Freezing of Gait in Parkinson's Disease Is Associated with Reduced 6-[(18)F]Fluoro-l-m-tyrosine Uptake in the Locus Coeruleus.
Freezing of gait (FOG) is a common disorder in Parkinson's disease (PD) and could be attributed to a reduction in brain noradrenaline. The aim of this study was to determine the relationship between aromatic l-amino acid decarboxylase (AADC) activity in the locus coeruleus (LC) and FOG in PD using high-resolution positron emission tomography with an AADC tracer, 6-[(18)F]fluoro-l-m-tyrosine (FMT). We assessed 40 patients with PD and 11 age-matched healthy individuals. PD was diagnosed based on the UK Brain Bank criteria by two movement disorder experts. FOG was directly observed by the clinician and assessed using a patient questionnaire. FMT uptake in the LC, caudate, and putamen was analyzed using PMOD software on coregistered magnetic resonance images. FOG was present in 30 patients. The severity of FOG correlated with the decrease of FMT uptake in the LC regardless of disease duration and the severity of other motor impairments, indicating dysfunction of the noradrenergic network in FOG.


Energy exponents of avalanches and Hausdorff dimensions of collapse patterns.
A simple numerical model to simulate athermal avalanches is presented. The model is inspired by the "porous collapse" process where the compression of porous materials generates collapse cascades, leading to power law distributed avalanches. The energy (E), amplitude (A_{max}), and size (S) exponents are derived by computer simulation in two approximations. Time-dependent "jerk" spectra are calculated in a single avalanche model where each avalanche is simulated separately from other avalanches. The average avalanche profile is parabolic, the scaling between energy and amplitude follows E∼A_{max}^{2}, and the energy exponent is ε = 1.33. Adding a general noise term in a continuous event model generates infinite avalanche sequences which allow the evaluation of waiting time distributions and pattern formation. We find the validity of the Omori law and the same exponents as in the single avalanche model. We then add spatial correlations by stipulating the ratio G/N between growth processes G (linked to a previous event location) and nucleation processes N (with new, randomly chosen nucleation sites). We found, in good approximation, a power law correlation between the energy exponent ε and the Hausdorff dimension H_{D} of the resulting collapse pattern H_{D}-1∼ɛ^{-3}. The evolving patterns depend strongly on G/N with the distribution of collapse sites equally power law distributed. Its exponent ɛ_{topo} would be linked to the dynamical exponent ε if each collapse carried an energy equivalent to the size of the collapse. A complex correlation between ɛ,ɛ_{topo}, and H_{D} emerges, depending strongly on the relative occupancy of the collapse sites in the simulation box.


Helicobacter pylori VacA suppresses Lactobacillus acidophilus-induced interferon beta signaling in macrophages via alterations in the endocytic pathway.
Helicobacter pylori causes chronic gastritis and avoids elimination by the immune system of the infected host. The commensal bacterium Lactobacillus acidophilus has been suggested to exert beneficial effects as a supplement during H. pylori eradication therapy. In the present study, we applied whole-genome microarray analysis to compare the immune responses induced in murine bone marrow-derived macrophages (BMDMs) stimulated with L. acidophilus, H. pylori, or both bacteria in combination. While L. acidophilus induced a Th1-polarizing response characterized by high expression of interferon beta (IFN-β) and interleukin 12 (IL-12), H. pylori strongly induced the innate cytokines IL-1β and IL-1α. In BMDMs prestimulated with L. acidophilus, H. pylori blocked the expression of L. acidophilus-induced IFN-β and IL-12 and suppressed the expression of key regulators of the Rho, Rac, and Cdc42 GTPases. The inhibition of L. acidophilus-induced IFN-β was independent of H. pylori viability and the virulence factor CagPAI; however, a vacuolating cytotoxin (vacA) mutant was unable to block IFN-β. Confocal microscopy demonstrated that the addition of H. pylori to L. acidophilus-stimulated BMDMs redirects intracellular processing, leading to an accumulation of L. acidophilus in the endosomal and lysosomal compartments. Thus, our findings indicate that H. pylori inhibits the development of a strong Th1-polarizing response in BMDMs stimulated with L. acidophilus by blocking the production of IFN-β in a VacA-dependent manner. We suggest that this abrogation is caused by a redirection of the endocytotic pathway in the processing of L. acidophilus. IMPORTANCE Approximately half of the world's population is infected with Helicobacter pylori. The factors that allow this pathogen to persist in the stomach and cause chronic infections have not yet been fully elucidated. In particular, how H. pylori avoids killing by macrophages, one of the main types of immune cell underlying the epithelium, remains elusive. Here we have shown that the H. pylori virulence factor VacA plays a key role by blocking the activation of innate cytokines induced by the probiotic Lactobacillus acidophilus in macrophages and suppresses the expression of key regulators required for the organization and dynamics of the intracellular cytoskeleton. Our results identify potential targets for the treatment of H. pylori infection and vaccination, since specific inhibition of the toxin VacA possibly allows the activation of an efficient immune response and thereby eradication of H. pylori in the host.


A procedure for tissue freezing and processing applicable to both intra-operative frozen section diagnosis and tissue banking in surgical pathology.
Different methods for snap freezing surgical human tissue specimens exist. At pathology institutes with higher work loads, solid carbon dioxide, freezing sprays, and cryostat freezing are commonly used as coolants for diagnosing frozen tissue sections, whereas for tissue banking, liquid nitrogen or isopentane cooled with liquid nitrogen is preferred. Freezing tissues for diagnostic and research purposes are therefore often time consuming, laborious, even hazardous, and not user friendly. In tissue banks, frozen tissue samples are stored in cryovials, capsules, cryomolds, or cryocassettes. Tissues are additionally embedded using freezing media or wrapped in plastic bags or aluminum foils to prevent desiccation. The latter method aggravates enormously further tissue handling and processing. Here, we describe an isopentane-based workflow which concurrently facilitates tissue freezing and processing for both routine intra-operative frozen section and tissue banking and satisfies the qualitative demands of pathologists, cancer researchers, laboratory technicians, and tissue bankers.


An ethically justified framework for clinical investigation to benefit pregnant and fetal patients.
Research to improve the health of pregnant and fetal patients presents ethical challenges to clinical investigators, institutional review boards, funding agencies, and data safety and monitoring boards. The Common Rule sets out requirements that such research must satisfy but no ethical framework to guide their application. We provide such an ethical framework, based on the ethical concept of the fetus as a patient. We offer criteria for innovation and for Phase I and II and then for Phase III clinical trials to improve the health of pregnant patients and of fetal patients and also criteria to responsibly manage the transition from investigation to clinical practice. Basing ethical criteria for research involving pregnant women on the ethical concept of the fetus as a patient insulates the proposed ethical framework and therefore research on pregnant women from the divisive and politicized concepts and discourses of personhood, fetal rights, and unborn child.


Transcriptional repression of estrogen receptor α signaling by SENP2 in breast cancer cells.
Estrogen receptors (ERs) are ligand-activated transcription factors involved in many physiological and pathological processes, including breast cancer. Their activity is fine-tuned by posttranslational modifications, notably sumoylation. In the present study, we investigated the role of the small ubiquitin-related modifier (SUMO) protease, SUMO1/sentrin/suppressor of Mif 2-specific peptidase 2 (SENP2), in the regulation of ERα activity. We first found SENP2 to significantly repress estradiol-induced transcriptional activity in breast cancer cells (MCF7 and T47D). This effect was observed with a reporter plasmid and on endogenous genes such as TFF1 and CTSD, which were shown to recruit SENP2 in chromatin immunoprecipitation experiments. Using glutathione S-transferase pull-down, coimmunoprecipitation and proximity ligation assays, SENP2 was found to interact with ERα and this interaction to be mediated by the amino-terminal region of the protease and the hinge region of the receptor. Interestingly, we demonstrated that ERα repression by SENP2 is independent of its SUMO protease activity and requires a transcriptional repressive domain located in the amino-terminal end of the protease. Using small interfering RNA assays, we evidenced that this domain recruits the histone deacetylase 3 (HDAC3), to be fully active. Furthermore, using both overexpression and knockdown strategies, we showed that SENP2 robustly represses estrogen-dependent and independent proliferation of MCF7 cells. We provided evidence that this effect requires both the proteolytic and transcriptional activities of SENP2. Altogether, our study unravels a new property for a SUMO protease and identifies SENP2 as a classical transcription coregulator.


Violence against Healthcare Workers during the COVID-19 Pandemic: A Review of Incidents from a Lower-Middle-Income Country.
Healthcare workers (HCWs) across the globe have met tremendous challenges during the COVID-19 (coronavirus disease 2019) pandemic, such as shortages of personal protective equipment, extensive work hours, and constant fear of catching the virus or transmitting it to loved ones. Adding on to the already existing burnout, an increase in incidents of violence and aggression against HCWs was seen in Pakistan and globally. Primarily to review cases of violence against HCWs in Pakistan, highlighting and comparing the instigating factors seen within the country and globally. Secondly, to enlist possible interventions to counter workplace violence in healthcare during a pandemic and in general. Incidents of violence towards HCWs in Pakistan during the COVID-19 pandemic occurring between April 7, 2020, and August 7, 2020, were included. The incidents reported from local newspapers were reviewed. A total of 29 incidents were identified, with perpetrators of violence most commonly being relatives of COVID-19 patients. Most frequent reasons included mistrust in HCWs, belief in conspiracy theories, hospitals' refusal to admit COVID-19 patients due to limited space, COVID-19 hospital policies, and the death of the COVID-19 patients. Protests by doctors and other HCWs for provision of adequate PPE, better quarantine conditions for doctors with suspected COVID-19, and better compensation for doctors on COVID-19 patient duty resulted in police violence towards HCWs. To avoid such incidents in the future, institutions, healthcare policymakers, media organisations, and law enforcement agencies must work together for widespread public awareness to counter misconceptions and to exhibit responsible journalism. In hospitals, measures such as de-escalation training and increased security must be implemented. Furthermore, law enforcement agencies must be trained in non-violent methods of crowd dispersal and control to manage peaceful protests by HCWs over legitimate issues.


Targeting PIM kinases in cancer therapy: An update on pharmacological small-molecule inhibitors.
PIM kinases, a serine/threonine kinase family with three isoforms, has been well-known to participate in multiple physiological processes by phosphorylating various downstream targets. Accumulating evidence has recently unveiled that aberrant upregulation of PIM kinases (PIM1, PIM2, and PIM3) are closely associated with tumor cell proliferation, migration, survival, and even resistance. Inhibiting or silencing of PIM kinases has been reported have remarkable antitumor effects, such as anti-proliferation, pro-apoptosis and resensitivity, indicating the therapeutic potential of PIM kinases as potential druggable targets in many types of human cancers. More recently, several pharmacological small-molecule inhibitors have been preclinically and clinically evaluated and showed their therapeutic potential; however, none of them has been approved for clinical application so far. Thus, in this perspective, we focus on summarizing the oncogenic roles of PIM kinases, key signaling network, and pharmacological small-molecule inhibitors, which will provide a new clue on discovering more candidate antitumor drugs targeting PIM kinases in the future.


Early detection of global cerebral anoxia: improved accuracy by high-b-value diffusion-weighted imaging with long echo time.
Early and accurate detection of global cerebral anoxia is important for determination of prognosis and further management. We evaluated whether accuracy in early detection of global cerebral anoxia was improved by high-b-value diffusion-weighted imaging (DWI) with long echo time (TE). Routine DWI (b = 1000 s/mm(2); TE = 139 ms), high-b-value DWI (b = 3000 s/mm(2); TE = 190 ms), T2-weighted imaging (T2WI), and fluid-attenuated inversion recovery (FLAIR) imaging were acquired in six patients who experienced cardiopulmonary arrest within 24 hours and six volunteers. Region of interest-based analysis was performed. Regions of interest of patients showing significant decrease in apparent diffusion coefficient (ADC) values than volunteers were considered abnormal. Three neuroradiologists independently assessed images of the patients for conspicuity of hyperintensity within regions of interest. Receiver operating characteristic (ROC) analysis was performed, and the area under the curve (Az) was compared among sequences and observers. Average contrast and contrast-to-noise ratios between abnormal regions of interest and regions of interest of normal surrounding parenchyma were calculated. For all observers, high-b-value DWIs achieved the largest Az, and FLAIR imaging the lowest Az. Az of routine DWI and T2WI were between these values. High-b-value DWI and FLAIR imaging showed no significant interobserver variation in Az, whereas routine DWI and T2WI did. High-b-value DWI also achieved the largest contrast and contrast-to-noise ratios. High-b-value DWI with long TE improved accuracy in early detection of global cerebral anoxia. Application of the sequence would facilitate early diagnosis.


Pharmacokinetics of interferon alpha-2b in healthy volunteers.
In a three-way crossover design, 12 healthy male volunteers received 5 X 10(6) IU/m2 body surface area interferon alpha-2b(IFN alpha-2b) by intravenous (IV) infusion over 30 minutes, intramuscular (IM) injections, and subcutaneous (SC) injections. Blood and urine samples were collected at specified times, and analysis of IFN alpha-2b concentrations was performed by immunoradiometric assay. "Flulike" symptoms were the most frequently reported adverse experiences and were independent of the route of administration. After a 30-minute IV infusion, IFN alpha-2b disappeared rapidly from serum, with distribution and elimination phase half-lives of 0.1 hour and 1.7 hours, respectively. Interferon alpha-2b was absorbed slowly after IM and SC administration, with similar absorption half-lives of 5.8 and 5.5 hours, respectively. The observed maximal concentrations after IM and SC administration were 42.1 IU/mL at six hours and 45.8 IU/mL at eight hours, respectively. Interferon alpha-2b was eliminated with half-lives of 2.2 hours after IM administration and 2.9 hours after SC administration. The areas under the serum concentration-time curves for the SC and IM doses were higher than those obtained for the IV infusion. Measurable amounts of IFN alpha-2b were not found in urine regardless of the route of administration.


Relationship of myocardial metabolism and coronary flow: dependence on extracellular calcium.
The hypothesis that extracellular [Ca2+] ([Ca2+]e) influences the transmission of vasoregulatory information from the heart to coronary smooth muscle was examined. The isolated, retrogradely perfused rat heart was used to evaluate vascular reactivity to transient infusion of either 0.88 mM Amytal (amobarbital), hypoxia (95% N2-5% CO2), or 12 microM adenosine in the presence and absence of 2.5 microM verapamil. Amytal alone enhanced coronary flow by 78% and lactate efflux by 10-fold, whereas it decreased O2 consumption by 40% and the ratio of the free cytosolic concentrations of ATP-to-ADP times the intracellular concentration of inorganic phosphate ([ATP]f/[ADP]f[Pi]) by 10-fold. These responses were attenuated by preperfusion with verapamil, i.e., flow and lactate efflux increased by 4% and fourfold, respectively, and O2 consumption and [ATP]f/[ADP]f/[Pi] decreased by 8% and threefold, respectively, as compared with values attained with verapamil at steady state. Washout of the calcium channel antagonist by perfusion for 55 min with a verapamil-free medium resulted in a return of metabolic variables to control levels. Subsequent infusion of Amytal induced metabolic changes that were similar to those obtained before exposure to the calcium antagonist, but flow rose only by 26%. Vascular responses to hypoxia were similarly modified, whereas those to adenosine were unaffected. Hearts perfused with media containing either low [Ca2+]e or 10 microM diltiazem and exposed to Amytal produced flow patterns analogous to those perfused with verapamil. These data, in part, distinguish changes in vascular resistance that occur in response to cardiac metabolism from those which result from agents with direct action on smooth muscle, in particular, adenosine.(ABSTRACT TRUNCATED AT 250 WORDS)


Glucose tolerance and insulin response in atherosclerosis.
Oral glucose tolerance tests were carried out on 51 men with atherosclerotic peripheral vascular disease, none of whom were known diabetics or had suffered recent myocardial infarction. The plasma insulin and blood sugar responses were compared with 47 age and sex-matched controls. There was no significant difference in obesity between the two groups. The patient group showed an increased plasma insulin response with a delay in return to fasting levels, and the blood sugar response was similar. These results suggest that hyperinsulinaemia and hyperglycaemia are often associated with atherosclerosis, and may have a role in its aetiology.


Fractional Flow on TOF-MRA as a Measure of Stroke Risk in Children with Intracranial Arterial Stenosis.
Conventional angiography is the criterion standard for measuring intracranial arterial stenosis. We evaluated signal intensity ratios from TOF-MRA as a measure of intracranial stenosis and infarct risk in pediatric stroke. A retrospective study was undertaken in children with intracranial arterial stenosis, who had TOF-MRA and conventional angiography performed within 6 months. Arterial diameters were measured for percentage stenosis. ROI analysis on TOF-MRA measured signal intensity in pre- and poststenotic segments, with post-/pre-signal intensity ratios calculated. The Pearson correlation was used to compare percentage stenosis on MRA with conventional angiography and signal intensity ratios with percentage stenosis; the point-biserial correlation was used for infarcts compared with percentage stenosis and signal intensity ratios. Sensitivity, specificity, and positive and negative predictive values were calculated for determining severe (≥70%) stenosis from MRA and signal intensity ratios against the criterion standard conventional angiography. P < .05 was considered statistically significant. Seventy stenotic segments were found in 48 studies in 41 children (median age, 11.0 years; range, 5 months to 17.0 years; male/female ratio, 22:19): 20/41 (48.8%) bilateral, 11/41 (26.8%) right, and 10/41 (24.4%) left, with the most common site being the proximal middle cerebral artery (22/70, 31%). Moyamoya disease accounted for 27/41 (65.9%). Signal intensity ratios and conventional angiography stenosis showed a moderate negative correlation (R = -0.54, P < .001). Receiver operating characteristic statistics showed an area under the curve of 0.86 for using post-/pre-signal intensity ratios to determine severe (≥70%) carotid stenosis, yielding a threshold of 1.00. Sensitivity, specificity, and positive and negative predictive values for severe stenosis were the following-MRA: 42.8%, 58.8%, 30.0%, and 71.4%; signal intensity ratio >1.00: 97.1%, 77.8%, 71.7%, and 97.4%; combination: 75.5%, 100%, 100%, and 76.8%, respectively. Signal intensity ratios decreased with increasing grade of stenosis (none/mild-moderate/severe/complete, P < .001) and were less when associated with infarcts (0.81 ± 0.52 for arteries associated with downstream infarcts versus 1.31 ± 0.55 for arteries without associated infarcts, P < .001). Signal intensity ratios from TOF-MRA can serve as a noninvasive measure of intracranial arterial stenosis and allow identification of high-risk lesions in pediatric stroke.


Dynamic association of RNA-editing enzymes with the nucleolus.
ADAR1 and ADAR2 are editing enzymes that deaminate adenosine to inosine in long double stranded RNA duplexes and specific pre-mRNA transcripts. Here, we show that full-length and N-terminally truncated forms of ADAR1 are simultaneously expressed in HeLa and COS7 cells owing to the usage of alternative starting methionines. Because the N-terminus of ADAR1 contains a nuclear export signal, the full-length protein localizes predominantly in the cytoplasm, whereas the N-terminally truncated forms are exclusively nuclear and accumulate in the nucleolus. ADAR2, which lacks a region homologous to the N-terminal domain of ADAR1, localizes exclusively to the nucleus and similarly accumulates in the nucleolus. Within the nucleolus, ADAR1 and ADAR2 co-localize in a novel compartment. Photobleaching experiments demonstrate that, in live cells, ADAR1 and ADAR2 are in constant flux in and out of the nucleolus. When cells express the editing-competent glutamate receptor GluR-B RNA, endogenous ADAR1 and ADAR2 de-localize from the nucleolus and accumulate at sites where the substrate transcripts accumulate. This suggests that ADAR1 and ADAR2 are constantly moving through the nucleolus and might be recruited onto specific editing substrates present elsewhere in the cell.


Randomized crossover study to examine the necessity of an injection-to-meal interval in patients with type 2 diabetes and human insulin.
Patients with diabetes and insulin therapy with human insulin were usually instructed to use an interval of 20-30 min between the injection and meal. We examined the necessity of the injection-to-meal interval (IMI) in patients with type 2 diabetes mellitus (T2DM) and flexible insulin therapy with human insulin. In this randomized, open crossover trial, 100 patients with T2DM (47% men, mean age = 66.7 years) were randomized to the IMI first group (phase 1, IMI 20 min; phase 2, no IMI) or IMI last group (phase 1, no IMI; phase 2, IMI 20 min). The main outcome measures were HbA1c, blood glucose profile, incidence of hypoglycemia, quality of life, treatment satisfaction, and patient preference. Forty-nine patients were randomized to the IMI first group and 51 patients to the IMI last group. Omitting the IMI only slightly increases HbA1c (average intraindividual difference = 0.08% [95% CI 0.01-0.15]). Since the difference is not clinically relevant, a therapy without IMI is noninferior to its application (P < 0.001). In the secondary outcomes, the incidence of mild hypoglycemia also did not differ between no IMI and IMI significantly (mean of differences = -0.10, P = 0.493). No difference in the blood glucose profile of both groups was found. Treatment satisfaction increased markedly, by 8.08, if IMI was omitted (P < 0.001). The total score of the quality of life measure did not show differences between applying an IMI or not. Insulin therapy without IMI was preferred by 86.5% of patients (P < 0.001). An IMI for patients with T2DM and preprandial insulin therapy is not necessary.


Asymmetric distribution of purinergic and adrenergic neurotransmission cooperates in the motor activity along the rat vas deferens.
The distribution of purinergic and adrenergic responses in the epididymal and prostatic segment of the rat vas deferens were studied in vitro. Prazosin antagonizes the twitch elicited by electrical stimulation mainly in the epididymal segment while alpha,beta-methyleneadenosine 5'-triphosphate (alpha,beta-mATP) preferentially inhibits the response of the prostatic segment. Using both prazosin plus alpha,beta-mATP, the response to field stimulation was completely inhibited. Concentration-response curves revealed that adrenergic compounds elicited a greater contraction in the epididymal portion than in the prostatic end of the ductus. Purinergic compounds caused a contraction of greater magnitude in the prostatic portion. The results suggest that adrenergic and purinergic mechanisms are asymmetrically distributed along the vas deferens reflecting a gradient of adrenergic and purinergic receptors along the ductus.


Correlation of P-glycoprotein expression and function in childhood acute leukemia: a children's cancer group study.
Clinical drug resistance may be attributed to the simultaneous selection and expression of genes modulating the uptake and metabolism of chemotherapeutic agents. P-glycoprotein (P-gp) functions as a membrane-associated drug efflux pump whose increased expression results in resistance to anthracyclines, epipodophyllotoxins, vinca alkaloids, and some alkylating agents. This type of resistance occurs as both de novo and acquired resistance to therapy for leukemia. We have studied P-gp expression and function in childhood acute leukemias by developing a series of doxorubicin- and vincristine-selected CEM, T-cell lymphoblastoid cell lines that recapitulate the low levels of expression and resistance seen clinically. These cell lines have been used to develop flow cytometric assays for the semiquantitative measurements of P-gp expression with the MRK16 monoclonal antibody and P-gp function using the enhanced retention of rhodamine 123 in the presence of verapamil, a resistance modulator. Kolmogorov-Smirnov statistics, represented by the D measurement, are used to determine the difference in level of P-gp expression by comparing MRK16 staining to an IgG2a isotype control. When D is > 0.09, there is an excellent correlation (R = 0.82) between P-gp expression and function. The evaluation of 107 bone marrow specimens from 84 children with lymphoblastic or myelogenous leukemia showed a statistically significant (P = .004) increase in P-gp function at relapse. P-gp expression at relapse, however, approached but did not reach a significant level (P = .097). Using this methodology, we can identify patients with levels of P-gp expression and function that we can define clinically, as well as children with discordant multidrug resistance phenotypes. This study supports the role of P-gp-mediated drug resistance in childhood leukemia and confirms that P-gp expression and function are measurable in their leukemic blasts. These assays provide the means for the in vitro testing of resistance modulators and the monitoring of in vivo response to treatment with these agents.


Dog Ownership and Survival After a Major Cardiovascular Event: A Register-Based Prospective Study.
Dog ownership is associated with increased physical activity levels and increased social support, both of which could improve the outcome after a major cardiovascular event. Dog ownership may be particularly important in single-occupancy households where ownership provides substitutive companionship and motivation for physical activity. We used the Swedish National Patient Register to identify all patients aged 40 to 85 presenting with an acute myocardial infarction (n=181 696; 5.7% dog ownership) or ischemic stroke (n=154 617; 4.8% dog ownership) between January 1, 2001 and December 31, 2012. Individual information was linked across registers for cause of death, sociodemographic, and dog ownership data. We evaluated all-cause mortality and risk of recurrent hospitalization for the same cause until December 31, 2012. Models were adjusted for socioeconomic, health, and demographic factors at study inclusion such as age, marital status, the presence of children in the home, area of residence, and income, as well as all registered comorbidities and hospitalization for cardiovascular disease in the past 5 years. Dog owners had a lower risk of death after hospitalization for acute myocardial infarction during the full follow-up period of 804 137 person-years, with an adjusted hazard ratio (HR) of 0.67 (95% CI, 0.61 to 0.75) for those who lived alone, and HR of 0.85 (95% CI, 0.80 to 0.90) for those living with a partner or a child. Similarly, after an ischemic stroke, dog owners were at lower risk of death during the full follow-up of 638 219 person-years adjusted HR of 0.73 (95% CI, 0.66 to 0.80) for those who lived alone and HR of 0.88 (95% CI, 0.83 to 0.93) for those living with a partner or a child. We further found an association of dog ownership with reduced risk of hospitalization for recurrent myocardial infarction (HR, 0.93; 95% CI, 0.87 to 0.99). We found evidence of an association of dog ownership with a better outcome after a major cardiovascular event. Although our models are adjusted for many potential confounders, there are also unmeasured confounders such as smoking that prevent us from drawing conclusions regarding a possible causal effect.


Modeling classical wavefront sensors.
We present an image formation model for deterministic phase retrieval in propagation-based wavefront sensing, unifying analysis for classical wavefront sensors such as Shack-Hartmann (slopes tracking) and curvature sensors (based on Transport-of-Intensity Equation). We show how this model generalizes commonly seen formulas, including Transport-of-Intensity Equation, from small distances and beyond. Using this model, we analyze theoretically achievable lateral wavefront resolution in propagation-based deterministic wavefront sensing. Finally, via a prototype masked wavefront sensor, we show simultaneous bright field and phase imaging numerically recovered in real-time from a single-shot measurement.


Non-infectious aggregates of the prion protein react with several PrPSc-directed antibodies.
The key event in the pathogenesis of prion diseases is the conformational conversion of the normal prion protein (PrP) (PrP(C)) into an infectious, aggregated isoform (PrP(Sc)) that has a high content of beta-sheet. Historically, a great deal of effort has been devoted to developing antibodies that specifically recognize PrP(Sc) but not PrP(C), as such antibodies would have enormous diagnostic and experimental value. A mouse monoclonal IgM antibody (designated 15B3) and three PrP motif-grafted monoclonal antibodies (referred to as IgG 19-33, 89-112, and 136-158) have been previously reported to react specifically with infectious PrP(Sc) but not PrP(C). In this study, we extend the characterization of these four antibodies by testing their ability to immunoprecipitate and immunostain infectious and non-infectious aggregates of wild-type, mutant, and recombinant PrP. We find that 15B3 as well as the motif-grafted antibodies recognize multiple types of aggregated PrP, both infectious and non-infectious, including forms found in brain, in transfected cells, and induced in vitro from purified recombinant protein. These antibodies are exquisitely selective for aggregated PrP, and do not react with soluble PrP even when present in vast excess. Our results suggest that 15B3 and the motif-grafted antibodies recognize structural features common to both infectious and non-infectious aggregates of PrP. Our study extends the utility of these antibodies for diagnostic and experimental purposes, and it provides new insight into the structural changes that accompany PrP oligomerization and prion propagation.


[The antiviral activity of peptide immunomodulator "Gepon" in experimental infections caused by herpes simplex viruses types 1 and 2].
Experimental data are reported on the antiviral activity of peptide immune-modulator "Gepon" in infections caused by herpes simplex virus (HSV), types 1 and 2, in vitro and in vivo. The drug proved to be non-toxic and did not possess any viricidal action. Its antiviral effect was registered in experiments with a multiple-infection cells' culture. The maximal effect was a 100-fold reduction of the viral titer when it was used in a concentration of 6.25 mcg/I as a preventive measure 24 hours before triggering the infection. The mentioned drug's effect was reliably higher than its use within a treatment scheme (1 hour after the infection onset). "Gepon" possessed the reliable protecting qualities (36% of protection with a mean increase of the infected mouse life by 1.9 days) in experiments with intraperitoneally infected mice (10 LD50/mouse of HSV, type 2), when the drug was administered in doses of 0.1 and 1 mcg/mouse.


Quantifying Replication Fork Progression at CTG Repeats by 2D Gel Electrophoresis.
Physical separation of branched DNA from linear molecules is based on the difference of mobility of linear versus branched DNA during two-dimensional agarose gel electrophoresis. Structured DNA migrates as slower species when compared to linear DNA of similar molecular weight. Metabolic processes such as S phase replication or double strand-break repair may generate branched DNA molecules. Trinucleotide repeats are naturally prone to form secondary structures that can modify their migration through an agarose gel matrix. These structures may also interfere in vivo with replication, by slowing down replication-fork progression, transiently stalling forks, possibly leading to secondary structure such as Holliday junctions or hemicatenanes. Alternatively, reversed replication forks may occur following fork stalling, disrupting replication dynamics and modifying DNA migration on agarose gel. So although two-dimensional agarose gel electrophoresis theoretically allows to resolve a mixture of structured DNA molecules and quantify them by radioactive hybridization, its practical application to trinucleotide repeats faces some serious technical challenges.


[Concepción inventory: Development of a screening instrument for depression in primary care in Chile].
Screening instruments are required for the detection of depressive disorders by primary care practitioners. To develop a screening instrument to detect depression, based on data gathered interviewing patients attending primary health care settings. The instrument was constructed with data about factors associated or triggering a depressive disorder obtained from 3,000 patients consulting for general morbidity. All patients answered the Composite International Diagnostic Interview, (version 2.1, section depression) and an inventory containing 39 risk factors for depression, obtained from the literature. A multiple imputation method using chained equations was carried out. Using a binary logistic regression with backward selection, an equation for depression screening was obtained. The c-index was calculated to estimate discriminating power of the model. A shrinkage factor was estimated to adjust the predictive model. Estimations were carried out with data from 2,552 patients with a median age of 47 years (73% women). Fifty five percent lived with a partner and 45% had basic studies. The method selected 14 significant predictors, with a shrinkage value of 0.861 and a c-index of 0.838 (95% confidence intervals 0.82-0.86). The instrument has adequate psychometric properties as a screening tool for depression in primary health care.


Promoters recognized by forkhead proteins exist for individual 21U-RNAs.
C. elegans 21U-RNAs are equivalent to the piRNAs discovered in other metazoans and have important roles in gametogenesis and transposon control. The biogenesis and molecular function of 21U-RNAs and piRNAs are poorly understood. Here, we demonstrate that transcription of each 21U-RNA is regulated separately through a conserved upstream DNA motif. We use genomic analysis to show that this motif is associated with low nucleosome occupancy, a characteristic of many promoters that drive expression of protein-coding genes, and that RNA polymerase II is localized to this nucleosome-depleted region. We establish that the most conserved 8-mer sequence in the upstream region of 21U-RNAs, CTGTTTCA, is absolutely required for their individual expression. Furthermore, we demonstrate that the 8-mer is specifically recognized by Forkhead family (FKH) transcription factors and that 21U-RNA expression is diminished in several FKH mutants. Our results suggest that thousands of small noncoding transcription units are regulated by FKH proteins.


Can the Clinical Utility of iBreastExam, a Novel Device, Aid in Optimizing Breast Cancer Diagnosis? A Systematic Review.
A portable, cost-effective, easy-to-use, hand-held Intelligent Breast Exam (iBE), which is a wireless, radiation-free device, may be a valuable screening tool in resource-limited settings. While multiple studies evaluating the use of iBE have been conducted worldwide, there are no cumulative studies evaluating the iBE's performance. Therefore this review aims to determine the clinical utility and applicability of iBE compared with clinical breast examinations, ultrasound, and mammography and discuss its strengths and weaknesses when performing breast-cancer screening. A systematic review was performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Four electronic databases were searched: PubMed, Cochrane Library, Web of Science, and Google Scholar. The review included 11 studies with a total sample size of 16,052 breasts. The mean age ranged from 42 to 58 years. The sensitivity and specificity of the iBE ranged from 34.3% to 86% and 59% to 94%, respectively. For malignant lesions, iBE demonstrated a moderate to higher diagnostic capacity ranging from 57% to 93% and could identify tumor sizes spanning from 0.5 cm to 9 cm. Our findings underscore the potential clinical utility and applicability of iBE as a prescreening and triaging tool, which may aid in reducing the burden of patients undergoing diagnostic imaging in lower- and middle-income countries. Furthermore, iBE has shown to diagnose cancers as small as 0.5 cm, which can be a boon in early detection and reduce mortality rates. However, the encouraging results of this systematic review should be interpreted with caution because of the device's low sensitivity and high false-positive rates.


Effect of asymptomatic infection with southern tomato virus on tomato plants.
Southern tomato virus (STV) is often found infecting healthy tomato plants (Solanum lycopersicum). In this study, we compared STV-free and STV-infected plants of cultivar M82 to determine the effect of STV infection on the host plant. STV-free plants exhibited a short and bushy phenotype, whereas STV-infected plants were taller. STV-infected plants produced more fruit than STV-free plants, and the germination rate of seeds from STV-infected plants was higher than that of seeds from STV-free plants. This phenotypic difference was also observed in progeny plants (siblings) derived from a single STV-infected plant in which the transmission rate of STV to progeny plants via the seeds was approximately 86%. These results suggest that the interaction between STV and host plants is mutualistic. Transcriptome analysis revealed that STV infection affects gene expression in the host plant and results in downregulation of genes involved in ethylene biosynthesis and signaling. STV-infected tomato plants might thus be artificially selected due to their superior traits as a crop.


An inconvenient sea truth: spread, steepness, and skewness of surface slopes.
Bréon and Henriot (BH) have collected eight million globally distributed satellite images of sunglitter, which yield a few simple, robust rules about the statistics of surface slopes: 1) constant angular spread, 2) linear steepness, and 3) sigmoid (near stepwise) skewness (all with respect to wind speed). Yet the information is sparse because it says nothing about time and space scales. The BH rules are an inconvenient sea truth, too fundamental to be ignored, too incomplete to be understood. With regard to BH rule 1 (BH:1), I suggest that the constant spread is associated with a wake-like geometry of the short gravities. Steepness linearity (BH:2) remains an enigma. Skewness (BH:3) appears to be correlated with a rather sudden onset of breaking for winds above 4 m s(-1). I do not think that skewness comes from parasitic capillaries. These are tentative conclusions; I look forward to intensive sea-going experiments over the next few years demolishing the proposed interpretations.


Asparagine-linked oligosaccharides of BHK cells treated with inhibitors of oligosaccharide processing.
Baby-hamster kidney (BHK) cells were labelled with [2-3H]mannose for 1-2 days in media containing 1-deoxynojirimycin, N-methyl-1-deoxynojirimycin or 1-deoxymannojirimycin. Glycopeptides obtained by Pronase digestion of disrupted cells were analysed by lectin affinity chromatography, by Bio-Gel P4 gel filtration and by paper chromatography of oligosaccharides released by endo-beta-N-acetylglucosaminidase H. Biosynthesis of complex-type oligosaccharides was diminished but not abolished, the greatest effect being obtained by continuous culture of cells with 1-deoxymannojirimycin. Under these conditions cells contained only 20-30% of the concentration of complex-type chains found in control cells and correspondingly increased amounts of oligomannose-type chains. Similar concentrations of asparagine-linked Man6-GlcNAc2 and Man5GlcNAc2 were present in 1-deoxymannojirimycin-treated cells and control cells, indicating that the inhibition of complex-type chain formation was not related simply to an inability of inhibitor-treated cells to carry out extensive mannosidase-catalysed processing. N-Methyl-1-deoxynojirimycin induced accumulation of oligomannose-type chains containing three glucose residues, and cells treated with 1-deoxynojirimycin contained oligosaccharides with one to three glucose residues. Cells cultured in the presence of the inhibitors retained sensitivity towards the galactose-binding lectins ricin and modeccin.


Polymer-Dispersed Cholesteric Liquid Crystal under Homeotropic Anchoring: Electrically Induced Structures with λ1/2-Disclination.
Orientational structures of polymer-dispersed cholesteric liquid crystal under homeotropic anchoring and their transformations under the action of an electric field are studied. The switching of cholesteric droplets between different topological states are experimentally and theoretically demonstrated. Structures with λ+1/2-disclination are found and considered. These structures are formed during the transformation of a twisted toroidal configuration induced by a decrease in the electric field when a relative chiral parameter N0>6.3. The transformation of the initial structure with a bipolar distribution of the helix axis into a twisted toroidal configuration and then into a structure with λ+1/2-disclination is investigated in detail. The behavior of these structures under the influence of an external electric field, as well as the appearance of structures with λ−1/2-disclination, are studied. Obtained results are promising for the development of optical materials with programmable properties.


Induction of autoimmune thyroiditis by depletion of CD4+CD25+ regulatory T cells in thyroiditis-resistant IL-17, but not interferon-gamma receptor, knockout nonobese diabetic-H2h4 mice.
Iodine-induced experimental autoimmune thyroiditis in the nonobese diabetic (NOD)-H2h4 mouse is a prototype of animal models of Hashimoto's thyroiditis in humans. Recent studies have shown the resistance to thyroiditis of NOD-H2h4 mice genetically deficient for either IL-17 or interferon (IFN)-γ, implicating both of T helper type 1 (Th1) and Th17 immune responses in disease pathogenesis. However, we hypothesized that robust induction of a single arm of effector T cells (either Th1 or Th17) might be sufficient for inducing thyroiditis in NOD-H2h4 mice. To address this hypothesis, enhanced immune responses consisting of either Th1 or Th17 were induced by anti-CD25 antibody-mediated depletion of regulatory T cells (Treg) in thyroiditis-resistant IL-17 knockout (KO) or IFN-γ receptor (IFN-γR) KO, respectively, NOD-H2h4 mice. Depletion of Treg in IL-17 KO mice (i.e. Th1 enhancement) elicited antithyroglobulin autoantibodies and thyroiditis. Immunohistochemical analysis of the thyroid glands revealed the similar intrathyroidal lymphocyte infiltration patterns, with CD4+ T and CD19+ B cells being dominant between the wild-type and Treg-depleted IL-17 KO mice. In contrast, Treg-depleted IFN-γR KO mice remained thyroiditis resistant. Intracellular cytokine staining assays showed differentiation of Th1 cells in IL-17 KO mice but not of Th17 cells in IFN-γR KO mice. Our findings demonstrate that a robust Th1 immune response can by itself induce thyroiditis in otherwise thyroiditis-resistant IL-17 KO mice. Thus, unlike Th17 cells in IFN-γR KO mice, Th1 cells enhanced by Treg depletion can be sustained and induce thyroiditis.


Sputum proteomics in inflammatory and suppurative respiratory diseases.
Markers of inflammatory activity are important for assessment and management of many respiratory diseases. Markers that are currently unrecognized may be more valuable than those presently believed to be useful. To identify potential biomarkers of suppurative and inflammatory lung disease in induced sputum samples. Induced sputum was collected from 20 healthy control subjects, 24 patients with asthma, 24 with chronic obstructive pulmonary disease, 28 with cystic fibrosis (CF), and 19 with bronchiectasis. Twelve patients with CF had sputum sampled before and after antibiotic therapy for an infective exacerbation. The fluid phase of induced sputum was analyzed by surface-enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectroscopy on three protein array surfaces. Some protein markers were selected for identification, and relevant ELISA assays sought. For 12 patients with CF, both SELDI-TOF and ELISA monitored changes in inflammatory responses during infective exacerbations. SELDI-TOF identified potential biomarkers that differentiated each of the disease groups from healthy control subjects: at a significance of P < 0.01, there were 105 for asthma, 113 for chronic obstructive pulmonary disease, 381 for CF, and 377 for bronchiectasis. Peaks selected for protein identification yielded calgranulin A, calgranulin B, calgranulin C, Clara cell secretory protein, lysosyme c, proline rich salivary peptide, cystatin s, and hemoglobin alpha. On treatment of an infective CF exacerbation, SELDI-TOF determined falls in levels of calgranulin A and calgranulin B that were mirrored by ELISA-measured falls in calprotectin (heterodimer of calgranulins A and B). Proteomic screening of sputum yields potential biomarkers of inflammation. The early development of a clinically relevant assay from such data is demonstrated.


A systematic review of the use of rifaximin for Clostridium difficile infections.
Clostridium difficile infection (CDI) is an increasingly common occurrence in the hospital setting, and it is associated with significant morbidity and mortality. In vitro studies have found that rifaximin, a nonabsorbable rifamycin antibiotic, displays potent antimicrobial activity against C. difficile. This systematic review thus aimed to examine the clinical role of rifaximin in CDI. Using the keywords [clostridium OR difficile OR colitis] AND [rifaximin OR xifaxan OR rifagut], a preliminary search on the PubMed, EMBASE, Medline, Clinicaltrials.gov and Google Scholar databases yielded 6210 papers published in English between 1-Jan-1988 and 1-Jul-2018. A total of eight clinical trials were systematically reviewed. Of these, only two were randomized, controlled trials. In the treatment of mild-moderate CDI, rifaximin is a viable alternative to existing therapies (metronidazole or vancomycin). More importantly, rifaximin has a potential role in conjunction with other therapies, showing to be efficacious in reducing the rate of CDI recurrence. However, clinical studies have reported a resistance rate in the range of 29.1-48.9%, with a geographical variance in the distribution of rifaximin-resistant C. difficile strains. With its unique eubiotic properties and positive modulation of the gut flora, rifaximin has a potential therapeutic role in the management of CDI, especially in CDI recurrences. As there is a paucity of randomized, controlled trials to support its use, studies with larger and more diverse populations should be conducted before the efficacy of rifaximin can be conclusively stated. Rifaximin is also a relatively expensive antimicrobial, further studies should include cost-benefit analyses.


Biochemical rationale and the cardiac response of patients with muscle disease to therapy with coenzyme Q10.
Cardiac disease is commonly associated with virtually every form of muscular dystrophy and myopathy. A double-blind and open crossover trial on the oral administration of coenzyme Q10 (CoQ10) to 12 patients with progressive muscular dystrophies and neurogenic atrophies was conducted. These diseases included the Duchenne, Becker, and limb-girdle dystrophies, myotonic dystrophy, Charcot-Marie-Tooth disease, and Welander disease. The impaired cardiac function was noninvasively and extensively monitored by impedance cardiography. Solely by significant change or no change in stroke volume and cardiac output, all 8 patients on blind CoQ10 and all 4 on blind placebo were correctly assigned (P less than 0.003). After the limited 3-month trial, improved physical well-being was observed for 4/8 treated patients and for 0/4 placebo patients; of the latter, 3/4 improved on CoQ10; 2/8 patients resigned before crossover; 5/6 on CoQ10 in crossover maintained improved cardiac function; 1/6 crossed over from CoQ10 to placebo relapsed. The rationale of this trial was based on known mitochondrial myopathies, which involve respiratory enzymes, the known presence of CoQ10 in respiration, and prior clinical data on CoQ10 and dystrophy. These results indicate that the impaired myocardial function of such patients with muscular disease may have some association with impaired function of skeletal muscle, both of which may be improved by CoQ10 therapy. The cardiac improvement was definitely positive. The improvement in well-being was subjective, but probably real. Likely, CoQ10 does not alter genetic defects but can benefit the sequelae of mitochondrial impairment from such defects. CoQ10 is the only known substance that offers a safe and improved quality of life for such patients having muscle disease, and it is based on intrinsic bioenergetics.


GCH1, BH4 and pain.
Understanding and consequently treating neuropathic pain effectively is a challenge for modern medicine, as unlike inflammation, which can be controlled relatively well, chronic pain due to nerve injury is refractory to most current therapeutics. Here we define a target pathway for a new class of analgesics, tetrahydrobiopterin (BH4) synthesis and metabolism. BH4 is an essential co-factor in the synthesis of serotonin, dopamine, epinephrine, norepinephrine and nitric oxide and as a result, its availability influences many systems, including neurons. Following peripheral nerve damage, levels of BH4 are dramatically increased in sensory neurons, consequently this has a profound effect on the physiology of these cells, causing increased activity and pain hypersensitivity. These changes are principally due to the upregulation of the rate limiting enzyme for BH4 synthesis GTP Cyclohydrolase 1 (GCH1). A GCH1 pain-protective haplotype which decreases pain levels in a variety of settings, by reducing the levels of endogenous activation of this enzyme, has been characterized in humans. Here we define the control of BH4 homeostasis and discuss the consequences of large perturbations within this system, both negatively via genetic mutations and after pathological increases in the production of this cofactor that result in chronic pain. We explain the nature of the GCH1 reduced-function haplotype and set out the potential for a ' BH4 blocking' drug as a novel analgesic.


Severe preeclampsia is characterized by increased placental expression of galectin-1.
Galectin-1 is a major anti-inflammatory protein expressed by the placenta and immune cells that can bias the character of inflammatory responses toward the Th2 type. Galectin-1 is expressed in immune privileged sites, it can facilitate immune tolerance and tumor immune escape, and it has been successfully used for the suppression of experimental autoimmune diseases as well as graft-versus-host disease in murine models. We propose that an abnormal immune response in some pregnancy complications may be associated with changes in placental expression of galectin-1. To test this hypothesis, we studied placental galectin-1 mRNA and protein expression and localization in women with preeclampsia (PE) and in those who delivered a small-for-gestational age (SGA) neonate. This cross-sectional study included pregnant women matched for gestational age at delivery in the following groups: (1) severe PE (n = 10), (2) severe PE complicated with SGA (n = 10), (3) SGA without PE (n = 10), and (4) controls (n = 10). Galectin-1 mRNA and protein were localized in placentas by in situ hybridization and immunofluorescence microscopy. Galectin-1 mRNA expression was determined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR), and galectin-1 protein content by Western blot. Non-parametric statistics were used for analysis. (1) In normal term placentas, galectin-1 mRNA or immunofluorescence signals were detected in the trophoblasts, villous stromal cells, Hofbauer cells, endothelial cells of the villous blood vessels, and the villous stroma. (2) Placental galectin-1 mRNA expression was significantly higher in severe PE (with or without SGA) than in controls (1.47-fold, p = 0.004; 1.44-fold, p = 0.003, respectively) and in SGA (1.68-fold, p = 0.001; 1.64-fold, p = 0.001, respectively). (3) Trophoblasts in placentas of patients with severe PE had the most intense galectin-1 immunostaining. (1) We report for the first time the placental expression and localization of galectin-1 mRNA and demonstrate that the protein is abundantly present in third trimester human placentas. (2) Placental galectin-1 expression is higher in severe PE than in normal pregnancy regardless of the presence of SGA. (3) However, it is not altered in SGA without PE. We propose that the increased placental expression of galectin-1 in patients with severe PE may represent a fetal response to an exaggerated systemic maternal inflammation; thus, galectin-1 may be implicated in maternal-fetal immune tolerance in humans.


Cyberbullying victimization and student engagement among adolescents: Does school climate matter?
Although the psychological impacts of cyberbullying victimization (CBV) have been documented, research is inconclusive about the role of contextual factors in the association between CBV and student engagement. Sampling 16,237 adolescents from 43 schools in Delaware, we used multilevel modeling to test how CBV was associated with emotional and cognitive-behavioral engagement at both the student and school levels, with the control for demographic factors and traditional bullying victimization (TBV). We also examined the moderating effects of school climate and grade level on the association between CBV and student engagement. CBV had a small but significant positive association with emotional engagement and a small but significant negative association with cognitive-behavioral engagement. School-level climate intensified the negative association between student-level CBV and cognitive-behavioral engagement and mitigated the positive association between student-level school climate and emotional engagement. The positive association between CBV and emotional engagement was stronger for high school than middle school students, whereas the negative association between CBV and cognitive-behavioral engagement was stronger for middle than high school students. The findings support the promotive role of positive school climate in student engagement promotion. The findings also support the healthy context paradox, which suggests that bullying victims' engagement in schools may be exacerbated in a social context with positive school climate perceived by the group members. (PsycINFO Database Record (c) 2020 APA, all rights reserved).


Middle cerebral artery occlusion in rats studied by magnetic resonance imaging.
Ischemia due to middle cerebral artery occlusion was studied in 29 rats from 1 to 24 hours after occlusion using magnetic resonance imaging. Images were made before and after the injection of a superparamagnetic iron oxide compound, AMI-25. Subtraction images demonstrated the region of perfusion deficit as early as 1 hour after occlusion, earlier than conventional T2-weighted images. The area of altered perfusion detected by this technique (subtraction imaging after AMI-25 administration) correlated with that demonstrated by iodoantipyrine autoradiography. Since this magnetic resonance technique can be used to serially estimate the location and size of the ischemic area, the technique can be an important adjunct to metabolic studies of focal ischemia using magnetic resonance spectroscopy. The technique may have clinical applications as well.


Full Modeling of High-Intensity Focused Ultrasound and Thermal Heating in the Kidney Using Realistic Patient Models.
High-intensity focused ultrasound (HIFU) therapy can be used for noninvasive treatment of kidney (renal) cancer, but the clinical outcomes have been variable. In this study, the efficacy of renal HIFU therapy was studied using nonlinear acoustic and thermal simulations in three patients. The acoustic simulations were conducted with and without refraction in order to investigate its effect on the shape, size, and pressure distribution at the focus. The values for the attenuation, sound speed, perfusion, and thermal conductivity of the kidney were varied over the reported ranges to determine the effect of variability on heating. Furthermore, the phase aberration was studied in order to quantify the underlying phase shifts using a second-order polynomial function. The ultrasound field intensity was found to drop on average 11.1 dB with refraction and 6.4 dB without refraction. Reflection at tissue interfaces was found to result in a loss less than 0.1 dB. Focal point splitting due to refraction significantly reduced the heating efficacy. Of all the tissue parameters, perfusion was found to affect the heating the most. Small changes in temperature were seen with varying attenuation and thermal conductivity, but no visible changes were present with sound speed variations. The aberration study revealed an underlying trend in the spatial distribution of the phase shifts. The results show that the efficacy of HIFU therapy in the kidney could be improved with aberration correction. A method is proposed by which patient specific pretreatment calculations could be used to overcome the aberration and therefore make ultrasound treatment possible.


Role of melatonin on diabetes-related metabolic disorders.
Melatonin is a circulating hormone that is mainly released from the pineal gland. It is best known as a regulator of seasonal and circadian rhythms, its levels being high during the night and low during the day. Interestingly, insulin levels are also adapted to day/night changes through melatonin-dependent synchronization. This regulation may be explained by the inhibiting action of melatonin on insulin release, which is transmitted through both the pertussis-toxin-sensitive membrane receptors MT(1) and MT(2) and the second messengers 3',5'-cyclic adenosine monophosphate, 3',5'-cyclic guanosine monophosphate and inositol 1,4,5-trisphosphate. Melatonin may influence diabetes and associated metabolic disturbances not only by regulating insulin secretion, but also by providing protection against reactive oxygen species, since pancreatic β-cells are very susceptible to oxidative stress because they possess only low-antioxidative capacity. On the other hand, in several genetic association studies, single nucleotide polymorphysms of the human MT(2) receptor have been described as being causally linked to an elevated risk of developing type 2 diabetes. This suggests that these individuals may be more sensitive to the actions of melatonin, thereby leading to impaired insulin secretion. Therefore, blocking the melatonin-induced inhibition of insulin secretion may be a novel therapeutic avenue for type 2 diabetes.


Electrophysiological studies in left bundle branch block as a result of procainamide therapy.
A patient of coronary heart disease with depressed intraventricular conduction who developed left bundle branch block upon administration of procainamide instead of the usual nonspecific widening of qrs is presented. The possible mechanism of genesis of this very rare side effect of procainamide is discussed and the review of the literature revealed this patient to be the first documented, electrophysiologically studied case with this side effect of procainamide.


Microbial community structures differentiated in a single-chamber air-cathode microbial fuel cell fueled with rice straw hydrolysate.
The microbial fuel cell represents a novel technology to simultaneously generate electric power and treat wastewater. Both pure organic matter and real wastewater can be used as fuel to generate electric power and the substrate type can influence the microbial community structure. In the present study, rice straw, an important feedstock source in the world, was used as fuel after pretreatment with diluted acid method for a microbial fuel cell to obtain electric power. Moreover, the microbial community structures of anodic and cathodic biofilm and planktonic culturewere analyzed and compared to reveal the effect of niche on microbial community structure. The microbial fuel cell produced a maximum power density of 137.6 ± 15.5 mW/m2 at a COD concentration of 400 mg/L, which was further increased to 293.33 ± 7.89 mW/m2 through adjusting the electrolyte conductivity from 5.6 mS/cm to 17 mS/cm. Microbial community analysis showed reduction of the microbial diversities of the anodic biofilm and planktonic culture, whereas diversity of the cathodic biofilm was increased. Planktonic microbial communities were clustered closer to the anodic microbial communities compared to the cathodic biofilm. The differentiation in microbial community structure of the samples was caused by minor portion of the genus. The three samples shared the same predominant phylum of Proteobacteria. The abundance of exoelectrogenic genus was increased with Desulfobulbus as the shared most abundant genus; while the most abundant exoelectrogenic genus of Clostridium in the inoculum was reduced. Sulfate reducing bacteria accounted for large relative abundance in all the samples, whereas the relative abundance varied in different samples. The results demonstrated that rice straw hydrolysate can be used as fuel for microbial fuel cells; microbial community structure differentiated depending on niches after microbial fuel cell operation; exoelectrogens were enriched; sulfate from rice straw hydrolysate might be responsible for the large relative abundance of sulfate reducing bacteria.


Regulation of G proteins by chronic opiate and clonidine treatment in the guinea pig myenteric plexus.
G proteins have been implicated in the development of opioid dependence of the guinea pig myenteric plexus as chronic fentanyl elevates G0/i alpha and pertussis toxin prevents this phenomenon. Therefore, the present study investigates G proteins more closely in this peripheral nerve plexus after chronic exposure to addictive drugs of the opiate and nonopiate type. After 6 days of treatment with either the mu receptor ligand fentanyl, the kappa- agonist U-50,488H or the alpha-2 adrenergic receptor ligand clonidine, at doses which render the myenteric plexus tolerant and dependent, the G protein subunits Go alpha and G beta were quantified by immunoblot analysis by using polyclonal antisera. Regardless of the drug used, these G proteins were found to be significantly increased in particulate membrane preparations linked to nerve somata and nerve terminals. This increase in G protein subunits is developed maximally after 6 days, is dose-dependent and reversible upon termination of the drug supply. The concentrations found elevated return to control levels within 4 to 5 days after commencing withdrawal. The common increase of Go alpha and G beta subunits observed after chronic opiate or clonidine exposure is associated with the phenomenon of cross-dependence among all drugs studied. The findings may suggest that in the guinea pig myenteric plexus multiple inhibitory receptor types make use of a common pool of G proteins.


Misplaced Cervical Screws Requiring Reoperation.
A multicenter, retrospective case series. In the past several years, screw fixation of the cervical spine has become commonplace. For the most part, this is a safe, low-risk procedure. While rare, screw backout or misplaced screws can lead to morbidity and increased costs. We report our experiences with this uncommon complication. A multicenter, retrospective case series was undertaken at 23 institutions in the United States. Patients were included who underwent cervical spine surgery from January 1, 2005, to December 31, 2011, and had misplacement of screws requiring reoperation. Institutional review board approval was obtained at all participating institutions, and detailed records were sent to a central data center. A total of 12 903 patients met the inclusion criteria and were analyzed. There were 11 instances of screw backout requiring reoperation, for an incidence of 0.085%. There were 7 posterior procedures. Importantly, there were no changes in the health-related quality-of-life metrics due to this complication. There were no new neurologic deficits; a patient most often presented with pain, and misplacement was diagnosed on plain X-ray or computed tomography scan. The most common location for screw backout was C6 (36%). This study represents the largest series to tabulate the incidence of misplacement of screws following cervical spine surgery, which led to revision procedures. The data suggest this is a rare event, despite the widespread use of cervical fixation. Patients suffering this complication can require revision, but do not usually suffer neurologic sequelae. These patients have increased cost of care. Meticulous technique and thorough knowledge of the relevant anatomy are the best means of preventing this complication.


Effects of dietary amines on small intestinal variables in neonatal pigs fed soy protein isolate.
Six litters of newborn crossbred piglets were utilized to examine 1) the effects of substituting 20% of the protein of an all-milk protein liquid diet with a soy protein isolate (milk-soy diet) on small intestinal variables and 2) the effects of supplementing this milk-soy diet with 25 g of either putrescine or ethylamine per kilogram diet on small intestinal variables. Small intestinal xylose absorption tended to increase from wk 1 to wk 2 of age in pigs fed the milk, putrescine and ethylamine diets, but not in pigs fed the milk, putrescine and ethylamine diets, but not in pigs fed the unsupplemented milk-soy diet. Crypt depth in pigs fed the milk-soy diet tended to be less (9.4%; P greater than .10) than the crypt depth in pigs fed the other diets, but mitotic index was not different (P greater than .10) among diets. Mucosal protein, DNA and RNA concentrations and mucosal brush border sucrase and cytosolic dipeptidase activities tended to be least in pigs fed the putrescine and ethylamine diets. Concentration of mucosal putrescine was greatest (P less than .002) in the distal regions of the small intestine of pigs fed putrescine. Mucosal ornithine decarboxylase activity was inhibited by putrescine (P less than .02), but it was not affected by the soybean protein isolate used in this study. Supplementing soy protein isolate diets with amines may enhance intestinal absorption and enterocyte proliferation.


The Impact of Non-normative Displays of Emotion in the Workplace: How Inappropriateness Shapes the Interpersonal Outcomes of Emotional Displays.
When it comes to evaluating emotions as either "good" or "bad," everyday beliefs regarding emotions rely mostly on their hedonic features-does the emotion feel good to the person experiencing the emotion? However, emotions are not only felt inwardly; they are also displayed outwardly, and others' responses to an emotional display can produce asymmetric outcomes (i.e., even emotions that feel good to the displayer can lead to negative outcomes for the displayer and others). Focusing on organizational settings, this manuscript reviews the literature on the outcomes of emotional expressions and argues that the evidence points to perceived (in)appropriateness of emotional displays as key to their consequences: emotional displays that are deemed inappropriate generate disadvantageous outcomes for the displayer, and at times also the organization. Drawing on relevant theoretical models [Emotions as Social Information (EASI) theory, the Dual Threshold Model of Anger, and Asymmetrical Outcomes of Emotions], the paper highlights three broad and interrelated reasons why emotion displays could be deemed unfitting and inappropriate: (1) characteristics of the displayer (e.g., status, gender); (2) characteristics of the display (e.g., intensity, mode); and (3) characteristics of the context (e.g., national or organizational culture, topic of interaction). The review focuses on three different emotions-anger, sadness, and happiness-which differ in their valence based on how they feel to the displayer, but can yield different interpersonal outcomes. In conclusion, the paper argues that inappropriateness must be judged separately from whether an emotional display is civil (i.e., polite and courteous) or uncivil (i.e., rude, discourteous, and offensive). Testable propositions are presented, as well as suggested future research directions.


Impaired health-related quality of life in children and adolescents with chronic conditions: a comparative analysis of 10 disease clusters and 33 disease categories/severities utilizing the PedsQL 4.0 Generic Core Scales.
Advances in biomedical science and technology have resulted in dramatic improvements in the healthcare of pediatric chronic conditions. With enhanced survival, health-related quality of life (HRQOL) issues have become more salient. The objectives of this study were to compare generic HRQOL across ten chronic disease clusters and 33 disease categories/severities from the perspectives of patients and parents. Comparisons were also benchmarked with healthy children data. The analyses were based on over 2,500 pediatric patients from 10 physician-diagnosed disease clusters and 33 disease categories/severities and over 9,500 healthy children utilizing the PedsQL 4.0 Generic Core Scales. Patients were recruited from general pediatric clinics, subspecialty clinics, and hospitals. Pediatric patients with diabetes, gastrointestinal conditions, cardiac conditions, asthma, obesity, end stage renal disease, psychiatric disorders, cancer, rheumatologic conditions, and cerebral palsy self-reported progressively more impaired overall HRQOL than healthy children, respectively, with medium to large effect sizes. Patients with cerebral palsy self-reported the most impaired HRQOL, while patients with diabetes self-reported the best HRQOL. Parent proxy-reports generally paralleled patient self-report, with several notable differences. The results demonstrate differential effects of pediatric chronic conditions on patient HRQOL across diseases clusters, categories, and severities utilizing the PedsQL 4.0 Generic Core Scales from the perspectives of pediatric patients and parents. The data contained within this study represents a larger and more diverse population of pediatric patients with chronic conditions than previously reported in the extant literature. The findings contribute important information on the differential effects of pediatric chronic conditions on generic HRQOL from the perspectives of children and parents utilizing the PedsQL 4.0 Generic Core Scales. These findings with the PedsQL have clinical implications for the healthcare services provided for children with chronic health conditions. Given the degree of reported impairment based on PedsQL scores across different pediatric chronic conditions, the need for more efficacious targeted treatments for those pediatric patients with more severely impaired HRQOL is clearly and urgently indicated.


c-myc reverses neu-induced transformed morphology by transcriptional repression.
Amplification or overexpression or both of either the c-myc or the human neu (C-erbB-2) gene are common events in many primary human tumors. Coamplification or overexpression or both of both genes have been reported in some breast cancers. The possibility of cooperation between the c-myc and the normal rat neu (c-neu) genes in transforming cells was examined. Surprisingly, the expression of c-myc in B104-1-1 cells, and activated rat neu oncogene (neu*)-transformed NIH 3T3 line, resulted in morphologic reversion. This reversion was found to be a consequence of a transcription-repressive action of c-myc on the neu gene via a 140-bp fragment on the neu gene promoter. The effective concentration of a positive factor(s) interacting with this fragment seemed to be lowered by the expression of c-myc. Our findings lend support to arguments concerning the long-suspected function of c-myc as a transcriptional modulator. They also imply that an oncogene such as c-myc, or possibly the rapidly explored class that encodes transcription factors, under certain conditions may act to reverse a transformed phenotype that is induced by another oncogene instead of contributing positively towards the transformation process. Therefore, the activity of an oncogene may depend on the environment in which it is expressed. In addition, we may have identified the neu gene as a cellular target gene of negative regulation by c-myc.


Participation of the cannabinoid system and the NO/cGMP/KATP pathway in serotonin-induced peripheral antinociception.
It has already been shown that serotonin can release endocannabinoids at the spinal cord level, culminating in inhibition of the dorsal horn. At the peripheral level, cannabinoid receptors modulate primary afferent neurons by inhibiting calcium conductance and increasing potassium conductance. Studies have shown that after the activation of opioid receptors and cannabinoids, there is also the activation of the NO/cGMP/KATP pathway, inducing cellular hyperpolarization. In this study, we evaluated the participation of the cannabinoid system with subsequent activation of the NO/cGMP/KATP pathway in the peripheral antinociceptive effect of serotonin. The paw pressure test of mice was used in animals that had their sensitivity to pain increased due to an intraplantar injection of PGE2 (2 μg). Serotonin (250 ng/paw), administered locally in the right hind paw, induced antinociceptive effect. CB1 and CB2 cannabinoid receptors antagonists, AM251 (20, 40 and 80 μg) and AM630 (25, 50 and 100 μg), respectively, reversed the serotonin-induced antinociceptive effect. MAFP (0.5 μg), an inhibitor of the FAAH enzyme that degrades anandamide, and JZL184 (3.75 μg), an inhibitor of the enzyme MAGL that degrades 2-AG, as well as the VDM11 (2.5 μg) inhibitor of anandamide reuptake, potentiated the antinociceptive effect induced by a low dose (62. 5 ng) of serotonin. In the evaluation of the participation of the NO/cGMP/KATP pathway, the antinociceptive effect of serotonin was reversed by the administration of the non-selective inhibitor of NOS isoforms L-NOarg (12.5, 25 and 50 μg) and by the selective inhibitor for the neuronal isoform LNPA (24 μg), as well as by the soluble guanylate cyclase inhibitor ODQ (25, 50 and 100 μg). Among potassium channel blockers, only Glibenclamide (20, 40 and 80 μg), an ATP-sensitive potassium channel blocker, reversed the effect of serotonin. In addition, intraplantar administration of serotonin (250 ng) was shown to induce a significant increase in nitrite levels in the homogenate of the plantar surface of the paw of mice. Taken together, these data suggest that the antinociceptive effect of serotonin occurs by activation of the cannabinoid system with subsequent activation of the NO/cGMP/KATP pathway.


On Demand Release and Retrieval of Bacteria from Microwell Arrays Using Photodegradable Hydrogel Membranes.
Microwell arrays are important tools for studying single cell behavior and cell-cell interactions, both in microbial and mammalian systems. However, retrieval of cells from microwell arrays with high spatial precision remains a major technical hurdle that prevents follow-up genetic and phenotypic characterization of cells within observed microwells. This work describes a new, material-based approach to grow and retrieve live bacterial cells from small (≥20 μm diameter) microwells in an array using the plant pathogen Agrobacterium tumefaciens as a model bacterium. Our approach uses a light-responsive, step-polymerized poly(ethylene glycol) hydrogel interface as a membrane that confines motile cells within microwells while allowing nutrient exchange and cell growth. The key design feature is the photodegradability of the membrane, as it enables individual wells of interest to be opened using patterned UV light for selective release and retrieval of cells. Extraction can occur in parallel from any number and combination of wells defined by the user. These advancements represent a new use for light-responsive hydrogels and the ability to retrieve cells from microwells with high spatial precision enables several applications that require the isolation and characterization of cells with rare phenotypes from heterogeneous populations.


Selected management of pancreatic pseudocysts: operative versus expectant management.
The aim of this study was to determine if nonoperative, noninterventional expectant management of pancreatic pseudocysts is warranted in selected patients. From 1980 to 1985, 114 patients with the diagnosis of pancreatic pseudocyst were evaluated. Forty-six patients underwent primary operative therapy, with 13% undergoing emergency operations for pseudocyst-related complications. Although no operative deaths occurred, significant morbidity occurred in 26% of patients (emergency operations, 67%; elective procedures, 10%). The remaining 68 patients were initially treated selectively with a nonoperative, expectant approach. Severe, life-threatening complications in this group followed up for a mean of 46 months occurred in only 6 patients (9%); 19 patients eventually underwent elective operation directed at either the pseudocyst or other complications related to pancreatitis. Overall, in patients managed by a nonoperative approach, resolution of the pseudocyst occurred in 57% of the 24 patients with satisfactory radiographic follow-up, with 38% resolving more than 6 months after diagnosis. Although patients eventually undergoing operation tended to have larger pancreatic pseudocysts than the patients managed successfully nonoperatively (6.9 vs 4.9 cm), no serious complications occurred in seven patients with pancreatic pseudocysts greater than 10 cm who were treated expectantly. A nonoperative, noninterventional, expectant approach is warranted in the management of selected patients with pancreatic pseudocysts.


Sildenafil (Viagra) in the treatment of male erectile dysfunction in Nairobi.
To evaluate the effectiveness of sildenafil (Viagra) in the treatment of male erectile dysfunction in Nairobi. Prospective open label extension study. Urology clinics at the Nairobi Hospital, Kenyatta National Hospital and the author's private clinic in Hurlingham, Nairobi. Two hundred and nineteen adult male patients with erectile dysfunction. The age range was 33-80 years with a mean of 62.5 years and a peak incidence in the 60-69 year age group. One hundred and nineteen patients (54.34%) had organic causes, 85 patients (38.81%) had psychogenic causes and 15 patients had mixed causes. Two hundred patients (91.32%) had improved sexual function after treatment with viagra. This improvement was sustained during the study period of sixteen weeks and included improved erectile and orgasmic functions and overall sexual satisfaction. One hundred and fifty seven of these patients responded to therapy with 50 mg of viagra; 40 patients with 25 mg and three patients with 100 mg of therapy. Nineteen patients (8.68%) had no improvement in sexual function after viagra administration. Seven patients (3.2%) had adverse effects which were mild and transient. They included mild headaches in three patients, mild dyspepsia in two patients and facial flushing and nausea and vomiting in one patient, respectively. Oral sildenafil (Viagra) is an effective well tolerated and simple treatment for male erectile dysfunction in the majority of cases. The cost of treatment at about ten United States dollars for the 50 mg tablet is prohibitive and may limit its wide use by many deserving patients in this locality.


Assortative mating in mixed swarms of the mosquito Anopheles gambiae s.s. M and S molecular forms, in Burkina Faso, West Africa.
The molecular form composition of Anopheles gambiae Giles s.s. (Diptera: Culicidae) mating swarms and the associated mating pairs (copulae) were investigated during two rainy seasons (July to October, 2005 and July to November, 2006) in the villages of Soumousso and Vallée du Kou (VK7). Although the habitats of these villages differ markedly, sympatric populations of M and S molecular forms of An. gambiae s.s. occur in both places periodically. The main aim was to assess the degree to which these molecular forms mate assortatively. In Soumousso, a wooded savannah habitat, the majority of swarm samples consisted of only S-form males (21/28), although a few M-form males were found in mixed M- and S-form swarms. In VK7, a rice growing area, the majority of swarm samples consisted of only M-form males (38/62), until October and November 2006, when there were nearly as many mixed-form as single-form swarms. Overall, ∼60% of M- and S-form swarms were temporally or spatially segregated; the two forms were effectively prevented from encountering each other. Of the remaining 40% of swarms, however, only about half were single-form and the rest were mixed-form. Of the 33 copulae collected from mixed-form swarms, only four were mixed-form pairs, significantly fewer than expected by random pairing between forms (χ(2) = 10.34, d.f. = 2, P < 0.01). Finally, all specimens of inseminated females were of the same form as the sperm contained within their spermatheca (n = 91), even for the four mixed-form copulae. These findings indicate that assortative mating occurs within mixed-form swarms, mediated most probably by close-range mate recognition cues.


Oscillatory shear stress induced calcium flickers in osteoblast cells.
The dynamic physical microenvironment of bone affects the activity of osteoblast cells, yet little is known about how osteoblast mechanotransduction depends on different features of a dynamic stimulus. Here we investigated the effect of physiologically relevant oscillatory flow shear stress on the calcium mobility in osteoblast cells within a microfluidic platform that mimics the confined environment of bone matrix. We characterized the spatiotemporal evolution of intracellular calcium 'flickers', an important signature of cell activation, in response to steady, pulsatile, and oscillatory shear stress. We found that oscillatory flow induces surprisingly higher flicker activity than other flow types. We could further attribute this phenomenon to the opening of a stretch activated ion channel, namely TRPM7. We also found that localization of TRPM7 within the cholesterol-enriched lipid raft domains of plasma membranes is essential for its activity. Collectively our findings elucidated a candidate mechanism for the flow mediated stimulation of osteoblast cells. They therefore have implications towards unveiling various facets of bone formation and remodelling in healthy and diseased conditions, including bone-metastasis of various cancer types, diabetes, and inflammatory autoimmune diseases.


Chromosome conformation capture-based studies reveal novel features of plant nuclear architecture.
Nuclear genome organization has recently received increasing attention due to its manifold functions in basic nuclear processes, such as replication, transcription, and the maintenance of genome integrity. Using technologies based on chromosome conformation capture, such as Hi-C, we now have the possibility to study the three-dimensional organization of the genome at unprecedented resolution, shedding light onto a previously unexplored level of nuclear architecture. In plants, research in this field is still in its infancy but a number of publications provided first insights into basic principles of nuclear genome organization and the factors that influence it. Apart from general aspects, newly discovered three-dimensional conformations, such as the KNOT, raise special interest on how nuclear organization may influence the function of the genome in previously unexpected ways.


Drug-Related Problems Among Ambulatory Heart Failure Patients on Follow-Up at Debre Berhan Comprehensive Specialized Hospital, Ethiopia.
The purpose of this study was to assess drug-related problems (DRPs) among ambulatory heart failure (HF) patients attending at medical referral clinic of Debre Berhan Comprehensive Specialized Hospital, Ethiopia. A hospital-based cross-sectional study was conducted among 344 HF patients. Drug-related problems were classified using modified Cipolle's DRP classification schemes and drug-drug interactions were assessed using Micromedex, up-to-date, and drug.com drug-drug interaction checkers. The data was entered into Epidata version 4.2.0 and analyzed using SPSS version 25.0 statistical software. Descriptive statistics were used to summarize patients' characteristics. Univariable and multivariable binary logistic regression analysis was performed to identify associated factors with dependent variables. P < 0.05 was considered statistically significant. The mean age of the study participants was 53.38 ± 18.84 years and nearly half (45%) were in the age group of 31-60 years. Drug-related problems were found in 80.8% of HF patients. A total of 416 DRPs were identified. Adverse drug reaction (35.58%) was the top DRPs identified followed by the need for additional drug therapy (30.53%) and ineffective drug therapy (26.9%), respectively. Diuretics (45%), beta-blockers (BBs) (12.42%), and angiotensin-converting enzyme inhibitors (ACEIs) (10%) were the commonly used drug classes by study participants. The presence of comorbidity (p ˂ 0.001) and level of education of study participants (p = 0.03) had a significant association with the occurrence of DRPs. The prevalence of DRPs among ambulatory HF patients was high. The presence of comorbidity and the educational level of study participants had a significant association with the occurrence of DRPs. Checking potential drug-drug interactions before starting a new therapy, monitoring adverse drug reactions, ensuring sustainable availability of medications, and regular education programs are recommended to minimize DRPs.


Management of pulmonary tuberculosis patients in an urban setting in Zambia: a patient's perspective.
Zambia continues to grapple with a high tuberculosis (TB) burden despite a long running Directly Observed Treatment Short course programme. Understanding issues that affect patient adherence to treatment programme is an important component in implementation of a successful TB control programme. We set out to investigate pulmonary TB patient's attitudes to seek health care, assess the care received from government health care centres based on TB patients' reports, and to seek associations with patient adherence to TB treatment programme. This was a cross-sectional study of 105 respondents who had been registered as pulmonary TB patients (new and retreatment cases) in Ndola District between January 2006 and July 2007. We administered a structured questionnaire, bearing questions to obtain individual data on socio-demographics, health seeking behaviour, knowledge on TB, reported adherence to TB treatment, and health centre care received during treatment to consenting respondents. We identified that respondents delayed to seek treatment (68%) even when knowledge of TB symptoms was high (78%) or when they suspected that they had TB (73%). Respondent adherence to taking medication was high (77%) but low adherence to submitting follow-up sputum (47%) was observed in this group. Similarly, caregivers educate their patients more often on the treatment of the disease (98%) and drug taking (100%), than on submitting sputum during treatment (53%) and its importance (54%). Respondent adherence to treatment was significantly associated with respondent's knowledge about the disease and its treatment (p < 0.0001), and with caregiver's adherence to treatment guidelines (p = 0.0027). There is a need to emphasise the importance of submitting follow-up sputum during patient education and counselling in order to enhance patient adherence and ultimately treatment outcome.


Clinical Results of Selective Laser Trabeculoplasty.
Purpose: Selective laser trabeculoplasty (SLT) is a new technique aimed to developed to impact pigmented trabecular cells selectively. Compared with ordinary argon laser trabeculoplasty, it is expected to have fewer complications with more efficacy for open-angle glaucoma. In this study we performed SLT on 17 eyes of 10 patients with primary open-angle glaucoma and 1 eye with capsular glaucoma.Methods: Follow-up period was up to 10 months. Average energy irrachieted was 28.14 mJ (0.47 mJ x 59 spots) against pigmented trabecular band over the half circumference of anterior chamber angle.Results: Preoperative mean intraocular pressure (IOP) was 22.8 mmHg and postoperative mean IOP was decreased significantly to 8.6, 17.3, and 16.1 mmHg at 1, 3, and 6 months after treatment, respectively. The average maximum IOP reduction was 8.8 (3 approximately 1 8) mmHg after SLT. Among 11 eyes showing transient IOP elevation, 6 eyes had an elevation of more than 6 mmHg. No remarkable postoperative complications were noted.Conclusion: SLT is a safe and effective modality for the treatment of open-angle glaucoma such as primary open-angle glaucoma (POAG) and capsular glaucoma.


Persistence of unmet need for care among people with systemic lupus erythematosus: a longitudinal study.
The extent and variability of unmet care needs over time of people with systemic lupus erythematosus (SLE) has not been previously reported. A prospective study was undertaken to determine the variability over time of such needs. A 97-item SLE needs questionnaire (SLENQ) was completed on two occasions 6 months apart by 233 members of a lupus support association. Levels of unmet need for each individual, and a mean symptom score were calculated. Overall level of unmet need increased, decreased or stayed the same for 18%, 37%, and 45% of participants respectively. Ninety-four percent of participants at time 1 and 95% of participants at time 2 reported an unmet need for care for at least one item. The prevalence of unmet need for care remained the same for 82 of the 97 items. The mean unmet need score declined significantly between time 1 (0.78) and time 2 (0.69). The mean unmet need scores were significantly correlated with mean symptom score. The results of the study suggest an unacceptable persistence of care needs not being met. There is an ongoing need to identify mechanisms to enhance care delivery so that the care needs of people with SLE are more effectively addressed.


The 97 kDa linear IgA bullous disease antigen is identical to a portion of the extracellular domain of the 180 kDa bullous pemphigoid antigen, BPAg2.
IgA autoantibodies from the sera of some patients with linear IgA bullous dermatosis (LABD) recognize a 97 kDa antigen (LABD97) located in the lamina lucida of the basement membrane zone. As LABD autoantibodies do not react with the 180 and 230 kDa proteins recognized by bullous pemphigoid autoantibodies, LABD97 has been thought to represent a separate lamina lucida protein. In this study, we purified LABD97 from the extract of human epidermis using a monoclonal antibody immunoaffinity column and analyzed the amino acid sequence of the N terminus of purified LABD97. This revealed a 16 amino acid sequence that was identical to a previously reported sequence of the 180 kDa antigen in bullous pemphigoid (BPAg2). The N terminus was located 41 amino acids downstream from the carboxyl end of the transmembrane domain of BPAg2 and 11 amino acids downstream from the MCW-1 domain, the predominant bullous pemphigoid epitope. Purified LABD97 was subsequently enzymatically digested with endoproteinase Arg C and separated by chromatography, which resulted in multiple peptide fractions. Fourteen of these fractions were subjected to amino acid sequencing. The amino acid sequence of the peptide fractions, totaling 205 amino acids, were identical to sequences contained within the extracellular domain of BPAg2. Whereas the predominant epitope identified with bullous pemphigoid sera is located in the noncollagenous region of this protein, the epitope recognized by LABD sera is either within or adjacent to the collagenous portion. We conclude that LABD97 represents a portion of the extracellular domain of BPAg2 and that the IgA autoantibodies are directed against an epitope within or adjacent to a collagenous domain.


Capabilities of simultaneous 193 nm - LIBS/LA-ICP-MS imaging for microplastics characterization.
Microplastics (MPs) are currently one of the major environmental challenges within our society. With the awareness of the impact of MPs on the environment increasing over the last years, the need for increased monitoring as well as comprehensive analysis to better understand the fate and impact of MPs has become more and more important. A major aspect of MP characterization is the assignment of the polymer type of individual particles. Here, per- and poly-fluoroalkyl substances (PFAS), originating from fluor-containing polymers, have gained a lot of attention due to the severe environmental impact. Additionally, quantitative analysis of the metal content is of great interest in the field, since MPs are prone to either leaching (in)organic additives into the environment or taking up and accumulating hazardous components (e.g., heavy metals). In this work we demonstrate the capabilities of a simultaneous LIBS/LA-ICP-MS setup for the analysis of MPs. In the first part, we demonstrate the potential of targeted LIBS analysis for the imaging of fluor-containing polymers. Using a laser spot size of 5 μm combined with highly sensitive ICCD detection enables analysis of particles in the low μm range. In the second part we combine the polymer-identification capabilities of LIBS with the high sensitivity of ICP-MS to perform matrix-matched quantification of the metal content of individual MPs. In this case we use a spot size of 50 μm facilitating polymer classification with a broadband spectrometer, resulting in detection limits of 0.72 μg/g for Pb and 9.5 μg/g for Sn simultaneously measured using ICP-MS.


[Epithelioid sarcoma. Immunohistochemical study. Therapy with recombinant alfa interferon].
A case of Epithelioid Sarcoma in a 26 years old woman is reported. Clinical aspects of this entity are commented. Immunomarcation with monoclonal and policlonal antibodies was made, the findings were compared to those of the literature. Supervivence of the patient for 6 years without agressive surgery is remarked. Treatment was ruled out with recombinant alfa interferon for the last 3 years.


Effects of endurance exercise on the morphology of mouse neuromuscular junctions during ageing.
It has been suggested that age changes in the morphology of the neuromuscular junction (NMJ) may reflect altered physical activity levels rather than the unique effects of ageing. Additionally, previous studies have indicated that the structure of the NMJ may be modulated with exercise. To investigate these questions, quantitative morphometry was determined on soleus and extensor digitorum longus (EDL) nerve terminals stained with zinc iodide-osmium from C57BL/6NNia mice under control and endurance exercised conditions at 12, 18 and 24 months of age. As previously observed, the area, perimeter, extent length and branch number of nerve terminals increased with age in both soleus and EDL. The changes were similar between the muscle types, although the changes were more pronounced in the phasic EDL. In 12-month-old animals, 2 months of endurance exercise resulted in significantly larger nerve terminals in both soleus and EDL, suggesting a functional adaptation. Exercised 18- and 24-month-old nerve terminals were smaller than corresponding controls, which indicated that exercise minimized or prevented further age-related nerve terminal elaboration. At all ages the exercised nerve terminals comprised a more homogeneous population than corresponding controls, which indicates that uniform physical activity can modulate NMJ morphometry. The magnitude of the changes suggests that subtle alterations in normal cage activity with advancing age do not have a significant effect on the morphology of nerve terminals. However, the morphology of the NMJ does change significantly in response to physical exercise training.