TrxR2 overexpression alleviates inflammation-mediated neuronal death via reducing the oxidative stress and activating the Akt-Parkin pathway.
Neuronal death caused by inflammatory cytokine-mediated neuroinflammation is being extensively explored. Thioredoxin reductase (TrxR) 2 is a novel mediator of inflammation response. In the current study, we focus on the mechanisms of TrxR2 overexpression in inflammation-mediated neuronal death. LPS was used to induce neuroinflammation in N2a cells in vitro. Adenovirus-loaded TrxR2 was transfected into N2a cells to up-regulate TrxR2 expression. Then, cell viability was determined via MTT assay and TUNEL assay. Apoptosis was measured via western blotting and ELISA. Oxidative stress was detected via ELISA and flow cytometry. A pathway inhibitor was used to verify the role of the Akt-Parkin pathway in the LPS-mediated N2a cell death in the presence of TrxR2 overexpression. With the help of immunofluorescence assay and western blotting, we found that TrxR2 expression was significantly reduced in response to LPS treatment, and this effect was associated with N2a cell death via apoptosis. At the molecular level, TrxR2 overexpression elevated the activity of the Akt-Parkin pathway, as evidenced by the increased expression of p-Akt and Parkin. Interestingly, inhibition of the Akt-Parkin pathway abolished the regulatory effect of TrxR2 on LPS-treated N2a cells, as evidenced by the decreased cell viability and increased apoptotic ratio. Besides, TrxR2 overexpression also reduced oxidative stress, inflammation factor transcription and mitochondrial apoptosis. However, inhibition of Akt-Parkin axis abrogated the protective effects of TrxR2 on redox balance, mitochondrial performance and cell survival. LPS-mediated neuronal death was linked to a drop in TrxR2 overexpression and the inactivation of the Akt-Parkin pathway. Overexpression of TrxR2 sustained mitochondrial function, inhibited oxidative stress, repressed inflammation response, and blocked mitochondrial apoptosis, finally sending a pro-survival signal for the N2a cells in the setting of LPS-mediated inflammation environment.


Racing-related factors and results of prerace physical inspection and their association with musculoskeletal injuries incurred in thoroughbreds during races.
To describe and compare data from Thoroughbreds that sustained musculoskeletal injuries while racing with data from matched control horses. Matched case-control study. 216 Thoroughbreds that sustained a musculoskeletal injury while racing and 532 horses from the same races that were not injured. Data regarding racing history, race-entrant characteristics, racing events determined by analysis of videotapes of races, and results of prerace physical inspections were determined for all horses. Injured horses were compared with control horses by using conditional logistic regression. Results of prerace inspection by regulatory veterinarians were significantly associated with injury. Odds of musculoskeletal injury, injury of the suspensory apparatus of the forelimb, and injury of the tendon of the superficial digital flexor muscle of the forelimb were 5.5 to 13.5 times greater among horses assessed to be at increased risk of injury by regulatory veterinarians on the basis of results of prerace inspection than for horses not considered to be at increased risk of injury. Odds of an abnormal finding in the suspensory ligament during prerace inspection were 3.4 times greater among horses that injured the suspensory apparatus than among control horses, and odds of an abnormal finding in the tendon of the superficial digital flexor muscle during prerace inspection were 15 times greater among horses that injured the tendon than among control horses. Regulatory veterinarians can identify horses during prerace physical inspection that have an increased risk of injury during races. Prerace physical inspections could be used to reduce the risk of injury to Thoroughbreds during races.


Halogen bond-assisted self-assembly of gold nanoparticles in solution and on a planar surface.
Halogen bonding (XB) has been shown to be a powerful tool for promoting molecular self-assembly in different fields. The use of XB for noncovalent assembly of inorganic nanoparticles (NP) is, instead, quite limited, considering how extensively other interactions (i.e., electrostatic forces, hydrophobic effect, hydrogen bonding, etc.) have been exploited to modulate and program NP self-assembly. Here, we designed and synthesized XB-capable organic ligands that were efficiently used to functionalize the surface of gold NPs (AuNPs). XB-assisted AuNP self-assembly was attained in solution mixing AuNPs bearing XB-donor ligands with ditopic XB-acceptor molecules and AuNPs functionalized with XB-acceptor moieties. Likewise, a preliminary study of XB-driven adsorption of these AuNPs on surface was performed via Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D), used as an in situ tool for measuring mass changes upon XB-driven self-assembly.


Human Flourishing and Integrated Care Models: The Development of the Flourish Index.
In evaluating integrated care models, traditional quality measures do not account for functional and quality of life factors, affecting older adults with multiple chronic conditions. The objective of this study was the development and validation of the Flourish Index (FI), an instrument to evaluate integrated care, using a determinants of health model. The study took place within the evaluation study of the Flourish Model (FM). The FM provides care coordination services using an integrated primary care and community-based services model. Baseline data from 70 older adults were used in the validation study. Twenty-seven quality of care indicators within six determinants of health, namely biological, psychological, individual health behaviors, health services, environmental, and social, formed part of the FI. Categorical principal components analysis showed a 5-dimensional structure with psychological determinants loading on the biological determinants of health. Internal consistency (Cronbach's alpha) for the determinants was as follows: biological/psychological = 0.73, individual = 0.58, environmental = 0.62, health services = 0.65, social = 0.67, total score = 0.97. Sensitivity to change was shown for the total FI score (F1,22 = 8.82, p = .01) and social (F1,22 = 5.82, p = .02), with a trend toward sensitivity for individual health behaviors (F1,22 = 3.95, p = .06) and health services (F1,22 = 3.26, p = .09). The preliminary analysis of the FI shows promise for the usability of the index to provide insight into the fundamental challenges of aging. It brings greater clarity in caring for older adults and supports quality evaluation of integrated care coordination models.


Ion-pair spectrophotometric estimation of gemifloxacin.
The main objective was to develop and validate a simple, accurate, precise, and sensitive ion-pair spectrophotometric extraction method for the assay of gemifloxacin mesylate (GFX) in pure, tablets and spiked human urine. The method is based upon the reaction of gemifloxacin with methyl orange, forming a yellow-colored complex in acidic medium, which is extracted in chloroform and analyzed. The extracted complexes showed absorbance maxima (λmax) found to be at 427 nm. Beer's law was obeyed for a wide concentration range, i.e., 10-80 μg/ mL as the extracted species seemed well defined and stable. The surface or an interphase adsorption phenomenon was not a problem. Optimization of the reaction was carried out with factors such as buffer strength, stability of complex, and molar ratio of drug: Dye and extraction time. The proposed method was validated as per the ICH guidelines. The recovery studies confirmed the accuracy and precision of the method. The above-mentioned method was a rapid tool for routine analysis of GFX in the bulk and pharmaceutical dosage forms.


Treatment of medical emergencies for the general practitioner.
Medical emergencies in the dental office are, fortunately, very infrequent. However, when they do happen, it pays to keep your head and stay focused so you, yourself, don't become the second emergency in your office that day. We have all sat through lectures and read articles as to what we should have in our emergency kit. We buy these medications, we renew them when they expire, and hope we never have the opportunity to use them. The purpose of this article is not simply to list what emergency medication to have in your office, but rather to help you appreciate the signs, symptoms and histories of your patients in order to avoid a medical emergency. This article is not geared for oral maxillofacial surgeons or practitioners with anesthesia experience, or advanced cardiac life support training. These specialists most likely will use their advanced armamentarium to manage medical emergencies in a different manner compared to the average general practitioner.


National patterns of care and predictors of neoadjuvant and concurrent chemotherapy use with definitive radiotherapy in the treatment of patients with oropharyngeal squamous cell carcinoma.
To the authors' knowledge, the patterns of care for the radiotherapy-based treatment of patients with stage III to IVB oropharyngeal squamous cell carcinoma (OPSCC) are poorly defined. The objective of the current study was to characterize the use and predictors of chemotherapy with radiotherapy for this population using the National Cancer Database. Patients in the National Cancer Database with AJCC (American Joint Committee on Cancer) stage III to IV OPSCC who were treated with radiotherapy between 2003 and 2012 were eligible for analysis. Treatment was defined as radiotherapy alone, concurrent chemoradiotherapy, or induction chemotherapy (IC). Multivariable regression with multilevel modeling was used to determine predictors of any chemotherapy use and, among patients receiving chemotherapy, the predictors of IC. The majority (90%) of the 30,875 eligible patients received chemotherapy, the majority of whom (71% of the total) were treated with definitive chemoradiotherapy; a sizeable percentage of patients received IC (19% of total). On multivariable regression, younger age, favorable comorbidity status, and more advanced tumor and lymph node disease were found to be independent predictors of any chemotherapy and IC use. Nonwhite patients (odds ratio [OR], 0.71; P<.0001), women (OR, 0.74; P<.0001), and individuals without private insurance were found to be significantly less likely to receive chemotherapy. Patients treated at higher-volume institutions were significantly less likely to receive IC (OR, 0.69; P = .0006). Human papillomavirus status did not appear to independently influence treatment choice. The vast majority of patients with stage III to IVB OPSCC who were treated with definitive radiotherapy received chemotherapy, which is consistent with high-level data and national recommendations. However, disparities with regard to race, sex, and insurance status emerged thereby requiring additional investigation. The frequent use of IC despite limited supportive evidence warrants research on physician and patient decision making and presents an opportunity to improve evidence-based treatment delivery. Cancer 2017;123:273-282. © 2016 American Cancer Society.


Evidence for recent invasion of the medaka fish genome by the Tol2 transposable element.
Tol2 is a transposable element of the terminal-inverted-repeat class, residing in the genome of the medaka fish Oryzias latipes. The genus Oryzias contains more than 10 species for which phylogenetic relationships have previously been estimated. To infer the history of Tol2 in this genus we performed genomic Southern blots and PCR analyses of 10 of the species. It was revealed that Tol2 occurs in 2 of the 10 species (O. curvinotus and O. latipes) and that the length and the restriction map structure of Tol2 are identical in the two cases. Further, sequencing analysis revealed an extremely low level of divergence compared with that in a nuclear gene. These results suggest recent incorporation of Tol2 into one or both of the two species, implying horizontal transfer of Tol2 from one species to the other or into them both from a common source.


All health care is not local: an evaluation of the distribution of Emergency Department care delivered in Indiana.
The Emergency Department (ED) delivers a major portion of health care - often with incomplete knowledge about the patient. As such, EDs are particularly likely to benefit from a health information exchange (HIE). The Indiana Public Health Emergency Surveillance System (PHESS) sends real-time registration information for emergency department encounters. Over the three-year study period, we found 2.8 million patients generated 7.4 million ED visits. The average number of visits was 2.6 visits/patient (range 1-385). We found more than 40% of ED visits during the study period were for patients having data at multiple institutions. When examining the network density, we found nearly all EDs share patients with more than 80 other EDs. Our results help clarify future health care policy decisions regarding optimal NHIN architecture and discount the notion that 'all healthcare is local'.


[Clinical course and treatment of Hodgkin's disease with concomitant bone marrow lesions].
Fifty patients with confirmed local multiple lesions of bone marrow were selected by 99mTc scintigraphy, magnetic resonance imaging and morphologically-supported trepan biopsy from 155 cases of Hodgkin's disease stage II-IVAB. Bone marrow lesions were relatively more common in younger patients 1-11 months after primary tumor detection (an average of 6.5 months). They were detected within 12-156 months (an average of 48 months) among relapsing patients with nodal sclerosis and mixed-cell tumors stage III-IVAB, mostly concomitant with anemia and lymphopenia. Standard combination chemotherapeutical regimens for primary patients (MOPP, ABVD and LOPP) and relapsing ones (CCNU-OPP and ABVD) were effective in those with bone marrow lesions. Side-effects (myelodepression) did not exceed normal levels, provided human recombinant interleukin-1 beta (beta-leukin) was administered for protection and leukopoietic stimulation.


Recombining Your Way Out of Trouble: The Genetic Architecture of Hybrid Fitness under Environmental Stress.
Hybridization between species can either promote or impede adaptation. But we know very little about the genetic basis of hybrid fitness, especially in nondomesticated organisms, and when populations are facing environmental stress. We made genetically variable F2 hybrid populations from two divergent Saccharomyces yeast species. We exposed populations to ten toxins and sequenced the most resilient hybrids on low coverage using ddRADseq to investigate four aspects of their genomes: 1) hybridity, 2) interspecific heterozygosity, 3) epistasis (positive or negative associations between nonhomologous chromosomes), and 4) ploidy. We used linear mixed-effect models and simulations to measure to which extent hybrid genome composition was contingent on the environment. Genomes grown in different environments varied in every aspect of hybridness measured, revealing strong genotype-environment interactions. We also found selection against heterozygosity or directional selection for one of the parental alleles, with larger fitness of genomes carrying more homozygous allelic combinations in an otherwise hybrid genomic background. In addition, individual chromosomes and chromosomal interactions showed significant species biases and pervasive aneuploidies. Against our expectations, we observed multiple beneficial, opposite-species chromosome associations, confirmed by epistasis- and selection-free computer simulations, which is surprising given the large divergence of parental genomes (∼15%). Together, these results suggest that successful, stress-resilient hybrid genomes can be assembled from the best features of both parents without paying high costs of negative epistasis. This illustrates the importance of measuring genetic trait architecture in an environmental context when determining the evolutionary potential of genetically diverse hybrid populations.


Targeting BCL-2 and ABL/LYN in Philadelphia chromosome-positive acute lymphoblastic leukemia.
Treatment of Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL) remains a challenge. Although the addition of targeted tyrosine kinase inhibitors (TKIs) to standard cytotoxic therapy has greatly improved upfront treatment, treatment-related morbidity and mortality remain high. TKI monotherapy provides only temporary responses and renders patients susceptible to the development of TKI resistance. Thus, identifying agents that could enhance the activity of TKIs is urgently needed. Recently, a selective inhibitor of B cell lymphoma 2 (BCL-2), ABT-199 (venetoclax), has shown impressive activity against hematologic malignancies. We demonstrate that the combination of TKIs with venetoclax is highly synergistic in vitro, decreasing cell viability and inducing apoptosis in Ph(+)ALL. Furthermore, the multikinase inhibitors dasatinib and ponatinib appear to have the added advantage of inducing Lck/Yes novel tyrosine kinase (LYN)-mediated proapoptotic BCL-2-like protein 11 (BIM) expression and inhibiting up-regulation of antiapoptotic myeloid cell leukemia 1 (MCL-1), thereby potentially overcoming the development of venetoclax resistance. Evaluation of the dasatinib-venetoclax combination for the treatment of primary Ph(+)ALL patient samples in xenografted immunodeficient mice confirmed the tolerability of this drug combination and demonstrated its superior antileukemic efficacy compared to either agent alone. These data suggest that the combination of dasatinib and venetoclax has the potential to improve the treatment of Ph(+)ALL and should be further evaluated for patient care.


Draft genomes of two rhizosphere associated bacterial isolates from Tims Branch, a heavy metal contaminated wetland.
Two bacterial isolates were recovered from wetland sediments from Tims Branch, a heavy metal contaminated wetland located at the Savannah River Site. Draft genomes of the two recovered isolates, Rhodoblastus strain 17X3 and Comamonas strain 17RB, were generated from Illumina MiSeq sequencing data.


Effects of stress and motivation on performing a spatial task.
Learning is ubiquitous in the animal kingdom but has been studied extensively in only a handful of species. Moreover, learning studied under laboratory conditions is typically unrelated to the animal's natural environment or life history. Here, we designed a task relevant to the natural behavior of male African cichlid fish (Astatotilapia burtoni), to determine if they could be trained on a spatial task to gain access to females and shelter. We measured both how successfully animals completed this task over time and whether and how immediate early gene and hormone expression profiles were related to success. While training fish in a maze, we measured time to task completion, circulating levels of three key hormones (cortisol, 11-ketotestosterone, and testosterone) and mRNA abundance of seven target genes including three immediate early genes (that served proxies for brain activity) in nine brain regions. Data from our subjects fell naturally into three phenotypes: fish that could be trained (learners), fish that could not be trained (non-learners) and fish that never attempted the task (non-attempters). Learners and non-learners had lower levels of circulating cortisol compared to fish that never attempted the task. Learners had the highest immediate early gene mRNA levels in the homologue of the hippocampus (dorsolateral telencephalon; Dl), lower cortisol (stress) levels and were more motivated to accomplish the task as measured by behavioral observations. Fish that never attempted the task showed the lowest activity within the Dl, high stress levels and little to no apparent motivation. Data from non-learners fell between these two extremes in behavior, stress, and motivation.


The phospholipid-binding protein SESTD1 is a novel regulator of the transient receptor potential channels TRPC4 and TRPC5.
TRPC4 and TRPC5 are two closely related members of the mammalian transient receptor potential cation channel family that have been implicated in important physiological functions, such as growth cone guidance and smooth muscle contraction. To further unravel the role of TRPC4 and TRPC5 in these processes in vivo, detailed information about the molecular composition of native channel complexes and their association with cellular signaling networks is needed. We therefore searched a human aortic cDNA library for novel TRPC4-interacting proteins using a modified yeast two-hybrid assay. This screen identified SESTD1, a previously uncharacterized protein containing a lipid-binding SEC14-like domain as well as spectrin-type cytoskeleton interaction domains. SESTD1 was found to associate with TRPC4 and TRPC5 via the channel's calmodulin- and inositol 1,4,5-trisphosphate receptor-binding domain. In functional studies, we demonstrate that SESTD1 binds several phospholipid species in vitro and is essential for efficient receptor-mediated activation of TRPC5. Notably, phospholipid binding to SESTD1 was Ca(2+)-dependent. Because TRPC4 and -5 conduct Ca(2+), SESTD1-channel signaling may be bidirectional and also couple TRPC activity to lipid signaling through SESTD1. The modulation of TRPC channel function by specific lipid-binding proteins, such as SESTD1, adds another facet to the complex regulation of these channels complementary to the previously described effects of direct channel-phospholipid interaction.


Clozapine Use in Patients with Early-Stage Schizophrenia in a Chinese Psychiatric Hospital.
Previous studies suggest that clozapine is commonly underutilized and that its initiation is delayed in patients with first-episode schizophrenia. Knowledge regarding clozapine use among Chinese patients with early-stage schizophrenia is limited. The aim of the present study was to investigate the point prevalence of and patterns and factors associated with clozapine use in patients with early-stage schizophrenia discharged from a psychiatric hospital in China. A retrospective study was conducted to analyze the prescriptions of 867 consecutive patients with early-stage schizophrenia who were admitted to the Affiliated Brain Hospital of Guangzhou Medical University between Jan 1, 2011 and Dec 31, 2016. At discharge from the hospital, 114 (13.1%) patients were prescribed clozapine. Among the patients taking clozapine, 93 patients (81.6%) were prescribed clozapine polypharmacy, and only 21 patients (18.4%) were prescribed clozapine monotherapy. None of the patients were prescribed an overdose of clozapine. The mean daily dosage of clozapine was 160.97 mg, 149.05 mg and 213.69 mg among all patients taking clozapine, patients taking clozapine polypharmacy and patients taking clozapine monotherapy, respectively. The antipsychotic most frequently combined with clozapine was risperidone. Logistic regression suggested that the length of hospital stay, high school education, lower benzodiazepine use and antipsychotic polypharmacy were independently and significantly associated with clozapine use (P<0.05). Although clozapine has been commonly used in China in recent years, the present study found that clozapine was not commonly used in patients with early-stage schizophrenia. An underutilization and delayed initiation of clozapine may exist in a portion of patients with early-stage schizophrenia. Given the unfavorable outcomes of underutilized and delayed clozapine use, future studies may be needed to assess and increase clozapine use in this population.


Evidence for a Role for NAD(P)H Dehydrogenase in Concentration of CO2 in the Bundle Sheath Cell of Zea mays.
Prior studies with Nicotiana and Arabidopsis described failed assembly of the chloroplastic NDH [NAD(P)H dehydrogenase] supercomplex by serial mutation of several subunit genes. We examined the properties of Zea mays leaves containing Mu and Ds insertions into nuclear gene exons encoding the critical o- and n-subunits of NDH, respectively. In vivo reduction of plastoquinone in the dark was sharply diminished in maize homozygous mutant compared to normal leaves but not to the extreme degree observed for the corresponding lesions in Arabidopsis. The net carbon assimilation rate (A) at high irradiance and saturating CO2 levels was reduced by one-half due to NDH mutation in maize although no genotypic effect was evident at very low CO2 levels. Simultaneous assessment of chlorophyll fluorescence and A in maize at low (2% by volume) and high (21%) O2 levels indicated the presence of a small, yet detectable, O2-dependent component of total linear photosynthetic electron transport in 21% O2 This O2-dependent component decreased with increasing CO2 level indicative of photorespiration. Photorespiration was generally elevated in maize mutant compared to normal leaves. Quantification of the proportion of total electron transport supporting photorespiration enabled estimation of the bundle sheath cell CO2 concentration (Cb) using a simple kinetic model of ribulose bisphosphate carboxylase/oxygenase function. The A versus Cb relationships overlapped for normal and mutant lines consistent with occurrence of strictly CO2-limited photosynthesis in the mutant bundle sheath cell. The results are discussed in terms of a previously reported CO2 concentration model [Laisk A, Edwards GE (2000) Photosynth Res 66: 199-224].


Early child neglect: does it predict obesity or underweight in later childhood?
Child neglect has been hypothesized to be a risk factor for both obesity and underweight in early childhood, although little research has examined the relation between neglect and body mass index (BMI). The current study examined the relation between neglect and BMI among 185 children (91 with a Child Protective Services [CPS] history of neglect) who were initially seen at ages 4-6 years and who were followed through ages 7-9 years. Neglected and comparison children were found to have similar BMIs, although both groups had BMIs that were significantly greater than Centers for Disease Control and Prevention (CDC) norms for age, gender, and ethnicity. Neglect chronicity did predict lower BMIs but only at ages 8 and 9 years. The current findings suggest that greater examination of moderators is needed to identify the specific contexts in which neglect is related to children's weight.


Palatal fistula resulting from cocaine abuse: a case report.
Cocaine blocks the reuptake of norepinephrine and dopamine of the sympathetic nervous system. It has a psychomotor stimulating effect that provokes euphoria, motor activity and an amplification of a well being sensation similar to the effect of amphetamines. Well known are the systemic effects of cocaine abuse (acute myocardial infarction, cardiac arrhythmias, aortic ruptures, cerebrovascular accidents). Several local effects are also documented involving above all palatal mucosa and central midface structures. The most common palatal lesions are fistulae resulting from ischemia caused by the constricting effect of cocaine on small vessels, mucosal breakdown, loss of palatal bone, and loss of nasal mucosa. IF cocaine use becomes chronic and compulsive, wider lesions may cause extensive destruction of the osteocartilagineous structures of the nose, sinus and palate, a syndrome called CIMDL (cocaine-induced midline destructive lesion). Many techniques have been described to treat these defects depending on the size and the involved structures of the face. These techniques range from local flaps to complex free flaps. The authors describe a case of a median hard and soft palatal fistula resulting from cocaine abuse treated with local bilateral palatal flaps, and analyze the most common therapeutic options described in literature.


Conflicting views on the molecular structure of the cancer antigen CA125/MUC16.
CA125 is a tumor antigen used to monitor the progression and regression of epithelial ovarian cancer. Despite the widespread use of CA125, the biochemical and molecular nature of this antigen is poorly understood. Analysis of the structure of CA125 is essential for determining the physiological role of this very significant tumor marker. Accumulated experimental evidence has shown that CA125 epitopes reside on a molecule of very complex architecture in terms of both protein backbone and oligosaccharide structures. It is not clear whether the heterogeneity of CA125 molecular characteristics are due to the variability of biological sources from which the molecule was isolated or to the different biophysical methods used for the characterization of all the oligosaccharides linked to CA125 or to the presence of glycoisoforms for this protein. This review attempts to summarize emerging data related to molecular characteristics of CA125 and to compare approaches undertaken to reach a better understanding of molecular features of this tumor marker.


Primary cultures of neuronal and non-neuronal rat brain cells secrete similar proportions of amyloid beta peptides ending at A beta40 and A beta42.
To determine the types of brain cells responsible for the production of amyloid beta peptides (A beta), as well as their carboxyl-terminal properties, we studied the secretion of A beta in rat neuronal, astrocytic, microglial and meningeal primary cell cultures. All four types of cells produced A beta, among which neurons secreted approximately 4 times more A beta than other cell types. The percentage of A beta42 ending at position 42 as a fraction of total A beta was similar between different cell types, ranging from 10 to 15%. These results suggest that neurons might be the most potent source for A beta production in the brain, although other non-neuronal type cells could also contribute to this process.


In vivo imaging of cardiac related impedance changes.
Electrical impedance tomography (EIT) produces cross-sectional images of the electrical resistivity distribution within the body, made from voltage or current measurements through electrodes attached around the body. The authors describe a gated EIT system to image the cardiogenic electrical resistivity variations and the results of in vivo studies on human subjects. It is shown that the sensitivity of EIT to tissue resistivity variations due to blood perfusion is good enough to image blood flow to the lungs; hence, abnormalities in pulmonary perfusion, such as pulmonary embolism, should appear in EIT images. In addition, more valuable information related to the cardiac activity can be gained from EIT images than from impedance cardiography. It is thus likely that a cardiac output index may be calculable from the average resistivity variations over the ventricles, but considerable research is required before the images can be understood in detail.


Appraising the "entourage effect": Antitumor action of a pure cannabinoid versus a botanical drug preparation in preclinical models of breast cancer.
Breast cancer is the second leading cause of death among women. Although early diagnosis and development of new treatments have improved their prognosis, many patients present innate or acquired resistance to current therapies. New therapeutic approaches are therefore warranted for the management of this disease. Extensive preclinical research has demonstrated that cannabinoids, the active ingredients of Cannabis sativa, trigger antitumor responses in different models of cancer. Most of these studies have been conducted with pure compounds, mainly Δ9-tetrahydrocannabinol (THC). The cannabis plant, however, produces hundreds of other compounds with their own therapeutic potential and the capability to induce synergic responses when combined, the so-called "entourage effect". Here, we compared the antitumor efficacy of pure THC with that of a botanical drug preparation (BDP). The BDP was more potent than pure THC in producing antitumor responses in cell culture and animal models of ER+/PR+, HER2+ and triple-negative breast cancer. This increased potency was not due to the presence of the 5 most abundant terpenes in the preparation. While pure THC acted by activating cannabinoid CB2 receptors and generating reactive oxygen species, the BDP modulated different targets and mechanisms of action. The combination of cannabinoids with estrogen receptor- or HER2-targeted therapies (tamoxifen and lapatinib, respectively) or with cisplatin, produced additive antiproliferative responses in cell cultures. Combinations of these treatments in vivo showed no interactions, either positive or negative. Together, our results suggest that standardized cannabis drug preparations, rather than pure cannabinoids, could be considered as part of the therapeutic armamentarium to manage breast cancer.


Prenatal bisphenol A exposure is associated with language development but not with ADHD-related behavior in toddlers from the Odense Child Cohort.
Bisphenol A (BPA) is a non-persistent chemical with endocrine disrupting abilities widely used in a variety of consumer products. The fetal brain is particularly sensitive to chemical exposures due to its rapid growth and complexity. Some studies have reported associationbetween maternal BPA exposure and behavior but few have assessed impact on cognitive development, and to our knowledge no studies have specifically assessed the impact on language development. We therefore assessed whether maternal urinary BPA concentration during pregnancy was associated with language development and attention-deficit and hyperactivity disorder (ADHD) symptoms in offspring aged 18-36 months in the prospective Odense Child Cohort. BPA was analyzed in 3rd trimester maternal fasting urine spot samples. Language development was addressed among 535 children using the Danish adaptation of the MacArthur-Bates Communicative Development Inventories at median age 21 months; ADHD traits were assessed by parents of 658 children using the Child Behavior Checklist for ages 1½-5 years at mean age 2.7 years. Associations were assessed using logistic regression models comparing children below the 15th percentile score for language and above the 85 percentiles score for ADHD with the other children while stratifying by sex and adjusting for maternal education, duration of breastfeeding and maternal urine phthalates. BPA was detected in 85.3% of the urine samples (median 1.2 ng/ml). Boys of mothers with BPA exposure in the highest tertile had an odds ratio of 3.70 (95% CI 1.34-10.21) of being in the lowest 15th percentile of vocabulary score compared to boys of mothers within the lowest tertile of BPA exposure after adjustment, whereas no association was found in girls. No clear dose-response relationship between maternal BPA and ADHD scores above the 85th percentile was found for either sex. Since early language development is a predictor of future reading skills and educational success, more epidemiological studies assessing BPA exposure and language skills are needed to confirm our findings.


The prognostic dilemma of nodal micrometastases in breast carcinoma.
The presence of axillary lymph node metastasis in patients with breast cancer is a major prognostic factor and also determines the use of adjuvant chemotherapy. Micrometastasis has been arbitrary defined as deposits of < 2 mm dimension. Earlier studies of micrometastases failed to demonstrate prognostic relevance. However, when larger numbers of patients were followed up for longer periods, micrometastasis was shown to be a significantly poor prognostic parameter with patients having a survival rate similar to those with macrometastasis or nodal disease. There are no compelling reasons to retain the term "micrometastasis" in the light of these findings and our understanding of tumor biology. Routine histological examination of axillary lymph nodes is a notoriously inaccurate method for the detection of metastases. When serial or multilevel sectioning and/or immunohistochemical staining for cytokeratin were employed, detection rates increased by as much as 33%. Reverse transcriptase-polymerase chain reaction and Southern blotting for CK19 may be a more accurate method of examination. However, there are inherent technical problems associated with this method, and the recent finding of a pseudogene with great homology to CK19 in normal peripheral blood nucleated cells further emphasises the need for caution in this approach. It is not cost-effective to employ serial sectioning and immunohistochemistry when examining the axillary contents. However, the introduction of sentinel-node biopsy may allow detailed examination of the single node most likely to harbour a metastatic tumor.


Seroprevalence of SARS-CoV-2 antibodies and knowledge, attitude and practice toward COVID-19 in the Republic of Srpska-Bosnia & Herzegovina: A population-based study.
The aim of the study was to assess the seroprevalence of SARS-CoV-2 in the Republic of Srpska, Bosnia and Herzegovina, and to analyse the knowledge, attitudes and practices of the population toward COVID-19. This population-based study was conducted in a group of 1,855 randomly selected individuals from all municipalities from 1 December 2020 to 15 January 2021. All individuals were asked to sign a consent form and to fill in a questionnaire, following which a blood samples were collected. Total anti-SARS-CoV-2 antibodies were determined in serum specimens using the total Ab ELISA assay. The overall seroprevalence rate was 40.3%. Subjects aged <65 years were 2.06 times more likely to be seropositive than those aged ≥65, and 30% of seropositive individuals presented no COVID-19 symptoms. The household members of seropositive individuals were 2.24 times more likely to develop COVID-19 symptoms than the household members of seronegative individuals. More than 95% of respondents believe that preventive measures are very important to control the infection transmission. Majority of respondents wear the masks properly, maintain the required physical distance whenever possible and wash hands with soap. Nearly 50% of individuals were of the opinion that the vaccine could prevent the infection. This study showed that an overall SARS-CoV-2 seropositivity rate by the middle of January 2021 was very high. Attitudes and practices regarding the COVID-19 indicate that additional efforts should be taken in order to improve the health education with a focus on preventive measures and vaccination.


An effectiveness trial of contingency management in a felony preadjudication drug court.
This study evaluated a contingency management (CM) program in a drug court. Gift certificates for compliance were delivered at 4- to 6-week intervals (total value = $390.00). Participants in one condition earned gift certificates that escalated by $5.00 increments. Participants in a second condition began earning higher magnitude gift certificates, and the density of reinforcement was gradually decreased. No main effects of CM were detected, which appears to be attributable to a ceiling effect from the intensive contingencies already delivered in the drug court and the low density of reinforcement. Preplanned interaction analyses suggested that participants with more serious criminal backgrounds might have performed better in the CM conditions. This suggests that CM programs may be best suited for more incorrigible drug offenders.


Calcium plus vitamin D supplementation and the risk of incident diabetes in the Women's Health Initiative.
Experimental and epidemiologic studies suggest that calcium and vitamin D may reduce the risk of developing diabetes. We examined the effect of calcium plus vitamin D supplementation on the incidence of drug-treated diabetes in postmenopausal women. The Women's Health Initiative Calcium/Vitamin D Trial randomly assigned postmenopausal women to receive 1,000 mg elemental calcium plus 400 IU of vitamin D3 daily, or placebo, in a double-blind fashion. Among 33,951 participants without self-reported diabetes at baseline, we ascertained by treatment assignment new diagnoses of diabetes treated with oral hypoglycemic agents or insulin. Effects of the intervention on fasting measurements of glucose, insulin, and insulin resistance were examined among a subset of participants. Over a median follow-up time of 7 years, 2,291 women were newly diagnosed with diabetes. The hazard ratio for incident diabetes associated with calcium/vitamin D treatment was 1.01 (95% CI 0.94-1.10) based on intention to treat. This null result was robust in subgroup analyses, efficacy analyses accounting for nonadherence, and analyses examining change in laboratory measurements. Calcium plus vitamin D3 supplementation did not reduce the risk of developing diabetes over 7 years of follow-up in this randomized placebo-controlled trial. Higher doses of vitamin D may be required to affect diabetes risk, and/or associations of calcium and vitamin D intake with improved glucose metabolism observed in nonrandomized studies may be the result of confounding or of other components of foods containing these nutrients.


Double Fano resonances in an individual metallic nanostructure for high sensing sensitivity.
In this paper, we report on the design and observation of double Fano resonances (DFRs) in an individual symmetry-reduced nanostructure and the induced high sensing sensitivity. Such a plasmonic nanostructure consists of a partially overlapped double-metallic nanotriangles with unequal sizes fabricated by using fast and low-cost angle-resolved nanosphere lithography. Symmetry breaking generates two narrow quadrupolar dark modes, which further enhance the coupling with fundamental bright dipole modes within the same structure, manifesting the effect of DFRs. The resonance wavelength and line shape of DFRs can be tailored by changing the degree of asymmetry as well as the size of the designed nanostructure. Based on DFRs, a high sensitivity to dielectric environment with a maximum figure of merit of 35 is measured. Due to a fast manufacturing process with high reproducibility and high structural tunability, the fabricated individual metallic nanostructure provides an opportunity for significant potential applications in localized surface plasmon resonance based single or double-wavelength sensors in the near-infrared region.


COVID-19: Prevention and control measures in community
On January 30, 2020, the WHO declared the COVID-19 outbreak a public health emergency of international concern and, in March 2020, began to characterize it as a pandemic in order to emphasize the gravity of the situation and urge all countries to take action in detecting infection and preventing spread. Unfortunately, there is no medication that has been approved by the FDA, gone through controlled studies and demonstrated an effect on the virus for this global pandemic. Although there are cures for illnesses and developments made by leaps and bounds in our day, the strongest and most effective weapon that society has against this virus that is affecting not just health but also economics, politics, and social order, is the prevention of its spread. The main points in preventing the spread in society are hand hygiene, social distancing and quarantine. With increased testing capacity, detecting more COVID-19 positive patients in the community will also enable the reduction of secondary cases with stricter quarantine rules.


Fast and slow inactivation kinetics of the Ca2+ channels ECaC1 and ECaC2 (TRPV5 and TRPV6). Role of the intracellular loop located between transmembrane segments 2 and 3.
The Ca(2+) channels ECaC1 and ECaC2 (TRPV5 and TRPV6) share several functional properties including permeation profile and Ca(2+)-dependent inactivation. However, the kinetics of ECaC2 currents notably differ from ECaC1 currents. The initial inactivation is much faster in ECaC2 than in ECaC1, and the kinetic differences between Ca(2+) and Ba(2+) currents are more pronounced for ECaC2 than ECaC1. Here, we identify the structural determinants for these functional differences. Chimeric proteins were expressed heterologously in HEK 293 cells and studied by patch clamp analysis. Both channels retained their phenotype after exchanging the complete N termini, the C termini, or even both N and C termini, i.e. ECaC1 with the ECaC2 N or C terminus still showed the ECaC1 phenotype and vice versa. The substitution of the intracellular loop between the transmembrane domains 2 and 3 of ECaC2 with that of ECaC1 induced a delay of inactivation. Three amino acid residues (Leu-409, Val-411 and Thr-412) present in this loop determine the fast inactivation behavior. When this intracellular loop between the transmembrane domains 2 and 3 of ECaC1 was exchanged with the TM2-TM3 loop of ECaC2, the ECaC1 kinetics were analogous to ECaC2. In conclusion, the TM2-TM3 loop is a critical determinant of the inactivation in ECaC1 and ECaC2.


Silver Nanoparticles Stabilized by Poly (Vinyl Pyrrolidone) with Potential Anticancer Activity towards Prostate Cancer.
Tumor necrosis factor (TNF-α) and inflammatory cytokine (IL-6) play a vital role in various cellular incidents such as the proliferation and death of cells during carcinogenesis. Hence, regulation of these biomarkers could be a promising tool for controlling tumor progression using nanoformulations. Silver nanoparticles-poly (vinyl pyrrolidone) (AgNPs-PVP) were prepared using the reduction of silver nitrate and stabilized with PVP. They are characterized through yield percentage, UV-VIS, FT-IR, size, charge, and morphology. The obtained AgNPs were tested for anticancer activity against prostate cancer (PC 3) and human skin fibroblast (HFS) cell lines. Moreover, biomarker-based confirmations like TNF-α and IL-6 were estimated. The synthesized AgNPs-PVP were stable, spherical in shape, with particle sizes of 122.33 ± 17.61 nm, a polydispersity index of 0.49 ± 0.07, and a negative surface charge of -19.23 ± 0.61 mV. In vitro cytotoxicity testing showed the AgNPs-PVP exhibited antiproliferation properties in PC3 in a dose-dependent manner. In addition, when compared to control cells, AgNPs-PVP has lower TNF-α with a significant value ( ∗ p < 0.05); the value reached 16.84 ± 0.71 pg/ml versus 20.81 ± 0.44 pg/ml, respectively. In addition, HSF cells showed a high level of reduction ( ∗∗∗ p < 0.001) in IL-6 production. This study suggested that AgNPs-PVP could be a possible therapeutic agent for human prostate cancer and anti-IL-6 in cancerous and noncancerous cells. Further studies will be performed to investigate the effect of AgNPs-PVP in different types of cancer.


Evaluation of results of linac-based radiosurgery for brain metastases from primary lung cancer.
The purpose of our review was to evaluate results of radiosurgery for patients with brain metastases from lung cancer. Lung cancer is the leading cause of death from cancer and the most common source of brain metastases. Radiosurgery allows the precise focal delivery of a high single radiation dose to brain metastases and results in high rates of local control. 83 patients were treated between 2006 and 2008. We evaluated local control and outcome after radiosurgery and identified prognostic factors. Median survival in the whole group was 7.8 months from radiosurgery and 11 months from diagnosis. Median survival in classes I, II and III was 13.2, 8.2 and 2.2 months. For 94% of patients symptoms improved or stabilised at the first follow-up visit and this status did not change during 7.1 months. According to the univariate analysis, factors associated with improved survival included: RPA class 1 compared with RPA 2 and 3, RPA class 2 compared with RPA 3, KPS > 70, control of the primary disease, radiosurgery performed more than once, level of haemoglobin >7 mmol/1, absence of extracranial metastases, volume of the biggest lesion <11 cm(3). The multivariate analysis confirmed a significant influence on survival for the following factors: RPA class 1 as compared with RPA 3, KPS > 70, absence of extracranial metastases, multiplicity of radiosurgery. Stereotactic radiosurgery is a safe and effective treatment. It proved to be effective and safe in older patients. Selection of patients who are likely to benefit most should be based on prognostic factors. KPS proved to be the most important prognostic factor. In the RPA III group (patients with KPS < 70) survival time was similar to that achieved after symptomatic medical management.


Healthcare Practitioners Knowledge of Shoulder Injury Related to Vaccine Administration (SIRVA).
Background: Vaccine pharmacovigilance is at the forefront of the public eye. Shoulder Injuries Related to Vaccine Administration (SIRVA) is a poorly understood Adverse Event Following Immunisation, with iatrogenic origins. Criteria for medicolegal diagnosis of SIRVA is conflicting, current literature and educational materials are lacking, and healthcare practitioner knowledge of the condition is unknown. Methods: A cross-sectional, convenience sampled survey, utilising a validated online questionnaire assessed practitioner knowledge of SIRVA, safe injecting, and upper limb anatomy, and preferred definition for SIRVA. Results: Mean scores were moderate for safe injecting knowledge (69%), and poor for knowledge of anatomy (42%) and SIRVA (55%). Non-immunising healthcare practitioners scored significantly (p = 0.01, and < 0.05, respectively) higher than immunising practitioners for anatomy (2.213 ± 1.52 vs. 3.12 ± 1.50), and safe injecting knowledge (6.70 ± 1.34 vs. 7.14 ± 1.27). Only 52% of authorised vaccinators accurately selected a 40 × 20 mm area recommended for safe injecting. Majority (91.7%) of respondents thought nerve injuries should be included in the diagnostic criteria for SIRVA. Discussion and conclusions: Greater education and awareness of SIRVA is needed in all healthcare disciplines. Consensus regarding SIRVA definition is paramount for accurate reporting and improved future understanding of all aspects of SIRVA.


[Septicaemic-endotoxic shock: problems of early diagnosis and treatment (author's transl)].
At least in its early stage, the syndrome of septicaemic-endotoxic shock gives rise to a heterogeneous clinical picture which often lacks definite clinical, haemodynamic and biochemical features. But even if this form of shock is merely suspected, adequate fluid supply and administration of appropriate antibiotics should be started, in addition to general supportive measures. The administration of vasoactive drugs and corticosteroids has provided favourable results. The low early mortality (10 among 50 patients) is probably due to the medical causes which precipitate shock in this group, and the early onset of treatment.


Delusional infestation: are you being bugged?
This case report documents a 58-year-old male who presented to the clinic with a 12-month history of a burrowing sensation in his eyelids that he attributed to a parasitic infestation. After being extensively investigated and reviewed by relevant specialties, no evidence of parasitic infestation was found. He was diagnosed with and treated for blepharitis. Psychiatric referral for presumed delusional infestation (DI) was recommended. Despite this, he remained insistent in his belief of infestation, and was inevitably lost to follow-up. DI, previously known as delusional parasitosis, is a rare delusional disorder where affected individuals have a fixed, false belief that they have a parasitic infestation. Diagnosis can be challenging. Practitioners need to evaluate between primary and secondary DI carefully, as management differs depending on the etiology. Despite this, patients diagnosed with primary DI tend to be resistant to psychiatric referral. This report aims to optimize management by giving the reader a guideline for appropriate investigations and advice on patient approach. It is important to recognize hallmark features of DI to minimize self-inflicted trauma and associated psychosocial consequences. Effective treatment for DI is available, and devastating consequences, including blindness, can be avoided.


In vitro self-assembly of gold nanoparticle-coated poly(3-hydroxybutyrate) granules exhibiting plasmon-induced thermo-optical enhancements.
Polyhydroxyalkanoate (PHA) synthase attached to gold nanoparticles (AuNP) produce poly(3-hydroxybutyrate) (PHB) upon the addition of 3-hydroxybutyrate-CoA, and then coalesce to form micrometer-sized AuNP-coated PHB granules. These AuNP-coated PHB granules are potential theranostic agents that have enhanced imaging capabilities and are capable of heating upon near-infrared laser irradiation. The AuNP-coated PHB exhibited 11-fold enhancement in surface-enhanced Raman scattering over particles prior polymerization. Stained AuNP-coated PHB exhibited a 6-fold enhancement in fluorescence intensity as well as a 1.3-fold decrease in photobleaching rate compared to PHB granules alone. The granules were also shown to emit heat when illuminated at 808 nm with a 3.9-fold increase in heating rate compared to particles alone.


Morphometric analysis of the C57BL/6J mouse brain.
Magnetic resonance microscopy (MRM), when used in conjunction with active staining, can produce high-resolution, high-contrast images of the mouse brain. Using MRM, we imaged in situ the fixed, actively stained brains of C57BL/6J mice in order to characterize the neuroanatomical phenotype and produce a digital atlas. The brains were scanned within the cranium vault to preserve the brain morphology, avoid distortions, and to allow an unbiased shape analysis. The high-resolution imaging used a T1-weighted scan at 21.5 microm isotropic resolution, and an eight-echo multi-echo scan, post-processed to obtain an enhanced T2 image at 43 microm resolution. The two image sets were used to segment the brain into 33 anatomical structures. Volume, area, and shape characteristics were extracted for all segmented brain structures. We also analyzed the variability of volumes, areas, and shape characteristics. The coefficient of variation of volume had an average value of 7.0%. Average anatomical images of the brain for both the T1-weighted and T2 images were generated, together with an average shape atlas, and a probabilistic atlas for 33 major structures. These atlases, with their associated meta-data, will serve as baseline for identifying neuroanatomical phenotypes of additional strains, and mouse models now under study. Our efforts were directed toward creating a baseline for comparison with other mouse strains and models of neurodegenerative diseases.


Glutamate pharmacology and the treatment of schizophrenia: current status and future directions.
Glutamate-containing neuronal terminals are ubiquitous in the central nervous system and their functional importance in mental activity is considerable. Therefore, the involvement of this neurotransmitter in the pathology of schizophrenia is being studied. Biochemical evidence has suggested that glutamatergic transmission may be regionally reduced in schizophrenia, although this evidence has never been completely consistent nor fully replicable. More striking has been the behavioral effects in humans of the antiglutamatergic drugs phencyclidine (PCP) and its congener ketamine. By historical report, PCP produces a 'schizophrenia-like' psychosis in normal humans and aggravates the psychosis in schizophrenics. More recently, ketamine has been shown to produce a mild psychotomimetic effect in normal volunteers, which has some schizophrenia-like features. We have studied the effects of ketamine in schizophrenic patients. Here, ketamine intensified each patient's specific underlying psychosis, an effect not blocked by haloperidol. Moreover, ketamine selectively increased cerebral blood flow (CBF) in the anterior cingulate cortex and reduced CBF in hippocampus and lingual gyrus. These data may be pertinent to the subject's psychosis exacerbation, especially because both cingulate and hippocampus have been previously implicated in schizophrenic psychosis. In addition, ketamine produced a distinctive dynamic time-course of regional CBF changes in different anatomic regions, with immediate (5-10 min) changes in cingulate, but somewhat more delayed changes (20-40 min) in the thalamus and cerebellum. Our immediate early gene (IEG) time-course data with c-fos and zif268 in rats following PCP suggest that a single dose of this antiglutamatergic compound can have an effect in some brain areas which lasts beyond 48 h, an effect which is distinct by IEG and by region. Together, these data suggest that glutamate-mediated neurotransmission has a strong influence in schizophrenia, although the specifics of this involvement have yet to be articulated.


DNA structure specificity of Rap endonuclease.
The Rap protein of phage lambda is an endonuclease that nicks branched DNA structures. It has been proposed that Rap can nick D-loops formed during phage recombination to generate splice products without the need for the formation of a 4-strand (Holliday) junction. The structure specificity of Rap was investigated using a variety of branched DNA molecules made by annealing partially complementary oligo-nucleotides. On Holliday junctions, Rap endonuclease shows a requirement for magnesium or manganese ions, with Mn(2+)supporting 5-fold more cleavage than Mg(2+). The location of endonuclease incisions was determined on 3'-tailed D-loop, bubble, flayed duplex, 5'-flap and Y junction DNA substrates. In all cases, Rap preferentially cleaves at the branch point of these molecules. With a flayed duplex, incisions are made in the duplex adjacent to the single-strand arms. Comparison of binding and cleavage specificities revealed that Rap is highly structure-specific and exhibits a clear preference for 4- and 3-stranded DNA over Y and flayed duplex DNA. Almost no binding or cleavage was detected with duplex, partial duplex and single-stranded DNA. Thus Rap endonuclease shows a bias for structures that resemble D-loop and Holliday junction recombination intermediates.


Rehabilitating the addicted brain with transcranial magnetic stimulation.
Substance use disorders (SUDs) are one of the leading causes of morbidity and mortality worldwide. In spite of considerable advances in understanding the neural underpinnings of SUDs, therapeutic options remain limited. Recent studies have highlighted the potential of transcranial magnetic stimulation (TMS) as an innovative, safe and cost-effective treatment for some SUDs. Repetitive TMS (rTMS) influences neural activity in the short and long term by mechanisms involving neuroplasticity both locally, under the stimulating coil, and at the network level, throughout the brain. The long-term neurophysiological changes induced by rTMS have the potential to affect behaviours relating to drug craving, intake and relapse. Here, we review TMS mechanisms and evidence that rTMS is opening new avenues in addiction treatments.


Measles resurgence in Belgium from January to mid-April 2011: a preliminary report.
From 1 January to 14 April 2011, a total of 155 measles cases were notified in Belgium, whereas throughout 2010, there were only 40. Of the 103 cases with known vaccination status, 87% had not been vaccinated with measles-mumps-rubella vaccine. The resurgence of measles is the consequence of insufficient vaccine coverage in previous years. Efforts to communicate the benefits of measles vaccination to the public and to advise health professionals on control measures and outbreak management are ongoing.


Affinity and hydrophobic interactions of penicillin amidase.
Binding of penicillin amidase from E. coli 436 to aniline-, benzylamine- and phenylethylamine-Sepharose was studied. Binding of the enzyme to aniline-Sepharose was exclusively due to hydrophobic interactions. Benzylamine-Sepharose binds the enzyme due to affinity interactions in the absence of ammonium sulphate and due to hydrophobic interactions in the presence of ammonium sulphate. A conformational change in the penicillin amidase molecule due to ammonium sulphate there by exposing the side chain binding site as a hydrophobic core is suggested.


Chemotaxis of Dictyostelium discoideum: collective oscillation of cellular contacts.
Chemotactic responses of Dictyostelium discoideum cells to periodic self-generated signals of extracellular cAMP comprise a large number of intricate morphological changes on different length scales. Here, we scrutinized chemotaxis of single Dictyostelium discoideum cells under conditions of starvation using a variety of optical, electrical and acoustic methods. Amebas were seeded on gold electrodes displaying impedance oscillations that were simultaneously analyzed by optical video microscopy to relate synchronous changes in cell density, morphology, and distance from the surface to the transient impedance signal. We found that starved amebas periodically reduce their overall distance from the surface producing a larger impedance and higher total fluorescence intensity in total internal reflection fluorescence microscopy. Therefore, we propose that the dominant sources of the observed impedance oscillations observed on electric cell-substrate impedance sensing electrodes are periodic changes of the overall cell-substrate distance of a cell. These synchronous changes of the cell-electrode distance were also observed in the oscillating signal of acoustic resonators covered with amebas. We also found that periodic cell-cell aggregation into transient clusters correlates with changes in the cell-substrate distance and might also contribute to the impedance signal. It turned out that cell-cell contacts as well as cell-substrate contacts form synchronously during chemotaxis of Dictyostelium discoideum cells.


Mycotic keratitis by Fusarium. A review and report of two cases.
World literature on the subject reports 112 cases of keratomycosis by Fusarium, including the two cases described in this paper and some personal communications. In most of these cases (60--70%) there is a history of trauma inflicted by vegetable matter or by foreign bodies introduced into the eye (metal, sand, stone, etc.). The different species of Fusarium isolated are F. episphaeria, F. dimerum, F. moniliforme, F. nivale, F. oxysporum, and F. solani. The treatment recommended in a 5-percent suspension or unguent of pimaricin (natamycin).


Selectivity of the highly preorganized tetradentate ligand 2,9-di(pyrid-2-yl)-1,10-phenanthroline for metal ions in aqueous solution, including lanthanide(III) ions and the uranyl(VI) cation.
Some metal ion complexing properties of DPP (2,9-Di(pyrid-2-yl)-1,10-phenanthroline) are reported with a variety of Ln(III) (Lanthanide(III)) ions and alkali earth metal ions, as well as the uranyl(VI) cation. The intense π-π* transitions in the absorption spectra of aqueous solutions of 10(-5) M DPP were monitored as a function of pH and metal ion concentration to determine formation constants of the alkali-earth metal ions and Ln(III) (Ln = lanthanide) ions. It was found that log K(1)(DPP) for the Ln(III) ions has a peak at Ln(III) = Sm(III) in a plot of log K(1) versus 1/r(+) (r(+) = ionic radius for 8-coordination). For Ln(III) ions larger than Sm(III), there is a steady rise in log K(1) from La(III) to Sm(III), while for Ln(III) ions smaller than Sm(III), log K(1) decreases slightly to the smallest Ln(III) ion, Lu(III). This pattern of variation of log K(1) with varying size of Ln(III) ion was analyzed using MM (molecular mechanics) and DFT (density functional theory) calculations. Values of strain energy (∑U) were calculated for the [Ln(DPP)(H(2)O)(5)](3+) and [Ln(qpy)(H(2)O)(5)](3+) (qpy = quaterpyrdine) complexes of all the Ln(III) ions. The ideal M-N bond lengths used for the Ln(III) ions were the average of those found in the CSD (Cambridge Structural Database) for the complexes of each of the Ln(III) ions with polypyridyl ligands. Similarly, the ideal M-O bond lengths were those for complexes of the Ln(III) ions with coordinated aqua ligands in the CSD. The MM calculations suggested that in a plot of ∑U versus ideal M-N length, a minimum in ∑U occurred at Pm(III), adjacent in the series to Sm(III). The significance of this result is that (1) MM calculations suggest that a similar metal ion size preference will occur for all polypyridyl-type ligands, including those containing triazine groups, that are being developed as solvent extractants in the separation of Am(III) and Ln(III) ions in the treatment of nuclear waste, and (2) Am(III) is very close in M-N bond lengths to Pm(III), so that an important aspect of the selectivity of polypyridyl type ligands for Am(III) will depend on the above metal ion size-based selectivity. The selectivity patterns of DPP with the alkali-earth metal ions shows a similar preference for Ca(II), which has the most appropriate M-N lengths. The structures of DPP complexes of Zn(II) and Bi(III), as representative of a small and of a large metal ion respectively, are reported. [Zn(DPP)(2)](ClO(4))(2) (triclinic, P1, R = 0.0507) has a six-coordinate Zn(II), with each of the two DPP ligands having one noncoordinated pyridyl group appearing to be π-stacked on the central aromatic ring of the other DPP ligand. [Bi(DPP)(H(2)O)(2)(ClO(4))(2)](ClO(4)) (triclinic, P1, R = 0.0709) has an eight-coordinate Bi, with the coordination sphere composed of the four N donors of the DPP ligand, two coordinated water molecules, and the O donors of two unidentate perchlorates. As is usually the case with Bi(III), there is a gap in the coordination sphere that appears to be the position of a lone pair of electrons on the other side of the Bi from the DPP ligand. The Bi-L bonds become relatively longer as one moves from the side of the Bi containg the DPP to the side where the lone pair is thought to be situated. A DFT analysis of [Ln(tpy)(H(2)O)(n)](3+) and [Ln(DPP)(H(2)O)(5)](3+) complexes is reported. The structures predicted by DFT are shown to match very well with the literature crystal structures for the [Ln(tpy)(H(2)O)(n)](3+) with Ln = La and n = 6, and Ln = Lu with n = 5. This then gives one confidence that the structures for the DPP complexes generated by DFT are accurate. The structures generated by DFT for the [Ln(DPP)(H(2)O)(5)](3+) complexes are shown to agree very well with those generated by MM, giving one confidence in the accuracy of the latter. An analysis of the DFT and MM structures shows the decreasing O--O nonbonded distances as one progresses from La to Lu, with these distances being much less than the sum of the van der Waals radii for the smaller Ln(III) ions. The effect that such short O--O nonbonded distances has on thermodynamic complex stability and coordination number is then discussed.


Strategies for screening for hereditary non-polyposis colorectal cancer.
Germline mutations in DNA mismatch repair genes (MLH1, MSH2, PMS1, PMS2, and MSH6) predispose to hereditary non-polyposis colorectal cancer (HNPCC). In the absence of pathognomonic clinical features, diagnosis of HNPCC is often based on family history. Microsatellite instability (MSI) analysis has successfully been used for screening colorectal cancer patients for HNPCC. The aim of this study was to evaluate the feasibility of a recently introduced logistical model based on family history data in detecting HNPCC patients with germline mutations. A series of 509 kindreds with a proband with colorectal cancer was studied. MSI analysis and subsequent germline mutation analysis (MLH1 and MSH2) in MSI positive patients had been performed previously. Of the 509 patients, 63 (12%) were MSI positive and 10 (2%) had a germline mutation in MLH1 or MSH2. The power of the logistical model was tested to determine its value in predicting the probability of a germline mutation. The model proposed a high probability in three out of 10 mutation positive cases when data on cancer in first degree relatives were considered (typically three generation pedigrees, consisting, on average, of eight people). Using extended pedigrees and family cancer data in the 10 mutation positive kindreds (an average of 38 family members available), the model suggested high probabilities in seven out of 10 mutation positive cases. We conclude that for the model to predict germline mutation cases, extensive pedigrees and family history data are required. When screening colorectal cancer patients for HNPCC, a model using a combination of family information and MSI has optimal specificity and sensitivity.


[Complication associated with celiac disease].
Enteropathy associated T cell lymphoma is the most serious complication of celiac sprue. The incidence of malignancy, in general, is approximately twofold greater in celiac disease than in the general population, but the risk of specific gastrointestinal malignancies is markedly increased. Lymphoma of the small intestine, comprises approximately 50% of the malignancies, and the most are of T-cell origin. We report a case of celiac disease associated T-cell Lymphoma of the jejunum in a woman. At the age of 33 the diagnosis of Celiac sprue was made, after institution of a gluten free diet the patient improved, but, fifteen year later, the patient begin with fever and abdominal pain. Laparotomy showed a perforation of the intestine by a tumour.


Novel assay method for mitoxantrone in plasma, and its application in cancer patients.
Mitoxantrone is an anthracene derivative that acts as a cytostatic in a variety of cancers. A quantitative analytical method has been established for the determination of mitoxantrone in plasma. The method employed C18 reversed-phase ion-pair chromatography with an isocratic mobile phase of 50.0% methanol in 10 mM phosphate buffer (pH 3.0) plus 0.09% 1-pentanesulphonic acid and ultraviolet detection. Sample preparation consisted of two extraction steps using same organic solvent system at different pH to remove plasma impurities efficiently. Potential adsorption of mitoxantrone onto glassware was considered. Silanization of all glassware with 5% dichlorodimethylsilane in chloroform increased the extraction recovery in plasma from 50 to 85% with high reproducibility. Mitoxantrone was unstable in human plasma. To maintain plasma sample integrity, each millilitre of sample should be fortified with 0.1 ml of 5% vitamin C (in citrate buffer) and kept frozen until analysis. Using this new method, the calibration curve of mitoxantrone in plasma in the range of interest (1-500 ng/ml) showed good linearity (r = 0.996) and precision (both between-day and within-day coefficients of variation less than 10%). The lower detection limit of this assay method was 1 ng. The application of this method allowed us to study the stability of mitoxantrone in plasma, and the pharmacokinetics of mitoxantrone in nasopharyngeal carcinoma patients receiving 12 mg/m2. The study revealed a prolonged terminal phase half-life for mitoxantrone.


Pharyngeal polyps in two feline siblings.
Pharyngeal polyps were diagnosed in 2 sibling kittens referred for evaluation of chronic upper respiratory tract disease. Surgical excision of the masses was performed in both kittens after skull radiography and oral examination. One kitten had a recurrence of pharyngeal polyps 5 weeks after surgery and was euthanatized and necropsied. The clinical and histologic findings in these 2 cases suggested a congenital origin of this disease.


A Horizon Scan of Emerging Issues for Global Conservation in 2019.
We present the results of our tenth annual horizon scan. We identified 15 emerging priority topics that may have major positive or negative effects on the future conservation of global biodiversity, but currently have low awareness within the conservation community. We hope to increase research and policy attention on these areas, improving the capacity of the community to mitigate impacts of potentially negative issues, and maximise the benefits of issues that provide opportunities. Topics include advances in crop breeding, which may affect insects and land use; manipulations of natural water flows and weather systems on the Tibetan Plateau; release of carbon and mercury from melting polar ice and thawing permafrost; new funding schemes and regulations; and land-use changes across Indo-Malaysia.


Effect of oxytocin on calcium influx and efflux in the rat myometrium.
45Ca was used to investigate the effect of oxytocin on calcium movements in uterine smooth muscle from estrogenized rats. Upon exposure to oxytocin the influx of 45Ca in myometrial strips increased rapidly. After 2 min of incubation with oxytocin (1 microM) there was a 30% increase over the control. Prolonged stimulation with oxytocin caused no further accumulation of 45Ca. The amount of Ca gained following oxytocin stimulation could be removed by washing with a lanthanum solution. The rate of 45Ca efflux from preloaded tissues was substantially increased by oxytocin. In the absence of external Ca, the stimulation of 45Ca efflux by oxytocin was much reduced. The data indicate that oxytocin-induced contraction of the myometrium is largely dependent on the influx of extracellular calcium. Oxytocin appears to be capable of releasing small amounts of calcium from intracellular storage sites. The data also provide evidence against the inhibition by oxytocin of the calcium extrusion pump.


Characteristics and prevalence within serogroup O4 of a J96-like clonal group of uropathogenic Escherichia coli O4:H5 containing the class I and class III alleles of papG.
The recent discovery of a geographically dispersed clonal group of Escherichia coli O4:H5 that includes prototypic uropathogenic strain J96 prompted us to determine the prevalence of J96-like strains within serogroup O4 and to further assess the characteristics of such strains. We used O:K:H;F serotyping, PCR-based genomic fingerprinting, pulsed-field gel electrophoresis (PFGE), multilocus enzyme electrophoresis (MLEE), and PCR detection of the three papG alleles and of the cytotoxic necrotizing factor 1 (cnf1) and aerobactin (aer) gene sequences to characterize the 15 O4 strains among 336 E. coli isolates from three clinical collections (187 from mixed-source bacteremia, 75 from urosepsis, and 74 from acute cystitis). J96-like strains constituted approximately half of the O4 strains, or 2% of the total population. In contrast to other O4 strains, the J96-like strains characteristically exhibited specific group III capsular antigens, the H5 flagellar and F13 fimbrial antigens, a distinctive PCR genomic fingerprint, the class III papG allele (plus, in 50% of strains, the enigmatic class I papG allele), and cnf1 but lacked aer. A subset of these strains was remarkably homogeneous with respect to all these characteristics and exhibited a distinctive PFGE fingerprint and MLEE pattern. These findings clarify the epidemiological relevance of J96 as a model extraintestinal pathogen, provide further evidence of the class I papG allele outside of strain J96, and offer insights into the evolution of E. coli serogroup O4.


Cutaneous lava migrans in the paediatric foot.
Dermatological referrals to paediatrics are a common occurrence often with a vast range of diagnoses made prior to referral to secondary care services. The authors report a case of cutaneous lava migrans infection in an 11-year-old girl in Northern England following her return from a family holiday in Jamaica. The authors report this case to raise awareness of this diagnosis. This girl initially received treatment for alternate diagnoses, and our aim is to encourage people to think of tropical dermatoses as a diagnosis in the returning traveller of all age groups.


Fresnel-propagated imaging for the study of human tooth dentin by partially coherent x-ray tomography.
Recent methods of phase imaging in x-ray tomography allow the visualization of features that are not resolved in conventional absorption microtomography. Of these, the relatively simple setup needed to produce Fresnel-propagated tomograms appears to be well suited to probe tooth-dentin where composition as well as microstructure vary in a graded manner. By adapting analytical propagation approximations we provide predictions of the form of the interference patterns in the 3D images, which we compare to numerical simulations as well as data obtained from measurements of water immersed samples. Our observations reveal details of the tubular structure of dentin, and may be evaluated similarly to conventional absorption tomograms. We believe this exemplifies the power of Fresnel-propagated imaging as a form of 3D microscopy, well suited to quantify gradual microstructural-variations in teeth and similar tissues.


Intrinsic disorder is a common feature of hub proteins from four eukaryotic interactomes.
Recent proteome-wide screening approaches have provided a wealth of information about interacting proteins in various organisms. To test for a potential association between protein connectivity and the amount of predicted structural disorder, the disorder propensities of proteins with various numbers of interacting partners from four eukaryotic organisms (Caenorhabditis elegans, Saccharomyces cerevisiae, Drosophila melanogaster, and Homo sapiens) were investigated. The results of PONDR VL-XT disorder analysis show that for all four studied organisms, hub proteins, defined here as those that interact with > or = 10 partners, are significantly more disordered than end proteins, defined here as those that interact with just one partner. The proportion of predicted disordered residues, the average disorder score, and the number of predicted disordered regions of various lengths were higher overall in hubs than in ends. A binary classification of hubs and ends into ordered and disordered subclasses using the consensus prediction method showed a significant enrichment of wholly disordered proteins and a significant depletion of wholly ordered proteins in hubs relative to ends in worm, fly, and human. The functional annotation of yeast hubs and ends using GO categories and the correlation of these annotations with disorder predictions demonstrate that proteins with regulation, transcription, and development annotations are enriched in disorder, whereas proteins with catalytic activity, transport, and membrane localization annotations are depleted in disorder. The results of this study demonstrate that intrinsic structural disorder is a distinctive and common characteristic of eukaryotic hub proteins, and that disorder may serve as a determinant of protein interactivity.


Variants in estrogen-related genes and risk of Parkinson's disease.
Incidence rates of Parkinson's disease are higher in men than in women at all ages, and these differences may be a result of the neuroprotective effects of estrogen on the nigrostriatal pathway. We investigated the association of common variants in 4 estrogen-related genes with Parkinson's disease. Tagging single-nucleotide polymorphisms in the CYP19A1, ESR1, ESR2, and PRDM2 genes were selected from the International Haplotype Map and genotyped in 1103 Parkinson's disease cases from the upper Midwest of the United States and in 1103 individually matched controls (654 unaffected siblings, and 449 unrelated controls from the same region). Of 137 informative single-nucleotide polymorphisms, 2 PRDM2 single-nucleotide polymorphisms were significantly associated with an increased risk of Parkinson's disease at the Bonferroni-corrected significance level of 0.0004 (rs2744690: OR, 1.54; SE(logOR), .109; 99.96% CI, 1.05-2.26; uncorrected P = .0001; rs2744687: OR, 1.53; SE(logOR), .113; 99.96% CI, 1.03-2.29, uncorrected P = .0002); the association was significant in the women-only stratum but not in the men-only stratum. An additional 6 single-nucleotide polymorphisms in PRDM2, 2 in ESR1, 1 in ESR2, and 1 in CYP19A1 had significant P values in the overall sample before Bonferroni correction. None of the single-nucleotide polymorphisms were significantly associated with age at onset of Parkinson's disease after Bonferroni correction. Our results confirm the association of PRDM2 variants with Parkinson's disease susceptibility, especially in women.


Low Plasma Testosterone Is Associated With Elevated Cardiovascular Disease Biomarkers.
The relation between testosterone (T) plasma concentration and cardiovascular (CV) risk is unclear, with evidence supporting increased risk in men with low and high T levels. Few studies have assessed CV risk as a function of plasma T levels using objective biomarkers. To determine the relation between T levels and high-sensitivity CV risk biomarkers. Ten thousand forty-one male patients were identified in the database of a commercial clinical laboratory performing biomarker testing. Patients were grouped by total T concentration and associations with the following biomarkers were determined: cardiac troponin I (cTnI), endothelin-1 (ET-1), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-17A, N-terminal pro-B-type natriuretic peptide (NTproBNP), high-density lipoprotein (HDL) cholesterol, high-sensitivity C-reactive protein (hs-CRP), hemoglobin A1c (HbA1c), and leptin. Association of CV risk markers with levels of T in men. The median age of the cohort was 58 years (interquartile range = 48-68), and the median plasma T level was 420 ng/dL (interquartile range = 304-565); T levels did not vary with patient age. An inverse relation between plasma T levels and CV risk was observed for 9 of 10 CV markers: cTnI, ET-1, IL-6, TNF-α, NTproBNP, HDL cholesterol, hs-CRP, HbA1c, and leptin. Even after adjusting for age, body mass index, HbA1c, hs-CRP, and HDL cholesterol levels, the CV markers IL-6, ET-1, NTproBNP, and leptin were significantly associated with a T level lower than 250 ng/dL. Men with low T levels could be at increased risk for increased CV disease as seen by increased CV risk markers. This study was performed in a group of 10,041 men and is the first study to examine CV risk associated with circulating T levels using a large panel of 10 objective biomarkers. This study is limited by an absence of clinical data indicating whether men had pre-existing CV disease or other CV risk factors. Men with low plasma T levels exhibit increases in CV risk markers, consistent with a potential increased risk of CV disease. Pastuszak AW, Kohn TP, Estis J, Lipshultz LI. Low Plasma Testosterone Is Associated With Elevated Cardiovascular Disease Biomarkers. J Sex Med 2017;14:1095-1103.


Variant alleles in factor V, prothrombin, plasminogen activator inhibitor-1, methylenetetrahydrofolate reductase and risk of thromboembolism in metastatic colorectal cancer patients treated with first-line chemotherapy plus bevacizumab.
Single-nucleotide polymorphisms (SNPs) related to hereditary thrombophilia were investigated as risk factors for thromboembolism in cancer patients. Their effect in metastatic colorectal cancer (mCRC) has never been explored so far. Our aim was to analyse the effect of coagulation factor V (FVL G1691A), prothrombin (PT G20210A), methylenetetrahydrofolate reductase (MTHFR C677T and A1298C) and plasminogen activator inhibitor type 1 (PAI-1 5G/4G) allelic variants in this setting. Fifty-two patients treated with first-line chemotherapy plus bevacizumab who developed a thromboembolic event in their lifetime were initially genotyped. A contemporary cohort of 127 patients who did not experience any thromboembolic event was also analysed. DNA was extracted from peripheral blood and genotypes were determined by real-time PCR, using LightSNiP (TIB MOLBIOL) on LightCcler 480 (Roche). The association between thromboembolism and SNPs was investigated by univariable and multivariable analyses. All SNPs were in Hardy-Weinberg equilibrium (χ2 test P>0.20). FVL G1691A and PT G20210A were present only in heterozygosis in 4 (2.2%) and 7 (3.9%) patients, respectively; MTHFR C677T in homozygosis in 29 (16.2%), MTHFR A1298C in homozygosis in 13 (7.3%); PAI-1 5G/4G in 98 (54.7%) and 4G/4G in 41 (23%) patients. At univariable analysis, treatment duration was significantly associated with thromboembolism (P<0.001), whereas gender, age, obesity, platelets count and chemotherapy backbone were not. Similarly, FVL G1691A and PT G20210A as well as MTHFR C677T and PAI-1 4G allele were significantly associated, whereas MTHFR A1298C was not. At multivariable model including PT G20210A, MTHFR C677T and PAI-1 4G (age, obesity, treatment duration and chemotherapy backbone were included as adjustment factors), the three SNPs were significantlty associated with higher risk of thromboembolism (P=0.025, <0.0001 and P=0.033, respectively). Further validation studies are warranted in order to design a prospective trial of thromboprophylaxis in mCRC patients with high-risk genotypes.


Effect of feeding a reduced-starch diet with or without amylase addition on lactation performance in dairy cows.
The objective of this study was to determine lactation performance responses of high-producing dairy cows to a reduced-starch diet compared with a normal-starch diet and to the addition of exogenous amylase to the reduced-starch diet. Thirty-six multiparous Holstein cows (51+/-22 DIM and 643+/-49kg of body weight at trial initiation) were randomly assigned to 1 of 3 treatments in a completely randomized design: a 3-wk covariate adjustment period during which the cows were fed the normal-starch diet, followed by a 12-wk treatment period during which the cows were fed their assigned treatment diets. The normal-starch TMR did not contain exogenous amylase (NS-). The reduced-starch diets, formulated by partially replacing corn grain with soy hulls, were fed without (RS-) and with (RS+) exogenous amylase added to the TMR. Starch and NDF concentrations averaged 27.1 and 30.6%, 21.8 and 36.6%, and 20.7 and 36.6% (dry matter basis) for the NS-, RS-, and RS+ diets, respectively. Dry matter intake for cows fed the RS- diet was 2.4 and 3.2kg/d greater than for cows fed the NS- and RS+ diets, respectively. Intake of NDF ranged from 1.19 to 1.52% of body weight among the treatments, with the RS- diet being 28% greater than the NS- diet and 13% greater than the RS+ diet. Milk yield averaged 50.4kg/d and was unaffected by treatment. Fat-corrected milk yield was 2.9kg/d greater for cows fed the RS- diet than for cows fed the NS- diet. Body weight and body condition score measurements were unaffected by treatment. Fat-, solids-, and energy-corrected milk feed conversions (kilograms/kilogram of DMI) were 12 to 13% greater for cows fed the RS+ diet than for cows fed the RS- diet. Dry matter and nutrient digestibilities were lowest for cows fed the NS- diet and greatest for cows fed the RS+ diet, and were greater for cows fed the RS+ diet than for cows fed the RS- diet, with the exception of starch digestibility, which was similar. Greater conversion of feed to milk for dairy cows fed reduced-starch diets that include exogenous amylase may offer potential for improving economic performance.


Susceptibilities to clarithromycin, amoxycillin and metronidazole of Helicobacter pylori isolates from the antrum and corpus in Tokyo, Japan, 1995-2001.
The aim of this study was to determine the susceptibilities to clarithromycin, amoxycillin and metronidazole of Helicobacter pylori isolates from the antrum and corpus of Japanese patients examined during the period 1995-2001. There was an increase, from 6.2% in 1995 to 22.1% in 2000-2001, in the proportion of patients infected with clarithromycin-resistant H. pylori. Of patients infected with clarithromycin-resistant H. pylori, 39.1% were infected with both clarithromycin-susceptible and -resistant H. pylori. Furthermore, the MIC90 of clarithromycin for H. pylori rose from < 1 mg/L in 1995-1998 to 8 mg/L in 1999. In contrast, the MIC90s of amoxycillin and metronidazole were < or = 0.125 and 4 mg/L, respectively, throughout the study period. The results showed that, while most H. pylori isolates were susceptible to amoxycillin and metronidazole, resistance to clarithromycin among H. pylori isolates increased markedly in Japan during 1995-2001. The results also indicated a need to test the susceptibility of H. pylori isolates from more than two samples obtained from two different sites in the stomach of a single patient in order to diagnose the presence of clarithromycin-resistant H. pylori correctly.


Neuronal SUMOylation: mechanisms, physiology, and roles in neuronal dysfunction.
Protein SUMOylation is a critically important posttranslational protein modification that participates in nearly all aspects of cellular physiology. In the nearly 20 years since its discovery, SUMOylation has emerged as a major regulator of nuclear function, and more recently, it has become clear that SUMOylation has key roles in the regulation of protein trafficking and function outside of the nucleus. In neurons, SUMOylation participates in cellular processes ranging from neuronal differentiation and control of synapse formation to regulation of synaptic transmission and cell survival. It is a highly dynamic and usually transient modification that enhances or hinders interactions between proteins, and its consequences are extremely diverse. Hundreds of different proteins are SUMO substrates, and dysfunction of protein SUMOylation is implicated in a many different diseases. Here we briefly outline core aspects of the SUMO system and provide a detailed overview of the current understanding of the roles of SUMOylation in healthy and diseased neurons.


Differential antibacterial activity against Pseudomonas aeruginosa by carbon monoxide-releasing molecules.
Carbon monoxide (CO) delivered in a controlled manner to cells and organisms mediates a variety of pharmacological effects to the extent that CO-releasing molecules (CO-RMs) are being developed for therapeutic purposes. Recently, ruthenium-based CO-RMs have been shown to posses important bactericidal activity. Here we assessed the effect of fast CO releasers containing ruthenium (Ru(CO)(3)Cl(glycinate) [CORM-3] and tricarbonyldichlororuthenium(II) dimer [CORM-2]) and a novel slow manganese-based CO releaser ([Me(4)N][Mn(CO)(4)(thioacetate)(2)] [CORM-371]) on O(2) consumption and growth of Pseudomonas aeruginosa (PAO1). We then compared these effects with the action elicited by sodium boranocarbonate (CORM-A1), which lacks a transition metal but liberates CO with a rate similar to CORM-371. CORM-2, CORM-3, and, to a lesser extent, CORM-371 exerted a significant bactericidal effect and decreased O(2) consumption in PAO1 in vitro. The effect appeared to be independent of reactive oxygen species production, but in the case of metal-containing compounds it was prevented by the thiol donor N-acetylcysteine. In contrast, CORM-A1 was bacteriostatic rather than bactericidal in vitro eliciting only a moderate and transient decrease in O(2) consumption. None of the tested CO-RMs was toxic to murine macrophages or human fibroblasts at the concentration impairing PA01 growth but only ruthenium-containing CO-RMs showed potential therapeutic properties by increasing the survival of mice infected with PA01. CO carriers inhibit bacterial growth and O(2) consumption in vitro, but transition metal carbonyls appear more powerful than compounds spontaneously liberating CO. The nature of the metal in CO-RMs also modulates the anti-bacterial effect, with ruthenium-based CO-RMs being efficacious both in vitro and in vivo.


[A lyophilized reagent for standardization of the activated partial thromboplastin time].
Native partial thromboplastin, prepared from cadaver brain thromboplastin (a reagent), is a cephalin analog used to detect disorders in the first phase of coagulation. It was lyophilized with the aim of its stabilization, potential commercial manufacture, and introduction into practical clinical laboratories. Native emulsion was distributed into 0.2 ml penicillin flasks and lyophilized for 18-20 hrs in IZ-9 (CSR) or similar equipment at the maximum temperature of the product 5 degrees C. Lyophilized reagent is a white or slightly cream-colored powder. It retains its activity for at least a year as evidenced by the activated partial thromboplastin time test. The range of activities within one lot is less if commercial lyophilized donor reference plasma is used in the test. The reagent is stored in a refrigerator at 4-8 degrees C.


Identification of 1600 replication origins in S. cerevisiae.
There are approximately 500 known origins of replication in the yeast genome, and the process by which DNA replication initiates at these locations is well understood. In particular, these sites are made competent to initiate replication by loading of the Mcm replicative helicase prior to the start of S phase; thus, "a site to which MCM is bound in G1" might be considered to provide an operational definition of a replication origin. By fusing a subunit of Mcm to micrococcal nuclease, a technique referred to as "Chromatin Endogenous Cleavage", we previously showed that known origins are typically bound by a single Mcm double hexamer, loaded adjacent to the ARS consensus sequence (ACS). Here we extend this analysis from known origins to the entire genome, identifying candidate Mcm binding sites whose signal intensity varies over at least 3 orders of magnitude. Published data quantifying the production of ssDNA during S phase showed clear evidence of replication initiation among the most abundant 1600 of these sites, with replication activity decreasing in concert with Mcm abundance and disappearing at the limit of detection of ssDNA. Three other hallmarks of replication origins were apparent among the most abundant 5,500 sites. Specifically, these sites (1) appeared in intergenic nucleosome-free regions that were flanked on one or both sides by well-positioned nucleosomes; (2) were flanked by ACSs; and (3) exhibited a pattern of GC skew characteristic of replication initiation. Furthermore, the high resolution of this technique allowed us to demonstrate a strong bias for detecting Mcm double-hexamers downstream rather than upstream of the ACS, which is consistent with the directionality of Mcm loading by Orc that has been observed in vitro. We conclude that, if sites at which Mcm double-hexamers are loaded can function as replication origins, then DNA replication origins are at least 3-fold more abundant than previously assumed, and we suggest that replication may occasionally initiate in essentially every intergenic region. These results shed light on recent reports that as many as 15% of replication events initiate outside of known origins, and this broader distribution of replication origins suggest that S phase in yeast may be less distinct from that in humans than is widely assumed.


Hypercholesterolemia and 5-year risk of development of coronary heart disease among university and school workers in Jeddah, Saudi Arabia.
A study was performed among university and school employees as a step in detecting the prevalence of coronary heart disease (CHD) risk factors among school and university staff and the relation of the various risk factors to hypercholesterolemia and to predict the 5-years probability of development of CHD. All university staff and a sample of school workers in Jeddah were included in the study. Data were collected by interview and self-administered questionnaires, as well as by direct observation and measurement of weight, height, blood pressure, and total blood cholesterol. Logistic regression analysis was performed to identify the impact of various risk factors on hypercholesterolemia. The 5-year risk of development of CHD was calculated to identify the proportion at highest risk. Some CHD risk factors were detected among the participants as 18.8% were current cigarette smokers, around half were classified as overweight, and 19.9% were classified as hypertensive. Hypercholesterolemia was present in 10.1% and was significantly related to older age (40 years and over) and place of work. The estimated risk of CHD in the coming half-decade for those classified in the fifth quintile was 0.068. The Q(5)/Q(1) ratio has shown that those classified in the fifth quintile were at 3.6 times more risk of developing CHD in the coming 5 years than those classified in the first quintile. CHD risk factors are becoming prevalent in our society. The risk of development of CHD in the coming decades is not trivial. Short- and long-term health strategies are recommended to decrease the risk of CHD and improve the quality of life.


Expression, purification and characterisation of full-length histidine protein kinase RegB from Rhodobacter sphaeroides.
The global redox switch between aerobic and anaerobic growth in Rhodobacter sphaeroides is controlled by the RegA/RegB two-component system, in which RegB is the integral membrane histidine protein kinase, and RegA is the cytosolic response regulator. Despite the global regulatory importance of this system and its many homologues, there have been no reported examples to date of heterologous expression of full-length RegB or any histidine protein kinases. Here, we report the amplified expression of full-length functional His-tagged RegB in Escherichia coli, its purification, and characterisation of its properties. Both the membrane-bound and purified solubilised RegB protein demonstrate autophosphorylation activity, and the purified protein autophosphorylates at the same rate under both aerobic and anaerobic conditions confirming that an additional regulator is required to control/inhibit autophosphorylation. The intact protein has similar activity to previously characterised soluble forms, but is dephosphorylated more rapidly than the soluble form (half-life ca 30 minutes) demonstrating that the transmembrane segment present in the full-length RegB may be an important regulator of RegB activity. Phosphotransfer from RegB to RegA (overexpressed and purified from E. coli) by RegB is very rapid, as has been reported for the soluble domain. Dephosphorylation of active RegA by full-length RegB has a rate similar to that observed previously for soluble RegB.