Salivary gland disease in human immunodeficiency virus-positive women from the WIHS study. Women's Interagency HIV Study.
To determine the prevalence of enlargement, tenderness, and absence of saliva on palpation as indicators of salivary gland disease in women who are human immunodeficiency virus (HIV)-positive. The study subjects are participants in the Women's Interagency HIV Study (WIHS), a multicenter study examining HIV-seropositive women and at-risk HIV-seronegative women. A total of 576 HIV-positive women and 152 HIV-negative women were examined at their baseline oral visit for clinical markers of salivary gland disease. Viral load levels, CD4 counts, and CD8 counts were obtained as part of the related core study. HIV-positive women had higher rates of salivary gland enlargement (4.3%), tenderness (6.9%), and absence of saliva on palpation (26.6%) compared with HIV-negative women, who had rates of 1.3%, 4.6%, and 13.2%, respectively. Absence of saliva was significantly different (P =. 001) between the 2 groups. When 2 of the 3 clinical findings were combined, comparisons between the HIV-positive women and HIV-negative women became significant at the P <.05 level for every combination, except for enlargement/tenderness for the submandibular/sublingual gland. For the HIV-positive women, the viral load was significantly related to enlargement (P =.019) and enlargement/absence of saliva on palpation (P =.037) for the parotids and enlargement (P =.046), absence of saliva (P =.043), and enlargement/absence of saliva (P =.022) for the submandibular/sublingual glands. Significant linear trends were found for increasing viral load and enlargement (P =.013) and enlargement/tenderness (P =.024) for the submandibular/sublingual glands. Significance was present for submandibular/sublingual absence of saliva and tenderness/absence of saliva for CD4 and CD8 medians. Serostatus is related to salivary gland disease as assessed by glandular enlargement, tenderness, and absence of saliva on palpation. Furthermore, our findings indicate that a multidimensional approach to gland assessment may provide a more complete and perhaps more adequate description of glandular involvement with HIV infection.


Methylphenidate-induced hallucinosis: case histories and possible mechanisms of action.
Published reports of toxic hallucinosis during treatment with methylphenidate are rare. Two cases are described and it is suggested that methylphenidate-induced hallucinosis is more common than has been appreciated. Symptoms may be understood in terms of dose-related effects at pre- and postsynaptic receptors. Two other hypotheses that may clarify underlying mechanisms of action that produce toxic effects relate to (1) noradrenergic facilitation transmission of information in the visual pathway, and (2) interactions among brain monoamine systems.


Lrp is a direct repressor of the dad operon in Escherichia coli.
Expression of the degradative D-amino acid dehydrogenase (dad) operon is known to be increased when Escherichia coli is grown in the presence of D- or L-alanine. Alanine is thought to act as an inducer to block the action of a postulated repressor. This operon is also believed to be regulated by catabolite repression. We have used in vivo and in vitro experiments that show that the dad repressor is the leucine-responsive regulatory protein (Lrp). dad expression in a dad-lacZ operon fusion strain was increased four- to sevenfold when cells were grown in minimal medium containing alanine or leucine. A strain lacking Lrp had high-level constitutive dad expression. Gel retardation and footprinting studies revealed that Lrp binds in vitro to multiple sites over a large area in the dad promoter region. This binding was reduced by alanine or leucine. In vitro transcription assays, using a plasmid template and primer extension analysis, identified three major dad transcripts (Tr1, Tr2, and Tr3). The formation of these transcripts was differentially regulated by cyclic AMP-cyclic AMP receptor protein complex, and each was strongly repressed by Lrp. Alanine or leucine completely (for Tr1 and Tr2) or partially (for Tr3) reversed Lrp inhibition. Site-directed mutagenesis of an Lrp binding site strongly reduced Lrp binding and prevented Lrp repression of dad transcription in vivo and in vitro. Taken together, these results strongly suggest that Lrp and alanine or leucine act directly to repress and induce, respectively, transcription of the dad operon.


A Novel Type of Neuron Within the Dorsal Striatum.
The striatum is predominantly composed of medium spiny projection neurons, with the remaining neurons consisting of several types of interneurons. Among the interneurons are a group of cells that express tyrosine hydroxylase (TH). Although the intrinsic electrical properties of these TH-expressing interneurons have been characterized, there is no agreement on the number of TH-expressing cell types and their electrical properties. Here, we have used transgenic mice in which YFP-tagged channelrhodopsin-2 (ChR2) was expressed in potential TH-expressing cells in a Cre-dependent manner. We found that the YFP+ neurons in the striatum were heterogeneous in their intrinsic electrical properties; unbiased clustering indicated that there are three main neuronal subtypes. One population of neurons had aspiny dendrites with high-frequency action potential (AP) firing and plateau potentials, resembling the TH interneurons (THIN) described previously. A second, very small population of labeled neurons resembled medium-sized spiny neurons (MSN). The third population of neurons had dendrites with an intermediate density of spines, showed substantial AP adaptation and generated prolonged spikes. This type of striatal neuron has not been previously identified in the adult mouse and we have named it the Frequency-Adapting Neuron with Spines (FANS). Because of their distinctive properties, FANS may play a unique role in striatal information processing.


Pharmacogenetics and human genetic polymorphisms.
The term pharmacogenetics was first used in the late 1950s and can be defined as the study of genetic factors affecting drug response. Prior to formal use of this term, there was already clinical data available in relation to variable patient responses to the drugs isoniazid, primaquine and succinylcholine. The subject area developed rapidly, particularly with regard to genetic factors affecting drug disposition. There is now comprehensive understanding of the molecular basis for variable drug metabolism by the cytochromes P450 and also for variable glucuronidation, acetylation and methylation of certain drugs. Some of this knowledge has already been translated to the clinic. The molecular basis of variation in drug targets, such as receptors and enzymes, is generally less well understood, although there is consistent evidence that polymorphisms in the genes encoding the beta-adrenergic receptors and the enzyme vitamin K epoxide reductase is of clinical importance. The genetic basis of rare idiosyncratic adverse drug reactions had also been examined. Susceptibility to reactions affecting skin and liver appears to be determined in part by the HLA (human leucocyte antigen) genotype, whereas reactions affecting the heart and muscle may be determined by polymorphisms in genes encoding ion channels and transporters respectively. Genome-wide association studies are increasingly being used to study drug response and susceptibility to adverse drug reactions, resulting in identification of some novel pharmacogenetic associations.


Advances in clinical research in Alzheimer's disease.
There has been an explosion of clinical research on Alzheimer's disease (AD) in recent years, shedding light on many aspects of the disease. These include the differentiation of AD from normal aging, appreciation of genetic aspects of the disease, characterization of its pathology and pathophysiology, and the beginnings of a rational approach to treatment. Alzheimer's disease (AD) is a clinicopathologic diagnosis. In the near future, it may be possible to confirm the clinical diagnosis by examination of spinal fluid or by skin biopsy, but the diagnosis of AD in 1991 is based on history, direct examination of the patient and, ultimately, confirmation by microscopic examination of the brain. In short, we stand very close to where Dr. Alzheimer stood when he reported a hitherto unrecognized form of dementing illness in a 51-year-old woman. Alzheimer's case was cited by the editor of the journal in which it was published as an example of the need to look closely at patients from both clinical and pathologic perspectives so that individual diseases might eventually be separated out from syndromes such as dementia. Since Alzheimer's report, many clinical studies have shed further light on this disease. Two important deterrents to Alzheimer's disease research were the presumption that dementia is either a normal age-related phenomenon or a psychotic disorder of old age. The assumption that dementia is a product of normal aging or a functional psychotic disorder of late life was first challenged by Corsellis, who found a strong correlation between the severity of cerebral degenerative change and clinical diagnosis of patients who died in a psychiatric hospital.(ABSTRACT TRUNCATED AT 250 WORDS)


Tumor necrosis factor alpha plays a central role in immune-mediated clearance of adenoviral vectors.
Adenovirus (Ad) gene transfer vectors are rapidly cleared from infected hepatocytes in mice. To determine which effector mechanisms are responsible for elimination of the Ad vectors, we infected mice that were genetically compromised in immune effector pathways [perforin, Fas, or tumor necrosis factor alpha (TNF-alpha)] with the Ad vector, Ad5-chloramphenicol acetyl transferase (CAT). Mice were sacrificed at 7-60 days postinfection, and the levels of CAT expression in the liver determined by a quantitative enzymatic assay. When the livers of infected mice were harvested 28 days postinfection, the levels of CAT expression revealed that the effectors most important for the elimination of the Ad vector were TNF-alpha > Fas > perforin. TNF-alpha did not have a curative effect on infected hepatocytes, as the administration of TNF-alpha to infected severe combined immunodeficient mice or to infected cultures in vitro had no specific effect on virus persistence. However, TNF-alpha-deficient mice demonstrated a striking reduction in the leukocytic infiltration early on in the infection, suggesting that TNF-alpha deficiency resulted in impaired recruitment of inflammatory cells to the site of inflammation. In addition, the TNF-deficient mice had a significantly reduced humoral immune response to virus infection. These results demonstrate a dominant role of TNF-alpha in elimination of Ad gene transfer vectors. This result is particularly important because viral proteins that disable TNF-alpha function have been removed from most Ad vectors, rendering them highly susceptible to TNF-alpha-mediated elimination.


Biochemical and genetic characterization of type I familial amyloidotic polyneuropathy.
Type I familial amyloidotic polyneuropathy is an autosomal dominant, inherited systemic amyloidosis characterized initially by dissociated sensory disturbance and autonomic dysfunction. The amyloid fibril protein seen in patients of Portuguese, Japanese, and Swedish descent in the U.S. mainly consists of a variant form of transthyretin (also called prealbumin) with the substitution of methionine for valine at position 30. Methods have been developed to detect this variant transthyretin in the serum, and to detect a base change in a mutated transthyretin gene. The biochemical and genetic abnormalities in transthyretin are completely linked to the clinical diagnosis of type I familial amyloidotic polyneuropathy. These diagnostic methods may allow early diagnosis and genetic counseling to avoid transmission of this intractable disorder to the next generation.


CALM Pregnancy: results of a pilot study of mindfulness-based cognitive therapy for perinatal anxiety.
Many women experience anxiety during pregnancy with potential negative effects on maternal, birth, and child outcomes. Because of potential risks of fetal exposure to psychotropic medications, efficacious non-pharmacologic approaches are urgently needed. However, no published studies of psychotherapeutic treatments for anxiety in pregnancy exist. Mindfulness-based cognitive therapy (MBCT) may substantially reduce anxiety and co-morbid symptoms in people with anxiety disorders. Coping with Anxiety through Living Mindfully (CALM) Pregnancy is an adaptation of MBCT designed to address anxiety in pregnant women. This study examined the feasibility, acceptability, and clinical outcomes of the CALM Pregnancy intervention in pregnant women anxiety. Twenty-four pregnant women with generalized anxiety disorder (GAD) or prominent symptoms of generalized anxiety participated in an open treatment trial of the CALM Pregnancy group intervention. Psychiatric diagnoses were determined by structured clinical interview, and self-report measures of anxiety, worry, depression, self-compassion, and mindfulness were completed at baseline and post-intervention. Qualitative feedback was elicited via questionnaire. Twenty-three participants completed the intervention with high attendance and good compliance with home practice. Completers showed statistically and clinically significant improvements in anxiety, worry, and depression, and significant increases in self-compassion and mindfulness. Of the 17 participants who met GAD criteria at baseline, only one continued to meet criteria post-intervention. Participants regarded their experience in the intervention to be overwhelmingly positive. MBCT in the form of the CALM Pregnancy intervention holds potential to provide effective, non-pharmacological treatment for pregnant women with anxiety. These promising findings warrant further testing of the intervention with a randomized controlled trial.


A Miniaturised, Fully Integrated NDIR CO2 Sensor On-Chip.
In this paper, we present a fully integrated Non-dispersive Infrared (NDIR) CO2 sensor implemented on a silicon chip. The sensor is based on an integrating cylinder with access waveguides. A mid-IR LED is used as the optical source, and two mid-IR photodiodes are used as detectors. The fully integrated sensor is formed by wafer bonding of two silicon substrates. The fabricated sensor was evaluated by performing a CO2 concentration measurement, showing a limit of detection of ∼750 ppm. The cross-sensitivity of the sensor to water vapor was studied both experimentally and numerically. No notable water interference was observed in the experimental characterizations. Numerical simulations showed that the transmission change induced by water vapor absorption is much smaller than the detection limit of the sensor. A qualitative analysis on the long term stability of the sensor revealed that the long term stability of the sensor is subject to the temperature fluctuations in the laboratory. The use of relatively cheap LED and photodiodes bare chips, together with the wafer-level fabrication process of the sensor provides the potential for a low cost, highly miniaturized NDIR CO2 sensor.


Wishart Deep Stacking Network for Fast POLSAR Image Classification.
Inspired by the popular deep learning architecture, deep stacking network (DSN), a specific deep model for polarimetric synthetic aperture radar (POLSAR) image classification is proposed in this paper, which is named Wishart DSN (W-DSN). First of all, a fast implementation of Wishart distance is achieved by a special linear transformation, which speeds up the classification of POLSAR image and makes it possible to use this polarimetric information in the following neural network (NN). Then, a single-hidden-layer NN based on the fast Wishart distance is defined for POLSAR image classification, which is named Wishart network (WN) and improves the classification accuracy. Finally, a multi-layer NN is formed by stacking WNs, which is in fact the proposed deep learning architecture W-DSN for POLSAR image classification and improves the classification accuracy further. In addition, the structure of WN can be expanded in a straightforward way by adding hidden units if necessary, as well as the structure of the W-DSN. As a preliminary exploration on formulating specific deep learning architecture for POLSAR image classification, the proposed methods may establish a simple but clever connection between POLSAR image interpretation and deep learning. The experiment results tested on real POLSAR image show that the fast implementation of Wishart distance is very efficient (a POLSAR image with 768 000 pixels can be classified in 0.53 s), and both the single-hidden-layer architecture WN and the deep learning architecture W-DSN for POLSAR image classification perform well and work efficiently.


Recruitment and selection of couples for intervention research: achieving developmental homogeneity at the cost of demographic diversity.
Demographic and relationship quality data were collected from 704 individuals recruited to participate in a randomized study of relationship enhancing interventions. Recruiting at bridal shows produced partners who were more satisfied, earlier in their relationships, and less likely to be parents, with a marginally higher proportion of Latino couples. Radio and television coverage produced more established couples with higher levels of relationship discord. Self-selection effects revealed that couples from demographic groups at greater risk for divorce (those who had not completed high school, those with children at marriage, and African American couples) were more likely to agree to participate. In contrast, imposing a set of common selection criteria served to exclude couples from demographic risk groups and selected for couples with higher marital quality. Implications for recruiting couples to participate in preventive interventions are outlined.


Lifestyle and psychological factors related to irritable bowel syndrome in nursing and medical school students.
Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder comprising abdominal pain, abdominal discomfort, and disordered defecation. The prevalence of IBS is 10-15% in the general population. This study investigated the prevalence of IBS and the relationship between IBS and stress, lifestyle, and dietary habits among nursing and medical school students. A blank self-administrated questionnaire was used to survey 2,639 students studying nursing or medicine. This questionnaire asked about IBS symptoms, lifestyle, dietary intake, life events, anxiety, and depression. The questionnaires were collected from 2,365 students (89.6%) and the responses of 1,768 students (74.8%) were analyzed. The prevalence of IBS was 35.5% as a whole, 25.2% in males and 41.5% in females. Significantly higher stress scores (anxiety and depression) and life events were found in the IBS group than in the non-IBS group. Sleep disorders and the time spent sitting were also higher in males with IBS. In the IBS group, females ate less fish, fruit, milk, and green-yellow vegetables, and more processed food products than the non-IBS group (p = 0.001, p = 0.002, p = 0.032, p = 0.037, p < 0.001). The rates of missed meals and irregular mealtimes were significantly higher in females in the IBS group (p = 0.001, p = 0.013). The prevalence of IBS was higher among nursing and medical students, and further interventional studies are needed to improve IBS symptoms.


Medroxyprogesterone acetate alters the vaginal microbiota and microenvironment in women and increases susceptibility to HIV-1 in humanized mice.
The hormonal contraceptive medroxyprogesterone acetate (MPA) is associated with increased risk of human immunodeficiency virus (HIV), via incompletely understood mechanisms. Increased diversity in the vaginal microbiota modulates genital inflammation and is associated with increased HIV-1 acquisition. However, the effect of MPA on diversity of the vaginal microbiota is relatively unknown. In a cohort of female Kenyan sex workers, negative for sexually transmitted infections (STIs), with Nugent scores <7 (N=58 of 370 screened), MPA correlated with significantly increased diversity of the vaginal microbiota as assessed by 16S rRNA gene sequencing. MPA was also significantly associated with decreased levels of estrogen in the plasma, and low vaginal glycogen and α-amylase, factors implicated in vaginal colonization by lactobacilli, bacteria that are believed to protect against STIs. In a humanized mouse model, MPA treatment was associated with low serum estrogen, low glycogen and enhanced HIV-1 susceptibility. The mechanism by which the MPA-mediated changes in the vaginal microbiota may contribute to HIV-1 susceptibility in humans appears to be independent of inflammatory cytokines and/or activated T cells. Altogether, these results suggest MPA-induced hypo-estrogenism may alter key metabolic components that are necessary for vaginal colonization by certain bacterial species including lactobacilli, and allow for greater bacterial diversity in the vaginal microbiota.This article has an associated First Person interview with the first author of the paper.


[Neoadjuvant chemo-radiotherapy in the treatment of rectal cancer: preliminary results].
Many studies in the literature have confirmed the role of combined therapy in the treatment of rectal neoplasms. Aim of the Authors' study was to evaluate the efficacy of preoperative neoadjuvant radiochemotherapy in these patients. The study group consisted of 15 patients, affected by rectal adenocarcinoma T < 4 M0 with age < 75 years, observed from January to August 1998, who underwent to RT of pelvis with three fields--total dose 32 Gy- in ten days associated with 5 FU(500 mg/m2) during the first five. After restaging the patients were operated on. Anterior resections (AR) were followed by mechanical colorectal anastomosis in 10 patients, coloendoanal anastomosis in 4, while in 1 case an abdominoperineal resection (APR) was performed. The protocol was well tolerated in every patient and perioperative complications were similar to those in the control group. A considerable reduction of the mass was obtained in a great percentage of patients studied, confirmed by pathologic examination like "down staging" of cancer. In no case a complete response to the therapy was observed (pT0N0). Survival in the patients operated on for rectal adenocarcinoma in about 50% and local relapse is the most important site of recurrent disease. The aim of neoadjuvant protocol is the control of lymphatic spread and reduction rectal neoplasm allowing coloanal anastomosis in the treatment of the 1/3 inferior rectal cancer. Literature data confirm a significant decrease of local relapse, probably with a better survival. The Authors study confirms that combined preoperative therapy is well tolerated in most of the cases and it represents the protocol of choice in the patient affected by rectal cancer.


The effect of transfusions on renal allograft survival in the cyclosporine era: a single center report.
The potential efficacy of prior transplant transfusions on graft survival in the cyclosporine era has been reported from several centers which are reconsidering their transfusion policy. The purpose of this study has been to evaluate the results obtained in a large series of first kidney transplant patients (N=284) treated with cyclosporine. Our past experience showed a beneficial effect of blood transfusions in cadaveric renal transplants on conventional immunosuppression. Forty-eight patients never received blood transfusions, 85 patients received 1-2 transfusions, 72 patients received 3-5 transfusions and 79 patients received more than 5 transfusions in their pre-transplant periods. We did not show an obvious beneficial effect of prior blood transfusions in improving 1- and 2-year patient and graft survival. In fact, no statistically significant differences were found among any of the groups depending upon the transfusion status. In the transfused group, graft survival rates were 81.4% (1 year) and 77.8% (2 years) while in the non-transfused group they were 81.2% (1 year) and 78.4% (2 years)(p=n.s.). The patients who rejected had a significantly lower graft survival in the transfused group as compared to the non-transfused group. On the other hand, the transfused patients without rejection episodes experienced the best graft survival, suggesting a beneficial transfusion effect. We conclude that since it is impossible to determine which patients will not reject, pre-transplant transfusions under certain circumstances might be harmful, or at least not beneficial. We would recommend restricting transfusions in cyclosporine-tested patients only in cases of therapeutic necessity.


[Major drug related problems leading to hospital admission in the elderly].
Drug related problems represent an important problem in geriatric patients, and contribute to hospitalization in 15 to 30% of the admissions. In most cases this concerns adverse drug reactions (normal dose and overdose) but also drug therapy failures (e.g. undertreatment) are common. The main pharmacological classes involved are cardiovascular drugs and drugs for the central nervous system. Drug related problems can be prevented by regular medication review to decide if all drugs have to be continued, and to check for adverse drug reactions. Clinical pharmacists in hospitals can detect drug related problems (over-, under- and misuse using a systematic approach) and recommend changes to improve pharmacotherapy. The community pharmacist can help by elaborating drug schemes, by screening for medication interactions, and by resolving practical problems.


Autism: the micro-movement perspective.
The current assessment of behaviors in the inventories to diagnose autism spectrum disorders (ASD) focus on observation and discrete categorizations. Behaviors require movements, yet measurements of physical movements are seldom included. Their inclusion however, could provide an objective characterization of behavior to help unveil interactions between the peripheral and the central nervous systems (CNSs). Such interactions are critical for the development and maintenance of spontaneous autonomy, self-regulation, and voluntary control. At present, current approaches cannot deal with the heterogeneous, dynamic and stochastic nature of development. Accordingly, they leave no avenues for real time or longitudinal assessments of change in a coping system continuously adapting and developing compensatory mechanisms. We offer a new unifying statistical framework to reveal re-afferent kinesthetic features of the individual with ASD. The new methodology is based on the non-stationary stochastic patterns of minute fluctuations (micro-movements) inherent to our natural actions. Such patterns of behavioral variability provide re-entrant sensory feedback contributing to the autonomous regulation and coordination of the motor output. From an early age, this feedback supports centrally driven volitional control and fluid, flexible transitions between intentional and spontaneous behaviors. We show that in ASD there is a disruption in the maturation of this form of proprioception. Despite this disturbance, each individual has unique adaptive compensatory capabilities that we can unveil and exploit to evoke faster and more accurate decisions. Measuring the kinesthetic re-afference in tandem with stimuli variations we can detect changes in their micro-movements indicative of a more predictive and reliable kinesthetic percept. Our methods address the heterogeneity of ASD with a personalized approach grounded in the inherent sensory-motor abilities that the individual has already developed.


tert-Butyl nitrite (TBN) as the N atom source for the synthesis of substituted cinnolines with 2-vinylanilines and a relevant mechanism was studied.
A green method to synthesize cinnolines by 6π electrocyclic reaction with alkenyl amines and TBN has been developed. TBN plays a dual role both as a nitrogen atom source and an oxidant in this procedure. Relevant mechanism experiments reveal that the reaction proceeds through electrocyclic reaction and with diazo hydroxide as a key intermediate.


Hepatitis C virus genotype and HIV coinfection affect cytokine mRNA levels in unstimulated PBMC but do not shift the T1/T2 balance.
Rapid progression of hepatitis C virus (HCV) disease in patients with HIV/HCV may reflect different cytokine responses and be influenced by HCV genotype. This is addressed by a study of patients with HIV/HCV coinfection and infection with HCV genotype 2 or 3 (2/3). They are compared with coinfected patients infected with genotype 1 and HCV monoinfected patients matched for HCV genotype. IFN-gamma, IL-10, IL-4 and IL-4delta2 mRNA were quantified by real-time PCR in unstimulated PBMC and after in vitro stimulation with HCV core or nonstructural 3/4A antigen. In unstimulated PBMC, levels of IFN-gamma and IL-4 mRNA were lowest in HIV/HCV genotype 1 patients, intermediate in HIV/HCV genotype 2/3 patients and highest in HCV genotype 2/3 patients. Neither HCV genotype nor HIV affected levels of IL-10 mRNA in unstimulated PBMC or IFN-gamma, IL-4 and IL-10 mRNA in PBMC stimulated with HCV antigens. Levels of IL-4 and IL-4delta2 mRNA correlated in mitogen-stimulated PBMC from all patient groups but both were low in HIV/HCV genotype 1 patients. Serum soluble CD30 levels (a putative marker of a T2 cytokine environment) did not differ between patient groups. The data do not suggest a shift in the T1/T2 balance driven by HIV coinfection or HCV genotype but either may affect IL-4 bioavailability.


Verification of MLC based real-time tumor tracking using an electronic portal imaging device.
The authors have developed a novel technique using an electronic portal imaging device (EPID) to verify the geometrical accuracy of delivery of dose-rate-regulated tracking (DRRT). This technique, called verification of real-time tracking with EPID (VORTE), can potentially be used for both on-line and off-line quality assurance (QA) of MLC-based dynamic tumor tracking. The shape and position of target as a function of time, which is assumed to be known, is projected onto the EPID plane. This projected sequence of apertures as a function of time (target motion) is then used as the reference. The accuracy of dynamic MLC tracking can then be assessed by how well the delivered beam follows this projected target motion without the use of a physical moving phantom. The beam apertures controlled by DRRT (aperture motion) is detected by the EPID as a function of time. The aperture motion is compared to the target motion to evaluate tracking error introduced by DRRT. The accuracy of VORTE was measured using film measurements of ten static fields. The VORTE for dynamic tumor tracking was tested with several target motions, including (1) rigid-body two-dimensional (2-D) cyclic motion in the superior-inferior direction with various period and amplitude; (2) the above 2-D cyclic motion plus cyclic deformation; and (3) 2-D cyclic motion with both deformation and rotation. For each target motion, the controlled aperture motion resulting from DRRT was acquired at approximately 8 Hz using EPID in the continuous-acquisition mode. Leaf positions in all captured frames were measured from the EPID and compared to their expected positions. The passing rate of 2 mm criteria for all leaves from all frames was calculated for each of the four patterns of tumor motion. Additionally, the root-mean-square (RMS) deviations of the centroid of the apertures between the designed and delivered beams were calculated for all three cases. The accuracy of MLC-leaf position determination by VORTE is 0.5 mm (1 standard deviation) by comparison to film measurements. With DRRT, the passing rates using the 2 mm criteria for all acquired frames are 100% for the 2-D displacement, 99% for the 2-D displacement with deformation, and 88% for the 2-D displacement combined with both deformation and rotation. The RMS deviations are 0.6 mm for the 2-D displacement, 1.0 mm for the 2-D displacement with deformation, and 1.1 mm for the 2-D displacement combined with both deformation and rotation. The VORTE can measure the accuracy of MLC-based tumor tracking without the necessity of employing a moving phantom. Moreover, it can be used for complex target motion (i.e., 2-D displacement combined with deformation and rotation) that is difficult to create with physical moving phantoms. Therefore, the VORTE and the novel QA process illustrated by this study have a great potential for verifying real-time tumor tracking.


Antituberculosis drug resistance acquired during treatment: an analysis of cases reported in California, 1994-2006.
To inform efforts to prevent antituberculosis drug resistance acquired during treatment, particularly multidrug-resistant (MDR) tuberculosis, we analyzed surveillance records from the US state with the highest morbidity. Surveillance data from the California tuberculosis registry of cases reported between 1994 and 2006 were examined retrospectively. Crude risks of acquired resistance were estimated. Multivariate logistic regression was used to estimate odds ratios of demographic, clinical, and case management characteristics associated with acquired drug resistance (ADR), and secular trends in the incidence of ADR were assessed. One in 688 patients acquired MDR tuberculosis, with crude risks varying greatly by initial drug susceptibility test results: 1 in 1909 if initially susceptible to isoniazid and rifampin, 1 in 113 if initially isoniazid resistant, and 1 in 23 if initially rifampicin resistant. Acquired isoniazid and rifampicin monoresistance occurred in 1 in 1018 and 1 in 1455 patients, respectively. Independent predictors of acquired MDR tuberculosis were initial isoniazid resistance (odds ratio [OR], 19.2; 95% confidence interval [CI], 8.25-44.7; P < .001), initial rifampicin resistance (OR, 35.9; 95% CI, 8.61-150; P < .001), human immunodeficiency virus (HIV) infection (OR, 5.07; 95% CI, 1.73-14.9; P = .003), and cavitary disease in the absence of directly observed therapy throughout therapy (OR, 2.65; 95% CI, 1.05-6.69; P = .04). The annual incidence of ADR declined over the study period. Although ADR is rare and declining in California, its costly consequences warrant improvements in treatment practices. Our findings suggest that we ensure DOT throughout the course of therapy for patients with baseline drug resistance, cavitary disease, or HIV infection.


Parallel Evolution of Host-Attachment Proteins in Phage PP01 Populations Adapting to Escherichia coli O157:H7.
The emergence of antibiotic resistance has sparked interest in phage therapy, which uses virulent phages as antibacterial agents. Bacteriophage PP01 has been studied for potential bio-control of Escherichia coli O157:H7, its natural host, but in the laboratory, PP01 can be inefficient at killing this bacterium. Thus, the goal of this study was to improve the therapeutic potential of PP01 through short-term experimental evolution. Four replicate populations of PP01 were serially passaged 21 times on non-evolving E. coli O157:H7 with the prediction that the evolved phage populations would adsorb faster and more efficiently kill the host bacteria. Dead-cell adsorption assays and in vitro killing assays confirmed that evolved viruses improved their adsorption ability on E. coli O157:H7, and adapted to kill host bacteria faster than the wildtype ancestor. Sequencing of candidate tail-fiber genes revealed that the phage populations evolved in parallel; the lineages shared two point mutations in gp38 that encodes a host recognition protein, and surprisingly shared a ~600 bp deletion in gp37 that encodes the distal tail fibers. In contrast, no mutations were observed in the gp12 gene encoding PP01's short tail fibers. We discuss the functional role of the observed mutations, including the possible adaptive role of the evolved deletions. This study demonstrates how experimental evolution can be used to select for viral traits that improve phage attack of an important bacterial pathogen, and that the molecular targets of selection include loci contributing to cell attachment and phage virulence.


Effects of particulate air pollution on systolic blood pressure: A population-based approach.
Given the hypothesis that particulate air pollution is associated with systolic blood pressure, the effect of daily concentrations of air pollution on blood pressure was assessed in 2612 elderly subjects in the urban area of Bordeaux, France. Blood pressure was measured by a digital monitor. Particle concentrations (PM10) were obtained from the AIRAQ association that operates a local monitoring network of the air quality. To represent the ambient urban air pollution, stations had to be sufficiently correlated (i.e., correlation >0.70) and to have sufficiently similar mean levels of pollution. Linear regression was used to model the association between concentrations of particles (PM10) and systolic blood pressure. We observed associations between the fifth lag hour and systolic blood pressure for an increase of 10 microg/m3 of PM10 (beta = -1.12, 95% confidence interval: [-1.90; -0.30]). Despite contradictory results, fine particles must be considered nowadays as a major component of atmospheric air pollution in which everything must be put into practice in terms of public health actions in order to protect the general population and particularly the elderly group.


Voltammetric determination of sinomenine in biological fluid using a glassy carbon electrode modified by a composite film of polycysteic acid and carbon nanotubes.
Polycysteic acid based electrochemical oxidation of L-cysteine (CySH) and carbon nanotubes (CNTs) formed a composite thin film material at a glassy carbon electrode (GCE) that was used a novel modifier for electroanalytical determination of sinomenine which is used for rheumatoid arthritis treatment. The determination of sinomenine at the composite modified electrode was studied by differential pulse voltammetry (DPV). The peak current obtained at + 0.632 V (vs SCE) from DPV was linearly dependent on the sinomenine concentration in the range of 1.0 x 10(-7) to 6.0 x 10(-5) M in a B-R buffer solution (0.04 M, pH 1.81) with a correlation coefficient of 0.998. The detection limit (S/N = 3) was 5.0 x 10(-8) M. The electrochemical reaction mechanism of sinomenine was also discussed. This new method was then applied to the high-throughput determination of sinomenine in human serum samples with satisfactory results. This polycysteic acid/CNTs composite film may be considered to be a promising, low-cost, durable, and biocompatible material for the modification of sensors in applications to pharmaceutical and biomedical analysis.


Prevalence of human T-cell lymphotropic virus I and II in Colombian blood donors, 2001-2014: Implications for transfusion safety.
The human T-cell lymphotropic virus (HTLV) 1 and 2 cause various clinical disorders associated with degenerative diseases. Blood transfusion is a primary mechanism of transmission that is associated with the use of cellular components such as red blood cells. To describe the epidemiology of HTLV 1 and 2 in blood donors in Colombia from 2001-2014. A retrospective analysis was performed using screening, reactivity and positivity for HTLV 1 and 2 data collected from 2001 to 2014 by Colombian blood banks and consolidated by the Instituto Nacional de Salud. Using this information, transfusion-associated infectivity was also estimated. From 2001 to 2014, 60.2% of blood collected in Colombia was screened for HTLV 1 and 2 and had a cumulative reactivity of 0.30%. This was 20 times higher in Chocó (6.28%), where blood collection ended in 2004. Blood screening for HTLV reached 94.9% in 2014 with a positive concordance of 14.7%, and an estimated 406 unscreened, potentially infectious blood units were released. The majority of the unscreened blood units (215 units, 53%) came from Antioquia, a non-endemic department. These results suggest that HTLV 1 and 2 infections are distributed in different areas of the country that were not previously classified as endemic. These findings support the importance of the universal screening of blood units to minimize the risk of infection through transfusion for this event.


Deficiency in induction but not expression of LTP in hippocampal slices from young rats.
In this study we examine developmental changes between postnatal day (PND) 4 and 14 in synaptic transmission and plasticity in the CA1 region of hippocampal slices. We confirm previous results that tetanus-induced long-term potentiation (LTP) in field recordings is diminished in slices from younger animals. LTP in whole-cell current-clamp recordings is also diminished in younger animals. However, robust LTP can be induced in young animals if sufficient postsynaptic depolarization is provided during LTP induction. Furthermore, we find differences in synaptic transmission between PND 4 and 14, suggesting that the depolarization during tetanic stimulation in young tissue is ineffective to produce LTP. These results indicate that the smaller potentiation in field recordings in slices from younger animals is attributable to insufficient postsynaptic depolarization during LTP induction rather than a defect in expression mechanisms.


Analysis of the early embryonic cell cycles of Xenopus; regulation of cell cycle length by Xe-wee1 and Mos.
In Xenopus, cdc2 tyrosine phosphorylation is detected in the first 60-75 minute cell cycle but not in the next eleven cell cycles (cycles 2-12) which are only 30 minutes long. Here we report that the wee1/cdc25 ratio increases before the first mitotic interphase. We show that the Xe-wee1 protein is absent in stage VI oocytes and is expressed from meiosis II until gastrulation. A dominant negative form of Xe-wee1 (KM wee1) reduced the level cdc2 tyrosine phosphorylation and length of the first cycle. However, the ratio of wee1/cdc25 did not decrease after the first cycle and therefore did not explain the lack of cdc2 tyrosine phosphorylation in, nor the rapidity of, cycles 2-12. Furthermore, there was no evidence for a wee1/myt1 inhibitor in cycles 2-12. We examined the role of Mos in the first cycle because it is present during the first 20 minutes of this cycle. We arrested the rapid embryonic cell cycle (cycle 2 or 3) with Mos and restarted the cell cycle with calcium ionophore; the 30 minute cycle was converted into a 60 minute cycle, with cdc2 tyrosine phosphorylation. In addition, the injection of a non-degradable Mos (MBP-Mos) into the first cycle resulted in a dramatic elongation of this cycle (to 140 minutes). MBP-Mos did not delay DNA replication or the translation of cyclins A or B; it did, however, result in the marked accumulation of tyrosine phosphorylated cdc2. Thus, while the wee1/cdc25 ratio changes during development, these changes may not be responsible for the variety of cell cycles observed during early Xenopus embryogenesis. Our experiments indicate that Mos/MAPK can also contribute to cell cycle length.


Analysis and discrimination of necrosis and apoptosis (programmed cell death) by multiparameter flow cytometry.
Necrosis and apoptosis are two distinct modes of cell death which differ in morphology, mechanism and incidence. Membrane disruptants, respiratory poisons and hypoxia cause ATP depletion, metabolic collapse, cell swelling and rupture leading to inflammation. These are typical features of necrosis. Apoptosis plays a crucial role in embryogenesis and development and is also prevalent in tumours. It is characterised by cell shrinkage, chromatin condensation and systematic DNA cleavage. Apoptotic cells are rapidly engulfed by phagocytes, thus preventing inflammatory reaction to degradative cell contents. In vivo, apoptosis is almost impossible to quantify due to problems of heterogeneity and the short half-life of an apoptotic cell. In vitro, mechanistic studies are further complicated by a late phase of apoptosis where the cell membrane becomes permeable to vital dyes and which occurs in the absence of phagocytes. Here we describe a novel and rapid multiparameter flow cytometric assay which discriminates and quantifies viable, apoptotic and necrotic cells via measurement of forward and side light scatter (proportional to cell diameter and internal granularity, respectively) and the DNA-binding fluorophores Hoechst 33342 and propidium. It is anticipated that mechanistic studies of apoptosis in a variety of cell types will greatly benefit from this mode of analysis.


Trajectories of mother-child and father-child relationships across middle childhood and associations with depressive symptoms.
Using a family systems perspective, we examined the trajectories of father-child and mother-child closeness and conflict across Grades 1, 3, 4, 5, and 6, and their associations with child depressive symptoms across middle childhood among 685 families in the Eunice Kennedy Shriver National Institute of Child Health and Human Development Study of Early Child Care and Youth Development (SECCYD). Father-child and mother-child relationship conflict increased, whereas relationship closeness decreased from Grades 1 to 6. Girls with more slowly increasing father-child conflict, and more slowly decreasing father-child closeness, were at lower risk for depressive symptoms. Boys with more slowly increasing mother-child conflict were at lower risk for depressive symptoms. These findings highlight the important roles of both father-child and mother-child relationships in children's emotional adjustment during middle childhood.


Patients above sixty years of age with Hodgkin's lymphoma treated with a new strategy.
In the Swedish National Care Programme for Hodgkin's lymphoma (HL) a less intensive chemotherapy regimen with individualized dosing (LVPP/OEPA) was introduced in 1989. In total, 139 patients, 77 between 1985 and 1988 and 62 between 1989 and 1992, were studied. Mean ages were 72 and 71 years, respectively. One hundred and nineteen patients were treated with curative intention, 63 (82%) between 1985 and 1988 vs. 56 (90%) between 1989 and 1992 (p = 0.11). All patients (13 vs. 20) treated with radiotherapy only achieved a complete remission (CR). The CR rates (67% vs. 65%) for patients treated with 6-8 cycles of chemotherapy were also similar in the two time periods. The 5-year survival rate was 45% in the period 1985-1988 and 48% in 1989-1992. The survival of elderly HL patients was thus not improved from 1985-1988 to 1989-1992. Thus efforts to improve the chemotherapy regimen with individualized dosing did not change the outcome. Many patients experienced myelosuppression and opportunistic infections that may have contributed to the poor treatment results.


Postoperative abdominal adhesions: pathogenesis and current preventive techniques.
Post-operative abdominal adhesions (PAA) are a condition that occurs in more than 90% of patients undergoing abdomen surgery; they can cause chronic abdominal pain, female infertility and repeated bowel obstruction, requiring repetitive surgical interventions causing morbidity and mortality, as well as high costs. The formation of the PAA is due to an imbalance between the fibrinogenesis and fibrinolysis in favor of the first, associated with tissue hypoxia secondary to aggression of the peritoneum, also due to the own inflammatory response and the increase in the population of adhesion fibroblasts which inhibit the degradation of the extracellular matrix and facilitate mature collagen and supporting connective tissue. The prevention of PAA will decrease secondary complications, as well as hospitalizations, surgeries and consequently, cost containment. The PAA pathophysiologic process allows establishing research strategies in order to prevent them.


Influence of TiO2 on prebiotic thermal synthesis of the Gly-Gln polymer.
The role of the titanium dioxide (rutile and anatase) with and without room light on the thermal synthesis of the glycine-L-glutamine (Gly-Gln) polymer is described. The efficiency in percentage of polymerization with room light was increased in 6% in the presence of rutile and in 23% in the presence of anatase. The thermal synthesis in the molten state was carried out in the absence and presence of both oxides. In all cases, the vibrational spectra showed characteristic group frequencies corresponding to a polypeptide structure. No spectral differences were observed by room light effect on the polymer on rutile. However, the polymer obtained in the presence of anatase and room light shows spectral changes associated with the formation of shorter new abundant and conformationally different species compared with the original polymer. The SEM-EDX characterization of the solid phase involved in the thermal synthesis showed that the morphology of the polypeptide is different in the presence of rutile compared to anatase. The SDS-PAGE and GPC results suggest that smaller chains are formed in the presence of both oxides and the distribution of the size and weight of each polymer molecule is completely different when the condensation is performed in the presence of anatase or rutile. Nuclear magnetic resonance analyses confirmed the incorporation of both Gly and Gln residues in the polymers, with a prevalence of Gly. Both possible sequences N-GlyGln-C and N-GlnGly-C were also detected.


[Immune-humoral response of water buffalo (Bubalus bubalis) against Anaplasma marginale (Theiler, 1910)].
The aim of the present study was to analyze the humoral-immune response of water buffalo (Bubalus bubalis) naturally infected against Anaplasma marginale. For this work, colostrums/milk and blood samples were sequentially collected from buffalo cows prior and after partum for a period of 335 days and from buffalo calves from birth to 365 days after. The antibodies in the colostrums/milk and serum samples of these animals were determined using an ELISA indirect method and the data were analyzed as a mean of a group of animals with the matched ages during the period of 1999/2000 or individually during the year of 2005. The data from animals analyzed in group showed that the antibodies against A. marginale were in low concentration (below the cut off point: D.O. = 0.265 and ELISA levels, EL < or =3), in the sera of buffalo, during the first 90 and 105 days, respectively for cows and calves. Then, the levels of antibodies in the serum samples of buffalo calves, slightly raised to above the cut-off point and kept in higher levels up to approximately 365 days after birth, indicating active acquired immunity. Furthermore, when the animals were individually examined, the buffalo cows showed high antibody levels in the colostrums, but low levels in the blood stream during the first seven days post-partum, suggesting antibody transference from blood to mammary gland. In addition to that, buffalo calves showed high antibody levels during the first 24 hours after suckling colostrum, indicating a colostral passive immunity. By conclusion, the buffaloes were able to arm a humoral immune response against A. marginale and were considered reservoir of this parasite.


Memory enhancing activity of naringin in unstressed and stressed mice: possible cholinergic and nitriergic modulation.
The present study was designed to evaluate the effect of Naringin on memory of unstressed and stressed Swiss young albino mice. Naringin (80 mg/kg, i.p.) and donepezil (10 mg/kg) were administered for 21 successive days to separate groups of unstressed and stressed mice. The nootropic activity was evaluated using elevated plus maze and Hebbs Williams Maze. Brain acetylcholinesterase (AChE), brain nitrite and plasma corticosterone levels were also estimated. unpredictable chronic mild stress was produced by using different stressors. Naringin (80 mg/kg) and donepezil significantly showed memory enhancing activity in both unstressed and stressed mice. Naringin significantly reduced brain AChE activity and brain nitrite levels in both unstressed and stressed mice. Naringin (80 mg/kg) significantly reversed scopolamine-induced amnesia in unstressed and stressed mice. 7-Nitroindazole [a neuronal nitric oxide synthase (NOS) inhibitor] and aminoguanidine (an inducible NOS inhibitor) significantly enhanced memory improving activity and brain nitrite decreasing effect of naringin in unstressed and stressed mice respectively. Plasma corticosterone levels were significantly decreased by naringin (80 mg/kg) in stressed mice as compared to its control. Thus, naringin showed memory enhancing activity in unstressed mice probably by decreasing brain AChE activity and by inhibition of neuronal NOS. The memory enhancing activity of naringin in stressed mice might be due to decrease in brain AChE activity, inhibition of inducible NOS and also by decreasing the elevated plasma corticosterone levels.


Molecular mechanisms by which dietary isoflavones potentially prevent atherosclerosis.
Dietary isoflavones are currently receiving much attention because of their potential role in preventing coronary artery disease and other chronic diseases. Accumulating evidence from cell culture and laboratory animal experiments indicates that isoflavones have the potential to prevent or delay atherogenesis. Suggested mechanisms of action include: a reduction in low-density lipoprotein (LDL) cholesterol and a potential reduction in the susceptibility of the LDL particle to oxidation; (2) an improvement in vascular reactivity; (3) an inhibition of pro-inflammatory cytokines, cell adhesion proteins and nitric oxide (NO) production; and (4) an inhibition of platelet aggregation. These mechanisms are consistent with the epidemiological evidence that a high consumption of isoflavone-rich soy products is associated with a reduced incidence of coronary artery disease. Biological effects of isoflavones are dependent on many factors, including dose consumed, duration of use, protein-binding affinity, and an individual's metabolism or intrinsic oestrogenic state. Further clinical studies are necessary to determine the potential health effects of isoflavones in specific population groups as we currently know little about age-related differences in exposure to these compounds and there are few guidelines on optimal dose for cardiovascular health benefits.


Neurotensin pulmonary metabolism in normal and asthmatic subjects.
The role of pulmonary metabolism of endogenous neurotensin (NT) in asthma is still unclear. Information on this subject in humans is scarce. Evaluation of the pulmonary metabolism of the endogenous NT in asthmatic subjects during symptom-free periods and after a methacoline challenge test and in healthy individuals. Ten asthmatic subjects (aged 34 to 70 years), diagnosed with extrinsic (n = 5), atopic (n = 3), and mixed asthma (n = 2), were compared to a group of 10 healthy individuals (aged 45 to 69 years). The asthmatic group of patients was evaluated with a PD20-FEV1 methacoline challenge test 3 days after a washout period from cessation of their regular medications. Two catheters were inserted in order to draw blood samples for the evaluation of NT concentration: one was inserted into the pulmonary artery and the other into the radial artery. The mean concentration of NT in pulmonary and systemic arterial blood, as well as the arteriovenous difference of NT and the absolute value of production rate [PR/m'], PR/m'/kg and PR/m'/m2 were calculated for each participant. The mean neurotensin concentration in normal subjects was higher in mixed venous blood (pulmonary artery) than in systemic arterial blood (p < 0001). Similarly, mean NT mixed venous levels in asthmatic subjects was shown to be higher than mean NT levels in systemic arterial blood, before and after the bronchoconstriction with methacoline (p = 0.05 and p = 0.02, respectively). In contrast, the arterovenous difference and the mean values of PR of NT were similar in both groups. Our findings suggest that (1) NT concentration in mixed venous blood changes in transit through the pulmonary parenchyma, indicating that the pulmonary parenchyma is an important site of NT metabolism; (2) Pulmonary clearance of NT is unaffected by cholinergic bronchoconstriction. Further clinical studies are needed in order to improve both the understanding and the therapeutic approach of the neurogenic process in asthmatic subjects.


Gross Negligence: Impacts of Microplastics and Plastic Leachates on Phytoplankton Community and Ecosystem Dynamics.
Plastic debris is an established environmental menace affecting aquatic systems globally. Recently, microplastics (MP) and plastic leachates (PL) have been detected in vital human organs, the vascular system, and in vitro animal studies positing severe health hazards. MP and PL have been found in every conceivable aquatic ecosystem─from open oceans and deep sea floors to supposedly pristine glacier lakes and snow covered mountain catchment sites. Many studies have documented the MP and PL impacts on a variety of aquatic organisms, whereby some exclusively focus on aquatic microorganisms. Yet, the specific MP and PL impacts on primary producers have not been systematically analyzed. Therefore, this review focuses on the threats posed by MP, PL, and associated chemicals on phytoplankton, their comprehensive impacts at organismal, community, and ecosystem scales, and their endogenous amelioration. Studies on MP- and PL-impacted individual phytoplankton species reveal the production of reactive oxygen species, lipid peroxidation, physical damage of thylakoids, and other physiological and metabolic changes, followed by homo- and heteroaggregations, ultimately eventuating in decreased photosynthesis and primary productivity. Likewise, analyses of the microbial community in the plastisphere show a radically different profile compared to the surrounding planktonic diversity. The plastisphere also enriches multidrug-resistant bacteria, cyanotoxins, and pollutants, accelerating microbial succession, changing the microbiome, and thus, affecting phytoplankton diversity and evolution. These impacts on cellular and community scales manifest in changed ecosystem dynamics with widespread bottom-up and top-down effects on aquatic biodiversity and food web interactions. These adverse effects─through altered nutrient cycling─have "knock-on" impacts on biogeochemical cycles and greenhouse gases. Consequently, these impacts affect provisioning and regulating ecosystem services. Our citation network analyses (CNA) further demonstrate dire effects of MP and PL on all trophic levels, thereby unsettling ecosystem stability and services. CNA points to several emerging nodes indicating combined toxicity of MP, PL, and their associated hazards on phytoplankton. Taken together, our study shows that ecotoxicity of plastic particles and their leachates have placed primary producers and some aquatic ecosystems in peril.


[Post traumatic ocular mycosis due to Penicillium oxalicum.].
An ocular fungal infection in a 33 year-old man is reported. The patient had a traumatic corneal ulcer with subsequent abscess and perforation. Antibiotics were administered at the beginning of the treatment. Two successive conjunctival flaps were performed but were unsuccessful, followed by a corneal transplant with unfavorable outcome and the appearance of endophthalmitis. Material from the vitreous and corneal ulcer margins was obtained and Penicillium oxalicum Currie & Thom was isolated in the Mycology laboratory. Local and systemic antifungal therapy was unsuccessful and the eyeball was enucleated.


Identification of TRAMs as sphingolipid-binding proteins using a photoactivatable and clickable short-chain ceramide analog.
Ceramide is a lipid molecule that regulates diverse physiological and pathological reactions in part through inverting the topology of certain transmembrane proteins. This topological inversion is achieved through regulated alternative translocation (RAT), which reverses the direction by which membrane proteins are translocated across the endoplasmic reticulum during translation. However, owing to technical challenges in studying protein-ceramide interaction, it remains unclear how ceramide levels are sensed in cells to trigger RAT. Here, we report the synthesis of pac-C7-Cer, a photoactivatable and clickable short-chain ceramide analog that can be used as a probe to study protein-ceramide interactions. We demonstrate that translocating chain-associated membrane protein 2 (TRAM2), a protein known to control RAT of transmembrane 4 L6 subfamily member 20, and TRAM1, a homolog of TRAM2, interacted with molecules derived from pac-C7-Cer. This interaction was competed by naturally existing long-chain ceramide molecules. We showed that binding of ceramide and its analogs to TRAM2 correlated with their ability to induce RAT of transmembrane 4 L6 subfamily member 20. In addition to probing ceramide-TRAM interactions, we provide evidence that pac-C7-cer could be used for proteome-wide identification of ceramide-binding proteins. Our study provides mechanistic insights into RAT by identifying TRAMs as potential ceramide-binding proteins and establishes pac-C7-Cer as a valuable tool for future study of ceramide-protein interactions.


Antioxidative and hypocholesterolemic activities of water-soluble puerarin glycosides in HepG2 cells and in C57 BL/6J mice.
Puerarin is an isoflavone derived from Kudzu roots and has antioxidant and hypocholesterolemic effects; however, its insolubility often limits its biological availability in vivo. Using a novel transglycosylation process, the solubility of puerarin glycosides was increased >100-fold, but it was not known whether these modified puerarin glycosides maintained biological activities. We found that water-soluble puerarin glycosides fully maintained antioxidant activities compared with puerarin assessed by radical scavenging activity, reducing power assay, superoxide dismutase activity, and non-site-specific hydroxyl radical scavenging activity. Both puerarin and its glycosides also significantly reduced low-density lipoprotein (LDL) oxidation. Mice fed with puerarin glycosides (0.1% w/w) showed significantly reduced plasma total cholesterol levels, thus, we further investigated their hypocholesterolemic mechanisms by assessing several key gene expressions both in vitro and in vivo. Puerarin and its glycosides induced multiple changes in hepatic cholesterol metabolism. The LDL receptor promoter activity was increased dose-dependently in puerarin glycosides-treated HepG2 cells. Accordingly, the expression of LDL receptor mRNA and protein were also significantly increased in HepG2 cells and mouse livers. The transcription and translation of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase were down-regulated both in vitro and in vivo. The cholesterol 7alpha-hydroxylase (CYP7A1) mRNA levels were not affected in vitro but significantly up-regulated in the mouse livers. Collectively, our results show that puerarin and its glycosides are biologically fully active isoflavone and have antioxidant and hypocholesterolemic effects in HepG2 cells and in C57BL/6J mice. In the livers, hypocholesterolemic effects of puerarin glycoside may be achieved by multiple mechanisms including increasing LDL uptake, reducing cholesterol biosynthesis, and possibly enhancing cholesterol degradation.


Field vaccination against hemorrhagic enteritis of turkeys by a cell-culture live-virus vaccine.
A cell-culture-propagated (CC) live-virus hemorrhagic enteritis (HE) vaccine was evaluated for efficacy and safety in two field trials conducted in North Carolina (NC) and Minnesota (MN). At 4 or 5 1/2 weeks of age, 9,839 poults in NC and 15,857 poults in MN were vaccinated with a CC HE vaccine administered via the drinking water. A comparable number of poults were maintained as unvaccinated controls. Vaccinated and unvaccinated poults were compared for seroconversion, response to laboratory challenge with a virulent HE virus at 3 weeks postvaccination, livability, percentage graded A, and average weight at marketing. In both trials, vaccination with the CC HE vaccine resulted in immunity against HE as indicated by seroconversion and by resistance to HE lesions following laboratory challenge with virulent HE virus. Compared with unvaccinated groups, vaccinated groups had a significantly higher percentage of turkeys graded A in the NC trial and in two of three flocks in the MN trial (P less than 0.005). Further, in the NC trial, livability was significantly higher (P less than 0.005) in vaccinated turkeys than in unvaccinated turkeys. These data indicate that the CC HE vaccine is efficacious and safe to use in the field.


Construction and next-generation sequencing analysis of a large phage-displayed VNAR single-domain antibody library from six naïve nurse sharks.
Background: Shark new antigen receptor variable domain (VNAR) antibodies can bind restricted epitopes that may be inaccessible to conventional antibodies. Methods: Here, we developed a library construction method based on polymerase chain reaction (PCR)-Extension Assembly and Self-Ligation (named "EASeL") to construct a large VNAR antibody library with a size of 1.2 × 1010 from six naïve adult nurse sharks (Ginglymostoma cirratum). Results: The next-generation sequencing analysis of 1.19 million full-length VNARs revealed that this library is highly diversified because it covers all four classical VNAR types (Types I-IV) including 11% of classical Type I and 57% of classical Type II. About 30% of the total VNARs could not be categorized as any of the classical types. The high variability of complementarity determining region (CDR) 3 length and cysteine numbers are important for the diversity of VNARs. To validate the use of the shark VNAR library for antibody discovery, we isolated a panel of VNAR phage binders to cancer therapy-related antigens, including glypican-3, human epidermal growth factor receptor 2 (HER2), and programmed cell death-1 (PD1). Additionally, we identified binders to viral antigens that included the Middle East respiratory syndrome (MERS) and severe acute respiratory syndrome (SARS) spike proteins. The isolated shark single-domain antibodies including Type I and Type II VNARs were produced in Escherichia coli and validated for their antigen binding. A Type II VNAR (PE38-B6) has a high affinity (Kd = 10.1 nM) for its antigen. Conclusions: The naïve nurse shark VNAR library is a useful source for isolating single-domain antibodies to a wide range of antigens. The EASeL method may be applicable to the construction of other large diversity gene expression libraries.


Biphasic presence of fibrocytes in a porcine hypertrophic scar model.
The duroc pig has been described as a promising animal model for use in the study of human wound healing and scar formation. However, little is known about the presence and chronology of the fibrocyte cell population in the healing process of these animals. Wounds known to form scar were created on red duroc swine (3" x 3") with a dermatome to a total depth of either 0.06 inches or 0.09 inches. These wounds were allowed to heal completely and biopsies were done at scheduled time points during the healing process. Biopsies were formalin fixed and paraffin embedded for immunohistochemical analysis. Porcine reactive antibodies to CD-45 and procollagen-1 and a human reactive antibody to LSP-1 were used to detect the presence of fibrocytes in immunohistochemistry, an immunocytochemistry. Initial immunohistochemical studies showed evidence of a biphasic presence of fibrocytes. Pigs with 0.06 inches deep wounds showed positive staining for CD-45 and LSP-1 within highly cellular areas at days 2 and 4 after wounding. Additional animals with 0.09 inches deep wounds showed positive staining within similar areas at days 56, 70, and 113 after wounding. There was no immunohistochemical evidence of fibrocytes in skin biopsies taken at days 14, 28, or 42. Procollagen-1 staining was diffused in all samples. Cultured cells were stained for CD-45, LSP-1, and procollagen-1 by immunocytochemistry. These data confirm that fibrocytes are indeed present in this porcine model. We conclude that these cells are present after initial wounding and later during scar formation and remodeling. We believe that this is an evidence of a biphasic presence of fibrocytes, first as an acute response to skin wounding followed by later involvement in the remodeling process, prompted by continued inflammation in a deep partial thickness wound.


Regulation of activator protein-1 by 8-iso-prostaglandin E2 in a thromboxane A2 receptor-dependent and -independent manner.
The thromboxane (TX) A(2) receptor (TP) encompasses two alternatively spliced forms, termed the platelet/placental (TP-P) and endothelial (TP-E) type receptors. Experimental evidence suggests that TP activity may be modulated by novel ligands, termed the isoprostanes, that paradoxically act as TP agonists in smooth muscle and TP antagonists in platelet preparations. Here we have investigated whether prototypical isoprostanes 8-iso-prostaglandin (PG)F(2 alpha) and 8-iso-PGE(2) regulate the activity of TP isoforms expressed in Chinese hamster ovary (CHO) cells using activator protein-1 (AP-1)-luciferase activity as a reporter. AP-1-luciferase activity was increased by a TP agonist [9,11-dideoxy-9 alpha,11 alpha-methanoepoxy PGF(2 alpha) (U46619)] in CHO cells transfected with the human TP-P and TP-E receptors, and this response was fully inhibited by TP antagonists [1S-[1 alpha,2 beta(Z),3 alpha,5 alpha]]-7-[3-[[4-iodophenyl)sulfonyl]amino]-6,6-dimethylbicyclo[3.1.1]hept-2-yl]-5-heptenoic acid (I-SAP) and [1S-[1 alpha,2 alpha(Z),3 alpha,4 alpha]]-7-[[2-[(phenylamino) carbonyl]hydrazino]methyl]-7-oxabicyclo[2.2.1] hept-2-yl]-5-heptenoic acid (SQ 29,548)]. AP-1-luciferase activity was potently (nanomolar concentrations) increased by 8-iso-PGE(2) in CHO TP-P and TP-E cells, and this response was partially inhibited by cotreatment of cells with TP antagonists, whereas 8-iso-PGF(2 alpha) was without effect. Cyclooxygenase inhibitors did not abolish 8-iso-PGE(2) mediated AP-1-luciferase activity, indicating that this response is not dependent on de novo TXA(2) biosynthesis. Interestingly, 8-iso-PGE(2)-mediated AP-1-luciferase activity was near maximal in naive cells between 1 and 10 nM concentrations, and this response was not inhibited by TP antagonist or reproduced by agonists for TP or EP(1)/EP(3) receptors. These observations 1) support a role for novel ligands in the regulation of TP-dependent signaling, 2) indicate that TP-P and TP-E couple to AP-1, 3) provide further evidence that isoprostanes function as TP agonists in a cell-type specific fashion, and 4) indicate that additional targets regulated by 8-iso-PGE(2) couple to AP-1.


Treatment for meralgia paraesthetica.
Meralgia paraesthetica is a clinical syndrome for which a number of treatments are in common use, including conservative measures, injection of corticosteroid with local anaesthetic and surgery. We aimed to examine the evidence for the relative efficacy of these interventions. This review was first published in 2008. Searches were updated in 2010 and 2012. To assess the relative efficacy of commonly used treatments for meralgia paraesthetica. We searched the Cochrane Neuromuscular Disease Group Specialized Register (1 October 2012), CENTRAL (2012, issue 9 in The Cochrane Library), MEDLINE (January 1966 to October 2012), EMBASE (January 1980 to October 2012) and CINAHL Plus (January 1937 to October 2012) for randomised controlled studies. Non-randomised studies were identified by searching MEDLINE (January 1966 to October 2012) and EMBASE (January 1980 to October 2012). We also inspected the reference lists of these studies. We were unable to identify any randomised controlled trials (RCTs) or quasi-RCTs. We therefore looked for high quality observational studies meeting the following criteria: (1) At least five cases of meralgia paraesthetica. (2) Follow-up of at least three months after intervention (if any). (3) At least 80% of cases followed up. Three authors independently extracted relevant data from each study meeting the selection criteria and transferred into a data extraction form. We found no RCTs or quasi-RCTs in the original review or updates in 20011 and 2012. Cure or improvement have been described in high quality observational studies: (1) A single study describes spontaneous improvement of meralgia paraesthetica in 20 (69%) of 29 cases. (2) Four studies evaluating the injection of corticosteroid and local anaesthetic found cure or improvement in 130 (83%) out of a combined total of 157 cases. (3) Surgical treatments have been found to be beneficial in 264 (88%) out of 300 cases treated with decompression (nine studies); and 45 (94%) out of 48 cases treated with neurectomy (three studies). (4) Ninety-nine (97%) out of 102 patients with iatrogenic meralgia paraesthetica recovered completely (three studies). In the absence of any published RCTs or quasi-RCTs, the objective evidence base for treatment choices in meralgia paraesthetica is weak. High quality observational studies report comparable high improvement rates for meralgia paraesthetica following local injection of corticosteroid and surgical interventions (either nerve decompression or neurectomy). However, a similar outcome has been reported without any intervention in a single natural history study.


Risk Factors for 1-Year Graft Loss After Kidney Transplantation: Systematic Review and Meta-Analysis.
With expansion of the pool of kidney grafts, through the use of higher-risk donors, and increased attention to donor management strategies, the 1-year graft survival rate is subject to change. It is, therefore, useful to elucidate 1-year graft survival rates by dissecting the characteristics of the low-risk and high-risk kidney transplant cases. The objective of our study was to evaluate factors purported to influence the risk of 1-year graft loss in kidney transplant recipients. We searched bibliographic databases from 2000 to 2017 and included observational studies that measured the association between donor, recipient, the transplant operation, or early postoperative complications, and 1-year death-censored graft loss. We identified 35 eligible primary studies, with 20 risk factors amenable to meta-analysis. Six factors were associated with graft loss, with moderate to high degree of certainty: donor age (hazard ratio [HR], 1.11 per 10-year increase; 95% confidence interval [95% CI], 1.04 to 1.18), extended criteria donors (HR, 1.35; 95% CI, 1.28 to 1.42), deceased donors (HR, 1.54; 95% CI, 1.32 to 1.82), number of HLA mismatches (HR, 1.08 per one mismatch increase; 95% CI, 1.07 to 1.09), recipient age (HR, 1.17 per 10-year increase; 95% CI, 1.09 to 1.25), and delayed graft function (HR, 1.89; 95% CI, 1.46 to 2.47) as risk factors for 1-year graft loss. Pooled analyses also excluded, with a high degree of certainty, any associations of cold ischemia time, recipient race, pretransplant body mass index, diabetes, and hypertension with 1-year graft loss. Recipient age, donor age, standard versus extended criteria donor, living versus deceased donor, HLA mismatch, and delayed graft function all predicted 1-year graft survival. The effect of each risk factor is small.


Alternative Activation of Macrophages in Mice Peritoneal Cavities and Diaphragms by Newborn Larvae of Trichinella spiralis.
Trichinellosis is a serious zoonosis with a worldwide distribution. Fecund adult worms in the intestine release newborn larvae (NBL) that enter the general circulation from 4 days post infection (dpi). Alternatively activated macrophages in the peritoneal cavities and the diaphragms in Trichinella spiralis infected mice have been reported. However, a role of newborn larvae is poorly understood. The total numbers of peritoneal macrophages in mice infected with 500 muscle-stage larvae were counted during early infection and then total RNA was extracted. Peritoneal macrophages from uninfected C57BL/6 mice were incubated with IL-4 or LPS as a control, or co-cultured with live NBL, and peritoneal macrophages were obtained from mice injected with live or frozen dead NBL into peritoneal cavity. Total RNA was extracted from these macrophages. Two types of gene expression, classical and alternative activation, were examined in the macrophages and diaphragms of the infected mice using semi-quantitative reverse transcription-PCR. The number of peritoneal macrophages in T. spiralis infected mice increased significantly. mRNA peak expression of alternative activation markers, Ym1 and arginase-1 (Arg1), was confirmed in the peritoneal macrophages and in diaphragm of mice around 15 dpi, while mRNA expression of classical activation markers, TNFα, IP-10, and iNOS was not detected. Injection of live NBL into the peritoneal cavities induced mRNA expression of Ym1 and Arg1 in the peritoneal macrophages of mice 9 dpi. However, dead NBL did not induce such gene expression. Alternative activation was not detected in the peritoneal macrophages co-cultured with NBL in vitro. Gene expression of alternative activation makers, Ym1 and Arg1, was confirmed in the peritoneal macrophages and diaphragms of mice infected with T. spiralis. However, gene expression of classical activation markers was not detected. Live NBL induced an alternative activation of peritoneal macrophages in vivo, but not in vitro.


Urea production by yeasts other than Saccharomyces in food fermentation.
Urea is an important intermediate in the synthesis of carcinogenic ethyl carbamate in various food fermentations. Identifying urea-producing microorganisms can help control or reduce ethyl carbamate production. Using Chinese liquor fermentation as a model system, we identified the yeasts responsible for urea production. Urea production was positively correlated to the yeast population (R = 0.523, P = 0.045), and using high-throughput sequencing, we identified 26 yeast species. Partial least squares regression and correlation analysis indicated that Wickerhamomyces anomalus was the most important yeast to produce urea (variable importance plot = 1.927; R = 0.719, P = 0.002). Besides, we found that in W. anomalus the CAR1 gene (responsible for urea production) was 67% identical to that of Saccharomyces cerevisiae. Wickerhamomyces anomalus produced more urea (910.0 μg L-1) than S. cerevisiae (300.1 μg L-1). Moreover, urea production increased to 1261.2 μg L-1 when the two yeasts were co-cultured in a simulated fermentation, where the transcription activity of the CAR1 gene increased by 140% in W. anomalus and decreased by 40% in S. cerevisiae. Our findings confirm that a yeast other than Saccharomyces, namely W. anomalus, contributes more to urea formation in a simulated sorghum fermentation. These findings provide the basis for strategies to control or reduce ethyl carbamate formation.


High incidence of malformation syndromes in a series of 1,073 children with cancer.
Constitutional molecular defects are known to play a role in oncogenesis, as shown by the increased incidence of embryonic cancers in children with Beckwith-Wiedemann syndrome (BWS) or of leukemia in children with Down syndrome. To establish the incidence and spectrum of malformation syndromes associated with childhood cancer we performed a clinical morphological examination on a series of 1,073 children with cancer. We diagnosed a syndrome in 42 patients (3.9%) and suspected the presence of a syndrome in another 35 patients (3.3%), for a total of 7.2%. This incidence of patients with a proven or suspected syndrome is high, and points to a possible association. We describe new syndrome-tumor associations in several entities: cleidocranial dysostosis (Wilms tumor), Bardet-Biedl syndrome (BBS) (acute lymphoblastic leukemia), Kabuki syndrome (neuroblastoma), LEOPARD syndrome (neuroblastoma), Poland anomaly (osteosarcoma; Hodgkin disease), and blepharophimosis epicanthus inversus syndrome (Burkitt lymphoma). Twenty of the 42 syndrome diagnoses were not recognized in the patients prior to this study, indicating that these diagnoses are commonly missed. We propose that all children with a malignancy should be examined by a clinical geneticist or a pediatrician skilled in clinical morphology to determine if the patients have a malformation syndrome.


Distinct functions of natural ADAM-15 cytoplasmic domain variants in human mammary carcinoma.
Adamalysins [a disintegrin and metalloproteinase (ADAM)] are a family of cell surface transmembrane proteins that have broad biological functions encompassing proteolysis, adhesion, and cell signal regulation. We previously showed that the cytoplasmic domain of ADAM-15 interacts with Src family protein tyrosine kinases and the adaptor protein growth factor receptor binding protein 2 (Grb2). In the present study, we have cloned and characterized four alternatively spliced forms of ADAM-15, which differ only in their cytoplasmic domains. We show that the four ADAM-15 variants were differentially expressed in human mammary carcinoma tissues compared with normal breast. The expression of the individual isoforms did not correlate with age, menopausal status, tumor size or grade, nodal status, Nottingham Prognostic Index, or steroid hormone receptor status. However, higher levels of two isoforms (ADAM-15A and ADAM-5B) were associated with poorer relapse-free survival in node-negative patients, whereas elevated ADAM-15C correlated with better relapse-free survival in node-positive, but not in node-negative, patients. The expression of ADAM-15A and ADAM-15B variants in MDA-MB-435 cells had differential effects on cell morphology, with adhesion, migration, and invasion enhanced by expression of ADAM-15A, whereas ADAM-15B led to reduced adhesion. Using glutathione S-transferase pull-down assays, we showed that the cytoplasmic domains of ADAM-15A, ADAM-15B, and ADAM-15C show equivalent abilities to interact with extracellular signal-regulated kinase and the adaptor molecules Grb2 and Tks5/Fish, but associate in an isoform-specific fashion with Nck and the Src and Brk tyrosine kinases. These data indicate that selective expression of ADAM-15 variants in breast cancers could play an important role in determining tumor aggressiveness by interplay with intracellular signaling pathways.


Patterns of antimicrobial use among nursing home residents with advanced dementia.
Nursing home residents with advanced dementia are at high risk of infections and antimicrobial exposure near the end of life. Detailed studies quantifying antimicrobial prescribing practices among these residents have not been performed. A cohort of 214 residents with advanced dementia from 21 Boston-area nursing homes were followed up prospectively for 18 months or until death. We analyzed antimicrobial use, including type, indication, and quantity, by days of therapy per 1000 resident-days. During an average of 322 days of follow-up, 142 residents (66.4%) with advanced dementia received at least 1 course of antimicrobial therapy (mean [SD] number of courses per resident, 4.0 [3.7]). The mean (SD) number of days of therapy per 1000 resident-days for the entire cohort was 53.0 (4.3). Quinolones and third-generation cephalosporins were the most commonly prescribed antimicrobials, accounting for 38.3% and 15.2%, respectively, of 540 prescribed antimicrobial therapy courses. A respiratory tract infection was the most common indication (46.7% of all antimicrobial therapy courses). Among 99 decedents, 42 (42.4%) received antimicrobials during the 2 weeks before death, of which 30 of 72 courses (41.7%) were administered via the parenteral route. The number of decedents receiving antimicrobials (P < .001), the number of antimicrobials prescribed (P = .01), and the days of therapy per 1000 resident-days (P < .001) increased significantly as subjects approached death. Persons with advanced dementia are frequently exposed to antimicrobials, especially during the 2 weeks before death. The implications of this practice from the perspective of the individual treatment burden near the end of life and its contribution to the emergence of antimicrobial resistance in the nursing home setting need further evaluation.


Radiographic alterations in short-stem total hip arthroplasty: a 2-year follow-up study of 216 cases.
In recent years a variety of short-stems have been introduced. Stable osteointegration is a key factor for a satisfactory long-term result. The purpose of this study was to evaluate postoperative radiological alterations and subsidence, as a result of using a newly developed device, over a 2-year follow-up. 216 short-stems were implanted in combination with a cementless cup. Patients were allowed full weight-bearing on the first day postoperatively. Pre- and postoperative x-rays were done using a standardised technique. Radiological alterations, such as bone resorption, radiolucency, osteolysis and cortical hypertrophy were detected and located using modified Gruen zones, and subsidence was measured via a conventional digital technique over a 2-year follow-up. In addition, Harris Hip Score (HHS), rest pain and load pain on the visual analogue scale (VAS) were assessed respectively. At 2-year follow-up 6 stems (2.9%) showed nonprogressive radiolucent lines with a maximum width of 2 mm. Resorption of femoral bone stock was detected in a total of 8 cases (3.9%). Femoral cortical hypertrophy was seen in a total of 9 hips (4.4%). No patient showed osteolysis. A measureable subsidence of at least 2 mm was observed in a total of 15.7% (32 cases) after 6 weeks, corresponding to an initial settlement given full weight-bearing ambulation. Only 1.1% (2 cases) showed further progression at the 6-month follow-up, whereas at the 1- and 2-year follow-ups no further subsidence was observed. After 2 years HHS was 98.1 (65.0-100.0), rest pain on the VAS was 0.2 (0.0-7.0) while load pain was 0.4 (0.0-7.0). The results of this radiographic analysis give support to the principle of using metaphyseal anchoring, calcar guided short-stems. The low incidence of bony alterations after a follow-up of 2 years indicates a physiological load distribution. After mild initial subsidence a stable osteointegration can be achieved over time.


Cystic diseases of the kidney: a perspective on medullary cystic disease.
Fifty-two cases of adequately documented cystic disease of the renal medulla (CDRM) were selected from available literature and analyzed for hereditary mode, age at onset, and duration. In 17, recessively inherited disease appeared at 9.4+/- 1.6(SE) and lasted 7.2+/- 1.7 years while in 21, dominantly inherited disease appeared at 29.3 +/- 2.3 and lasted 2.4+/- 0.4 years. In 14 isolated cases similar data were intermediate. These findings support an hypothesis that at least two genotypes are responsible for the phenotypic expression of CDRM.


Berberine modulates ASK1 signaling mediated through TLR4/TRAF2 via upregulation of miR-23a.
The current study was designed to explore the underlying therapeutic effect of berberine (BBR), an alkaloid compound against LPS (1 μg/ml)/TNFα (10 ng/ml) mediated apoptosis signal-regulating kinase 1 (ASK1) signaling in RAW 264.7 macrophages and adjuvant-induced arthritic synovial macrophages (AA-SM) with relation to miR-23a levels. LPS and TNFα stimulation abrogated the expression of miR-23a resulting in TLR4/TRAF2 mediated ASK1 activation and downstream phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK). BBR (25-75 μM) treatment ameliorated the gene expression levels of TLR4, TRAF2, TNFα, IL-6, and IL-23 through the upregulation of miR-23a. Subsequently, BBR suppressed the levels of TLR4/TRAF2 mediated phosphorylation of ASK1/p38 and attenuated the expression of various pro-inflammatory cytokines (TNFα, IL-6 & IL-23) in RAW 264.7 macrophages and AA-SM cells. BBR was able to counteract these factors through activation of miR-23a levels in LPS/TNFα stimulated RAW 264.7 macrophages and AA-SM cells. NQDI1 (30 μM) treatment inhibited ASK1 activation resulting in basal levels of miR-23a, owing to the conclusion that ASK1 activation downregulates miR-23a levels inside the cells. Overall, our current findings predict that BBR is a potential candidate for therapeutic targeting of TLR4/TRAF2 mediated ASK1 activation in inflammatory and in RA pathogenesis possibly through post-transcriptional gene silencing via upregulation of miR-23a.


Animating ground water levels with Excel.
This note describes the use of Microsoft Excel macros (programs written in Excel's internal language, Visual Basic for Applications) to create simple onscreen animations of transient ground water data within Excel. Compared to many specialized visualization software packages, the use of Excel macros is much cheaper, much simpler, and can rapidly be learned. The Excel macro can also be used to create individual GIF files for each animation frame. This series of frames can then be used to create an AVI video file using any of a number of graphics packages, such as Corel PhotoPaint. The technique is demonstrated through a macro that animates changes in the elevation of a water table along a transect over several years.


Research criteria versus clinical criteria for interstitial cystitis.
In March 2003 experts on interstitial cystitis (IC) from around the world met in Kyoto, Japan for a series of workshops intended to define areas of agreement and difference in the approach to this disease. This workshop explored the diagnostic criteria IC in the clinical and research settings. The NIDDK criteria are used almost universally for research but there is tremendous disparity in the clinics and no consensus at all as to the best practice in diagnosing IC for clinical purposes. Throughout the world many physicians use the NIDDK research criteria for clinical diagnosis; in some countries this is standard. Undoubtedly, this mandates under-diagnosis and denies treatment to many patients. Still, a majority feel that one diagnostic system is mandatory to promote clarity in communication. The group called for two initiatives to help advance this area of controversy: (1) a large-scale, prospective study critically examining the role of cystoscopy, bladder distention, and biopsy in the diagnosis of IC; (2) a concerted effort to develop a single evidence-based guideline or algorithm that would be flexible enough to meet the needs of clinicians, researchers, and patients.


[Challenges of inter-sector collaboration in public policies: the dilemma between nutritional supplementation and the promotion of healthy eating in schools].
The study aimed to identify and analyze the development of inter-sector arrangements between the health and education sectors in the framework of the implementation of the NutriSUS strategy in a municipality (county) in the state of Rio de Janeiro, Brazil. The study was conducted from March 2015 to March 2017 and used policy analysis as the analytical reference, with the following analytical dimensions: operational design; ideas on the nutritional supplementation; institutional context; and inter-sector collaboration. Semi-structured interviews were held with health professionals and administrators and schoolteachers and school system administrators at the three levels of government, in addition to document analysis. The NutriSUS strategy was implemented in five preschool units and required establishing agreements between health professionals and teachers that were affected by the following: the design based on the preventive approach to diseases with emphasis on action by the health sector; mechanisms of induction and convergence of interests between the federal and local governments; management mechanisms and institutional spaces including different stages in the implementation process, that is, the NutriSUS management centered in the health sector but operationalized in the school system; and the intersection between NutriSUS and the National School Food Program which increased the visibility of competing concepts concerning supplementation as a preventive strategy and different approaches to promote healthy eating. Thus, the factors identified by the study challenge the development of inter-sector collaboration oriented by shared processes in the entire chain of implementation of inter-sector programs for schools.


[The role of biofilms in pathogenesis of chronic bacterial rhinosinusitis].
The objective of the present work was to study the contribution of biofilms to the development of chronic bacterial rhinosinusitis. A total of 50 patients with this pathology were available for the examination. Mucosal swabs were taken from the middle nasal passages of all the patients to be used for the detection of biofilms by luminescence microscopy. The lactoferrine content in mucosal secretion from the nasal cavity was determined by an immunoenzymatic assay. Two groups of the patients presenting with bacterial rhinosinusitis were distinguished in the study of impression smears from nasal cavity mucosa by luminescence microscopy; one of them was comprised of biofilm-positive patients the other of biofilm-negative ones (56% and 44% respectively). The patients showing biofilms over nasal cavity mucosa had the lactoferrine content in mucosal secretion on the order of 0.0033±0.0008 mg/l compared with 0.0068±0.00075 mg/l in the biofilm-negative patients and 0.55±0.0005 mg/l in the healthy volunteers (controls). In other words, the biofilm-positive patients presenting with chronic bacterial rhinosinusitis had two times lower content of lactoferrine in mucosal secretion from the middle nasal passages than those in the biofilm negative group and 126 times lower content of lactoferrine than in the control group.


Is the Organization of the Primary Motor Cortex in Low Back Pain Related to Pain, Movement, and/or Sensation?
Primary motor cortex (M1) organization differs between individuals with and without chronic low back pain (CLBP), in parallel with motor and sensory impairments. This study investigated whether movement behaviour and tactile/pain sensation are related to M1 organisation in CLBP. Transcranial magnetic stimulation (TMS) was used to map the M1 representation of the erector spinae and multifidus muscles in 20 participants with and without CLBP. Cortical organisation was quantified by: map volume; center of gravity (CoG); number of peaks; and primary and secondary peak location. Movement behaviour was assessed as the ability to dissociate lumbar from thorax motion and sensory function as two-point discrimination, pressure pain thresholds, and pain intensity (visual analogue scale). People with CLBP showed more anterior location of the CoG than controls. Map peaks were more numerous in CLBP participants who performed the movement task good than those with poor performance. In CLBP, smaller map volume correlated with greater pain during the movement task. Movement behaviour was not linearly correlated with M1 features. This study confirms that M1 maps differ between people with and without CLBP, but these changes are variable within the CLBP group and are not related to motor and sensory features in a simple manner.


Community ecology of invasions: direct and indirect effects of multiple invasive species on aquatic communities.
With many ecosystems now supporting multiple nonnative species from different trophic levels, it can be challenging to disentangle the net effects of invaders within a community context. Here, we combined wetland surveys with a mesocosm experiment to examine the individual and combined effects of nonnative fish predators and nonnative bullfrogs on aquatic communities. Among 139 wetlands, nonnative fish (bass, sunfish, and mosquitofish) negatively influenced the probability of occupancy of Pacific treefrogs (Pseudacris regilla), but neither invader correlated strongly with occupancy by California newts (Taricha torosa), western toads (Anaxyrus boreas), or California red-legged frogs (Rana draytonii). In mesocosms, mosquitofish dramatically reduced the abundance of zooplankton and palatable amphibian larvae (P. regilla and T. torosa), leading to increases in nutrient concentrations and phytoplankton (through loss of zooplankton), and rapid growth of unpalatable toad larvae (through competitive release). Bullfrog larvae reduced the growth of native anurans but had no effect on survival. Despite strong effects on natives, invaders did not negatively influence one another, and their combined effects were additive. Our results highlight how the net effects of multiple nonnative species depend on the trophic level of each invader, the form and magnitude of invader interactions, and the traits of native community members.


Detection of novel biomarkers for ovarian cancer with an optical nanotechnology detection system enabling label-free diagnostics.
Ovarian carcinoma has the highest lethality rate of gynecologic tumors, largely attributed to the late-stage diagnosis of the disease. Reliable tools for both accurate diagnosis and early detection of disease onset are lacking, and presently less than 20% of ovarian cancers are detected at an early stage. Protein biomarkers that allow the discrimination of early and late stages of ovarian serous carcinomas are urgently needed as they would enable monitoring pre-symptomatic aspects of the disease, disease progression, and the efficacy of intervention therapies. We compare the absolute and relative protein levels of six protein biomarkers for ovarian cancer in five different established ovarian cancer cell lines, utilizing both quantitative immunoblot analysis and a guided-mode resonance (GMR) bioassay detection system that utilizes a label-free optical biosensor readout. The GMR sensor approach provided highly accurate, consistent, and reproducible quantification of protein biomarkers as validated by quantitative immunoblotting, as well as enhanced sensitivity, and is therefore suitable for quantification and detection of novel biomarkers for ovarian cancer. We identified fibronectin, apolipoprotein A1, and TIMP3 as potential protein biomarkers for the differential diagnosis of primary versus metastatic ovarian carcinoma. Future studies are needed to confirm the suitability of protein biomarkers tested herein in patient samples.