Application of Ligand- and Structure-Based Prediction Models for the Design of Alkylhydrazide-Based HDAC3 Inhibitors as Novel Anti-Cancer Compounds.
Histone deacetylases (HDAC) represent promising epigenetic targets for several diseases including different cancer types. The HDAC inhibitors approved to date are pan-HDAC inhibitors and most show a poor selectivity profile, side effects, and in particular hydroxamic-acid-based inhibitors lack good pharmacokinetic profiles. Therefore, the development of isoform-selective non-hydroxamic acid HDAC inhibitors is a highly regarded field in medicinal chemistry. In this study, we analyzed different ligand-based and structure-based drug design techniques to predict the binding mode and inhibitory activity of recently developed alkylhydrazide HDAC inhibitors. Alkylhydrazides have recently attracted more attention as they have shown promising effects in various cancer cell lines. In this work, pharmacophore models and atom-based quantitative structure-activity relationship (QSAR) models were generated and evaluated. The binding mode of the studied compounds was determined using molecular docking as well as molecular dynamics simulations and compared with known crystal structures. Calculated free energies of binding were also considered to generate QSAR models. The created models show a good explanation of in vitro data and were used to develop novel HDAC3 inhibitors.


The effects of soy protein on chronic kidney disease: a meta-analysis of randomized controlled trials.
There is a growing body of evidence to indicate that soy protein consumption may have a beneficial effect on kidney function. We performed a meta-analysis to evaluate the effects of soy protein consumption compared with animal protein consumption in patients with pre-dialysis chronic kidney disease (CKD). We conducted a structured electronic search of the databases PubMed, EMBASE, Cochrane Library and Chinese Biological Medicine for randomized controlled trials published up to March 2014. The outcome measures were serum creatinine (SCR), triglyceride (TG), total cholesterol (TC), calcium (Ca) and phosphorus concentrations. Weighted or standard mean differences were calculated for net changes using random-effects models. The meta-analysis consisted of nine trials, comprising 197 subjects. Soy protein intake significantly reduced SCR and serum phosphorus concentrations. The mean difference was -6.231 μmol/l (95% confidence interval (CI): -11.109, -1.352 μmol/l) for SCR (P=0.012) and -0.804 (95% CI: -1.143, -0.464 μmol/l) for serum phosphorus (P=0.00). It also significantly reduced serum TG, with a pooled estimated change of -0.223 mmol/l (95% CI: -0.396, -0.051 mmol/l; P=0.011) after the exclusion of one trial indicated by sensitivity analyses. No statistically significant effects were observed for TC (-0.135 mmol/l (95% CI: -0.289, 0.019 mmol/l)) or Ca (0.023 mmol/l (95% CI: -0.016, 0.062 mmol/l)). The meta-analysis suggested a protective effect of soy protein consumption on SCR and serum phosphorus concentrations in pre-dialysis CKD patients. It may also have a significant effect on lowering serum TG concentrations. However, nonsignificant effects on TC and Ca were observed. Evidence was limited because of the relatively small number of available trials and subjects.


Neuropsychological correlates of decision making in patients with bulimia nervosa.
In addition to the core psychopathology of bulimia nervosa (BN), patients with BN often show impulsive behavior that has been related to decision making deficits in other patient groups, such as individuals with anorexia nervosa and pathological gamblers. However, it remains unclear whether BN patients also show difficulties in decision making. In this study, 14 patients with BN and 14 healthy comparison subjects, matched for age, gender, education, body mass index, and intelligence, were examined with the Game of Dice Task (M. Brand, E. Fujiwara, et al., 2005), a gambling task that has fixed winning probabilities and explicit rules for gains and losses, as well as with a neuropsychological test battery and personality questionnaires. On the task, the patients with BN chose the disadvantageous alternatives more frequently than did the comparison subjects. Performance on the Game of Dice Task was related to executive functioning but not to other neuropsychological functions, personality, or disease-specific variables in the BN group. Thus, in patients with BN, decision making abnormalities and executive reductions can be demonstrated and might be neuropsychological correlates of the patients' dysfunctional everyday-life decision making behavior. Neurocognitive functions should be considered in the treatment of BN.


The Impact of Modern Dietary Practices on Cancer Risk and Progression: A Systematic Review.
Cancer is a leading cause of mortality around the world, despite continued advancements in the management of cancer. Recent research efforts have shifted to evaluating the role that modifiable risk factors play in cancer risk and development, as diet and nutrition have been found to play a significant role in the onset and progression of cancer. As a result, there has been an increasing focus on the impact of dietary modifications on preventing the onset, progression, and reoccurrence of cancer. In this systematic review, data were collected on three common diets, the Mediterranean diet (MD), ketogenic diet (KD), and plant-based diet, to gain insight into the application of these three dietary modification approaches for risk prevention and limitation of cancer burden. Initially, 4,397 articles were identified from three databases (Ovid, Web of Science, and CINHAL). After removing studies based on the exclusion criteria, only 23 studies were eligible to be included in the systematic review of which 15 evaluated the MD, four assessed the ketogenic diet, and four evaluated the plant-based diet. Each article was considered for its methods, procedures, and findings. The findings indicate that dietary interventions may effectively reduce the odds of cancer development and the advancement of diagnosed cancers. With the introduction of the MD, KD, and plant-based diets, significant improvements in lowering cancer development, recurrence-free status, and limiting tumor growth were noted across numerous cancer types. Currently, the MD has been extensively studied in the literature, and amongst the widest variety of cancer types. Additional information and evaluation are required on the ketogenic and plant-based diets to fully understand their impact on the cancer burden across a wider subset of cancers. Clinicians should evaluate and recommend nutritional adaptations to their patients to limit the development of specific cancers and as an adjunctive therapy to traditional pharmacological treatment options for patients with diagnosed cancers.


Validation of a measure of family experience of patients with serious illness: the QUAL-E (Fam).
Family members of seriously ill patients experience significant burden as they advocate with providers and participate in key decisions for loved ones. Most assessments focus on patient experience, yet family members' own quality of experience is central to comprehensive care. This study validated a measure of the quality of family experience, the QUAL-E (Fam), in palliative care. We enrolled family members of terminally ill patients admitted to general medicine services at two hospitals. Items were based on foundational work originally done for the patient instrument, QUAL-E, and interviews of family members of terminally ill patients. Cognitive interviewing was used to refine items, which then underwent formal testing factor analysis. In the first sample, we assessed factor structure. A subsequent sample established predictive validity and test-retest reliability. The initial item pool was reduced to a four-domain, 35-item scale and administered to the validation sample. Further analyses produced a final brief scale comprising 17 items, demonstrating appropriate convergent and divergent validity. Test-retest reliability demonstrated expected levels of stability in a highly changeable population. The scale provides an assessment of family experience and includes subscales assessing relationship with health care provider and completion. Additional scale items assess symptom experience and issues of preparation. The QUAL-E (Fam) is a companion instrument to the patient QUAL-E measure of quality of life at the end of life and is part of a package of assessment tools that can help evaluate the entire patient experience and contribute to quality care.


A sialosyl sulfonate as a potent inhibitor of influenza virus replication.
A new direction for influenza virus sialidase inhibitor development was identified using a sulfonate congener of 2-deoxy-2-β-H N-acetylneuraminic acid. Sialosyl sulfonates can be synthesised efficiently in four steps from N-acetylneuraminic acid via a microwave assisted decarboxylation. The presence of the sulfonate group significantly increases inhibition of influenza virus sialidase and viral infection when compared to the carboxylate congener, and also to the benchmark sialidase inhibitor 2,3-dehydro-2-deoxy-N-acetylneuraminic acid, Neu5Ac2en.


Multistate outbreak of Salmonella infections associated with peanut butter and peanut butter-containing products--United States, 2008-2009.
On November 25, 2008, an epidemiologic assessment began of a growing cluster of Salmonella serotype Typhimurium isolates that shared the same pulsed-field gel electrophoresis (PFGE) pattern in PulseNet. As of January 28, 2009, 529 persons from 43 states and one person from Canada had been reported infected with the outbreak strain. This report is an interim summary of results from ongoing epidemiologic studies and recall and control activities by CDC, the Food and Drug Administration (FDA), and state and local public health agencies. Confirmed, reported onset of illness dates have ranged from September 1, 2008, to January 16, 2009. A total of 116 patients were reported hospitalized, and the infection might have contributed to eight deaths. Sequential case-control studies have indicated significant associations between illness and consumption of any peanut butter (matched odds ratio [mOR] = 2.53), and specific brands of prepackaged peanut butter crackers (mOR = 12.25), but no association with national brand jarred peanut butter sold in grocery stores. Epidemiologic and laboratory findings indicate that peanut butter and peanut paste produced at one plant are the source of the outbreak. These products also are ingredients in many foods produced and distributed by other companies. This outbreak highlights the complexities of "ingredient-driven" outbreaks and the importance of rapid outbreak detection and investigation. Consumers are advised to discard and not eat products that have been recalled.


A Phase I study of docetaxel plus cyclophosphamide in solid tumors followed by a Phase II study as first-line therapy in metastatic breast cancer.
In Phase I, the purpose was to determine the maximum tolerated dose and pharmacokinetics of docetaxel plus cyclophosphamide (DC) with and without granulocyte colony-stimulating factor in the treatment of patients with solid tumors. For Phase II, the purpose was to determine the safety and efficacy of this combination as first-line treatment in patients with metastatic breast cancer (MBC). In Phase I (45 patients), docetaxel was escalated from 60 mg/m(2) to 85 mg/m(2), and cyclophosphamide from 600 mg/m(2) to 800 mg/m(2). Pharmacokinetic evaluation of docetaxel was performed in 19 patients with MBC. In Phase II (34 patients), patients received cyclophosphamide (600 mg/m(2)) followed by docetaxel (75 mg/m(2)), i.v. In Phase I, the dose-limiting toxicity was neutropenia-related events. The maximum tolerated dose for DC was 75 mg/m(2)/700 mg/m(2) in solid tumor patients treated previously and 75 mg/m(2)/800 mg/m(2) for patients not treated previously for MBC. Dose escalation of docetaxel >75 mg/m(2) was not tolerated, despite prophylactic granulocyte colony-stimulating factor treatment. In Phase II, 71% of patients received prior anthracycline therapy. Neutropenic fever requiring i.v. antibiotics occurred in 6 patients (19%). One patient had grade 3 neuropathy. There was no cardiotoxicity. The overall Phase II intent-to-treat objective response rate was 65% (complete responses, 12%). The median overall survival was 22 months, and the median time to progression was 6 months. DC combination therapy is an active regimen with acceptable toxicity and is appropriate regardless of prior anthracycline therapy. In view of the high activity and lack of cardiotoxicity, this combination warrants additional investigation in early stage breast cancer and in combination with trastuzumab.


Enzyme-Catalyzed oxidation of cholesterol in physically characterized water-in-oil microemulsions.
The enzymatic conversion of cholesterol to cholestenone by cholesterol oxidase (Brevibacterium sp.)in reversed micelles in a system composed of AOT/isooctane/water/cholesterol has been examined. The catalytic activity of the enzyme was correlated with the physicochemical properties of water in water-in-oil (w/o) microemulsion systems. In a system consisting of 3 wt % AOT in isooctane, reversed micelles started to form as the [H(2)O]/[AOT] (e.g., the w(0)) ratio increased above 4-5. The formation of reversed micelles with a core of neat (bulk) water was verified from determinations of both the partial molar volume of water and the scissors vibration of water [with Fourier transform infrared (FTIR) spectroscopy] in the w/o microemulsion systems. A plot of enzyme activity vs. w(0) indicated that the hydration of enzyme molecules per se was not sufficient to give rise to catalytic activity. Instead, it appeared that the formation of an aqueous micellar core was necessary for full activation of the enzyme. Based on micelle size distribution analysis, it was estimated that about one micelle per one thousand contained an enzyme molecule. Since the apparent reaction rate could be markedly enhanced by increasing the enzyme/water ratio, we conclude that the number of enzyme-containing micelles was an important rate-limiting factor in the system.


Hyperbaric oxygen for persistent post-concussive symptoms: long-term follow-up.
We report results of an observational cohort study investigating long-term follow-up in participants from two completed United States military trials of hyperbaric oxygen (HBO₂) for persistent post-concussive symptoms (PCS), as well as challenges in recruitment and retention in active-duty military personnel. After informed consent, participants completed an electronic survey assessing PCS, post-traumatic stress disorder (PTSD), anxiety, depression and quality of life. Of 132 HBO₂ study participants, 40 (30%) completed the survey (42 could not be contacted; 50 were lost to follow-up or declined). All were male, age 28.1 ±6.6 years (mean ±1SD). Time to follow-up was 39.2 ±6.1 months. At follow-up, participants reported continued symptoms of PTSD, depression, anxiety and reduced quality of life. Among DARPA/VCU study participants, total PCS scores worsened in the 1.5 atmospheres absolute (ATA) equivalent HBO₂ group (mean change 7.4 ±15.8) and improved in the sham (-8.0 ±7.7) and 2.0 atmospheres absolute equivalent HBO₂ groups (-3.3 ±7.4). Individual changes varied widely, range -23 to +28 points. In participants from the HOPPS study, total PCS scores worsened in all groups: local care (10.5 ±8.7), sham (7.9 ±11.9) and 1.5 ATA HBO₂ (1.0 ±19.4). In this limited, cross-sectional sample, PCS and PTSD symptoms did not appear to improve over time by descriptive analyses. Low participation rates and potential response bias limit our ability to perform statistical hypothesis testing and to draw conclusions from these data. Future studies should prospectively plan longitudinal follow-up and regular engagement with participants to minimize attrition.


[Scientificity of traditional commodity grade of Chinese medicinal materials--taking Lonicerae Japonicae Flos as example].
Taking Lonicerae Japonicae Flos as an example, the method of "expert consensus of different regions" was used to screen the representative samples and evaluate their commodity grades. The correlation analysis, hierarchical cluster analysis and partial least squares discriminant analysis(PLS-DA) of "commodity grade-appearance characteristic-component content" were carried out to reveal the scientificity of traditional commodity grade of Chinese medicinal material. By referring to the existing literature and the grade investigation from the sample collection regions, 78 "initial grade" samples were screened out from 118 collected samples. Authoritative experts from four regions(n=4) including Linyi(Shangdong province), Bozhou(Anhui province), Anguo(Hebei province) and Beijing were organized to evaluate their commodity grades, separately. Based on the grade consistency rate(R_i≥70%), 69 "local grade" samples were screened out from the "initial grade" samples. Based on the average grade consistency rate ■ "authoritative grade" samples were screened out from the "local grade" samples, including15 first-grade samples, 9 second-grade samples, 11 third-grade samples and 17 fourth-grade samples. For these "authoritative grade" samples, the main appea-rance characteristics were quantified and the contents of 13 components were determined by ultra performance liquid chromatography(UPLC). Furthermore, the total contents of 6 phenolic acids, 4 flavonoids and 3 iridoids were calculated, respectively. The results of correlation analysis showed that 4 appearance characteristics indices were correlated with the commodity grades: color, rate of yellow bars(including blooming flowers), rate of black heads(including black bars), and rate of stems and leaves(including bud debris). Five component content indices were correlated with the commodity grades: chlorogenic acid, isochlorogenic acid C, sweroside, loganin and the total contents of six phenolic acids. Furthermore, chlorogenic acid, loganin and the total contents of six phenolic acids showed significantly negative correlation with the main appearance characteristics, indicating that the appearance characteristics of Lonicerae Japonicae Flos can reflect its internal quality, and these 3 indices can be used as quality markers(Q-markers). The results of hierarchical cluster analysis showed that the samples of four grades were classified into four categories, and the samples with the same grades and the same categories accounted for 80.8% of the total samples, while the samples with the different grades were obviously classified into different categories. The results of PLS-DA analysis showed that the samples of different grades showed obvious intragroup aggregation and intergroup dispersion. The above results indicated that it was feasible to evaluate the traditional commodity grade of Lonicerae Japonicae Flos by the method of "expert consensus of different regions". For the evaluation of traditional commodity grade of Chinese medicinal material, the samples should be representative, expert conclusions should have enough consensuses, and grade determination should be authoritative. As the crystallization of clinical experience, traditional commodity grade can scientifically reflect the internal quality of Chinese medicinal material.


Viral zoonotic risk is homogenous among taxonomic orders of mammalian and avian reservoir hosts.
The notion that certain animal groups disproportionately maintain and transmit viruses to humans due to broad-scale differences in ecology, life history, and physiology currently influences global health surveillance and research in disease ecology, virology, and immunology. To directly test whether such "special reservoirs" of zoonoses exist, we used literature searches to construct the largest existing dataset of virus-reservoir relationships, consisting of the avian and mammalian reservoir hosts of 415 RNA and DNA viruses along with their histories of human infection. Reservoir host effects on the propensity of viruses to have been reported as infecting humans were rare and when present were restricted to one or two viral families. The data instead support a largely host-neutral explanation for the distribution of human-infecting viruses across the animal orders studied. After controlling for higher baseline viral richness in mammals versus birds, the observed number of zoonoses per animal order increased as a function of their species richness. Animal orders of established importance as zoonotic reservoirs including bats and rodents were unexceptional, maintaining numbers of zoonoses that closely matched expectations for mammalian groups of their size. Our findings show that variation in the frequency of zoonoses among animal orders can be explained without invoking special ecological or immunological relationships between hosts and viruses, pointing to a need to reconsider current approaches aimed at finding and predicting novel zoonoses.


Investigational drug tracking: phases I-III and NDA submissions--Part II.
The author catalogs over 800 investigational drugs/biologicals currently in Phase I, II or III clinical trials or drugs/biologicals submitted to the FDA as new drug applications. Part I of this article appeared in the September issue of Hospital Pharmacy. The list assists in predicting when new drugs will be marketed. The entries include generic/chemical name, investigational drug number, synonyms, trade names, manufacturers, clinical trial status, predicted approval year, indications or drug class, whether the drug has been developed through biotechnology, and references. Entries were gleaned from medical journals, stock market analysis publications, and the Pharmaceutical Manufacturers Association's Medicines in Development Series. The list is alphabetized by the generic/chemical name or investigational drug number and cross-indexed by the trade name and synonyms. The list reflects those drugs which were not FDA approved as of April 15, 1994. Part I concludes with the remaining alphabetical listing by generic/chemical name or investigational drug number.


[Diagnostics and therapy of depressive symptoms in schizophrenic patients].
The occurrence of depressive symptoms in schizophrenia is fairly common. In contrast to earlier assumptions, they usually have unfavorable consequences on the course of the disease. The response of depressive symptoms to therapy is well documented by empirical evidence and studies. In reality, however, the treatment of depressive symptoms in cases of schizophrenia occurs too rarely. This lack of medical treatment is partially explained by the clinicians' apprehension that antidepressive therapy could provoke a flare up of the psychosis. Nonetheless, this situation is often founded on the incorrect assessment of the depressive symptoms as negative symptoms that, in turn, are regarded by many clinicians as having limited treatability. In regards to this circumstance, a rethinking is desirable and, as a matter of principle, necessary.


Hypothalamic obesity, brown adipose tissue, and sympathoadrenal activity in rats.
Obesity-producing, hypothalamic knife cuts and ventromedial hypothalamic (VMH) lesions in ad libitum-fed adult rats increased intake of a high-fat diet (123 and 130%) and energy retention (880 and 1,099%) during the 4-wk period postsurgery; even when pair fed to control rats, energy retention of the knife-cut and lesioned rats was still elevated (105 and 155%). Thermogenic capacity of brown adipose tissue (BAT), estimated from guanosine diphosphate (GDP) binding to BAT mitochondria, was unchanged in hyperphagic knife-cut and VMH-lesioned rats and was reduced approximately 50% when these rats were pair fed to controls. Urinary excretion of norepinephrine (NE) was approximately twofold higher in ad libitum-fed, knife-cut, and lesioned rats than in control rats; restriction of energy intake decreased NE excretion to control values. Rates of NE turnover in heart paralleled urinary NE excretion, whereas NE turnover in BAT was generally not increased in the hyperphagic rats. Urinary epinephrine excretion, an index of adrenal medullary activity, was depressed in all knife-cut and VMH-lesioned rats. Hyperphagia coupled with a lack of increased heat production in BAT causes gross obesity in ad libitum-fed, knife-cut, and VMH-lesioned rats, whereas obesity in pair-fed rats develops in part at least as a result of reduced heat production by BAT.


In vitro studies of DNA condensation by bridging protein in a crowding environment.
The macromolecules of the bacterial cell occupy 20-40% of the total cytosol volume, and crowded environments have long been known to compact and stabilize DNA. Nevertheless, investigations on DNA-protein binding are generally performed in the absence of crowding, which may yield an incomplete understanding of how nucleoid-assembling proteins work. A family of such proteins, abundant in Gram-negative bacteria, is the histone-like nucleoid structuring proteins (H-NS). Herein, the synergistic role of macromolecular crowding (mimicked using polyethylene glycol, PEG) and H-NS was investigated using fluorescence correlation spectroscopy (FCS) and enzyme protection assays. We show that crowding enhances the binding of H-NS to the AT-rich tracks of the DNA, where it preferentially binds to, protecting these tracks towards enzyme digestion, inducing some DNA condensation, and inhibiting the biological function of DNA. We further suggest that the looping of DNA chains, induced by H-NS, contributes to the synergistic effect of DNA-binding protein and crowding agents, on DNA condensation.


The molecular and magnetic structure of carbon-enclosed and partially covered Fe55 particles.
The structure and magnetic moment distribution are studied for an iron nanoparticle with varying degree of carbon adatom coverage. The limiting models of the study are the clean icosahedral Fe55 particle and the iron particle completely enclosed in carbon cages. Between the two extrema, partially covered particles are considered. The iron cluster with partial coverage of carbon adatoms represents a model of active catalysts in the chemical vapor deposition synthesis of carbon nanotubes. The investigated structures are the bare Fe55 cluster, Fe55N4C(x) (x = 27, 37, 47, 54, 65), and Fe55 encapsulated inside C180 and C240. The two latter are extreme examples of an iron particle completely enclosed in a carbon network. Fe55@C180 and Fe55@C240 present novel structures resembling the endohedral metallofullerenes. Two structural isomers of the Fe55@C180 are considered. Enclosing the Fe55 cluster inside C180 and C240 fullerenes gives rise to changes in the Fe-Fe bond lengths. This alters the magnetic structure of the iron cluster considerably. The interaction between the fullerenes and the enclosed iron cluster is reflected in a charge transfer of 8-13 electrons in the considered endohedral complexes. The localization of atomic charges on the C180 and C240 cages suggests site-selective reactivity of the endohedral complexes. The total magnetic moments of the Fe55N4C(x) nanoparticles vary with the degree of adatom coverage. The magnetic moments of individual Fe atoms depend strongly on the element of the nearest-neighbor atoms and on the coordination number and carry therefore information about the local chemistry.


Antitumor Effect of Poplar Propolis on Human Cutaneous Squamous Cell Carcinoma A431 Cells.
Propolis is a gelatinous substance processed by western worker bees from the resin of plant buds and mixed with the secretions of the maxillary glands and beeswax. Propolis has extensive biological activities and antitumor effects. There have been few reports about the antitumor effect of propolis against human cutaneous squamous cell carcinoma (CSCC) A431 cells and its potential mechanism. CCK-8 assays, label-free proteomics, RT-PCR, and a xenograft tumor model were employed to explore this possibility. The results showed that the inhibition rate of A431 cell proliferation by the ethanol extract of propolis (EEP) was dose-dependent, with an IC50 of 39.17 μg/mL. There were 193 differentially expressed proteins in the EEP group compared with the control group (p < 0.05), of which 103 proteins (53.37%) were upregulated, and 90 proteins (46.63%) were downregulated. The main three activated and suppressed Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were extracellular matrix (ECM)-receptor interaction, amoebiasis, cell adhesion molecules (CAMs), nonalcoholic fatty liver disease (NAFLD), retrograde endocannabinoid signaling, and Alzheimer's disease. The tumor volume of the 100 mg/kg EEP group was significantly different from that of the control group (p < 0.05). These results provide a theoretical basis for the potential treatment of human CSCC A431 cell tumors using propolis.


Current approaches to flexible loop modeling.
Loops are key components of protein structures, involved in many biological functions. Due to their conformational variability, the structural investigation of loops is a difficult topic, requiring a combination of experimental and computational methods. This paper provides a brief overview of current computational approaches to flexible loop modeling, and presents the main ingredients of the most standard protocols. Despite great progress in recent years, accurately modeling the conformational variability of long flexible loops remains a challenging problem. Future advances in this field will likely come from a tight coupling of experimental and computational techniques, which would enable a better understanding of the relationships between loop sequence, structural flexibility, and functional roles. In fine, accurate loop modeling will open the road to loop design problems of interest for applications in biomedicine and biotechnology.


Frontal cortical tissue composition in abstinent cocaine abusers: a magnetic resonance imaging study.
Cocaine abusers exhibit impairment of executive cognitive functions that are mediated by the frontal cortex. This work tested for structural (i.e., tissue composition) abnormalities that may underlie such performance deficits. Research participants were cocaine abusers (n = 14) abstinent for 20 days and a non-drug-using comparison group (n = 11), who underwent magnetic resonance imaging (T1-weighted scans of the brain). Gray matter and white matter tissue densities were determined using voxel-based morphometry with small volume correction based on a priori hypotheses derived from functional imaging of the same subjects. Cocaine abusers had significantly lower gray matter tissue density than did the non drug users in 10 of 13 small volumes analyzed in the frontal cortex [bilateral anterior cingulate gyrus (infragenual and perigenual regions) and medial orbitofrontal cortex and the lateral orbitofrontal cortex and middle/dorsal cingulate gyrus in the right hemisphere]. No group differences were found in white matter density of the frontal cortex. These results extend our previous findings of defective frontal cortical activation (indexed by cerebral blood flow) in cocaine abusers to include abnormalities in gray matter tissue density in the same frontal cortical regions.


Differences in striatal dopamine transporter density between tremor dominant and non-tremor Parkinson's disease.
Parkinson's disease (PD) can manifest with a tremor-dominant or a non-tremor (akinetic-rigid) phenotype. Although the tremor-dominant subtype may show a better prognosis, there is limited information on the phenotypic differences regarding the level of striatal dopamine transmission. The present study investigated striatal dopamine transporter (DAT) binding characteristics in a large sample of patients with and without tremor. [(123)I]FP-CIT SPECT scans of 231 patients with a clinical diagnosis of PD and abnormal FP-CIT binding (157 with tremor, 74 without tremor) and 230 control patients with normal FP-CIT binding (148 with tremor, 82 without tremor) were analysed using an automated region-of-interest analysis of the scans (BRASS). Specific striatal binding ratios were compared between phenotypes and groups using age, sex, and symptom duration, predominant side of symptoms, dopaminergic medications and scanner as covariates. Patients with PD had 28.1 - 65.0 % lower binding in all striatal regions compared to controls (p < 0.001). The mean FP-CIT caudate nucleus uptake and the left caudate nucleus uptake were higher in PD patients with tremor than in PD patients without tremor (mean 9.0 % higher, left 10.5 % higher; p < 0.05), whereas there were no differences between tremor and non-tremor control patients. No significant effects of tremor on DAT binding were observed in the anterior or posterior putamen. The motor phenotype is associated with the extent of caudate dopamine terminal loss in PD, as dopamine function is relatively more preserved in tremor patients. Symptom type is related to caudate dopamine function only in association with Parkinsonian dopaminergic degeneration, not in intact dopamine systems in patients with non-PD tremor.


Is the masked continuous glucose monitoring system clinically useful for predicting hemoglobin A1C in type 1 diabetes?
The masked continuous glucose monitoring system (Masked-CGMS) differs from standard CGMSs in three ways: (1) there is no feedback to the user so that no immediate regimen changes can be made; (2) it can only be worn for up to 5 days; and (3) there are no alarms to warn of hyperglycemia or hypoglycemia. Since 2008 masked-CGMS has become popular for identifying reasons that a patient's hemoglobin A1C does not correlate closely with his or her capillary blood glucose measurements. To date only one study addressing the clinical utility of Masked-CGMS for improving A1C in diabetes has been published. No studies are available specifically examining the variability and correlation of Masked-CGMS and A1C. We performed 156 Masked-CGMS studies (40 patients studied sequentially a maximum of four times each) in type 1 diabetes patients. We then analyzed the resulting interstitial glucose levels obtained from the Masked-CGMS compared with an A1C measurement performed within 1 week of the Masked-CGMS study. There was a very low correlation between the A1C and the Masked-CGMS-derived mean interstitial glucose level. This statistic did not provide sufficiently predictive information to be clinically useful for changing an individual patient's intensive insulin therapy regimen. Our data demonstrate that a very weak correlation exists between 5 days of masked CGMS and a concurrently measured A1C level. For the individual type 1 diabetes patient, this relationship would unlikely to be clinically useful in altering the individual's treatment regimen.


Diversity and pathogenecity of Vibrio species in cultured bivalve molluscs.
Shellfish production is seriously affected by bacterial pathogens that cause high losses in hatcheries and in the aquaculture sector. A number of Vibrio species are considered important pathogens and have provoked severe mortality outbreaks. The pathologies caused by vibrios in bivalves have been described since the 1960s; however, over recent years, successive episodes of high mortality have been recorded due to these microorganisms. The present work provides an updated overview of the different studies performed in relation with the diversity of Vibrio spp. associated to bivalves. Special attention is given to the main Vibrio diseases and implicated species affecting the different life stages of cultured bivalves.


Changes in the contractile activity of the rabbit myocardium during burn shock.
As revealed on the papillary muscles of the rabbit heart in vitro, contractile function of the myocardium proved to be disturbed in 50% of cases an hour after the infliction of burn to these animals. This could serve as one of the causes of a sharp cardiac output derangement found in these experiments.


Minor Salivary Gland Acinic Cell Carcinoma With Orbital Extension.
An 80-year-old male presented with painless blurred vision from the right eye following endoscopic biopsy of a right maxillary sinus mass with the otolaryngology service. The patient admitted to progressive right-sided proptosis over the past few weeks. Past medical history was significant for acinic cell adenocarcinoma of buccal space origin with resection 3 years prior. Orbital CT revealed a large, poorly circumscribed destructive mass in the right masticator space with extension to the paranasal sinuses, inferior and medial orbit, and middle cranial fossa. Histopathology confirmed the mass to represent a recurrent acinic cell carcinoma. The patient was treated with chemotherapy, and at last follow-up 2 years after presentation had stable painless globe dystopia and visual acuity. The authors describe a very rare case of a recurrent minor salivary gland acinic cell carcinoma with secondary orbital extension.


Use of WATCH antibiotics prior to presentation to the hospital in rural Burkina Faso.
In low- and middle-income countries, the prevalence of antimicrobial resistance (AMR) is increasing. To control AMR, WHO recommends monitoring antibiotic use, in particular Watch antibiotics. These are critically important antibiotics, with restricted use because at risk of becoming ineffective due to increasing AMR. We investigated pre-hospital antibiotic use in rural Burkina Faso. During 2016-2017, we collected data from patients aged > 3 months presenting with severe acute fever to the rural hospital of Nanoro Health District, Burkina Faso, including antibiotic use in the two weeks prior to consultation or hospitalization. We analysed reported antibiotic use by applying the WHO Access, Watch, Reserve classification. Of 920 febrile participants (63.0% ≤ 14 years), pre-hospital antibiotic use was reported by 363 (39.5%). Among these 363, microbiological diagnoses were available for 275 (75.8%) patients, of whom 162 (58.9%) were non-bacterial infections. Use of more than one antibiotic was reported by 58/363 (16.0%) participants. Of 491 self-referred patients who did not previously visit a primary health care center, 131 (26.7%) reported antibiotic use. Of 424 antibiotics reported, 265 (62.5%) were Access and 159 (37.5%) Watch antibiotics. Watch antibiotic use was more frequent among patients > 14 year olds (51.1%) compared to those 0-14 year old (30.7%, p < 0.001) and among referrals from the primary health care centers (42.2%) compared to self-referred patients (28.1%, p = 0.004). Most frequently reported Watch antibiotics were ceftriaxone (114, 71.7%) and ciprofloxacin (32, 20.1%). The reported frequent use of Watch group antibiotics among febrile patients prior to presentation to the hospital in rural Burkina Faso highlights the need to develop targeted interventions to improve antibiotic use in community settings as part of strengthening antibiotic stewardship in low- and middle-income countries. This should include facilitating referral, access to qualified prescribers and diagnostic tools in rural primary health care centers. Trial registration ClinicalTrials.gov identifier: NCT02669823. Registration date was February 1, 2016.


The outcome of patients with lupus nephritis and the impact of cardiovascular risk factors.
Systemic lupus erythematosus (SLE) is the prototype of autoimmune connective tissue diseases. Renal disease is a frequent manifestation of SLE that influences the outcome of the patients. The aim of the current study was to determine and analyze the clinical features and subsequent outcome of 70 patients with LN, followed in our department over the past 5 years, focusing on the impact of cardiovascular risk factors in the renal outcome and mortality. Our prospective study included 70 patients with SLE and LN and 70 patients with SLE without signs of renal involvement, all patients fulfilled the revised ACR (American College of Rheumatology) criteria for the classification of SLE. Demographical data, risk factors and comorbidities were recorded. Patients with lupus nephritis had a mean age of 37 years (range 15-65, SD 1.8). During the study, we had a rate of drop off of 15 patients with lupus nephritis (21%) and 19 patients without nephritis (26%). Patients with LN had a higher prevalence of positive anti-dsDNA antibodies (85.4% vs 49%, p<0.001, RR=2.2) and a lower percent of rheumatoid factor (FR) positive (5.45% vs 15.68%, p=0.03, RR=0.34) compared with the controls, a higher prevalence of corticosteroid treatment (65.45% vs 7.83%, p<0.001, RR=2.1) and immunosuppressive treatment (AZA 27.27% vs 3.92%, p=0.01, RR=1.71, CFM 34.54% vs 0%, p<0.001, RR=2.16), a higher frequency of hypertension (47.27% vs 9.8%, p<0.001, RR=2.4), hyperlipidaemia (49.09% vs 1.96%, p<0.001, RR=1.81) and anti-PL antibodies (49.09% vs 20%, p=0.001, RR=2.70),and a higher mortality (16% vs 2%, p=0.02, RR=1.76). 20 patients (36.36%) from the survival group (55 patients), evoluated to renal failure, 9.09% of these with end -stage renal failure, results that are similar with the ones in other studies. The study reveals the fact that cardiovascular risk factors such as hypertension, hyperlipidaemia and antiphospholipid syndrome are associated with a higer rate of mortality and an evolution to end-stage renal disease.


[Epidemiological features of maternal deaths occurred in Recife, PE, Brazil (2000-2006)].
This was a cross-sectional study that aimed to describe the epidemiological characteristics of maternal deaths among women living in Recife, PE, Brazil that occurred between 2000 and 2006. The data source consisted of investigation files on maternal deaths. To analyze the data, the EpiInfo 6.04d software was used. The analysis considered 111 deaths, corresponding to a maternal death ratio of 65.99/100,000 live births. The obstetric data showed that these women had had fewer than six prenatal consultations, between one and four previous pregnancies, cesarean deliveries and hospital admission in a severe condition. Hypertensive disorders were the main cause of death. Most of the deaths were considered avoidable. The results indicate the need to improve the healthcare for pregnant women prenatally, at delivery and during the puerperium.


Therapeutic engagement: a proposed model of engagement in medical rehabilitation.
A major goal of physical medicine and rehabilitation is the recovery of function after an injury or the underlying medical condition that has caused impairment in one's daily functioning. The rehabilitation process involves a complex interplay of many factors that influence how well a person benefits from medical rehabilitation. There is burgeoning evidence that for patients to maximize rehabilitation benefits, they must be actively involved or engaged in the process. In this article, the construct of therapeutic engagement is defined, and a theoretical model is proposed to illustrate how and why individuals engage in medical rehabilitation treatment. It is hoped that such a model can form the basis for further research on therapeutic engagement that facilitates the understanding of the process variables that play a role in rehabilitation outcomes. Areas for future research and the implications for clinical application are discussed.


Chronic fatigue syndrome and the immune system: Where are we now?
Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is characterised by multiple symptoms including fatigue, headaches and cognitive impairment, which have a significantly adverse effect on the normal functioning and well-being of the individual. These symptoms are often triggered or worsened following physical or mental exertion. ME/CFS has long been thought of as having a significant immunological component, but reports describing changes in immune function are often inconsistent between study groups. Although the wide range of physical, neurocognitive and autonomic symptoms reported have seriously hampered attempts to understand pathophysiological pathways, investment in biomedical research in ME/CFS is finally increasing with a number of novel and promising investigations being published. The onset of ME/CFS may often be linked to (viral) infections which would be consistent with a variety of alterations in natural killer (NK) cell function as described by a number of different groups. Consistency in cytokine data has been lacking so far, although recently more sophisticated approaches have led to more robust data from large patient cohorts. New hope has also been given to sufferers with the possibility that therapies that deplete B cells can result in clinical improvement. To understand the pathogenic mechanism in this complex condition, it is important to consider repeated analysis in different cohorts. In this review, we will discuss the potential of different components of the immune system to be involved in the pathogenesis of ME/CFS.


In vitro cultivation of epithelial cells derived from tumors of the human urinary tract.
Finely minced explants from 54 TCC2 of the human urinary tract were cultured in vitro in an attempt to establish cell lines. Cells with epithelial morphology grew out from 48 tumor explants, and long-term cell cultures were established from 10. Six of the cell cultures have been maintained for over 18 months with 50 to 70 transfers and, therefore, are considered permanent cell lines. The epithelial cells in the established cultures are small, exhibit rapid doubling time, and show multilayering. The cells were examined both microscopically and by cultivation techniques, and they were found to be free from contaminating microorganisms, including Mycoplasma. The established cultures grow rapidly in roller bottles and, therefore, can be produced in large quantities. These cells also remain viable after being stored for 3 years in liquid nitrogen.


A common mutation in the surfactant protein C gene associated with lung disease.
To determine the contribution of the surfactant protein C (SP-C) I73T mutation to lung disease. Genomic DNA was obtained from 116 children with interstitial lung disease (ILD) or chronic lung disease of unclear cause and from 166 control subjects and was screened for the I73T mutation using an allele-specific polymerase chain reaction assay. The I73T mutation was found on 7 of 232 SP-C alleles from 7 unrelated children with ILD but was not found on 332 control SP-C alleles ( P < .01, Fisher exact test). The I73T mutation segregated with lung disease in one kindred with familial ILD. The I73T mutation was found in an asymptomatic parent from two different families with affected children consistent with variable penetrance, but it was not found in either asymptomatic parent of two other unrelated affected children consistent with a de novo mutation. Analysis of single nucleotide polymorphisms indicated diverse genetic backgrounds of the I73T alleles. Immunohistochemical analysis of lung tissue from an infant with the I73T mutation demonstrated normal staining patterns for proSP-B, SP-B, and proSP-C. These findings support the hypothesis that the I73T mutation predisposes to or causes lung disease.


Role of glutathione in cancer progression and chemoresistance.
Glutathione (GSH) plays an important role in a multitude of cellular processes, including cell differentiation, proliferation, and apoptosis, and disturbances in GSH homeostasis are involved in the etiology and progression of many human diseases including cancer. While GSH deficiency, or a decrease in the GSH/glutathione disulphide (GSSG) ratio, leads to an increased susceptibility to oxidative stress implicated in the progression of cancer, elevated GSH levels increase the antioxidant capacity and the resistance to oxidative stress as observed in many cancer cells. The present review highlights the role of GSH and related cytoprotective effects in the susceptibility to carcinogenesis and in the sensitivity of tumors to the cytotoxic effects of anticancer agents.


Adaptation to metabolic acidosis by turtle urinary bladder.
We utilized the turtle urinary bladder to study the mechanisms responsible for adaptation to metabolic acidosis. Bladders removed from acidotic turtles had a higher rate of H+ secretion in vitro than bladders from control turtles, despite identical extracellular pH. HCO3 secretion, however, was not different between the two groups. The increase in H+ secretion could be mediated by a decrease in intracellular pH and/or by an increase in the number of cells thought to be responsible for H+ secretion. To study this issue, we measured intracellular pH with the fluorescent dye 6-carboxyfluorescein diacetate and quantified the number of cells by fluorescence microscopy utilizing acridine orange, rhodamine 123, and 6-carboxyfluorescein diacetate in turtles receiving different acid loads. Urinary acidification measured in vivo was increased in turtles fed a low-acid load for 48 h and in turtles fed a high-acid load for 24-48 h. Intracellular pH was lower in bladders from turtles fed a high-acid load for 48 h but it was not different from controls in the other groups, indicating that intracellular pH cannot account for the adaptive increase in H+ secretion. Bladders from all groups fed an acid load had a higher number of cells with positive staining for acridine orange compared with controls. Double labeling with acridine orange and the mitochondrial stain rhodamine 123 or 6-carboxyfluorescein showed a significant increase in the number of mitochondria-rich cells between control and bladders from turtles fed an acid load. The increase in the number of rhodamine 123- or 6-carboxyfluorescein-positive cells was lower than the increase in acridine orange-positive cells, suggesting that the apparent increase in the number of acridine orange-positive cells is due to an increase in the number of acidic vesicles in the mitochondria-rich cells and in the granular cells rather than solely to an increase in the number of mitochondria-rich cells. Plasma membrane fraction prepared from control and acidotic bladders failed to disclose an increase in the putative H+-ATPase as assessed by enzymatic activity and transport studies. In conclusion, the present study suggests that the adaptive increase in H+ secretion in metabolic acidosis is associated both with an increase in the number of mitochondria-rich cells as well as with an increase in the number of acidic vesicles in these cells.


Telomere tales: chromatin, telomerase and telomere function in Saccharomyces cerevisiae.
Unusual chromatin structures underlie epigenetic effects at the silent mating-type loci and telomeres in yeast. Many of the same genes appear to function in transcriptional silencing observed at both the silent mating-type loci and at telomeres. The observation that these loci are united by a requirement for shared factors suggests that the structure of chromatin at these regions is similar. Alteration of telomeric chromatin components affects regulation of transcription, telomeric length, recombination and chromosomal stability. Mutations in TLC1 and EST2, which both encode components of telomerase, cause identical phenotypes: progressive shortening of telomeric DNA, increased chromosome loss and eventually cell death. In this review, we examine the relationship between telomeric chromatin and telomere replication and discuss the possibility that telomerase itself is an integral part of telomeric chromatin structure.


Automatic Detection and Classification of Convulsive Psychogenic Nonepileptic Seizures Using a Wearable Device.
Epilepsy is one of the most common neurological disorders and patients suffer from unprovoked seizures. In contrast, psychogenic nonepileptic seizures (PNES) are another class of seizures that are involuntary events not caused by abnormal electrical discharges but are a manifestation of psychological distress. The similarity of these two types of seizures poses diagnostic challenges that often leads in delayed diagnosis of PNES. Further, the diagnosis of PNES involves high-cost hospital admission and monitoring using video-electroencephalogram machines. A wearable device that can monitor the patient in natural setting is a desired solution for diagnosis of convulsive PNES. A wearable device with an accelerometer sensor is proposed as a new solution in the detection and diagnosis of PNES. The seizure detection algorithm and PNES classification algorithm are developed. The developed algorithms are tested on data collected from convulsive epileptic patients. A very high seizure detection rate is achieved with 100% sensitivity and few false alarms. A leave-one-out error of 6.67% is achieved in PNES classification, demonstrating the usefulness of wearable device in the diagnosis of PNES.


Expression of p40 in Primary Cutaneous Mucinous Carcinoma Versus Primary Mucinous Carcinomas of the Breast and Colon.
The transcription factor p63 is a homolog of p53, expressed in basal layers of epithelia and myoepithelial cells. Some studies have suggested that p63 may provide utility in differentiating primary versus metastatic mucinous carcinoma of the skin, given its preferential expression in primary adnexal neoplasms. There have been few studies investigating the expression patterns of p40, an isoform of p63, in primary cutaneous mucinous carcinomas. An immunohistochemical panel including p40, CK7, CK20, estrogen receptor, and progesterone receptor was applied to primary mucinous carcinomas of the skin, breast, and colon. Only a small subset (25%) of primary cutaneous mucinous carcinomas displayed focal positive staining with p40, similar to what has been reported in the literature for p63. All primary mucinous carcinomas of skin and breast labeled positively with CK7, estrogen receptor, and progesterone receptor. Primary colon mucinous carcinomas were only positive for CK20. Based on these results, p40 seems to be an insufficient marker for distinguishing primary versus metastatic mucinous carcinoma due to its low rate of positivity in primary cutaneous mucinous carcinomas.


Physiotherapy management for COVID-19 in the acute hospital setting: clinical practice recommendations.
This document outlines recommendations for physiotherapy management for COVID-19 in the acute hospital setting. It includes: recommendations for physiotherapy workforce planning and preparation; a screening tool for determining requirement for physiotherapy; and recommendations for the selection of physiotherapy treatments and personal protective equipment. It is intended for use by physiotherapists and other relevant stakeholders in the acute care setting caring for adult patients with confirmed or suspected COVID-19.


The effect of artificial antigen-presenting cells with preclustered anti-CD28/-CD3/-LFA-1 monoclonal antibodies on the induction of ex vivo expansion of functional human antitumor T cells.
Adoptive cell therapy with ex vivo expanded autologous antitumor cytotoxic T lymphocytes represents an important therapeutic option as an anticancer strategy. In order to identify a reliable method for producing adequate amounts of functional antitumor cytotoxic T lymphocytes with a potentially long in vivo lifespan, we tested the T-cell expansion efficiency of a new artificial antigen-presenting cell-based system. Our artificial antigen-presenting cells were generated with activating (anti-CD3), co-stimulating (anti-CD28) and adhesion (anti-LFA-1) biotinylated monoclonal antibodies preclustered in microdomains held on a liposome scaffold by neutravidin rafts. The co-localization of T-cell ligands in microdomains and the targeting of an adhesion protein, increasing the efficiency of immunological synapse formation, represent the novelties of our system. The activity of our artificial antigen-presenting cells was compared with that of anti-CD3/-CD28 coated immunomagnetic microbeads and immobilized anti-CD3 monoclonal antibody (OKT3 clone), the only two commercially available artificial systems. Our artificial antigen-presenting cells expanded both polyclonal T cells and MART-1-specific CD8(+) T cells in a more efficient manner than the other systems. Stimulation with artificial antigen-presenting cells allows for the generation of viable T cells displaying an immunophenotype consistent with in vivo potential for persistence, without increasing the frequency of regulatory T cells. The starting specificity of anti MART-1 CD8(+) T cells was preserved after stimulation with artificial antigen-presenting cells and it was statistically greater when compared to the activity of the same cells expanded with the other systems. Finally, our artificial antigen-presenting cells proved to be suitable for large-scale application, minimizing the volume and the costs of T-cell expansion. Our artificial antigen-presenting cells might represent an efficient tool to rapidly obtain a sufficient number of functional T cells for adoptive immunotherapy in patients with cancer.


Radical resection of intracavernous chondrosarcoma using an intradural keyhole approach: how I do it.
Skull base chondrosarcoma is rare, arising from the clivus or petroclival junction, and usually presents as ocular motility disorders. Endonasal transsphenoidal surgery may be performed in some cases of midline clivus chondrosarcoma. Chondrosarcoma is located within the cavernous sinus and presents a softer/gelatinous mass and can be removed with suctions and curettage. We have been using a simple intradural keyhole transcavernous approach, avoiding a complex extradural transcavernous dissection. The intracavernous chondrosarcoma was removed via a 5 mm keyhole opening over the Parkinson's triangle using a standard frontotemporal intradural approach. Minimally invasive keyhole surgical resection can be performed to eradicate skull base chondrosarcomas, avoiding complex extradural cranial base approaches.


Substitution of foreign protein sequences into a chimeric toxin composed of transforming growth factor alpha and Pseudomonas exotoxin.
TGF alpha-PE40 is a chimeric toxin made by replacing domain Ia of Pseudomonas exotoxin (PE) with transforming growth factor alpha (TGF alpha). We have now replaced a portion of domain Ib of PE with different polypeptides or an extra domain III of PE in transforming growth factor alpha-PE40 and maintained cell killing. Thus, TGF alpha-PE40 can be used to transport foreign protein sequences into the cytosol of cells.


Small invasive colonic cancer occurring in a hyperplastic polyp.
We report a case of advanced colon cancer which was supposed to have arisen from a hyperplastic polyp in a 68-year-old man. Colonoscopy revealed a depressed reddish area with a surrounding elevated lesion that was of a faded color compared with the normal mucosa. After the mucosal surface had been sprayed with crystal violet dye, magnifying colonoscopy showed an amorphous area in the central depression and the surrounding, slightly elevated lesion had an asteroid pattern. The depressed area was therefore considered to be a colonic cancer surrounded by a hyperplastic polyp. Endoscopic ultrasonography showed that the lesion was infiltrating further than the deep submucosal layer and it was therefore decided to treat the patient by laparoscopically assisted right hemicolectomy. The depressed lesion was found to be a well-differentiated adenocarcinoma invading the muscularis propria (diagnosed as IIc + IIa-like advanced adenocarcinoma). The surrounding flat elevated lesion was found to be hyperplastic mucosa. No adenomatous lesions were found. There have been few reported cases in which a preoperative diagnosis of carcinoma in a hyperplastic polyp has been made, but the possibility of carcinogenesis from hyperplastic polyps has come under consideration recently. This case was considered to be important because it raises the possibility that nonpolypoid cancer can develop from a hyperplastic polyp.


The cost of flares among patients with systemic lupus erythematosus with and without lupus nephritis in the United States.
Assess healthcare costs associated with systemic lupus erythematosus (SLE) flares among patients with and without lupus nephritis (LN). This retrospective cohort study used medical and pharmacy claims data from the United States-based Optum Clinformatics database to identify adults with SLE between 1 January 2016, and 31 December 2018. Index was the date of a patient's earliest SLE diagnosis claim during the identification period. Patients were categorized based on ICD-9/-10 diagnosis codes into one of two cohorts: SLE with LN (LN) and SLE without LN (non-LN). Baseline characteristics were assessed in the 12 months preceding index (baseline period). The presence, severity, and healthcare costs (in 2019 US dollars) of flares were determined in the 12 months following index (follow-up period). Overall, 11,663 patients with SLE were included (LN, n = 2916; non-LN, n = 8747). During the baseline period, a greater proportion of patients in the LN cohort versus non-LN cohort had a Charlson Comorbidity Index score ≥4 (72.5% vs 13.7%) and inpatient stays (41.0% vs 17.0%). A total of 12,190 flares were identified during the follow-up period (LN, 3494; non-LN, 8696). A greater proportion of flares experienced by patients with LN versus those without LN were moderate (61.2% vs 53.6%) and severe (10.6% vs 5.4%). The mean (standard deviation [SD]) number of moderate and severe flares per patient was greater among the LN cohort than the non-LN cohort (moderate: LN, 1.8 [1.2] and non-LN, 1.4 [1.2]; severe: LN, 0.2 [0.6] and non-LN, 0.1 [0.3]). The mean (SD) total healthcare costs associated with SLE flares of any severity were greater for patients with LN (LN, $5842 [9604]; non-LN, $2600 [4249]). The mean (SD) cost per flare increased with severity (mild: LN, $2753 [4640] and non-LN, $1606 [2710]; moderate: LN, $4561 [7156] and non-LN, $2587 [3720]; severe: LN, $29,148 [27,273] and non-LN, $14,829 [19,533]). Patients with SLE with LN have greater healthcare costs than those without LN. Flares among patients with LN were more frequent, severe, and costly than among patients without LN. This highlights the need for treatments that prevent or reduce flares among patients with SLE, both with and without LN.


Immune recovery after starting ART in HIV-infected patients presenting and not presenting with tuberculosis in South Africa.
We studied the immune response after starting antiretroviral treatment (ART) in 15,646 HIV-infected patients with or without tuberculosis (TB) at presentation in 3 ART programs in South Africa between 2003 and 2010. Patients presenting with TB had similar increases in CD4 cells compared with all other patients (adjusted difference 4.9 cells/µL per 6 months, 95% confidence interval: 0.2 to 9.7). Younger age, advanced clinical stage, female sex, and lower CD4 cell count at ART start were all associated with steeper CD4 slopes. In South Africa, HIV-infected patients presenting with TB experience immune recovery after starting ART that is no worse than in other patients.


Development of the Sri Lankan early teenagers' violence inventory: an instrument to measure peer violence in schools.
This study was designed to develop an inventory to measure peer violence among early teens (13-15 years of age) in schools in Sri Lanka. Development of SLETVI was carried out in two phases. In phase I, development of an operational definition for peer violence, identification, and finalizing violent acts for inventory was done by a combination of qualitative methods: a comprehensive literature review, focus group discussions among 13-15-year-old adolescents, their teachers and parents, and consultative meetings with experts in the field. Inventory was then pretested. In phase II, elaboration of SLETVI was carried out by administering it to a sample of 1700 adolescents (13-15 years old). Exploratory factor analysis using principal component analysis was performed separately for experiences of victimization and perpetration. Test-retest reliability of SLETVI was assessed. SLETVI included 37 items in three factors: "less severe violence," "severe physical," and "severe relational" violence. Combined use of qualitative and quantitative methods enabled development of a culturally valid and reliable operational inventory to assess early teenagers' peer violence in Sri Lankan and other South Asian schools.


Detection of preprocholecystokinin and preproenkephalin A mRNAs in rat brain by hybridization histochemistry using complementary RNA probes.
The distributions of mRNAs encoding preprocholecystokinin and preproenkephalin A in the rat brain were examined by in situ hybridization histochemistry. Complementary RNA probes 0.5-1.0 kilobase in length were synthesized in vitro with 35S-ribonucleotides using SP6 polymerase and a transcription vector containing the SP6 promotor. Following hybridization to fixed tissue sections, signals were detected by autoradiography after relatively short exposure times. These studies demonstrate that neuropeptide mRNAs can be detected within specific cells in the brain and that the patterns of hybridization match the localization of the peptides visualized by immunohistochemistry. Minimum numbers of mRNA copies encoding the peptides were determined by quantitative autoradiography.


Isolation and characterization of the pigment-protein complexes of Rhodopseudomonas sphaeroides by lithium dodecyl sulfate/polyacrylamide gel electrophoresis.
When purified photosynthetic membranes from Rhodopseudomonas sphaeroides were treated with lithium dodecyl sulfate and subjected to polyacrylamide gel electrophoresis at 4 degrees C, up to 11 pigment-protein complexes were resolved. Absorption spectra revealed that the smallest complex contained reaction center pigments and the others contained the antenna components B850 and B875 in various proportions. Of these antenna complexes, the largest was almost entirely B850 and the smallest contained only B875. After solubilization at 100 degrees C and electrophoresis on polyacrylamide gradient gels, the B850 complex gave rise to two polypeptide components migrating with apparent Mr of 10,000 and 8000, whereas with the B875 complex, two components were observed with apparent Mr of 12,000 and 8000. The reaction center complex gave rise to only the 24 and 21 kilodalton polypeptide subunits. Fluorescence emission spectra showed maxima at 872 and 902 nm for B850 and B875, respectively. Analyses of bacteriochlorophyll a and carotenoids indicated that, in the B875 complex, two molecules of each of these pigments are associated with the two polypeptides. The associations of B850 and B875 in large and small complexes obtained by lithium dodecyl sulfate treatment are consistent with models of their organization within the membrane.


Stimulation of pentose phosphate pathway dehydrogenase enzyme activities in ethionine-treated mice.
1. Mice treated with ethionine (intraperitoneally, 5mg./day for 4 days or 10mg./day for 3 days) showed a profound loss of hepatic glycogen, a decrease of glycogen synthetase activity, a development of hypoglycaemia, a two- to five-fold increase in the activity of glucose 6-phosphate dehydrogenase but no change in 6-phosphogluconate dehydrogenase and an earlier manifestation of the solubilization of phosphorylase as compared with glycogen synthetase. The administration of ATP did not prevent these effects. 2. During the early post-injection period (2-3 days) there was a further enhancement of the activity of glucose 6-phosphate dehydrogenase (tenfold) in the liver and a clear elevation of 6-phosphogluconate dehydrogenase activity (twofold). Subsequently, the glycogen concentration was restored, followed by an earlier reassociation of glycogen particle with phosphorylase than with glycogen synthetase, along with a disappearance of ethionine effect at about the eighteenth day. 3. Glucose 6-phosphate dehydrogenase from both control and ethionine-treated animals showed a marked preference for glucose 6-phosphate as substrate rather than for galactose 6-phosphate, whose rate of oxidation was only 10% of that of the glucose 6-phosphate. 4. Since actinomycin D, puromycin, 5-fluorouracil and dl-p-fluorophenylalanine failed to block the ethionine-enhanced glucose 6-phosphate dehydrogenase activity, the possibility that new enzyme protein synthesis is responsible for the effect is doubtful.


Bisphosphonates and risk of atrial fibrillation: a meta-analysis.
Bisphosphonates are the most commonly used drugs for the prevention and treatment of osteoporosis. Although a recent FDA review of the results of clinical trials reported no clear link between bisphosphonates and serious or non-serious atrial fibrillation (AF), some epidemiologic studies have suggested an association between AF and bisphosphonates. We conducted a meta-analysis of non-experimental studies to evaluate the risk of AF associated with bisphosphonates. Studies were identified by searching MEDLINE and EMBASE using a combination of the Medical Subject Headings and keywords. Our search was limited to English language articles. The pooled estimates of odds ratios (OR) as a measure of effect size were calculated using a random effects model. Seven eligible studies with 266,761 patients were identified: three cohort, three case-control, and one self-controlled case series. Bisphosphonate exposure was not associated with an increased risk of AF [pooled multivariate OR 1.04, 95% confidence interval (CI) 0.92-1.16] after adjusting for known risk factors. Moderate heterogeneity was noted (I-squared score = 62.8%). Stratified analyses by study design, cohort versus case-control studies, yielded similar results. Egger's and Begg's tests did not suggest an evidence of publication bias (P = 0.90, 1.00 respectively). No clear asymmetry was observed in the funnel plot analysis. Few studies compared risk between bisphosphonates or by dosing. Our study did not find an association between bisphosphonate exposure and AF. This finding is consistent with the FDA's statement.


Abnormalities of the S-T segment--Part I.
The usual normal S-T segment of the surface electrocardiogram (ECG) contributes little to the diagnostic procedure. When the S-T segment is too long or too short, or when it is displaced upward or downward, it is commonly abnormal. Under such circumstances, an S-T segment abnormality usually contributes considerable diagnostic information. When the Grant method of ECG interpretation is used, it is possible to perceive abnormalities of the S-T segment that may otherwise be ignored or misinterpreted. This paper describes the method of identification and the significance of primary and secondary S-T segment abnormalities. When a mean vector constructed for the S-T segment displacement seen in 12 ECG leads is relatively parallel with a mean vector representing the T wave, it is, with a few exceptions, part of the repolarization process and is therefore part of the T wave. This may be called a secondary S-T segment abnormality. When a mean vector constructed for the S-T segment displacement seen in 12 ECG leads is not relatively parallel with a mean vector representing the T wave, it is, with a few exceptions, not part of the repolarization process and is therefore not part of the T wave. This may be called a primary S-T segment abnormality.


An unexpected cause of acute liver failure.
A 66-year-old man presented with decompensated acute liver failure with no clear etiology. Eventually, a bone marrow biopsy revealed hemophagocytic cells, and he was diagnosed with hemophagocytic lymphohistiocytosis. This is an uncommon condition in adults, with high morbidity and mortality, and it is often indistinguishable from other forms of acute liver failure. Early clinical suspicion is critical to making the diagnosis, based on meeting at least four of nine specified criteria.


A new, vitamin D-based, multidimensional nomogram for the diagnosis of primary hyperparathyroidism.
To refine the diagnostic criteria for primary hyperparathyroidism (1°HPT) to identify atypical patients, in whom serum calcium, parathyroid hormone (PTH), or both are within the "normal" range. Total serum calcium, intact PTH, and 25-hydroxyvitamin D [25(OH)D] levels were measured in patients with 1°HPT and healthy patient groups. Multivariate analysis of healthy patient data first identified factors that significantly affected PTH levels and defined a new PTH reference range with a mathematical model. That nomogram was then validated for prediction of atypical 1°HPT in patients with surgically confirmed disease. On multivariate analysis, calcium (P = .0002), 25(OH)D (P<.0001), and age (P = .015) independently affected PTH. With these variables, we created a 4-dimensional nomogram that distinguished normal patients from those with hyperparathyroid states. Mathematically, this nomogram predicts 1°HPT when the measured serum PTH value is higher than PTH calculated by the following formula: PTH (pg/mL) = 120 - [6 × calcium (mg/dL)] - [0.52 × 25(OH)D (ng/mL)] + [0.26 × patient age (years)]. When applied to our surgical group of patients, this nomogram successfully identified 100% of patients (238 of 238) with classic 1°HPT, 84% (64 of 76) with normocalcemic 1°HPT, and 54% (20 of 37) with 1°HPT and normal PTH. This study uniquely defines a patient-specific upper limit of normal for PTH based on the readily available variables of serum calcium, 25(OH)D, and patient age. Our nomogram may allow for more rapid definitive diagnosis and treatment of 1°HPT in patients with atypical presentations.


[The function of the hypothalamo-hypophyseal-gonadal system in patients with Friedreich's ataxia].
Both the decrease of reproductive adaptation and effect of reproductive compensation were revealed in 19 cases of Friedreich's ataxia (FA). Retardation of sexual development was observed, the degree of which correlated with the age of disease manifestation. By means of radioimmunoassay basal and stimulated secretion of hypophysial and peripheral sex hormones were examined in 6 males and 9 females (17-30-years). The gonadal hypofunction was observed as the consequence of hypothalamo-hypophysial insufficiency. The decrease of follicle stimulating hormone and prolactin levels was found. The conclusion was made that pathological process in FA involves hypothalamus that causes in turn insufficiency of norepinephrine and epinephrine hypothalamic neurotransmission.


Brunn: an open source laboratory information system for microplates with a graphical plate layout design process.
Compound profiling and drug screening generates large amounts of data and is generally based on microplate assays. Current information systems used for handling this are mainly commercial, closed source, expensive, and heavyweight and there is a need for a flexible lightweight open system for handling plate design, and validation and preparation of data. A Bioclipse plugin consisting of a client part and a relational database was constructed. A multiple-step plate layout point-and-click interface was implemented inside Bioclipse. The system contains a data validation step, where outliers can be removed, and finally a plate report with all relevant calculated data, including dose-response curves. Brunn is capable of handling the data from microplate assays. It can create dose-response curves and calculate IC50 values. Using a system of this sort facilitates work in the laboratory. Being able to reuse already constructed plates and plate layouts by starting out from an earlier step in the plate layout design process saves time and cuts down on error sources.


[Calcified bladder squamous carcinoma: radiologic considerations].
To report a case of squamous cell carcinoma of the bladder, an uncommon tumor type, that presented with wide calcified areas. The literature on calcified bladder tumors is reviewed with special reference to the diagnostic aspects. The plain film, US and CT findings are presented. All the diagnostic imaging techniques demonstrated calcifications on the tumor surface and the characteristic curvilinear shape of this lesion. Calcified bladder carcinoma is rare and occurs in only 0.5% of the cases evaluated by conventional radiology. All calcifications probably arising in the bladder detected on conventional radiological evaluation should be studied further with other imaging techniques. If its etiology is unclear, cystoscopy with biopsy and/or a microbiological study should be performed.


The gene for the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase is located close to the gene for the large subunit in the cyanobacterium Anacystis nidulans 6301.
The gene for the small subunit (SS) of ribulose-1,5-bisphosphate carboxylase/oxygenase from a cyanobacterium, Anacystis nidulans 6301, has been cloned and subjected to sequence analysis. The SS coding region is located close to and downstream from the large subunit (LS) coding region on the same DNA strand. The spacer region between the LS and the SS coding regions contains 93 base pairs (bp), and has no promoter-like sequences. The coding region of A. nidulans SS gene contains 333 bp (111 codons). The deduced amino acid sequence of the A. nidulans SS protein shows 40% homology with those of higher plants.


Blockade of endothelial enzymes: new therapeutic targets.
Nitric oxide (NO) is the principal vasoactive substance produced by the vascular endothelium with antitrombotic, antiatherogenic and vasodilator actions. The loss of these functions is now known as endothelial dysfunction (ED) and it has been proposed that it is the final common pathway in cardiovascular disease. At the moment there is an important body of evidence that supports the proposal that ED is a consequence of an imbalance between the free radicals, NO, superoxide (O(-)(2)) and peroxynitrate (ONOO(-)). This imbalance is the result of the actions of well known risk factors associated with an inappropriate diet and infection-inflammation. Angiotensin-converting enzyme (ACE) inhibitors are highly effective against a variety of cardiovascular disorders. Experimental and clinical studies have demonstrated a beneficial effect of ACE inhibition on endothelial function. This action is mainly due to an increase in the concentration of bradykinin, which stimulates NO production. ACE inhibitors also block the formation of angiotensin II that results in a lower production of O(-)(2). These effects lead to improve the imbalance between NO and O(-)(2) observed in cardiovascular disease. This proposal is supported by different clinical trials that have shown that the ACE inhibitors with higher affinity by the tissular ACE, such as quinapril, are the most effective in reversing ED principally by accumulating bradykinin. Recently, the HOPE study conducted in patients at a high risk of cardiovascular events, showed how ramipril, an ACE inhibitor with high affinity by tissular ACE, decreased the mortality rate due to cardiovascular disease independently of changes in blood pressure.


Efficacy and safety concerns regarding Complementary and Alternative Medicine use among diabetes patients.
There has been an ever increasing rise in the use of Complementary and Alternative Medicinal (CAM) practices among general populations during past few decades. Individuals with diabetes being prone to an array of related health complications, demand special attention concerning their interest towards different CAM practices. Apart from clinical practitioner and patient based awareness programmes regarding the safety and efficacy implications of CAM therapies, stringent regulations should also be imposed on these unconventional health practices so as to strike off any major adverse events. This review is an effort to bring up the issues concerning these scientifically unproven practices with a focus on populations with diabetes.


Purification of stearidonic acid (18:4(n-3)) and hexadecatetraenoic acid (16:4(n-3)) from algal fatty acid with lipase and medium pressure liquid chromatography.
Stearidonic acid (18:4(n-3)) and hexadecatetraenoic acid (16:4(n-3)) are included in some edible marine algae such as Undaria pinnatifida and Ulva pertusa with relatively high compositions (up to 40%) of total fatty acids. In order to prepare 16:4(n-3) and 18:4(n-3) enriched fatty acid concentrates, we screened for a suitable lipase which concentrates these acids by the removal of other fatty acids in the selective esterification reaction reported by Shimada et al. (Shimada et al. (1997), J. Am. Oil Chem. Soc., 74, 1465-1470). In combination with the lipase reaction and reversed-phase medium pressure liquid chromatography, we purified 18:4(n-3) and 16:4(n-3) to more than 95% purity.


Mechanisms of human melanoma cell growth and tumor suppression by chromosome 6.
Breaks and deletions of chromosome 6 are among the most frequent karyotypic abnormalities that appear in human malignant melanoma cells, and chromosome transfer experiments have provided functional evidence for the presence of a melanoma tumor suppressor locus on chromosome 6[J.M. Trent et al., Science (Washington DC), 247: 568-571, 1990]. We have investigated the genetic mechanism of this suppression. We have found that the suppression of tumorigenicity that follows the introduction of a normal copy of chromosome 6 into the UACC 903 human melanoma cell line is correlated with increased chromosome 6 dosage, rather than with the presence of the transferred normal copy of the chromosome. Transfer of chromosome 6 into another human melanoma cell line, MelJuSo, does not result in suppression of primary tumor formation, although the additional copy of chromosome 6 does reduce the cell growth rate in vitro. Finally, we have identified a substantial portion of the chromosome that is evidently not involved in tumor suppression. We have observed that a copy of chromosome 6 derived from a normal cell but with a deletion involving chromosome bands 6q22-6q24 suppresses primary tumor formation in UACC 903 cells as effectively as the intact chromosome. We have thus provided additional information about the chromosomal location of this tumor suppressor and about its mode of action.


Morphology and morphometry of adult nematodes on Sumatran elephants (Elephas maximus sumatranus) in Way Kambas National Park area, Indonesia.
Worms from nematodes are the most numerous and the most detrimental in elephants. Most adult worms are located in the digestive tract. Nematode infection is at higher risk in young elephants, which caused several cases such as anemia, hypoalbuminemia, enteritis, and even death. This study aimed to determine the morphology and morphometry of adult nematodes on Sumatran elephants in Way Kambas National Park area. Nematode samples were obtained from Sumatran elephants' feces (Elephas maximus sumatranus) in Way Kambas National Park, Lampung Province, after being given Kalbazen® containing albendazole 1000 mg at a dose of 10 mg/kg by the veterinarian in charge of the National Park area. For the morphological and morphometric examinations, we used an Olympus BX 51 microscope equipped with Olympus DP 12 camera and were conducted at the Parasitology Laboratory, Faculty of Veterinary Medicine, Universitas Gadjah Mada. The scanning electron microscopic (SEM) analysis was carried out at the Biology Research Center of the Indonesian Institute of Sciences (Lembaga Ilmu Pengetahuan Indonesia). The results of macroscopic observations of the obtained nematodes showed that the nematodes which were found have the characteristics of round, slim, and white color. The size of a female worm was larger than a male worm. Microscopic examination in four anterior papillae indicated that the dorsal lobe in the copulatory bursa was longer than lateral lobe. The result of inspection with the SEM showed a leaf crown consisting of 10 elements, a pair of amphids laterally, and two pairs of papilla in a submedian region. Based on our morphology and morphometry examinations of adult nematodes in Sumatran elephant (E. maximus sumatranus) in Way Kambas National Park area, the adult nematodes which were found are species of Quilonia travancra.


Induction of the human heat shock promoter HSP70B by nutritional stress: implications for cancer gene therapy.
We designed and tested, in vitro, an adenoviral construct containing the feline interleukin-12 (IL-12) gene under control of the heat-inducible promoter HSP70B. This construct, AdhspfIL12, was used in a phase I trial in feline soft tissue sarcomas. During the course of our experiments, we noted that IL-12 was being produced in the transfected Crandell Feline Kidney (CrFK) cells under certain conditions even in the absence of hyperthermia. This observation was further explored to identify the cause of this unintended HSP70B induction. We used real-time PCR as a sensitive method to quantitatively detect the presence of even small amounts of IL-12 mRNA. This served as a surrogate indicator of HSP70B induction. Various conditions were tested to induce the heat shock promoter, including nutritional deprivation, radiation and changes in pH. Nutritional stresses, specifically the absence of glucose and glutamine, could induce the heat shock promoter, thus, resulting in production of the downstream gene product. Other factors known to trigger the heat shock response, pH change, and reactive oxygen species production were also studied but were not found to contribute to heat shock promoter induction in our setting. The human heat shock promoter (HSP70B) is reported to be an efficient and tightly regulated promoter. We discovered, using sensitive real-time PCR techniques, that it can also be induced in response to cellular nutrient stresses. The pros and cons of this phenomenon and its implications for cancer gene therapy are discussed.


The association between recreational physical activity and depression in the short sleep population: a cross-sectional study.
Short sleep is more common in the modern society. Recreational physical activity (RPA) like exercise brings both mental and physiological benefits to depression; paradoxically, lack of sleep is harmful. Evidence on the association between RPA and depression in the short sleep population is limited. Participants with short sleep condition from the National health and Nutrition Examination Surveys (NHANES 2007-2018) were included in the present study. Short sleep condition was defined as ≤ 7 h per night. Sleep duration and RPA status were self-reported in NHANES by the Physical Activity Questionnaire using a 7-day recall method. Multivariable logistic regression was applied to evaluate the association between RPA and depression. Additionally, the non-linear relationship between RPA and depression was evaluated using the threshold effect analysis and restricted cubic spline. This cross-sectional study comprised 6,846 adults' data, and the weighted participants were 52,501,159. The weighted prevalence of depression was higher in females, which took up 65.85% of all depression patients. In fully adjusted models, sufficient volume of RPA was associated with lower depression risks, with OR (95% CI) =0.678 (0.520, 0.883). Further analysis revealed a U-shaped association between RPA and incident depression, and the inflection point was 640 MET-minutes/week. When RPA <640 MET-minutes/week, increased RPA was associated with lower risk of incident depression, with OR (95% CI) = 0.891 (0.834, 0.953). When RPA ≥ 640 MET-minutes/week, the benefits of RPA seemed to be not significant, with OR (95% CI) = 0.999 (0.990, 1.009). Our findings observed associations between RPA condition and incident depression in the short sleep population. Moderate RPA was beneficial to maintain mental health and associated with lower incidence of depression for short sleepers, but excessive RPA might increase the risk of depression. For general short sleepers, keeping the RPA volume approximately 640 MET-minutes/week was beneficial to lower risks of depression. Gender difference should be considered as an important factor for further studies to examine these relationships and explore mechanisms.


Using genetics to detangle the relationships between red cell distribution width and cardiovascular diseases: a unique role for body mass index.
Red cell distribution width (RDW) is an enigmatic biomarker associated with the presence and severity of multiple cardiovascular diseases (CVDs). It is unclear whether elevated RDW contributes to, results from, or is pleiotropically related to CVDs. We used contemporary genetic techniques to probe for evidence of aetiological associations between RDW, CVDs, and CVD risk factors. Using an electronic health record (EHR)-based cohort, we built and deployed a genetic risk score (GRS) for RDW to test for shared genetic architecture between RDW and the cardiovascular phenome. We also created GRSs for common CVDs (coronary artery disease, heart failure, atrial fibrillation, peripheral arterial disease, venous thromboembolism) and CVD risk factors (body mass index (BMI), low-density lipoprotein, high-density lipoprotein, systolic blood pressure, diastolic blood pressure, serum triglycerides, estimated glomerular filtration rate, diabetes mellitus) to test each for association with RDW. Significant GRS associations were further interrogated by two-sample Mendelian randomisation (MR). In a separate EHR-based cohort, RDW values from 1-year pre-gastric bypass surgery and 1-2 years post-gastric bypass surgery were compared. In a cohort of 17 937 subjects, there were no significant associations between the RDW GRS and CVDs. Of the CVDs and CVD risk factors, only genetically predicted BMI was associated with RDW. In subsequent analyses, BMI was associated with RDW by multiple MR methods. In subjects undergoing bariatric surgery, RDW decreased postsurgery and followed a linear relationship with BMI change. RDW is unlikely to be aetiologically upstream or downstream of CVDs or CVD risk factors except for BMI. Genetic and clinical association analyses support an aetiological relationship between BMI and RDW.


Congenital pigmented nevi of the auricle: clinical experience and approach to treatment.
Congenital pigmented nevi of the auricle are uncommon. The authors' approach is to excise these nevi and perform reconstruction because of the risk of malignant transformation and the aesthetic and psychological effects these nevi can have on the child. This study presents the authors' experience in treating congenital nevi of the ear and suggests treatment principles and guidelines for the reconstructive surgeon. Fourteen patients with congenital nevus of the ear were treated from October of 1992 to September of 2008 by the senior surgeon (B.S.B.). Nevi involving the more stable areas such as the concha can be resected and grafted early; the antihelix, scapha, and triangular fossa area can be resected and grafted next; and the helical rim, having the most easily distorted cartilage, should be treated last. Lobule reconstruction requires combined flaps and a dermal fat graft or a postauricular fascial fat flap. Successful reconstruction was achieved in 10 patients. Three patients require final revision procedures (lobule reconstruction). One patient, early in our series, developed a deformed helical rim resulting from skin grafting at age 16 months, before the cartilage was firm enough to withstand the contraction forces of the skin graft. All subsequent patients with helical rim involvement had treatment delayed until the ear was at or near completion of its growth. Congenital nevi of the ear present a challenging reconstruction surgeon. The authors developed a treatment plan that breaks the ear down to aesthetic units and considers the location of the nevus, patient age, and the firmness of the cartilage.


Offline recognition of unconstrained handwritten texts using HMMs and statistical language models.
This paper presents a system for the offline recognition of large vocabulary unconstrained handwritten texts. The only assumption made about the data is that it is written in English. This allows the application of Statistical Language Models in order to improve the performance of our system. Several experiments have been performed using both single and multiple writer data. Lexica of variable size (from 10,000 to 50,000 words) have been used. The use of language models is shown to improve the accuracy of the system (when the lexicon contains 50,000 words, the error rate is reduced by approximately 50 percent for single writer data and by approximately 25 percent for multiple writer data). Our approach is described in detail and compared with other methods presented in the literature to deal with the same problem. An experimental setup to correctly deal with unconstrained text recognition is proposed.


Manganese-dependent polioviruses caused by mutations within the viral polymerase.
Viral RNA-dependent RNA polymerases exhibit great sequence diversity. Only six core amino acids are conserved across all polymerases of positive-strand RNA viruses of eukaryotes. While exploring the function of one of these completely conserved residues, asparagine 297 in the prototypic poliovirus polymerase 3D(pol), we identified three viable mutants with noncanonical amino acids at this conserved position. Although asparagine 297 could be replaced by glycine or alanine in these mutants, the viruses exhibited Mn(2+)-dependent RNA replication and viral growth. All known RNA polymerases and replicative polymerases of bacterial, eukaryotic, and viral organisms are thought to be magnesium dependent in vivo, and therefore these mutant polioviruses may represent the first viruses with a requirement for an alternative polymerase cation. These results demonstrate the extreme functional flexibility of viral RNA-dependent RNA polymerases. Furthermore, the finding that strictly conserved residues in the nucleotide binding pocket of the polymerase can be altered in a manner that supports virus production suggests that drugs targeting this region of the enzyme will still be susceptible to the problem of drug-resistant escape mutants.


Persistence of Japanese encephalitis virus is associated with abnormal expression of the nonstructural protein NS1 in host cells.
Persistent infection with Japanese encephalitis virus (JEV) was established in murine neuroblastoma N18 cells, and the persistency has been maintained in cell culture for over 6 months. From the persistently infected cells, a clone named C2-2 was selected and expanded to form a stable cell line. The vast majority of C2-2 cells showed viral protein staining by immunofluorescence and continuously produced low levels of virus (10(3) to 10(4) PFU/ml) without marked cytopathic effects or cyclic variations. In addition to the wild-type viral proteins, truncated forms of the viral nonstructural protein 1 (NS1) as well as its derivative NS1' were produced in C2-2 cells. Both truncated NS1 and NS1' contain deletions at their N-termini; however, the analyses by RT-PCR and direct sequencing of the viral RNA failed to detect any truncations or mutations within the NS1 region, suggesting that NS1 truncation was a result of a unique posttranslational proteolytic cleavage of NS1 in the persistently infected cells. Similar but not identical truncation of NS1 was also observed in two other persistently infected cell lines established in Vero and DBT (murine astrocytoma) cells. However, viruses released from C2-2 cells did not produce truncated NS1 upon infection of N18 cells, suggesting that NS1 truncations were the result of virus-cell interaction in persistently infected cells. These data indicate a strong association between abnormal NS1 expression and JEV persistency. A probable involvement of dysfunctional NS1 in the establishment and/or maintenance of JEV persistency in tissue culture is discussed.


Leptomeningeal Neural Organoid (LMNO) Fusions as Models to Study Meninges-Brain Signaling.
Neural organoids derived from human induced pluripotent stem cells (iPSCs) provide a model to study the earliest stages of human brain development, including neurogenesis, neural differentiation, and synaptogenesis. However, neural organoids lack supportive tissues and some non-neural cell types that are key regulators of brain development. Neural organoids have instead been co-cultured with non-neural structures and cell types to promote their maturation and model interactions with neuronal cells. One structure that does not form de novo with neural organoids is the meninges, a tri-layered structure that surrounds the CNS and secretes key signaling molecules required for mammalian brain development. Most studies of meninges-brain signaling have been performed in mice or using two-dimensional (2D) cultures of human cells, the latter not recapitulating the architecture and cellular diversity of the tissue. To overcome this, we developed a co-culture system of neural organoids generated from human iPSCs fused with fetal leptomeninges from mice with fluorescently labeled meninges ( Col1a1-GFP ). These proof-of-concept studies test the stability of the different cell types in the leptomeninges (fibroblast and macrophage) and the fused brain organoid (progenitor and neuron), as well as the interface between the organoid and meningeal tissue. We test the longevity of the fusion pieces after 30 days and 60 days in culture, describe best practices for preparing the meninges sample prior to fusion, and examine the feasibility of single or multiple meninges pieces fused to a single organoid. We discuss potential uses of the current version of the LMNO fusion model and opportunities to improve the system.


Immunological protection against mycobacterium tuberculosis infection.
Mycobacterium tuberculosis (Mtb) invades the lung via an airborne pathway, and the bacteria reach the lung alveoli and are then engulfed by alveolar macrophages in innate immunity, which uses germline-encoded pattern recognition receptors such as the mannose receptor and Toll-like receptor for recognition of the pathogen. Mtb can survive in phagosomes of macrophages by escaping from lysosomal enzyme and reactive nitrogen intermediate toxicity. Dendritic cells transport Mtb from lung mucosa to draining lymph nodes and present the mycobacterial antigens to CD4+ and CD8+ T cells in adaptive immunity, which uses antigen-specific receptors formed by somatic gene rearrangement. Natural memory T cells such as gammadelta T cells and CD 1-restricted T cells are also stimulated and serve to shape the subsequent development of T-helper 1-dominant immunity, which induces granuloma at the initial focus of infection, is sufficient to prevent acute tuberculosis, but is unable to eradicate the infection completely. Once the host's defense becomes compromised, reactivation of the disease, which is called latent infection with Mtb, occurs. Recent progress in elucidating adaptive immune responses to acute and latent infection with Mtb may lead to the establishment of effective therapeutic and prophylactic approaches for controlling Mtb infection.