialylated glycans with multiple lactosamine units appeared to determine their binding to T-cells.


Biotransformation of progesterone by Aspergillus nidulans VKPM F-1069 (wild type).
Biotechnological transformation of steroids using enzyme systems of microorganisms is often the only possible method to modify the molecule in the industrial production of steroid drugs. Filamentous fungus Aspergillus nidulans has been little studied as a steroid-transforming microorganism. We studied the ability of the A. nidulans VKPM F-1069 strain to transform progesterone (PG) for the first time. This strain converts PG into 3 main products: 11α-hydroxy-PG, 11α-acetoxy-PG and 6β,11α-dihydroxy-PG. It has been established that in the first stage, the hydroxylation of PG occurs into C11α position, then the formed 11α-hydroxy-PG is modified into 11α-acetoxy-PG and 6β,11α-dihydroxy-PG. It was found that changes in the composition of the growth medium, aeration and the duration of the mycelium cultivation do not affect the qualitative composition of PG transformation products, but their ratios have changed. Under conditions of limited aeration, the direction of secondary modification of 11α-hydroxy-PG is shifted towards the formation of 11α-acetoxy-PG.


Reversible pure red cell aplasia associated with diphenylhydantoin therapy.
Pure red cell aplasia (PRCA) is characterised by an anaemia with reticulocytopenia but with normal leukocyte and platelet counts, and a bone marrow with the selective absence of erythroid precursor cells. Drug-induced PRCA is a rare cause of secondary erythroid aplasia, but distinct from the primary and most secondary forms, it is usually acute and fully reversible upon withdrawal of the causative drug. We report a 36 year-old Chinese man who developed diphenylhydantoin associated PRCA two months after commencing the treatment. Reappearance of reticulocytes was observed six days following the cessation of diphenylhydantoin therapy and the haemoglobin level rose to normal one month later. The extreme rarity of this adverse reaction to a drug used so widely strongly suggests an individual predisposition.


The stability of attributional style and its relation to psychological distress.
A group of yound adults completed the Attributional Style Questionnaire and measures of depressive affect and hopelessness on two occasions separated by a time interval of three years. Attributional style was demonstrated to be relatively stable over this time period, and was correlated with the measures of psychological well-being. Specifically, those who scored highest on depressive affect and hopelessness attributed good outcomes more externally and less stably, and attributed bad outcomes to relatively more stable and global causes. In contrast to the hopelessness model of depression, however, multiple regression analyses showed that depressive attributions were not antecedent to increased psychological distress; nor were they a consequence. Furthermore, negative life-events did not contribute to the prediction. It was concluded that the data are most consonant with Brewin's (1985) symptom model, in which depressive attributions are a concomitant or symptom of depression but have no causal impact on the onset or course of the disorder.


Educational inclusion of deaf children: current policy, practices, and future possibilities.
This paper examines policies and practices of inclusive education for young deaf children in Ghana. Though existing Government policy promotes the principles and implementation of educational inclusion of all children in Ghana, caregivers often encounter significant challenges ensuring access to early years' education for their deaf children. We used Bronfenbrenner's (2005) bioecological systems theory and interviewed 15 educators of deaf children and 9 hearing-health clinicians, to capture different knowledge, experience, and perspectives on policy, practice, and inclusive approaches to deaf children's education. We ask: (a) How do teachers, clinicians, and caregivers connect to promote early childhood care education (ECCE) for young deaf preschoolers in Ghana? and (b) What resources and precarities are available to educators of deaf children and hearing-health clinicians in sustaining inclusive educational practices across urban and rural contexts in Ghana? We address these unanswered questions about the potential pathway to bring about changes in policy and practice in promoting successful inclusive education for deaf children in Ghana. Implications for national and international policy and practice of ECCE for deaf children are discussed and recommendations are made.


Glycoprotein IIIa Pl(A1/A2) polymorphism and sudden cardiac death.
We studied the association of the Pl(A1/A2) polymorphism with coronary thrombosis, myocardial infarction (MI) and sudden cardiac death (SCD) in autopsied victims of sudden death. Sudden cardiac death is one of the leading symptoms of coronary heart disease in early middle age. Platelet glycoprotein (GP)IIb/IIIa fibrinogen receptors play a key role in coronary thrombosis and MI. Pl(A1/A2) polymorphism of the gene for GPIIIa has been previously studied in hospital MI patients. Significance of the Pl(A1/A2) polymorphism in victims of SCD is not known. The Pl(A1/A2) polymorphism was studied in the Helsinki Sudden Death Study comprising 700 autopsied middle-aged white Finnish men (33 to 70 years, mean 53 years) who suffered sudden or violent out-of-hospital death. Prevalence of the A2 allele decreased with age in the series. This decrease was observed among victims of SCD (n = 281) but not in men who died violently (n = 258) or of other diseases (n = 127). Of SCD victims below 50 years, 39.7% were carriers of the A2 allele compared with 28.3% among men under 50 who died of other causes (odds ratio [OR] 2.5, p = 0.01). Men with acute fatal coronary thrombosis (n = 39) were more often (OR 3.4, p < 0.01) carriers of the A2 allele than were men (n = 242) with SCD in the absence of acute coronary thrombosis (48.7% vs. 24.4%, respectively). In addition, men with MI and recent or old thrombosis (n = 67) were more often (OR 3.6, p = 0.005) carriers of the A2 allele than were men (n = 123) with MI in the absence of thrombosis (44.8% vs. 20.3%, respectively). These associations were especially strong in men under 60. Our results suggest that the A2 allele of the Pl(A1/A2) polymorphism of GPIIIa is a major risk factor of coronary thrombosis and may be one important predictor of SCD in early middle age.


Exclusion of T4 phage by the hok/sok killer locus from plasmid R1.
The hok (host killing) and sok (suppressor of killing) genes (hok/sok) efficiently maintain the low-copy-number plasmid R1. To investigate whether the hok/sok locus evolved as a phage-exclusion mechanism, Escherichia coli cells that contain hok/sok on a pBR322-based plasmid were challenged with T1, T4, T5, T7, and lambda phage. Upon infection with T4, the optical density of cells containing hok/sok on a high-copy-number plasmid continued to increase whereas the optical density for those lacking hok/sok rapidly declined. The presence of hok/sok reduced the efficiency of plating of T4 by 42% and decreased the plaque size by approximately 85%. Single-step growth experiments demonstrated that hok/sok decreased the T4 burst size by 40%, increased the time to form mature phage (eclipse time) from 22 to 30 min, and increased the time to cell lysis (latent period) from 30 to 60 min. These results further suggest that single cells exhibit altruistic behavior.


Associations between schizophrenia genetic risk, anxiety disorders and manic/hypomanic episode in a longitudinal population cohort study.
Studies involving clinically recruited samples show that genetic liability to schizophrenia overlaps with that for several psychiatric disorders including bipolar disorder, major depression and, in a population study, anxiety disorder and negative symptoms in adolescence.AimsWe examined whether, at a population level, association between schizophrenia liability and anxiety disorders continues into adulthood, for specific anxiety disorders and as a group. We explored in an epidemiologically based cohort the nature of adult psychopathology sharing liability to schizophrenia. Schizophrenia polygenic risk scores (PRSs) were calculated for 590 European-descent individuals from the Christchurch Health and Development Study. Logistic regression was used to examine associations between schizophrenia PRS and four anxiety disorders (social phobia, specific phobia, panic disorder and generalised anxiety disorder), schizophrenia/schizophreniform disorder, manic/hypomanic episode, alcohol dependence, major depression, and - using linear regression - total number of anxiety disorders. A novel population-level association with hypomania was tested in a UK birth cohort (Avon Longitudinal Study of Parents and Children). Schizophrenia PRS was associated with total number of anxiety disorders and with generalised anxiety disorder and panic disorder. We show a novel population-level association between schizophrenia PRS and manic/hypomanic episode. The relationship between schizophrenia liability and anxiety disorders is not restricted to psychopathology in adolescence but is present in adulthood and specifically linked to generalised anxiety disorder and panic disorder. We suggest that the association between schizophrenia liability and hypomanic/manic episodes found in clinical samples may not be due to bias.Declarations of interestNone.


Indoor air quality modeling for PM 10, PM 2.5, and PM 1.0 in naturally ventilated classrooms of an urban Indian school building.
Assessment of indoor air quality (IAQ) in classrooms of school buildings is of prime concern due to its potential effects on student's health and performance as they spend a substantial amount of their time (6-7 h per day) in schools. A number of airborne contaminants may be present in urban school environment. However, respirable suspended particulate matter (RSPM) is of great significance as they may significantly affect occupants' health. The objectives of the present study are twofold, one, to measure the concentrations of PM(10) (<10 microm), PM(2.5) (<2.5 microm), and PM(1.0) (<1.0 microm) in naturally ventilated classrooms of a school building located near a heavy-traffic roadway (9,755 and 4,296 vehicles/hour during weekdays and weekends, respectively); and second, to develop single compartment mass balance-based IAQ models for PM(10) (NVIAQM(pm10)), PM(2.5) (NVIAQM(pm2.5)), and PM(1.0) (NVIAQM(pm1.0)) for predicting their indoor concentrations. Outdoor RSPM levels and classroom characteristics, such as size, occupancy level, temperature, relative humidity, and CO(2) concentrations have also been monitored during school hours. Predicted indoor PM(10) concentrations show poor correlations with observed indoor PM(10) concentrations (R (2) = 0.028 for weekdays, and 0.47 for weekends). However, a fair degree of agreement (d) has been found between observed and predicted concentrations, i.e., 0.42 for weekdays and 0.59 for weekends. Furthermore, NVIAQM(pm2.5) and NVIAQM(pm1.0) results show good correlations with observed concentrations of PM(2.5) (R(2) = 0.87 for weekdays and 0.9 for weekends) and PM(1.0) (R(2) = 0.86 for weekdays and 0.87 for weekends). NVIAQM(pm10) shows the tendency to underpredict indoor PM(10) concentrations during weekdays as it does not take into account the occupant's activities and its effects on the indoor concentrations during the class hours. Intense occupant's activities cause resuspension or delayed deposition of PM(10). The model results further suggests conductance of experimental and physical simulation studies on dispersion of particulates indoors to investigate their resuspension and settling behavior due to occupant's activities/movements. The models have been validated at three different classroom locations of the school site. Sensitivity analysis of the models has been performed by varying the values of mixing factor (k) and newly introduced parameter R(c). The results indicate that the change in values of k (0.33 to 1.00) does not significantly affect the model performance. However, change in value of R(c) (0.001 to 0.500) significantly affects the model performance.


New generation genetic testing entering the clinic.
New generation sequencing (NGS) genetic testing is a powerful diagnostic tool and is increasingly used in the clinical workup of patients, especially in unusual presentations or where a positive family history suggests heritable disease. This review addresses the NGS technologies Targeted sequencing (TS), Whole exome sequencing (WES), Whole genome sequencing (WGS), and the use of gene panels or gene lists for clinical diagnostic purposes. These methods primarily assess nucleotide sequence but can also detect copy number variants and many tandem repeat expansions, greatly simplifying diagnostic algorithms for movement disorders. Studies evaluating the efficacy of NGS in diagnosing movement disorders have reported a diagnostic yield of up to 10.1% for familial and 15.7% for early-onset PD, 11.7-37.5% for dystonia, 12.1-61.8% for ataxia/spastic paraplegia and 11.3-28% for combined movement disorders. Patient selection and stringency in the interpretation of the detected variants and genotypes affect diagnostic yield. Careful comparison of the patient's or family's disease features with the previously reported phenotype associated with the same variant or gene can avoid false-positive diagnoses, although some genes are implicated in various phenotypes. Moving from TS to WES and WGS increases the number of patients correctly diagnosed, but for many patients, a genetic cause cannot be identified today. However, new genetically defined entities are discovered at rapid pace, and genetic databases and our knowledge of genotype-phenotype correlations expand steadily. We discuss the need for clear communication of genetic results and suggest a list of aspects to consider when reporting neurogenetic disorders using NGS testing.


A bit or a lot on the side? Observational study of the energy content of starters, sides and desserts in major UK restaurant chains.
Our objective was to examine the kilocalorie (kcal) content of starters, sides and desserts served in major UK restaurant chains, comparing the kcal content of these dishes in fast-food and full-service restaurants. Observational study. Menu and nutritional information provided online by major UK restaurant chains. During October to November 2018, we accessed websites of restaurant chains with 50 or more outlets in the UK. Menu items that constituted starters, sides or desserts were identified and their kcal content was extracted. Accompanying beverages were not included. We used multilevel modelling to examine whether mean kcal content of dishes differed in fast-food versus full-service restaurants. The mean kcal content of dishes and the proportion of dishes exceeding public health recommendations for energy content in a main meal (>600 kcal). A total of 1009 dishes (212 starters, 318 sides and 479 desserts) from 27 restaurant chains (21 full-service, 6 fast-food) were included. The mean kcal content of eligible dishes was 488.0 (SE=15.6) for starters, 397.5 (SE=14.9) for sides and 430.6 (SE=11.5) for desserts. The percentage of dishes exceeding 600 kcal was 26.4% for starters, 21.7% for sides and 20.5% for desserts. Compared with fast-food chains, desserts offered at full-service restaurants were on average more calorific and were significantly more likely to exceed 600 kcal. The average energy content of sides, starters and desserts sold in major UK restaurants is high. One in four starters and one in five sides and desserts in UK chain restaurants exceed the recommended energy intake for an entire meal.


[Morphology of skeletal muscles after long-term hypokinesia and laser acupuncture].
Hypokinesia (Hk) lasting for 150 days has been modelling on Wistar male rats by means of their holding in special boxes. After decapitation in 10, 20, 50, 100 and 150 days the muscles of their extremities have been studied, using stereological methods. At Hk, especially for 50 days, inhibition of morphogenesis of ultrastructures and development of certain pathological processes are noted in the muscles with their preponderance in red fibers. Laser puncture stimulates formation of the ultrastructures responsible for contraction and energy supply and sharply decreases development of the destructive processes in muscle fiber, thus providing a satisfactory endurance of hypokinesia.


A paper based, all organic, reference-electrode-free ion sensing platform.
We present a reference-electrode free, all organic K+ sensitive ion sensing platform fabricated by simplest means on a plain sheet of paper. This platform is based on two identical ion selective electrodes (ISEs) which are assembled by bonding a polymeric ion selective membrane (ISM) directly onto a drop-casted PEDOT:PSS electrode. Taking full advantage of the so called pulsetrode concept, a current pulse is used to measure the concentration of the targeted ion. The current forces an ion flux out of the first ISE, through the sample and into the second ISE. This flux leads to a well-defined potential jump at the second ISE, as soon as the target ion locally depletes within the analyte, whereas the current induced potential change at the first ISE does not depend noticeably on the type of background electrolyte. Hence, the potential difference between the ISEs, required to apply the current is directly related to the ion concentration within the sample. This concept allows for a 20-fold sensitivity enhancement compared to classical potentiometric measurements in physiological backgrounds. As mutual potential drifts of the ISEs cancel out, the sensor response showed excellent stability and did not change during multiple measurements over three months.


Classical swine fever in wild boar (Sus scrofa)--experimental infections and viral persistence.
A pregnant wild boar and two wild boar weaners were inoculated intranasally with a field isolate of classical swine fever virus (CSFV) recently derived from a diseased domestic pig. The clinical, pathological and haematological findings noted in the young wild boars were comparable to those in domestic weaner pigs inoculated with the same virus isolate. Both wild boars showed the acute haemorrhagic form of CSF, one animal died 18 days post inoculation (p. i.) and the second one had to be euthanized when moribund two days later. The wild boar sow did not show any signs of illness p. i. but seroconversion was noticed. Twenty-eight days p. i. birth was given to six clinically healthy offsprings. One of the newborn proved to be viraemic until death when 39 days of age. Except for poor growth no other symptoms were noticed in this piglet. The non-viraemic litter mates remained healthy, although they had close contact to the persistently infected piglet. High titres of neutralizing antibodies against CSFV were measured in the serum samples of these offsprings. All findings were more or less in accordance with observations previously made in domestic pigs when infected with CSFV around 85 to 90 days of gestation. The wild boar was calculated to have been inoculated at about 87 to 92 days of gestation.


Colonization of Vitis vinifera by a green fluorescence protein-labeled, gfp-marked strain of Xylophilus ampelinus, the causal agent of bacterial necrosis of grapevine.
The dynamics of Xylophilus ampelinus were studied in Vitis vinifera cv. Ugni blanc using gfp-marked bacterial strains to evaluate the relative importance of epiphytic and endophytic phases of plant colonization in disease development. Currently, bacterial necrosis of grapevine is of economic importance in vineyards in three regions in France: the Cognac, Armagnac, and Die areas. This disease is responsible for progressive destruction of vine shoots, leading to their death. We constructed gfp-marked strains of the CFBP2098 strain of X. ampelinus for histological studies. We studied the colonization of young plants of V. vinifera cv. Ugni blanc by X. ampelinus after three types of artificial contamination in a growth chamber and in a greenhouse. (i) After wounding of the stem and inoculation, the bacteria progressed down to the crown through the xylem vessels, where they organized into biofilms. (ii) When the bacteria were forced into woody cuttings, they rarely colonized the emerging plantlets. Xylem vessels could play a key role in the multiplication and conservation of the bacteria, rather than being a route for plant colonization. (iii) When bacterial suspensions were sprayed onto the plants, bacteria progressed in two directions: both in emerging organs and down to the crown, thus displaying the importance of epiphytic colonization in disease development.


Characterization of luteinizing hormone secretion in the primiparous, lactating sow: relationship to blood metabolites and return-to-estrus interval.
Lactating, primiparous Landrace x Yorkshire sows were used to characterize LH secretion during lactation in sows that experienced an early (less than 9 d; n = 14) or late (greater than 15 d; n = 9) return to estrous postweaning and to evaluate the relationship between LH secretion and blood metabolites. Twenty-three sows were fed one of nine corn-soybean meal diets to achieve a matrix of lysine (15 to 45 g/d) and energy (6.5 to 16.5 Mcal of ME/d) intakes and a range in metabolite concentrations and return-to-estrus intervals. Blood samples for LH analysis were collected every 15 min for 6 h on d 0, 7, 14, 21, and 28 of lactation. Circulating concentrations of glucose, amino acids, insulin, triglycerides, urea N, and nonesterified fatty acids also were measured on d 7 and 21. Mean LH concentrations were .27 and .42 ng/mL at farrowing for sows with an early and late return to estrus, respectively, but decreased (P less than .01) to .12 ng/mL by d 7 in both early and late groups. Mean LH and number of LH peaks per 6 h increased linearly (P less than .01) from d 7 to 28 for early sows. Early sows had a higher LH mean and more LH peaks per 6 h on d 14, 21, and 28 than did late sows (P less than .05). Early sows had higher serum insulin on d 7 (P less than .05) and d 21 (P less than .01) than did late sows. Concentrations of other metabolites did not differ (P less than .10) between early and late sows.(ABSTRACT TRUNCATED AT 250 WORDS)


Contrarian clade confirms the ubiquity of spatial origination patterns in the production of latitudinal diversity gradients.
The latitudinal diversity gradient (LDG), wherein the number of species and higher taxa peaks in the tropics and decreases toward the poles, is the best-documented large-scale diversity pattern on Earth, but hypotheses explaining the standard LDG must also account for rare "contrarian" taxa that show diversity maxima outside of the tropics. For marine bivalves, one of the few groups that provide spatially explicit temporal data on a global scale, we show that a major contrarian group, the Anomalodesmata, unexpectedly exhibits the same large-scale dynamics as related clades having normal LDGs in two key respects. First, maxima in standing genus diversity and genus origination rates coincide spatially. Second, the strength of a clade's present-day LDG is significantly related to the proportion of its living genera that originated in the tropics during the late Cenozoic, with the contrarian gradient strength at both species and genus level predicted quantitatively by the values for the other clades. Geologic age distributions indicate that the anomalous LDG results from origination that is damped in the tropics rather than heightened in the temperate zones. The pervasive role of spatial origination patterns in shaping LDGs, regardless of the position of their diversity maxima, corroborates hypotheses based on clades showing standard gradients and underscores the insights that contrarian groups can provide into general principles of diversity dynamics.


Operationally realistic validation for prediction of cocoa sensory qualities by high-throughput mass spectrometry.
The potential of analytical chemistry to predict sensory qualities of food materials is a major current theme. Standard practice is cross-validation (CV), where a set of chemical and associated sensory data is partitioned so chemometric models can be developed on training subsets, and validated on held-out subsets. CV demonstrates prediction, but is an unlikely scenario for industrial operations, where concomitant data acquisition for model development and test materials would be unwieldy. We evaluated cocoa materials of diverse provenance, and analyzed on different dates to those used in model development. Liquor extracts were analyzed by flow-injection electrospray-mass spectrometry (FIE-MS), a novel method for sensory quality prediction. FIE-MS enabled prediction of sensory qualities described by trained human panelists. Optimal models came from the Weka data-mining algorithm SimpleLinearRegression, which learns a model for the attribute giving minimal training error, which was (-)-epicatechin. This flavonoid likewise dominated partial least-squares (PLS)-regression models. Refinements of PLS (orthogonal-PLS or orthogonal signal correction) gave poorer generalization to different test sets, as did support vector machines, whose hyperparameters could not be optimized in training to avoid overfitting. In conclusion, if chemometric overfitting is avoided, chemical analysis can predict sensory qualities of food materials under operationally realistic conditions.


Development of topical chitosan/ β-glycerophosphate-based hydrogel loaded with levofloxacin in the treatment of keratitis: An ex-vivo study.
Staphylococcus species are responsible for most cases of post-operative endophthalmitis. Topical ocular drug was applied for post-operative infection prevention, but the way of delivery encounters many challenges in terms of patient's compliance, drug efficacy, and drug penetration. We used the levofloxacin-loaded chitosan/gelatin/β-glycerophosphate hydrogel sustained releasing system with good in vitro anti-bacterial efficacy and biocompatibility, which we had previously designed, for ex vivo keratitis model to test the preclinical drug efficacy and to determine drug level in the anterior chamber of the eye. The result showed that the ex-vivo corneal keratitis model with S. aureus infection revealed mild opacity over the central cornea with stromal infiltrate, but without obvious stromal infiltration post levofloxacin-loaded hydrogel treatment after 24 h of infection. Quantification of viable bacteria showed a significant anti-bacterial activity. The histological evidence also showed no visible S. aureus after levofloxacin-loaded hydrogel treatment, with a significant anti-inflammatory effect. We also examined the drug concentration in the aqueous humor 24 h after instilling one drop of the levofloxacin-loaded hydrogel. The concentration achieved to a desired drug level. These results suggested that by the ex-vivo model, levofloxacin-loaded hydrogel can be applied for treatment in post-operative endophthalmitis or keratitis after the ophthalmic surgery.


Tigecycline: a critical analysis.
Tigecycline (GAR-936) is the first glycylcycline antibiotic to be approved by the US Food and Drug Administration (FDA). The drug overcomes the 2 major resistance mechansisms of tetracycline: drug-specific efflux pump acquisition and ribosomal protection. Tigecycline is active against many gram-positive and -negative organisms, including methicillin-resistant Staphylococcus aureus, vancomycin-intermediate and -resistant enterococci, and extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae. It is also active against many anaerobic bacteria, as well as atypical pathogens, including rapidly growing, nontuberculous mycobacteria. Tigecycline is concentrated in cells and is eliminated primarily via biliary excretion. Diminished renal function does not significantly alter its systemic clearance. Furthermore, tigecycline does not interfere with common cytochrome P450 enzymes, making pharmacokinetic drug interactions uncommon. It provides parenteral therapy for complicated skin/skin-structure and intra-abdominal infections. The only prominent adverse effects are associated with tolerability, most notably nausea and vomiting. Tigecycline will be most useful as empirical therapy for polymicrobial infections, especially in cases in which deep tissue penetration is needed or in which multidrug-resistant pathogens are suspected.


Weight, weight gain, activity, and major illnesses: the Nurses' Health Study.
The Nurses' Health Study was initiated in 1976 to study the relation between oral contraceptives and breast cancer. Subsequent funding was available to follow the cohort to address relations between cigarette smoking, postmenopausal hormones, hair dyes, and a range of cancers including breast, endometrial, ovarian, and lung cancer. The 121,700 participating nurses are followed up every 2 years via mail questionnaire to update exposure information to identify incident cancers and other illnesses. Follow-up through 1994 has achieved 90% response from living cohort members. Over the course of the study, additional exposures have been added and refined, including weight at age 18, current weight, height, waist and hip measurements and history of major voluntary weight loss among others. Our focus has been on the health effects of weight gain during middle age. The results relating to diabetes, coronary heart disease, certain types of cancer and total mortality are reviewed. Our primary analytic tools have been multiple logistic regression and Cox proportional hazards models. These methods allow for flexibility in defining the exposures of interest as well as determining their relative importance while controlling for key risk factors. Our models show that even moderate weight gain after age 18 increases risk of each condition. The benefits of physical activity include reduced risk of disease. Issues in the measurement and validation of weight and activity highlight the complexities that are inherent in observational studies addressing the health consequences of lifestyles and anthropometric variables. Our experience of working with repeated measures of body weight and recreational activity are described. Recreational activity has been ascertained in various ways on several questionnaires and may be subject to misclassification. For both weight and activity it may be the pattern of these values that is of importance (e.g. "weight cycling") for some outcomes or conditions. Our research in this area is ongoing. These issues regarding longitudinal measurement will never be completely resolved because weight and activity are intrinsically complex concepts. For these and other such variables, the primary solution is to minimize the problems associated with longitudinal studies. This is best accomplished by developing and maintaining a very strong study design/protocol, including: careful consideration of the sample frame and sample size; maintenance of a high response rate; and continuous monitoring and improvement of the survey/interview instrument(s).


Evaluation of AGP Fucosylation as a Marker for Hepatocellular Carcinoma of Three Different Etiologies.
A mass spectrometric analysis platform has been developed to determine whether glycosylation patterns of alpha-1 acid glycoprotein (AGP) could be used as a marker for early detection of hepatocellular carcinoma (HCC) in different etiologies, i.e. non-alcoholic steatohepatitis (NASH), alcoholic liver disease (ALC), and hepatitis C virus (HCV). MALDI-MS profiling of N-glycans of AGP purified from 20 μL of patient serum in HCC (n = 72) and liver cirrhosis (n = 58) showed that a unique trifucosylated tetra-antennary glycan (m/z 3490.76) was predominantly identified in HCCs but was absent in healthy subjects and the majority of cirrhosis patients. Receiver operation characteristic (ROC) curve analysis showed that the trifucosylated N-glycan of AGP (triFc_AGP) could differentiate HCC from cirrhosis with an area under the curve (AUC) of 0.707, 0.726 and 0.751 for NASH, ALC and HCV, respectively. When combining triFc_AGP with INR and AFP, the panel had the greatest benefit in detection of NASH-related HCCs, with a significantly improved AUC of 0.882 for all NASH HCCs and 0.818 for early NASH HCCs compared to AFP alone (0.761 and 0.641, respectively). Moreover, triFc_AGP could serve as a potential marker for monitoring AFP-negative HCC patients.


[Bone metastasis versus insufficiency fractures due to pelvic radiotherapy for gynecologic neoplasm].
Five cases of patients with gynecological neoplasm (four cervix carcinoma and one endometrial sarcoma) who underwent pelvic external radiotherapy and intracavitary brachytherapy in whom pathologic pelvic uptake was found in the bone scan are presented. The diagnosis was pelvic insufficiency fractures due to radiotherapy adverse effects on the skeletal system confirmed by CT and by the favorable scintigraphy and clinical outcome. Both bone metastases and insufficiency fractures must be considered in the differential diagnosis of bone pain in irradiated pelvises. The bone scintigraphy detects these insufficiency fractures early and can show a typical symmetric uptake pattern. In asymmetric lesions, the CT and clinical follow-up as well as the scintigraphic evolution of the lesions should confirm the findings of the bone scintigraphies.


Application of two-dimensional band-target entropy minimization to fluorescence data: implications for the recovery of patterns arising from only bilinear and not trilinear structures.
Pattern recognition in two-dimensional (2D) spectroscopy, without recourse to spectral libraries, etc., has a number of important potential applications. In the present contribution, two blind source separation techniques (spectral reconstruction) are applied to sets of 2D fluorescence data possessing both Rayleigh scattering and Raman scattering. The two methods used are (1) two-dimensional band-target entropy minimization (2D-BTEM), which models data as a bilinear form (in terms of a weighted sum of 2D patterns) and (2) parallel factor analysis (PARAFAC), which models data as a trilinear form. In addition, an a priori estimate of the number of patterns present is not required by 2D-BTEM but is required in PARAFAC. Both 2D-BTEM and PARAFAC are successfully applied to the real three-component data, and good 2D spectral reconstructions of the three amino acids are achieved. Moreover, 2D-BTEM was also able to recover the 2D Raman scattering directly, whereas PARAFAC did not recover the 2D Raman scatter (the Raman scatter does not possess a trilinear form). The present results suggest that 2D-BTEM can be useful in a wide range of spectroscopic applications for the recovery of underlying 2D patterns.