Ganglion cell loss in early glaucoma, as assessed by photopic negative response, pattern electroretinogram, and spectral-domain optical coherence tomography.
To evaluate discrimination ability of pattern electroretinography (PERG) and photopic negative response (PhNR) between early glaucoma and healthy controls, and their relationship with structural measurements using spectral-domain optical coherence tomography (SD-OCT). Cross-sectional study with 34 patients with ocular hypertension (n = 7), suspect glaucoma (n = 17), and early glaucoma (n = 10), plus 24 age-matched controls. The following parameters were analyzed: P50 and N95 amplitude of the PERG, PhNR amplitude and PhNR/b-wave ratio, peripapillary retinal and macular nerve fiber layer (NFL) thicknesses, and ganglion cell complex (GCC) thickness. Data from only one eye per individual were included in the statistical analysis. Descriptive statistics, ANOVA, receiver operating characteristics (ROC) curves, and correlation tests were used for analysis of the variables. PERG N95 and PhNR amplitudes were significantly reduced in suspect and early glaucoma eyes versus controls. Significant differences across ocular hypertensive, suspect, and early glaucoma eyes were found for macular NFL and GCC thickness, but not for any of the ERG parameters. The mean PhNR amplitude did not differ across these groups and was already reduced on average by 46% in ocular hypertensive and early glaucoma eyes and by 52% in suspect glaucoma eyes. The P50 and N95 amplitudes showed similar reduction in suspect and early glaucoma eyes on average by 15 and 26%, respectively. Of the ERG parameters, PhNR amplitude distinguished best between glaucoma and control groups, with an area under ROC curve of 0.90 for suspect glaucoma, and 0.86 for early glaucoma. PhNR/b-wave ratio showed strongest association in suspect glaucoma eyes with peripapillary retinal (r, 0.61) and macular NFL (r, 0.76) thicknesses. In eyes with early glaucoma, peripapillary retinal NFL thickness correlated best with PhNR amplitude (r, 0.71) and PERG P50 amplitude (r, 0.67). In eyes with suspect glaucoma, important decrease in PhNR amplitude is associated with small changes in peripapillary retinal and macular NFL thicknesses. These findings suggest that PhNR may be a useful and sensitive test in eyes with diagnostic dilemma, although further follow-up of such eyes is required for definitive confirmation.


Genetic and environmental influences on the junior temperament and character inventory in a preadolescent twin sample.
This study evaluated the genetic and environmental structure of personality variables from the Junior Temperament and Character Inventory (JTCI), in 605 pairs of 9- and 10-year old twins. There is a paucity of information on the biometric structure of temperament and character traits in preadolescent children. Latent factor models were fit to the subscales/items of each trait as a method of estimating genetic and environmental effects on true score variance, especially since internal consistency and reliability were moderate or low for some scales (particularly Reward Dependence and Persistence). Shared environmental influences on Cooperativeness were substantial. Significant heritability estimates were obtained for Self-directedness and Harm Avoidance, but not Novelty Seeking, Reward Dependence or Persistence. With the exception of Harm Avoidance, each of the scales failed to show measurement invariance with respect to sex, suggesting these scales may differ in meaning for boys and girls at this age.


Age- and risk factor-based serologic screening for Hepatitis C virus among an Urban, high-risk population.
Hepatitis C virus (HCV) screening among individuals born between 1945 and 1965 (ie birth cohort) may augment risk factor-based screening. We assessed HCV seropositivity among injection drug users (IDUs) and birth cohort members from New York City. We assessed HCV risk factors and seropositivity in 7722 participants from community health, HIV prevention, syringe exchange and drug treatment programmes. A total of 26.6% were HCV seropositive, 55.8% were born between 1945 and 1965, and 82.2% had ever injected drugs. Among all participants, HCV seropositivity was higher among IDUs compared to non-IDUs (60.5% versus 7.7%, odds ratio (OR) = 18.5, 95% confidence interval (CI) [16.2, 21.1], P < .0001) and among birth cohort members compared to non-birth cohort members (31.3% versus 22.3%, OR = 1.6, 95%CI [1.4, 1.8], P < .0001). Within the birth cohort, HCV seroprevalence among IDUs was 68.5% versus 11.8%, OR = 16.2, 95%CI [13.7, 19.3]. After adjustment, HCV seroprevalence was higher in IDUs, previously incarcerated, whites (<42 years) and 'other races' (versus blacks), HIV-infected, those who snorted heroin, those with liver disease history, and those who had sex with an HCV-seropositive partner. HCV seroprevalence among IDU, birth cohort members, was considerably higher than among the general population. In this high-risk, urban population, the association between IDU and HCV seropositivity was approximately ten times that between birth cohort membership and HCV seropositivity.


Critical thinking in African American mothers who care for adult children with HIV: a cultural analysis.
This research study defines critical thinking in nursing and examines the thinking processes revealed by 15 African American mothers who are caregivers to adult children with HIV. The purpose of this cultural analysis was to compare the mothers' decision-making processes with their critical-thinking processes. Their culture, heritage, faith, and value of family influenced caregivers in this study. Their testimony revealed the patterns of creating a different path of care, weaving together resources, choosing among negative alternatives, and selecting stories to tell. Mothers' decisions were based on complex and holistic knowledge of their situations and culture and could be termed multilogical, a type of thinking considered necessary for managing complex situations. Health providers can benefit from an understanding of these decision-making processes.


Occurrence of brominated persistent organic pollutants (PBDD/DFs, PXDD/DFs, and PBDEs) in Baltic wild salmon ( Salmo salar ) and correlation with PCDD/DFs and PCBs.
The contamination profiles of polybrominated dibenzo-p-dioxins (PBDDs), dibenzofurans (PBDFs), diphenyl ethers (PBDEs), and mixed monobromo/chloro dibenzo-p-dioxins (PXDDs) and dibenzofurans (PXDFs) were determined in the tissue of Baltic wild salmon and compared with those of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and biphenyls (PCBs). Out of the analyzed PXDD/DFs, only the 3-B-2,7,8-triCDF was detected (in the concentration range of 0.039-0.075 pg g(-1) fresh weight (f.w.)). The toxic equivalents (TEQs) for analyzed PBDD/DFs (0.074-0.142 pg TEQ g(-1) f.w.) were found to contribute on average 2.1% to the total PCDD/DF-PBDD/DF-TEQ. The total concentrations of 27 PBDE congeners were in the range of 1.3-5.6 ng g(-1) f.w., with an average of 3.3 ng g(-1) f.w. The levels of PCDD/DFs and PCBs were found to be in the range of 4.53-14.6 pg WHO(2005)-PCDD/DF-PCB-TEQ g(-1) f.w., and concentrations of these compounds in most of the analyzed samples were above the maximum levels specified in Commission Regulation (EU) No. 1259/2011. Good correlation was observed between WHO(2005)-PCDD/DF-TEQ and WHO(2005)-PCB-TEQ (r(2) = 0.98), and these parameters were well correlated with the total sum of PBDE concentrations (r(2) = 0.91 and r(2) = 0.94, respectively). The results suggest that the consumption of Baltic wild salmon has no crucial impact on the average POP intake for typical Latvians.


An RF-based wearable sensor system for indoor tracking to facilitate efficient healthcare management.
To understand the utilization of clinical resources and improve the efficiency of healthcare, it is often necessary to accurately locate patients and doctors in a healthcare facility. However, existing tracking methods, such as GPS, Wi-Fi and RFID, have technological drawbacks or impose significant costs, thus limiting their applications in many clinical environments, especially those with indoor enclosures. This paper proposes a low-cost and flexible tracking system that is well suited for operating in an indoor environment. Based on readily available RF transceivers and microcontrollers, our wearable sensor system can facilitate locating users (e.g., patients or doctors) or objects (e.g., medical devices) in a building. The strategic construction of the sensor system, along with a suitably designed tracking algorithm, together provide for reliability and dispatch in localization performance. For demonstration purposes, several simplified experiments, with different configurations of the system, are implemented in two testing rooms to assess the baseline performance. From the obtained results, our system exhibits immense promise in acquiring a user location and corresponding time-stamp, with high accuracy and rapid response. This capability is conducive to both short- and long-term data analytics, which are crucial for improving healthcare management.


Acetylcholinesterase activity changes on visceral organs of VMH lesion-induced obese rats.
It is accepted that the tone of the parasympathetic nervous system increases after VMH lesion, whereas the sympathetic tone decreases. To reinforce investigations over outcomes from disturbances of the hypothalamic neuronal systems on peripheral autonomic nerve activity this study determined the acetylcholinesterase (AchE) activity in visceral organs, known as vagal targets, from VMH-lesioned obese rats. It was found that AchE activity was significantly increased in liver, pancreas, and stomach from these animals. However, it was not changed in kidneys, being decreased in spleen. The results suggest that AchE activity is enhanced in vagus innervated tissues to following up the unbalance of the autonomic nervous system as observed in VMH lesion-induced obesity.


[Effects of snake venom cystatin gene on gene expression profiles in mouse melanoma cell line B16F1].
Previously, we have demonstrated that snake venom cystatin (sv-cystatin) plays an important role in tumor invasion and metastasis. This study was to investigate the effects of sv-cystatin on the gene expression profile of mouse melanoma B16F1 cells. pcDNA3.1/sv-cystatin plasmid was constructed and transfected into B16F1 using lipofectamine. Differentially expressed genes between B16F1 transfected with pcDNA3.1/sv-cystatin and pcDNA3.1 were analysed by high throughput microarray technique. Five up-regulated genes and five down-regulated genes were confirmed by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Out of 1218 transcript species, 45 showed altered expressions: 21 were up-regulated and 24 were down-regulated. These altered genes are involved in cell adhesion and migration, cell immunomodulation, proliferation, differentiation, apoptosis, as well as gene transcription and intra-cellular signal transduction. RT-PCR results of 10 alerted genes were in accordance with the microarray data. Sv-cystatin not only inhibits the extracellular matrix, but may also possess other diverse biological functions, including cell immunomodulation, proliferation and differentiation, apoptosis, gene transcription and intra-cellular signal transduction.


DMclust, a Density-based Modularity Method for Accurate OTU Picking of 16S rRNA Sequences.
Clustering 16S rRNA sequences into operational taxonomic units (OTUs) is a crucial step in analyzing metagenomic data. Although many methods have been developed, how to obtain an appropriate balance between clustering accuracy and computational efficiency is still a major challenge. A novel density-based modularity clustering method, called DMclust, is proposed in this paper to bin 16S rRNA sequences into OTUs with high clustering accuracy. The DMclust algorithm consists of four main phases. It first searches for the sequence dense group defined as n-sequence community, in which the distance between any two sequences is less than a threshold. Then these dense groups are used to construct a weighted network, where dense groups are viewed as nodes, each pair of dense groups is connected by an edge, and the distance of pairwise groups represents the weight of the edge. Then, a modularity-based community detection method is employed to generate the preclusters. Finally, the remaining sequences are assigned to their nearest preclusters to form OTUs. Compared with existing widely used methods, the experimental results on several metagenomic datasets show that DMclust has higher accurate clustering performance with acceptable memory usage.


Modelling cancer progression using Mutual Hazard Networks.
Cancer progresses by accumulating genomic events, such as mutations and copy number alterations, whose chronological order is key to understanding the disease but difficult to observe. Instead, cancer progression models use co-occurrence patterns in cross-sectional data to infer epistatic interactions between events and thereby uncover their most likely order of occurrence. State-of-the-art progression models, however, are limited by mathematical tractability and only allow events to interact in directed acyclic graphs, to promote but not inhibit subsequent events, or to be mutually exclusive in distinct groups that cannot overlap. Here we propose Mutual Hazard Networks (MHN), a new Machine Learning algorithm to infer cyclic progression models from cross-sectional data. MHN model events by their spontaneous rate of fixation and by multiplicative effects they exert on the rates of successive events. MHN compared favourably to acyclic models in cross-validated model fit on four datasets tested. In application to the glioblastoma dataset from The Cancer Genome Atlas, MHN proposed a novel interaction in line with consecutive biopsies: IDH1 mutations are early events that promote subsequent fixation of TP53 mutations. Implementation and data are available at https://github.com/RudiSchill/MHN. Supplementary data are available at Bioinformatics online.


Development of a common carp (Cyprinus carpio) pregnane X receptor (cPXR) transactivation reporter assay and its activation by azole fungicides and pharmaceutical chemicals.
In mammals, the pregnane X receptor (PXR) is a transcription factor with a key role in regulating expression of several genes involved in drug biotransformation. PXR is present in fish and some genes known to be under its control can be up-regulated by mammalian PXR ligands. Despite this, direct involvement of PXR in drug biotransformation in fish has yet to be established. Here, the full length PXR sequence was cloned from carp (Cyprinus carpio) and used in a luciferase reporter assay to elucidate its role in xenobiotic metabolism in fish. A reporter assay for human PXR (hPXR) was also established to compare transactivation between human and carp (cPXR) isoforms. Rifampicin activated hPXR as expected, but not cPXR. Conversely, clotrimazole (CTZ) activated both isoforms and was more potent on cPXR, with an EC50 within the range of concentrations of CTZ measured in the aquatic environment. Responses to other azoles tested were similar between both isoforms. A range of pharmaceuticals tested either failed to activate, or were very weakly active, on the cPXR or hPXR. Overall, these results indicate that the cPXR may differ from the hPXR in its responses and/or sensitivity to induction by different environmental chemicals, with implications for risk assessment because of species differences.


Failed medial patellofemoral ligament reconstruction: Causes and surgical strategies.
Patellar instability is a common clinical problem encountered by orthopedic surgeons specializing in the knee. For patients with chronic lateral patellar instability, the standard surgical approach is to stabilize the patella through a medial patellofemoral ligament (MPFL) reconstruction. Foreseeably, an increasing number of revision surgeries of the reconstructed MPFL will be seen in upcoming years. In this paper, the causes of failed MPFL reconstruction are analyzed: (1) incorrect surgical indication or inappropriate surgical technique/patient selection; (2) a technical error; and (3) an incorrect assessment of the concomitant risk factors for instability. An understanding of the anatomy and biomechanics of the MPFL and cautiousness with the imaging techniques while favoring clinical over radiological findings and the use of common sense to determine the adequate surgical technique for each particular case, are critical to minimizing MPFL surgery failure. Additionally, our approach to dealing with failure after primary MPFL reconstruction is also presented.


Development of Decarboxylative Cyanation Reactions for C-13/C-14 Carboxylic Acid Labeling Using an Electrophilic Cyanating Reagent.
Degradation-reconstruction approaches for isotope labeling synthesis have been known for their remarkable efficiency, but applications are scarce due to some fundamental limitations of the chemistries developed to date. The decarboxylative cyanation reaction, as a degradation-reconstruction approach, is especially useful in rapid carboxylic acid carbon isotope labeling, however development toward its application as a widespread technique has stalled at the early stages due to numerous limitations which include somewhat narrow applicability. Employing the electrophilic cyanating reagent N-cyano-N-phenyl-p-toluenesulfonamide (NCTS) as the cyano source, efficient decarboxylative cyanation chemistry has been developed for aryl and alkyl carboxylic acids respectively with two rationally designed reaction pathways. The reactions provided good yields of nitrile products from carboxylic acids, with complete retention of isotopic purity from the [13CN]-NCTS used. The reaction conditions are relatively mild requiring no oxidant and no excess toxic heavy metal and the reagent [13/14CN]-NCTS is a stable, easy-to-handle crystalline solid that can be prepared quickly and effectively from the readily available [13/14C]-KCN. The following work describes this novel and efficient method for alkyl and aryl carboxylic acid isotopic labeling using a single reagent.


Rework Potential of Soy and Pea Protein Isolates in High-Moisture Extrusion.
High-moisture extrusion (HME) is an effective process to make fibrous products that can be used as meat analogues. In this study, the effect of extrusion of already extruded products (i.e., re-extrusion) was tested with the aim to explore the potential of rework in HME. The rework of material is important because it is a route to reduce waste, which is always produced, for example during the start or at the end of a production run. Pea and soy protein isolates (PPI and SPI) were first extruded, then freeze-dried and ground, and extruded again. The visual and textural properties of the fibrous products were evaluated. Also, the rheological properties, solubility, and water-holding capacity (WHC) of the ingredients and the products after the first and second extrusion were quantified. The obtained freeze-dried powders after the first HME cycle had a reduction in solubility of 15% for PPI and 74% for SPI. Furthermore, WHC was reduced by 65% and 17% for PPI and SPI, respectively. After the second HME cycle, the reduction in solubility and WHC was augmented to 22% and 90% for PPI, and 79% and 63% for SPI. No effect on stock and loss moduli after heating and cooling were found, even after two HME cycles. SPI fibrous products did not differ in cutting strength, anisotropy index, or visual appearance after re-extrusion. Only, a decrease in hardness was detected, from 62.0 N to 51.1 N. For PPI, re-extrusion did reduce the cutting force and hardness but not the anisotropy index. It was concluded that even though HME induces a loss of solubility and WHC, this did not affect the fibrous texture formation of the protein. This means that the texture formed during HME does not depend on the process history and that rework is thus possible for fibrous products.


Quantitative measurement of oxidative damage in erythrocytes as indicator in benzene intoxications.
The metabolism of aromatic hydrocarbons by the organism forms products that cause cell death depending on the type of exposure. Benzene exposure has been linked to oxidative stress, hepatic damage, aplastic anemia, and hematopoietic cancer as lymphoid and myeloid leukemia. However, there are not fast methods to evaluate chronic benzene exposure in human blood. The objective of this work was the evaluation of the correlation between oxidative damage with benzene exposure and the level of cellular plasma membrane stability (CPMS) in erythrocytes to use it as a future indicator to determine the grade of benzene intoxications. CPMS in vitro assays were used to evaluate damage for benzene, toluene, and xylene. Erythrocytes CPMS assays in vitro shows a progressive reduction with benzene, toluene, and xylene suggesting that aromatic hydrocarbons complexity favors CPMS damage. Eight groups of Wistar rats (n = 5) were used to study the level of damage on CPMS by acute and chronic benzene administration. Enzymatic, metabolic, histological, and oxidative damage tests were performed. Acute administration (100 μL/100 g/single dose) showed a decrease of 66.7% in CPMS, while 63.6% for chronic administration (5 μL/100 g/every 2 days/3 months) showing a correlation with liver damage principally (transaminases activity increase, glycogen level decrease, and high oxidative damage). Tissue damage was observed in bone marrow, kidney, spleen, and lungs. Benzene produces damage on CPMS depending on the exposure time and dose. The CPMS technique could be used as an important aromatic hydrocarbons intoxication indicator.


Molecular markers linked to the Rf2 fertility restorer gene in cotton.
Cytoplasmic male sterility (CMS) is a maternally inherited trait in which plants do not produce viable pollen. Fertility in plants with CMS can be recovered by nuclear restorer genes. Most restorer genes cloned so far are members of the pentatricopeptide repeat (PPR) protein family. The objective of our study was to use the CMS-D8 and restoration (Rf2) system of cotton (Gossypium hirsutum L.) to develop more DNA markers for the Rf2 gene. In a backcross population with 112 plants, segregation of male fertility was 1 fertile : 1 sterile. Three new RAPD markers were identified for Rf2, one of which was converted to a CAPS marker. In addition, 2 AFLP markers and 1 SSR marker were identified to be linked to the fertility restorer gene (Rf2). PPR motif primers were designed based on the conserved PPR motifs and used in combination with AFLP primers to test the mapping population, and 1 PPR-AFLP marker was identified. A linkage map with 9 flanking markers including 1 from a previous study was constructed.


Robot-assisted laparoscopic surgery versus conventional laparoscopic surgery in randomized controlled trials: A systematic review and meta-analysis.
This review provides a comprehensive comparison of treatment outcomes between robot-assisted laparoscopic surgery (RLS) and conventional laparoscopic surgery (CLS) based on randomly-controlled trials (RCTs). We employed RCTs to provide a systematic review that will enable the relevant community to weigh the effectiveness and efficacy of surgical robotics in controversial fields on surgical procedures both overall and on each individual surgical procedure. A search was conducted for RCTs in PubMed, EMBASE, and Cochrane databases from 1981 to 2016. Among a total of 1,517 articles, 27 clinical reports with a mean sample size of 65 patients per report (32.7 patients who underwent RLS and 32.5 who underwent CLS), met the inclusion criteria. CLS shows significant advantages in total operative time, net operative time, total complication rate, and operative cost (p < 0.05 in all cases), whereas the estimated blood loss was less in RLS (p < 0.05). As subgroup analyses, conversion rate on colectomy and length of hospital stay on hysterectomy statistically favors RLS (p < 0.05). Despite higher operative cost, RLS does not result in statistically better treatment outcomes, with the exception of lower estimated blood loss. Operative time and total complication rate are significantly more favorable with CLS.


Effects of exercise therapy on painful temporomandibular disorders.
Temporomandibular disorders (TMD) are common chronic musculoskeletal pain conditions among orofacial pain. Painful TMD condition such as myalgia and arthralgia can be managed by exercise therapy. However, as it is hard to access actual effect of each modality that is included in an exercise therapy programme due to multiple choice of the management modality, their efficacy remains controversial. Therefore, this review focused on the effects of exercise therapy for the management of painful TMD. The aims of this review were to summarise the effects of exercise therapy for major symptoms of painful TMD and to establish a guideline for the management of painful TMD, resulting in higher quality and reliability of dental treatment. In this review, exercise modalities are clearly defined as follows: mobilisation exercise, muscle strengthening exercise (resistance training), coordination exercise and postural exercise. Furthermore, pain intensity and range of movements were focused as outcome parameters in this review. Mobilisation exercise including manual therapy, passive jaw mobilisation with oral appliances and voluntary jaw exercise appeared to be a promising option for painful TMD conditions such as myalgia and arthralgia. This review addressed not only the effects of exercise therapy on various clinical conditions of painful TMD shown in the past, but also an urgent need for consensus among dentists and clinicians in terms of the management of each condition, as well as terminology.


Characteristic and functional specificity of anti-human BAT (brain associated thymocyte antigen) serum.
A rabbit antiserum to human fetal brain after multiple absorption reacted with 100% of thymocytes, 55% of peripheral blood lymphocytes and 90% of enriched T lymphocytes, but not significantly with B lymphocytes. Spontaneous SRBC rosette formation was inhibited by anti-BAT pretreatment, but EAC-rosette formation remained unaffected. The antiserum was itself highly stimulatory. However, cells treated with the antiserum and complement exhibited marked inhibition of responsiveness to Con A, little effect with PHA and no alteration with PWM. The MLC reaction was inhibited only when the responder cells were treated with the antiserum and complement. Treatment of sensitized lymphocytes with the antiserum and complement caused a dose-dependent suppression of blastogenic response to both PPD and n-DNA. No effect, however, was noted in MIF producing cells. Con A induced suppressor function of lymphocytes was abolished by treatment with the antiserum and complement. These results indicate that the anti-BAT serum obtained by us can be utilized for the isolation of T lymphocyte subsets.


Perspectives on effect of spleen in ischemic stroke.
Despite decades of research, stroke therapies are limited to recanalization therapies that can only be used on <10% of stroke patients; the vast majority of stroke patients cannot be treated by these methods. Even if recanalization is successful, the outcome is often poor due to subsequent reperfusion injury. A major damage mechanism operating in stroke is inflammatory injury due to excessive pro-inflammatory cascades. Many studies have shown that, after stroke, splenic inflammatory cells, including neutrophils, monocytes/macrophages, and lymphocytes, are released and infiltrate the brain, heightening brain inflammation, and exacerbating ischemia/reperfusion injury. Clinical studies have observed spleen contraction in acute stroke patients where functional outcome improved with the gradual recovery of spleen volume. These observations are supported by stroke animal studies that have used splenectomy- or radiation-induced inhibition of spleen function to show spleen volume decrease during the acute phase of middle cerebral artery occlusion, and transfer of splenocytes to stroke-injured brain areas. Thus, activation and release of splenic cells are upstream of excessive brain inflammation in stroke. The development of reversible means of regulating splenic activity offers a therapeutic target and potential clinical treatment for decreasing brain inflammation and improving stroke outcomes.


Dual Neutral Sphingomyelinase-2/Acetylcholinesterase Inhibitors for the Treatment of Alzheimer's Disease.
We report the discovery of a novel class of compounds that function as dual inhibitors of the enzymes neutral sphingomyelinase-2 (nSMase2) and acetylcholinesterase (AChE). Inhibition of these enzymes provides a unique strategy to suppress the propagation of tau pathology in the treatment of Alzheimer's disease (AD). We describe the key SAR elements that affect relative nSMase2 and/or AChE inhibitor effects and potency, in addition to the identification of two analogs that suppress the release of tau-bearing exosomes in vitro and in vivo. Identification of these novel dual nSMase2/AChE inhibitors represents a new therapeutic approach to AD and has the potential to lead to the development of truly disease-modifying therapeutics.


Degenerative myelopathy in a German shepherd.
A 9-year-old male German Shepherd had marked stumbling, staggering and weakness in both rear limbs. Pelvic radiographs revealed only mild hip dysplasia, while survey spinal radiographs and a myelogram revealed only areas of possible pachymeningitis. Results of CSF analysis were normal. Degenerative myelopathy was suspected, but a mitogen response assay failed to confirm this diagnosis. Another mitogen response assay, performed several months later, again failed to indicate degenerative myelopathy as the cause of clinical signs. The dog's condition worsened and the animal was euthanized. At necropsy, classic histopathologic lesions of degenerative myelopathy were noted in the thoracic spinal cord.


Toward solving the folding pathway of barnase: the complete backbone 13C, 15N, and 1H NMR assignments of its pH-denatured state.
The structures of the major folding intermediate, the transition state for folding, and the folded state of barnase have been previously characterized. We now add a further step toward a complete picture of the folding of barnase by reporting the backbone 15N, 13C, and 1H NMR assignments for barnase unfolded at pH 1.8 and 30 degrees C. These assignments, which were obtained from a combination of heteronuclear magnetization transfer and backbone triple-resonance NMR experiments, constitute the first stage in the structural characterization of this denatured state by NMR. Interresidue nuclear Overhauser effect contacts and deviations from 1H random-coil chemical shifts provide evidence for stable residual structure. The structured regions span residues in the native protein that contain its major alpha-helix and central strands of the beta-sheet. Earlier experiments have shown that these regions are predominantly intact in the major folding intermediate and that their docking is partly rate determining in folding.


Primary gastrointestinal lymphoma--a clinicopathologic study.
Gastrointestinal tract is the most common site for the development ofextra nodal lymphoma. This study was done to analyze clinical and pathological features as well as the treatment outcome of this disease. We carried out analysis of25 cases of primary gastrointestinal (GI) lymphomas during period from March 2001 to February 2003 at Gujarat Cancer & Research Institute. Out of 25 cases of primary GI lymphoma, nine cases of gastric lymphoma, nine cases of small intestinal lymphoma and seven cases of large intestinal lymphoma were identified. A male to female ratio of 2.6:1 was observed. Peak incidence was observed infirst and second decades of life (range 4-63 years). Abdominal pain and abdominal lump were the two most common presenting symptoms. Diffuse large B-cell type and Burkitt's lymphoma were the most common histologic variants, accounting for equal proportions (36% each). All the patients were treated with either surgery alone or in combination of surgery, chemotherapy and radiotherapy depending on the site, stage and histology. Anti H-pylori kit was used in early stage GI maltomas. 18 cases of GI lymphoma were evaluable, and out of these, 66.6% (11 cases) attained complete remission with a median follow up time of 12 months. The disease free survival was 50% (9 cases), and the overall survival was 72.2% (13 cases). In conclusion, although there are considerable therapeutic controversies, surgery with adjuvant chemotherapy and radiotherapy yield good survival. Clinical and histopathologic characteristics and prognosis of our cases with primary gastrointestinal lymphoma were usually similar to the cases in western countries with some differences in the incidence and histologic subtypes.


Development of valid methods to measure insulin-like growth factors-I and -II in bone cell-conditioned medium.
A variety of human cells and biological fluids have been shown to produce or contain insulin-like growth factor (IGF)-specific binding proteins (BPs). The existence of these BPs in serum and conditioned medium of cell and organ cultures has complicated radioligand assays for measurement of IGFs. Various strategies have been proposed to avoid interference of BPs with these assays, including acid-ethanol precipitation of BPs and acid-gel filtration. Many of these procedures are time consuming, exhibit low recoveries, and do not completely eliminate BP artifacts. In this study we have investigated interference of inhibitory IGF-BP (In-IGF-BP) purified from bone cell-conditioned medium in an IGF-II RRA and IGF-I RIA and developed methods to neutralize In-IGF-BP artifacts in IGF assays. In the IGF-II RRA, purified In-IGF-BP competed for [125I]IGF-II binding to H-35 cells in a dose-dependent manner and, thus, increased the apparent value for IGF-II in the medium. Fifty percent inhibition of [125I]IGF-II binding to H-35 cells was seen at 12.2 and 5.7 ng/ml unlabeled IGF-II and IN-IGF-BP, respectively. In-IGF-BP also competed for [125I]IGF-I in the IGF-I RIA; however, the interference was much less in the IGF-I RIA compared to the IGF-II RRA. Fifty percent displacement of [125I]IGF-I binding was seen at 0.25 and 5 ng/ml unlabeled IGF-I and In-IGF-BP, respectively. Our approach to eliminate BP artifacts was as follows. We knew that In-IGF-BP showed comparatively equal binding affinities with both IGF-I and IGF-II, and binding of these ligands to cell receptors (IGF-II) and antibodies (IGF-I) was very specific (2% and 0.5% cross-reactivity for IGF-I and IGF-II, respectively). Therefore, in the IGF-I RIA we blocked the In-IGF-BP artifacts by adding an excess of IGF-II, and in the IGF-II RRA we blocked the In-IGF-BP artifacts by adding an excess of IGF-I. By incubating purified In-IGF-BP with different amounts of IGF-I, we found that 30-min preincubation of 5 ng In-IGF-BP with 10 ng IGF-I completely blocked BP artifacts in the IGF-II RRA. Similarly, preincubation of In-IGF-BP with IGF-II blocked BP artifacts in the IGF-I RIA.(ABSTRACT TRUNCATED AT 250 WORDS)


The simulator for icy world interiors: a 700 MPa pressure system for impulsive stimulated scattering and other optical measurements, with thermal control from -20 to 100 degrees C.
The simulator for icy world interiors (SIWI) was developed for sound velocity measurements in simulated extraterrestrial ocean materials by the method of impulsive stimulated scattering (ISS). The design and operation of SIWI are described. Optical measurements at low temperature in corrosive solution were enabled by enclosing sample fluid in a stoppered spectrophotometric cuvette. To maintain minimum optical density it was necessary to limit contact of the dioctyl sebacate hydraulic fluid with nitrile o-rings, which caused discoloring enhanced absorption of the 532 nm ISS probe pulses. Dilution of hydraulic fluid with kerosene in the amount of 5% by volume prevented clouding at high pressures and at low temperatures. Pure kerosene was found to attenuate the ISS signal, and so was deemed unsatisfactory at low temperatures where the lower thermal expansion of water leads to weaker density contrasts between heated and unheated regions in the sample fluid. When 50 cS silicone oil was used as a hydraulic medium, clouding was observed at pressures above 400 MPa when temperatures were raised above approximately 50 degrees C and lowered to less than approximately 10 degrees C. To our knowledge, such clouding has not been reported previously for dioctyl sebacate, nor for silicone at such low pressures.


A review of methods for nematode identification.
Nematodes are non-segmented roundworms found in soil, aquatic environment, plants, or animals. Either useful or pathogenic, they greatly influence environmental equilibrium, human and animal health, as well as plant production. Knowledge on their taxonomy and biology are key issues to answer the different challenges associated to these organisms. Nowadays, most of the nematode taxonomy remains unknown or unclear. Several approaches are available for parasite identification, from the traditional morphology-based techniques to the sophisticated high-throughput sequencing technologies. All these techniques have advantages or drawbacks depending on the sample origin and the number of nematodes to be processed. This review proposes an overview of all newly available methods available to identify known and/or unknown nematodes with a specific focus on emerging high-throughput molecular techniques.


Generation of Noninvasive, Quantifiable, Orthotopic Animal Models for NF2-Associated Schwannoma and Meningioma.
Schwannomas and meningiomas are nervous system tumors that can occur sporadically or in patients with neurofibromatosis type 2 (NF2). Mutations of the Neurofibromatosis 2 (NF2) gene are frequently observed in these tumors. Schwannomas and meningiomas cause significant morbidities, and an FDA-approved medical therapy is currently not available. The development of preclinical animal models that accurately capture the clinical characteristics of these tumors will facilitate the evaluation of novel therapeutic agents for the treatment of these tumors, ultimately leading to more productive clinical trials. Here, we describe the generation of luciferase-expressing NF2-deficient schwannoma and meningioma cells and the use of these cells to establish orthotopic tumor models in immunodeficient mice. The growth of these tumors and their response to treatment can be measured effectively by bioluminescence imaging (BLI) and confirmed by small-animal magnetic resonance imaging (MRI). These and other animal models, such as genetically-engineered models, should substantially advance the investigation of promising therapies for schwannomas and meningiomas.


Early coagulopathy of major burns.
The pathophysiology and time-course of coagulopathy post major burns are inadequately understood. The aims of this study were to review the incidence of acute coagulopathy post major burns, potential contributing factors associated with this coagulopathy and outcome of patients who developed early coagulopathy. A retrospective review of all patients with major burns (≥20% total body surface area (TBSA)) presenting to a tertiary burns referral centre was conducted. Data on demographic, injury characteristics and fluid resuscitation practices were recorded and tested for association with coagulopathy (INR>1.5 or aPTT>60 s) at hospital presentation and within 24 h of burns injury. Mortality, intensive care unit (ICU) admission, mechanical ventilation and blood and blood product usage were primary endpoints. There were 99 patients who met the inclusion criteria with 36 (16) %TBSA burns. Coagulopathy was present in only three patients on presentation, but 37 (37%) patients developed early onset (within 24 h of injury) coagulopathy. Early onset coagulopathy was independently associated with %TBSA burnt (p<0.001) and volume of fluid administered (p=0.005). Early onset coagulopathy was associated with higher volumes of blood and blood product administration, ICU admission and prolonged mechanical ventilation. Post major burns, a very low proportion of patients presented with coagulopathy, but a substantial proportion of patients developed coagulopathy within 24 h. This and the association of coagulopathy with the volume of fluid resuscitation suggest dilution as a major cause of the early coagulopathy of major burns.


Long-term monitoring to detect atrial fibrillation with the indwelling implantable cardiac monitors.
An increasing number of detection tools are available and several detection strategies have been described to pursue the diagnosis of atrial fibrillation to prevent ischemic stroke. Monitoring tools include standard electrocardiography, snapshot single-lead recordings with professional or personal devices (e.g. smartphone-based), Holter monitor, external devices with long-term recording capabilities, and cardiac implantable electronic devices, including pacemakers, implantable cardioverter defibrillators and insertable cardiac monitors. Insertable cardiac monitors have shown high sensitivity and specificity for the detection of atrial fibrillation, allow up to three years of continuous monitoring, do not require cooperation of the patient, are well tolerated, have a short device-related time delay between detection of atrial fibrillation and notification to the physician, provide information on atrial fibrillation burden and are minimally invasive. On the other hand, insertable cardiac monitors require a considerable use of resources to process the recordings and have a significant initial cost. Studies conducted with insertable cardiac monitors on patients with prior stroke and on patients with risk factors for stroke but no prior cerebrovascular events or atrial fibrillation have consistently shown a measurable incidence of atrial fibrillation at follow-up. However, the effectiveness of oral anticoagulations in reducing the incidence of ischemic stroke in patients with atrial fibrillation lasting less than 24 h, though reasonable, is currently unproven. The future of atrial fibrillation detection tools and atrial fibrillation detection strategies will be influenced by ongoing studies exploring whether oral anticoagulations reduce the incidence of stroke in patients with atrial fibrillation burden lower than 24 h.


[Familial nephrogenic diabetes insipidus with chronic hypernatremia and cerebral calcifications].
The authors report on the unusual evolution of two brothers suffering from nephrogenic diabetes insipidus after a 25-year follow-up. The polyuro-polydipsic syndrome was clinically well tolerated, without acute complications even during infancy. However, despite the lack of water restriction and the presence of thirst sensation, the children remained chronically hypernatremic. An intellectual deficiency was present, with a dysmorphic syndrome unrelated to a biochemically determined central nervous system disorder, or an X fragility. Calcifications of basal ganglia and frontal lobes were discovered on a cerebral computed tomography performed at 16 and 18 years. This syndrome may represent a specific entity.


Isolation and analysis of the gene and cDNA for a human Mu class glutathione S-transferase, GSTM4.
We have isolated the human gene and cDNA that encode the fourth member of the Mu class glutathione S-transferases (GSTs). The complete gene sequence (GenBank M96233) and the cDNA sequence (M96234) are reported and named GSTM4. The gene, located on chromosome 1, is comprised of eight exons with an organization similar to that of other Mu class genes. GSTM4 differs from a partial gene sequence, termed GSTmu2 (Taylor, J. B., Oliver, J., Sherrington, R., and Pemble, S. E. (1991) Biochem. J. 274, 587-593), in exons 3 through 5 by only 11 nucleotides within introns. The cDNA for GSTM4 was isolated from a library derived from the cervical carcinoma cell line, HeLa. Northern blots demonstrate the presence of GSTM4 mRNA in human heart, placenta, lung, brain, liver, skeletal muscle, pancreas, testis, cerebral cortex, uterus, ovary, a lymphoblastoid cell line, and four carcinoma cell lines. The deduced amino acid sequence of GSTM4 is 87% (GSTM1), 83% (GSTM2), and 70% (GSTM3) identical to the previously described human Mu class GSTs. The polypeptide encoded by the GSTM4 cDNA, produced in a bacterial expression system, conjugates 1-chloro-2,4-dinitrobenzene to GSH with a specific activity of 1.39 +/- 0.21 mumol/min/mg.


Quantification of long-term effects of botulinum injection in a case of cerebral palsy affecting the upper limb movement.
The aim of this work was to put into evidence the long-lasting modification induced by botulinum toxin injection and rehabilitative treatment on motor control. In this contribution, we report the case of a female child showing hemiplegia, due to cerebral palsy. She underwent botulinum injection, followed by physical and occupational therapy. We quantified the biomechanical, cerebral and occupational aspects of her impaired upper limb, also dynamically, with respect to her pre- and post-treatment condition. Small long-lasting improvements--induced on biomechanics by botulinum injection--triggered wide cerebral modification, well reflected in improved contextual movements and motor strategy. These results provide evidences that small modifications in the end-effector performance often imply cerebral modifications and improvement in finalized motor strategy.


Experiential Learning Through Local Implementation of a National Chief Resident in Quality and Patient Safety Curriculum.
The Clinical Learning Environment Review was created to evaluate quality improvement and patient safety (QIPS) beginning in 2013. Little guidance has been offered on implementing QIPS curricula for residency education. The aim was to provide a model QIPS residency curriculum from VA Boston Healthcare System (VABHS), wherein a chief resident in quality and patient safety (CRQS) participates in a national curriculum implementing skills and concepts locally. The CRQS mentors a patient safety resident with faculty oversight. The program involves case investigations, educational conferences, and experiential learning. Participants are residents from Beth Israel Deaconess Medical Center, Boston Medical Center, and Brigham and Women's Hospital and medical students from Boston University Medical School and Harvard Medical School. Local and national CRQS programs are evaluated. The patient safety rotation is evaluated locally. The local curriculum at VABHS augments the national curriculum and deploys a patient safety education that develops experiential learning skills.


Social Risk Factors and Desire for Assistance Among Patients Receiving Subsidized Health Care Insurance in a US-Based Integrated Delivery System.
Because social conditions such as food insecurity and housing instability shape health outcomes, health systems are increasingly screening for and addressing patients' social risks. This study documented the prevalence of social risks and examined the desire for assistance in addressing those risks in a US-based integrated delivery system. A survey was administered to Kaiser Permanente members on subsidized exchange health insurance plans (2018-2019). The survey included questions about 4 domains of social risks, desire for help, and attitudes. We conducted a descriptive analysis and estimated multivariate modified Poisson regression models. Of 438 participants, 212 (48%) reported at least 1 social risk factor. Housing instability was the most common (70%) factor reported. Members with social risks reported more discomfort being screened for social risks (14.2% vs 5.4%; P = .002) than those without risks, although 90% of participants believed that health systems should assist in addressing social risks. Among those with 1-2 social risks, however, only 27% desired assistance. Non-Hispanic Black participants who reported a social risk were more than twice as likely to desire assistance compared with non-Hispanic White participants (adjusted relative risk [RR] 2.2; 95% CI, 1.3-3.8). Athough most survey participants believed health systems have a role in addressing social risks, a minority of those reporting a risk wanted assistance and reported more discomfort being screened for risk factors than those without risks. Health systems should work to increase the comfort of patients in reporting risks, explore how to successfully assist them when desired, and offer resources to address these risks outside the health care sector.VISUAL ABSTRACT.


Interventions for treating depression after stroke.
Depressive and anxiety disorders following stroke are often undiagnosed or inadequately treated. This may reflect difficulties with the diagnosis of abnormal mood among older people with stroke-related disability, but may also reflect uncertainty about the effectiveness of such therapies in this setting. To determine whether pharmacological, psychological, or electroconvulsive treatment (ECT) of depression in patients with stroke can improve outcome. The Cochrane Stroke Group Trials Register (last searched June 2003). The Cochrane Central Register of Controlled Trials (The Cochrane Library, Issue 3, 2002), MEDLINE (1966 to September 2002), EMBASE (1980 to September 2002), CINAHL (1982 to September 2002), PsychINFO (1967 to September 2002), Applied Science and Technology Plus (1986 to September 2002), Arts and Humanities Index (1991 to September 2002), Biological Abstracts (1969 to September 2002), General Science Plus (1994 to September 2002), Science Citation Index (1992 to September 2002), Social Sciences Citation Index (1991 to September 2002), and Sociofile (1974 to September 2002). Reference lists from relevant articles and textbooks were searched, and authors of known studies and pharmaceutical companies who manufacture psychotropic medications were contacted. Randomised and quasi-randomised controlled trials comparing different types of pharmaceutical agents with placebo, or various forms of psychotherapy with standard care (or attention control), in patients with recent, clinically diagnosed, acute stroke, where treatment was explicitly intended of treat depression. Primary analyses focussed on the prevalence of diagnosable depressive disorder at the end of treatment. Secondary outcomes included depression or mood scores on standard scales, disability or physical function, death, recurrent stroke, and adverse effects. We did not pool the data for summary scores. We performed meta-analysis for only some binary endpoints and data on adverse events. Nine trials, with 780 participants, were included in the review. Data were available for seven trials of pharmaceutical agents, and two trials of psychotherapy. There were no trials of ECT. The analyses were complicated by the lack of standardised diagnostic and outcome criteria, and differing analytic methods. There was no strong evidence of benefit of either pharmacotherapy or psychotherapy in terms of a complete remission of depression following stroke. There was evidence of a reduction (improvement) in scores on depression rating scales, and an increase in the proportion of participants with anxiety at the end of follow up. This review found no evidence to support the routine use of pharmacotherapeutic or psychotherapeutic treatment for depression after stroke. More research is required before recommendations can be made about the most appropriate management of depression following stroke.


Phospholipase Activity of Candida Species Isolated from Diabetic Patients.
Candidiasis is one of the prevalent fungal infections caused by the genus Candida. The clinical manifestation ranges from mucocutaneous colonization to disseminated and fatal infections such as candidemia. Diabetes mellitus is one of the significant predisposing factors for this fungal infection. Candida spp. may release many exoenzymes such as phospholipase to debilitate the immune system and facilitate adherence and invasion of the fungus to the host cells. The aim of the present study is evaluation of phospholipase activity of Candida species isolated from candidemia and gastroesophageal candidiasis (GEC) among diabetic patients. Eighty-three Candida isolates were evaluated for enzyme activity by phenotypic (the precipitation zone around the colonies) and molecular methods (detection of phospholipase genes using duplex polymerase chain reaction with specific primers). Eight out of eighty-three clinical isolates (9.6%) were negative for phospholipase production. All phospholipase producers among candidemia and GEC isolates were categorized in high production group. Our findings revealed no differences in phospholipase activity among isolates obtained from different body sites (blood, oesophagus and stomach); however, non-albicans Candida species had less phospholipase activity.


Colour tuneability in sol-gel nano-glass-ceramics comprising Yb(3+)-Er(3+)-Tm3+ Co-doped NaYF4 nanocrystals.
White light can be obtained by up-conversion processes in a cheap and efficient way, being of great interest for lighting, displays, and photonic integrated devices. Here we report a generation of white light under 980 nm laser infrared excitation in Y(b3+)-Er(3+)-Tm3+ triply doped sol-gel derived nano-glass-ceramics containing NaYF4 nanocrystals where co-dopants ions are partitioned. Efficient simultaneous blue, green, and red up-conversion emissions give rise to a balanced white overall emission and corresponding up-conversion mechanisms have been analyzed. Changes in pump power intensity or heat treatment temperature widely vary the ratio of emission bands, resulting in a colour tuneability which has been analyzed and quantified in terms of CIE standard chromaticity diagram.


The endophenotype project and the validation theory: integration of neurobiology and psychiatry.
In this paper we set forth the major meta-empirical concepts for mental disorders determination. We especially focus on the methodological prerequisites and particular compounds and the endophenotype construct in a case study of bipolar disorders. It is discussed as a mediating level between the genetic causation and the environmental influences in the mental disorder causation. This epigenetic area includes such constructs as cognitive neuropsychological markers, neurotrophic proteins impairment, instability of circadian rhythms (other chronobiological disturbances as well), deregulation of the motivation and reward mechanisms, changes in the GABA metabolism, symptom provocation with "psychotomymetic" agents, etc. Eventually the core ideas of the original validation theory in comparison with the endophenotype concept are shown as milestones of the novel integration of neurobiology and psychiatry. The major issue posed in the endophenotype debate concerns "the independence from the state". According to the formulation of Hassler and Drevets this notion indicates the presentation of the endophenotype regardless of the activity of the disorder, i.e., its presence even in the pre-morbid and remission periods. In the present study we argue that there is another dimension of "state independence" to be assessed. The validation theory in particular defines the entire space-and-time sequence disposition of the endophenotype indicators, e.g. cognitive-psychological and neuroimaging. It is well known that mental states, whether normal or pathological, are very dynamical, just as their neurophysiological correlates are. By this reason it is necessary to explore their relationships simultaneously.


CD20 as a target for therapeutic type I and II monoclonal antibodies.
The last decade has seen the monoclonal antibody (mAb), rituximab, transform clinical management of many non-Hodgkin lymphomas and more recently provide new opportunities for controlling autoimmune conditions, such as rheumatoid arthritis. Although not yet fully determined, the explanation for this success appears to lie with the inherent properties of its target, CD20, which allow rituximab to recruit potent cytotoxic effectors with unusual efficiency. In this review we detail the properties of CD20 that make it such an effective therapeutic target and describe how different mAbs change the membrane distribution and internalization of CD20 and have distinct modes of cytotoxic activity.


Chemical composition and biological activities of Calamintha officinalis Moench essential oil.
Calamintha officinalis Moench essential oil is used in cooking as an aromatic herb and also to improve the flavor and fragrance of several pharmaceutical products. The essential oil, obtained by hydrodistillation (5 mL/kg), was analyzed by gas chromatography-mass spectrometry and gas chromatography-flame ionization detection. Sixty-four components were identified, constituting 99.7% of the total oil. The major component was found to be carvone (38.7%), followed by neo-dihydrocarveol (9.9%), dihydrocarveol acetate (7.6%), dihydrocarveol (6.9%), 1,8 cineole (6.4%), cis-carvyl acetate (6.1%), and pulegone (4.1%). The essential oil showed antifungal and antimicrobial activity against Gram-positive bacteria. In addition, it presented a very low toxicity both in vivo (50% lethal dose >100 mg/kg) and in vitro in the Artemia salina test (50% lethal concentration >500 μL/mL). C. officinalis essential oil, in rodents, produces the typical effects in behavior of a nonselective central nervous system-depressant drug; it potentiates the hypnotic effects of sodium pentobarbital, decreasing the induction time and enhancing the sleeping time. Moreover, it produces a decrease in body temperature and a protection against pentylenetetrazole-induced convulsions.


Update: Exertional hyponatremia, active component, U.S. Armed Forces, 2001-2016.
From 2001 through 2016, there were 1,519 incident diagnoses of exertional hyponatremia among active component service members (incidence rate: 6.9 cases per 100,000 person-years [p-yrs]). The incidence rate in 2016 (6.6 cases per 100,000 p-yrs) represented a decrease of 23.3% from 2015. Compared to their respective counterparts, overall incidence rates of exertional hyponatremia were higher among females, those aged 19 years or younger, and recruit trainees. The overall incidence rate during the surveillance period was highest in the Marine Corps, intermediate in the Army and Air Force, and lowest in the Navy. Overall incidence rates were lowest among black, non-Hispanic service members and highest among white, non-Hispanic and Asian/Pacific Islander service members. Service members (particularly recruit trainees) and their supervisors must be vigilant for early signs of heat-related illnesses and must be knowledgeable of the dangers of excessive water consumption and the prescribed limits for water intake during prolonged physical activity (e.g., field training exercises, personal fitness training, recreational activities) in hot, humid weather.


Outbreak of ampicillin/piperacillin-resistant Klebsiella pneumoniae in a neonatal intensive care unit (NICU): investigation and control measures.
Klebsiella pneumoniae is a frequent cause of infectious outbreaks in Neonatal Intensive Care Units (NICUs). The aim of this paper is to describe an outbreak occurred in a 13-bed NICU and the control measures adopted in order to interrupt the chain of transmission. We described the microbiological investigations, the NICU staff compliance to the infection control measures by means of a specifically designed check-list and the control measures adopted. Six cases of primary bloodstream infections sustained by ampicillin/piperacillin-resistant Klebsiella pneumoniae were observed over a two-month period. One culture obtained from a 12% saccarose multiple-dose solution allowed the growth of Klebsiella pneumoniae. During the inspections performed by the Hospital Infection Control Team, using the check-list for the evaluation of the NICU staff compliance to the infection control measures, several breaches in the infection control policy were identified and control measures were adopted. In our case the definition of a specific check-list led to the adoption of the correct control measures. Further studies would be helpful in order to develop a standard check-list able to identify critical flows in the adhesion to the guidelines. It could be used in different NICUs and allow to obtain reproducible levels of infection control.


Developmental changes of BKCa channels depend on differentiation status in cultured podocytes.
The podocyte is a remarkable cell type, which encases the capillaries of the kidney glomerulus. Podocytes are of keen interests because of their key roles in kidney development and disease. Large-conductance Ca(2+)-activated K(+) channels (BKCa channels) are important ion channels located in podocytes and play the essential role in regulating calcium homeostasis cell signaling. In this research, we studied the undergoing developmental changes of BKCa channels and their contribution to functional maturation of podocytes. Our results showed that the distribution of BKCa channels changed with the maturity of differentiation in a conditionally immortalized mouse podocyte cell line. Additionally, the increase of BKCa channel protein expression was detected by immunofluorescence staining with confocal microscopy in podocytes, which was consistent with the increase in the current density of BKCa channels examined by whole-cell patch-clamp technique. Our results suggested that the developmental changes of BKCa channels may help podocytes adapt to changes in pressure gradients occurring in physiological conditions. Those findings may have implications for understanding the physiology and development of kidney and will also serve as a baseline for future studies designed to investigate developmental changes of ion channel expression in podocytes.


Ecotoxicologic impacts of agricultural drain water in the Salinas River, California, USA.
The Salinas River is the largest of the three rivers that drain into the Monterey Bay National Marine Sanctuary in central California (USA). Large areas of this watershed are cultivated year-round in row crops, and previous laboratory studies have demonstrated that acute toxicity of agricultural drain water to Ceriodaphnia dubia is caused by the organophosphate (OP) pesticides chlorpyrifos and diazinon. We investigated chemical contamination and toxicity in waters and sediments in the river downstream of an agricultural drain water input. Ecological impacts of drain water were investigated by using bioassessments of macroinvertebrate community structure. Toxicity identification evaluations were used to characterize chemicals responsible for toxicity. Salinas River water downstream of the agricultural drain was acutely toxic to the cladoceran Ceriodaphnia dubia, and toxicity to C. dubia was highly correlated with combined toxic units (TUs) of chlorpyrifos and diazinon. Laboratory tests were used to demonstrate that sediments in this system were acutely toxic to the amphipod Hyalella azteca, a resident invertebrate. Toxicity identification evaluations (TIEs) conducted on sediment pore water suggested that toxicity to amphipods was due in part to OP pesticides; concentrations of chlorpyrifos in pore water sometimes exceeded the 10-d mean lethal concentration (LC50) for H. azteca. Potentiation of toxicity with addition of the metabolic inhibitor piperonyl butoxide suggested that sediment toxicity also was due to other non-metabolically activated compounds. Macroinvertebrate community structure was highly impacted downstream of the agricultural drain input, and a number of macroinvertebrate community metrics were negatively correlated with combined TUs of chlorpyrifos and diazinon, as well as turbidity associated with the drain water. Some macroinvertebrate metrics were also correlated with bank vegetation cover. This study suggests that pesticide pollution is the likely cause of ecological damage in the Salinas River, and this factor may interact with other stressors associated with agricultural drain water to impact the macroinvertebrate community in the system.


Acquisition of habitual visual attention and transfer to related tasks.
Extensive research has shown that statistical learning affects perception, attention, and action control; however, few studies have directly linked statistical learning with the formation of habits. Evidence that learning can induce a search habit has come from location probability learning, in which people prioritize locations frequently attended to in the past. Here, using an alternating training-testing procedure, we demonstrated that the initial attentional bias arises from short-term intertrial priming, whereas probability learning takes longer to emerge, first reaching significance in covert orienting (measured by reaction times) after about 48 training trials, and in overt orienting (measured by eye movements) after about 96 training trials. We further showed that location probability learning is persistent after training is discontinued, by transferring from a letter search task to a scene search task-emulating another characteristic feature of habits. By identifying the onset of probability learning and investigating its task specificity, this study provides evidence that probability cuing can induce habitual spatial attention.


Noise exposure during alpine helicopter rescue operations.
We estimated the noise exposure of crews working in alpine helicopter rescue systems. Noise levels of the the helicopters used (Alouette III, Alouette II 'Lama', Ecureuil and BK 117) were measured with a device according to class 2 DIN IEC 651. These data were combined with the flight data of the personnel to evaluate the equivalent noise level according to DIN 45645-2. While the risk to patients should be limited to temporary threshold shifts the crew members are regularly exposed to equivalent noise levels of >85 dB(A) and, therefore, are at risk of permanent threshold shifts. Consequences for crew fitness to fly and for noise prevention (crew and patients) are discussed.


Immunomodulatory Nonstructural Proteins of Influenza A Viruses.
Influenza epidemics and pandemics still represent a severe public health threat and cause significant morbidity and mortality worldwide. As intracellular parasites, influenza viruses are strongly dependent on the host cell machinery. To ensure efficient production of progeny viruses, viral proteins extensively interfere with cellular signalling pathways to inhibit antiviral responses or to activate virus-supportive functions. Here, we review various functions of the influenza virus nonstructural proteins NS1, PB1-F2, and PA-X in infected cells and how post-transcriptional modifications of these proteins affect the viral life cycle. Furthermore, we discuss newly discovered interactions between these proteins and the antiviral interferon response.


Evidence for rheumatic valve disease in patients with severe tricuspid regurgitation long after mitral valve surgery: the role of 3D echo reconstruction.
Although severe tricuspid regurgitation (TR) is a well-recognized, long-term complication of rheumatic mitral valve replacement that impairs the functional results of surgery, its exact basis remains unclear and its management is unsatisfactory. The study aim was to obtain a detailed assessment of tricuspid valve morphology and function using 2D transesophageal echocardiography (TEE) with 3D reconstruction, and to determine long-term clinical outcome in patients after surgery for rheumatic mitral valve disease. A total of 42 patients (mean age 50 +/- 10 years) was followed up; 39 patients had mitral replacement and three had valvotomy. Thirty patients had developed impaired exercise tolerance, fluid retention and echocardiographic evidence of severe TR at 8.2 +/- 2.6 years after surgery; the remainder had mild regurgitation. Follow up showed greater mortality in the severe TR group, with approximately 50% survival at 60 months after diagnosis compared with mild TR. None of the patients with severe TR had a dysfunctional mitral prosthesis. In these patients, transthoracic echo-Doppler showed enlarged right atrium and right ventricle, a mean transtricuspid retrograde pressure drop of 15 +/- 4 mmHg and apparently normal leaflet anatomy. Twenty patients (15 with severe TR) underwent a TEE and 3D reconstruction study for further evaluation. Abnormal leaflet anatomy was demonstrated in all patients with severe TR, with restricted leaflet motion in 10, leaflet shortening and thickening in the remainder, and dilatation of tricuspid valve annular insertion suggestive of rheumatic involvement. Although diastolic transtricuspid velocities were increased (peak flow 0.8 +/- 0.1 m/s) in these patients due to increased stroke volume, significant tricuspid stenosis was present in only two cases (mean gradient 4 and 3 mmHg respectively). Histopathology confirmed the presence of leaflet vascularization and extensive fibrosis in two patients who underwent tricuspid valve replacement. Rheumatic leaflet involvement contributes to severe TR occurring long after mitral valve replacement, though overt stenosis is uncommon. Knowledge of the structural basis of this condition may thus improve its long-term management, possibly with early tricuspid valve repair.


k-Nearest neighbors optimization-based outlier removal.
Datasets of molecular compounds often contain outliers, that is, compounds which are different from the rest of the dataset. Outliers, while often interesting may affect data interpretation, model generation, and decisions making, and therefore, should be removed from the dataset prior to modeling efforts. Here, we describe a new method for the iterative identification and removal of outliers based on a k-nearest neighbors optimization algorithm. We demonstrate for three different datasets that the removal of outliers using the new algorithm provides filtered datasets which are better than those provided by four alternative outlier removal procedures as well as by random compound removal in two important aspects: (1) they better maintain the diversity of the parent datasets; (2) they give rise to quantitative structure activity relationship (QSAR) models with much better prediction statistics. The new algorithm is, therefore, suitable for the pretreatment of datasets prior to QSAR modeling.


Fungal Screening on Olive Oil for Extracellular Triacylglycerol Lipases: Selection of a Trichoderma harzianum Strain and Genome Wide Search for the Genes.
A lipolytic screening with fungal strains isolated from lignocellulosic waste collected in banana plantation dumps was carried out. A Trichoderma harzianum strain (B13-1) showed good extracellular lipolytic activity (205 UmL-1). Subsequently, functional screening of the lipolytic activity on Rhodamine B enriched with olive oil as the only carbon source was performed. The successful growth of the strain allows us to suggest that a true lipase is responsible for the lipolytic activity in the B13-1 strain. In order to identify the gene(s) encoding the protein responsible for the lipolytic activity, in silico identification and characterization of triacylglycerol lipases from T. harzianum is reported for the first time. A survey in the genome of this fungus retrieved 50 lipases; however, bioinformatic analyses and putative functional descriptions in different databases allowed us to choose seven lipases as candidates. Suitability of the bioinformatic screening to select the candidates was confirmed by reverse transcription polymerase chain reaction (RT-PCR). The gene codifying 526309 was expressed when the fungus grew in a medium with olive oil as carbon source. This protein shares homology with commercial lipases, making it a candidate for further applications. The success in identifying a lipase gene inducible with olive oil and the suitability of the functional screening and bioinformatic survey carried out herein, support the premise that the strategy can be used in other microorganisms with sequenced genomes to search for true lipases, or other enzymes belonging to large protein families.


Acute Coronary Syndrome: Current Treatment.
Acute coronary syndrome continues to be a significant cause of morbidity and mortality in the United States. Family physicians need to identify and mitigate risk factors early, as well as recognize and respond to acute coronary syndrome events quickly in any clinical setting. Diagnosis can be made based on patient history, symptoms, electrocardiography findings, and cardiac biomarkers, which delineate between ST elevation myocardial infarction and non-ST elevation acute coronary syndrome. Rapid reperfusion with primary percutaneous coronary intervention is the goal with either clinical presentation. Coupled with appropriate medical management, percutaneous coronary intervention can improve short- and long-term outcomes following myocardial infarction. If percutaneous coronary intervention cannot be performed rapidly, patients with ST elevation myocardial infarction can be treated with fibrinolytic therapy. Fibrinolysis is not recommended in patients with non-ST elevation acute coronary syndrome; therefore, these patients should be treated with medical management if they are at low risk of coronary events or if percutaneous coronary intervention cannot be performed. Post-myocardial infarction care should be closely coordinated with the patient's cardiologist and based on a comprehensive secondary prevention strategy to prevent recurrence, morbidity, and mortality.


Bacterial resistance mechanisms as therapeutic targets.
In the 50 years since antimicrobial agents were first introduced, bacteria have acquired a wide variety of mechanisms which have enabled them to resist the effects of these drugs. One way of overcoming this problem is to administer an antibiotic with an agent which counteracts the mechanism of resistance to that antibiotic; an example of such an approach which has already been successfully implemented is the combination of a beta-lactam antibiotic with a beta-lactamase inhibitor. This review describes antibiotic resistance mechanisms which might lend themselves to an inhibitor approach and the potential therapeutic applications of such a strategy.


Epidemiologic study of bluetongue viruses in Central America and the Caribbean: 1986-1988. Regional Bluetongue Team.
Results of a prospective serologic and virologic study of ruminant livestock in Central America and the Caribbean islands revealed bluetongue virus (BTV) to be enzootic in the 9 countries participating in the study. Bluetongue virus serotypes 1, 3, 6, and 12 were isolated from sentinel animals. To the authors' knowledge, these are the first isolations of BTV from the region studied and the first isolations of these serotypes in the Western Hemisphere. Clinical disease attributable to BTV infection was not observed in sentinel animals. The incidence pattern, with respect to age and geographic location, was determined. The need to evaluate the epizootiologic features or arthropod-borne viruses (arboviruses) on a regional ecologic basis is stressed.


[The origin of critical medicine and its expansion in Venezuela].
A historical review on the evolution of the Critical Medicine, from its origin in Scandinavian, 1952, and development in other countries of Europe and the United States, to its introduction in Venezuela, 1965, and later acknowledgement as a medical specialty by the Venezuelan Federation of Physicians is presented after fixing its synonymous in different countries.


Optimization of polyhydroxybutyrate production by mixed cultures submitted to aerobic dynamic feeding conditions.
Activated sludge submitted to aerobic dynamic feeding conditions showed a good and stable capacity to store polyhydroxybutyrate (PHB). The system, working for 2 years, selected a microbial population with a high PHB storage capacity. The influence of carbon and nitrogen concentrations on the PHB accumulation yield was studied in a range of 15-180 Cmmol/l for acetate and 0-2.8 Nmmol/l for ammonia. Low ammonia concentrations favored PHB accumulation. The maximum PHB content, 67.5%, was obtained for 180 Cmmol/l of acetate supplied in one pulse. However, such high substrate concentration proved to be inhibitory for the storage mechanism, causing a slowdown of the specific PHB storage rate. In order to avoid substrate inhibition, 180 Cmmol/l of acetate was supplied in different ways: continuously fed and in three pulses of 60 Cmmol/l each. In both cases the specific PHB storage rate increased and the PHB content obtained were 56.2% and 78.5%, respectively. The latter value of PHB content is similar to that obtained by pure cultures and was never reported for mixed cultures. Addition of acetate by pulses controlled by the oxygen concentration was kept for 16 days, the PHB content being always above 70% of cell dry weight.


Synthesis, biological activities, and future perspectives of steroidal monocyclic pyridines.
Steroidal pyridines are a class of compounds that have been the subject of extensive research in recent years due to their potential biological activities. The introduction of a pyridine ring into the steroid skeleton can significantly alter the chemical and biological properties of the compound, making it more potent and/or selective for a particular target. Different synthetic methods have been developed for the preparation of steroidal pyridines. This review provides an overview of the synthesis, biological activities, and future perspectives of steroidal monocyclic dihydropyridines, tetrahydropyridines, and pyridines from 2005 to the present. The different synthetic methods that have been developed for the preparation of these steroids are discussed, as well as the proposed mechanisms and the biological activities that have been reported. Finally, the potential of steroidal monocyclic pyridines for the development of new drugs is discussed. This review is intended to provide a comprehensive overview of the field of steroidal monocyclic pyridines for researchers and scientists who are interested in this area of research. It is also hoped that this review will stimulate further research into the synthesis and biological activities of steroidal pyridines to develop new and improved drugs for the treatment of diseases.


Cloning of an E. coli RecA and yeast RAD51 homolog, radA, an allele of the uvsC in Aspergillus nidulans and its mutator effects.
An E. coli RecA and yeast RAD51 homolog from Aspergillus nidulans, radA, has been cloned by screening genomic and cDNA libraries with a PCR-amplified probe. This probe was generated using primers carrying the conserved sequences of eukaryotic RecA homologs. The deduced amino acid sequence revealed two conserved Walker-A and -B type nucleotide-binding domains and exhibited 88%, 60%, and 53% identity with Mei-3 of Neurospora crassa, rhp51+ of Schizosaccharomyces pombe, and Rad51 of Saccharomyces cerevisiae, respectively. radA null mutants constructed by replacing the whole coding region with a selection marker showed high methyl methanesulfonate (MMS) sensitivity. Heterozygous diploids of radA disruptant with the uvsC114 mutant failed to complement with respect to MMS-sensitivity, indicating that radA is an allele of uvsC. In selecting spontaneous forward selenate resistant mutations, mutator effects were observed in radA null mutants similarly to those shown in uvsC114 mutant strains.


Pravastatin at 10 mg/day does not decrease plasma levels of either amyloid-beta (Abeta) 40 or Abeta 42 in humans.
It has been assumed that statins work as a preventative drug for Alzheimer's disease (AD). Although some epidemiological observations raise doubts to the effectiveness of statins for AD, many in vitro and clinical studies insist on the effectiveness of statins decreasing amyloid-beta (Abeta) levels in medium or blood. To explore the effect of pravastatin on Abeta production, we followed the longitudinal plasma levels of both Abeta 40 and Abeta 42 during the allocation of pravastatin in 46 patients with hyperlipidemia. We found no correlation between plasma cholesterol levels or the decreasing values of total cholesterol and those of Abeta 40 or Abeta 42. Patients having Apolipoprotein E4 (ApoE4) had higher low-density lipoprotein levels and lower Abeta 40 levels in plasma, suggesting ApoE4 seems to influence plasma Abeta levels via cholesterol metabolism.


Concussion education perceptions among secondary school athletic trainers.
Describe concussion education perceptions among secondary school athletic trainers and determine the relationship between education program characteristics and perceived effectiveness. Two hundred and three participants completed at least one survey item (age = 35.2 ± 9.4 years; male = 27.6%). We used descriptive statistics to describe concussion education characteristics and point out biserial correlations to determine if relationships existed between concussion education characteristics and perceived effectiveness. The most frequently used mandated concussion education programs were informational handouts created by the state interscholastic association, the Centers for Disease Control and Prevention's (CDC) Heads Up materials, and the National Federation of State High School Athletic Association's materials. The CDC's Heads Up materials, handouts created by the state interscholastic association, and in-person presentations created by respondents were the most frequent concussion educational programs utilized by respondents who indicated no specific program mandated. Educational programs that were engaging increased perceived effectiveness (n = 131, rpb = 0.31,p < 0.001). Our results highlight that materials created by state interscholastic association and the CDC's Heads Up program among others are utilized to satisfy mandates and are perceived to be moderately effective, especially when perceived to be engaging. These results can help guide future studies to examine specific educational tools and administration strategies to determine effectiveness on concussion disclosure.


T lymphocyte cytokine production patterns in hashimoto patients with elevated calcitonin levels and their relationship to tumor initiation.
The aim of the study was to evaluate the possible changes in CD4+ and CD8+ T-cell cytokine production patterns in Hashimoto's thyroiditis (HT) with elevated calcitonin (CT). Fourteen consecutive patients with verified HT were included in the present study. Patients were divided into two groups. Group I: 7 HT patients with elevated CT levels (>10 pg/ml); Group II: 7 HT patients with CT levels <10 pg/ml). All patients underwent intracellular cytokine detection in CD4+ and CD8+ T-cells of peripheral blood mononuclear cells (PBMC) by flow cytometry. Patients with elevated CT levels (group I) had significantly higher percentages of CD8+ cells producing IFN-gamma compared to healthy donors. A detailed analysis of cytokine production patterns revealed that this was accompanied by an increased frequency of single IFN-gamma positive cells, i.e., cells not expressing most of the other cytokines tested. Similarly, patients in group I also showed higher percentages of CD8+ TNF-alpha positive cells than healthy donors. In this case, cells co-expressing TNF-alpha and IFN-gamma were found at significantly higher frequencies. No increase in Th1 type cytokines, such as IFN-gamma or TNF-alpha, was detectable in CD4+ T-cells. In contrast, CD4+ T-cells from group I patients showed significantly higher percentages of cells producing Th2 cytokines, such as IL-4 or IL-13. The lack of increased Th1 cytokine production accompanied by an increased Th2 cytokine production seems to be a special feature of HT patients with elevated CT levels that may reflect a pathogenetic mechanism for tumor initiation.


Autonomic nervous system modulation and clinical outcome after pulmonary vein isolation using the second-generation cryoballoon.
The intrinsic cardiac autonomic nervous system (ANS) plays a significant role in atrial fibrillation (AF) mechanisms. This study evaluated the incidence and impact of intraprocedural vagal reactions and ANS modulation by pulmonary vein isolation (PVI) using second-generation cryoballoons on outcomes. One hundred three paroxysmal AF patients underwent PVI with one 28-mm second-generation balloon. The median follow-up was 15.0 (12.0-18.0) months. ANS modulation was defined as a >20% cycle length decrease on 3-minute resting electrocardiograms at 1, 3, 6, and 12 months postindex procedure relative to baseline if sinus rhythm was maintained. Marked sinus arrests/bradycardia and atrioventricular block (intraprocedural vagal reaction) occurred in 14 and 2 patients, and all sinus arrest/bradycardia occurred in 44 patients with left superior pulmonary veins (PVs) targeted before right PVs. ANS modulation was identified in 66 of 95 (69.5%) patients, and it persisted 12-month postprocedure in 36 (37.9%) patients. Additional β-blocker administration was required in 9 patients for sinus tachycardia. ANS modulation was similarly observed in patients with and without intraprocedural vagal reactions (P = 0.443). Forty-eight (46.6%) patients experienced early recurrences, and the single procedure success at 12 months was 72.7%. Neither intraprocedural vagal reactions nor ANS modulation predicted AF freedom within or after the blanking period. Thirty-three patients underwent second procedures, and reconnections were detected in 39 of 130 (30.0%) PVs among 23 (69.7%) patients. The incidence of reconnections was similar in patients with and without ANS modulation. Increased heart rate persisted in 37.9% of patients even at 12-month post-second-generation cryoballoon PVI. Neither intraprocedural vagal reactions nor increased heart rate predicted a single procedure clinical outcome.


Systematic review of the literature on triclosan and health outcomes in humans.
The ability of epidemiologic evidence to inform regulatory decisions is largely dependent on the coherence and quality of the published literature. This systematic review examines the quality and consistency of studies assessing health outcomes associated with exposure to triclosan (TCS), an antimicrobial chemical with a short physiologic half-life. We used elements of the Biomonitoring, Environmental Epidemiology, and Short-Lived Chemicals instrument to evaluate aspects of study quality. Each methodological domain - overall design, exposure assessment, and data analysis - was categorized according to three tiers where Tier 1 indicated the highest quality. We also examined consistency of methods, results and reporting as considerations for weight of evidence (WOE) assessment. Studies were considered sufficiently comparable if they addressed the same or similar research questions. Forty-two studies met the criteria for inclusion. Only one randomized cross-over clinical trial of TCS was assigned to Tier 1 for all three domains. Most other studies were assigned to Tier 3 for at least one domain. Although the available literature examined more than 100 different health endpoints and reported hundreds of different measures of association, few studies were considered comparable. For reported measures of association, most were not significantly different from the null; the few statistically significant results represented isolated findings without a discernable across- or within-study pattern. We conclude that the current body of epidemiologic literature does not allow a meaningful WOE assessment due to methodological limitations of individual studies and lack of inter-study consistency. On the other hand, methodologically stronger studies may be used to inform future research.


Is receptor activation involved in the mechanism by which (+)-fenfluramine and (+)-norfenfluramine deplete 5-hydroxytryptamine in the rat brain?
1 The effects of (+)-fenfluramine, (+)-norfenfluramine and reserpine on the concentrations of 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in brainstem and telencephalon were studied in rats treated with methergoline, a 5-HT antagonist. 2 Methergoline significantly reduced the effect of (+)-norfenfluramine (5 mg/kg) on 5-HT levels in telencephalon and brainstem but did not modify the effect of (+)-norfenfluramine (2.5 mg/kg). 3 Neither the effect of (+)-fenfluramine on 5-HT levels nor the decrease of 5-HT metabolism caused by (+)-fenfluramine and (+)-norfenfluramine was significantly modified by methergoline treatment. 4 Methergoline potentiated the effects of reserpine on brain indoles. The effects was particularly evident on 5-HIAA levels in the brainstem, although significant effects were found on 5-HT in the brainstem and 5-HIAA in the telencephalon depending on the dose reserpine used. 5 The results show that postsynaptic receptor activation may partially contribute to the depletion of brain 5-HT caused by (+)-norfenfluramine in the rat. This mechanism does not seem to play a significant role in the effect of (+)-fenfluramine.


Evaluation of Preexisting Anti-Hemagglutinin Stalk Antibody as a Correlate of Protection in a Healthy Volunteer Challenge with Influenza A/H1N1pdm Virus.
Influenza virus hemagglutinin (HA) surface glycoprotein is currently the primary target of licensed influenza vaccines. Recently, broadly reactive antibodies that target the stalk region of the HA have become a major focus of current novel vaccine development. These antibodies have been observed in humans after natural infection with influenza A virus, but the data are limited. Using samples and data from the uniquely controlled setting of an influenza A/H1N1 virus human challenge study of healthy volunteers, we performed a secondary analysis that for the first time explores the role of anti-HA stalk antibody as a human correlate of protection. An anti-HA stalk antibody enzyme-linked immunosorbent assay (ELISA) was performed on samples from 65 participants challenged with a 2009 H1N1pdm virus. Pre- and postchallenge anti-HA stalk titers were then correlated with multiple outcome measures to evaluate anti-HA stalk antibody titer as a correlate of protection. Anti-HA stalk antibody titers were present before challenge and rose in response to challenge in 64% of individuals. Those individuals with higher titers at baseline were less likely to develop shedding, but not less likely to develop symptoms. Similar to the hemagglutination inhibition (HAI) titer, the baseline anti-HA stalk antibody titer did not independently predict a decrease in the severity of influenza disease, while the antineuraminidase (neuraminidase inhibition [NAI]) titer did. As a correlate of protection, the naturally occurring anti-HA stalk antibody titer is predictive of a reduction of certain aspects of disease similar to HAI titer, but the NAI titer is the only identified correlate that is an independent predictor of a reduction of all assessed influenza clinical outcome measures.IMPORTANCE This is the first study to evaluate preexisting anti-HA stalk antibodies as a predictor of protection. We use a healthy volunteer influenza challenge trial for an examination of the role such antibodies play in protection. This study demonstrates that anti-HA stalk antibodies are naturally generated in response to an infection, but there is significant variability in response. Similar to antibodies that target the HA head, baseline anti-HA stalk antibody titer is a correlate of protection in terms of reduced shedding, but it is not a predictor of reduced clinical disease or an independent predictor of disease severity. These results, in the context of the limited data available in humans, suggest that vaccines that induce anti-HA stalk antibodies could play a role in future vaccine strategies, but alone, this target may be insufficient to induce a fully protective vaccine and overcome some of the issues identified with current vaccines.


Intracytoplasmic sperm injection after total conventional in-vitro fertilization failure.
This study assessed the value of delayed intracytoplasmic sperm injection after failure of initial in-vitro fertilization in patients with normal seminology. A case series analysis was conducted on 82 couples who underwent delayed intracytoplasmic sperm injection after failure of initial in-vitro fertilization at Fakeeh Hospital, Jeddah, Kingdom of Saudi Arabia between January 1995 and January 2001. Seventy-eight couples had 616 unfertilized metaphase II mature oocytes after initial in-vitro fertilization. Second day "rescue" reinsemination by intracytoplasmic sperm injection (ICSI) resulted in 174 embryos that were suitable for transfer. This resulted in 4 clinical pregnancies (5.1% per started rescue ICSI cycle). The limited success rate in this study provides confirmatory data that rescue ICSI is not an efficient adjuvant laboratory procedure.


Contribution of the p38MAPK signalling pathway to proliferation in human cultured airway smooth muscle cells is mitogen-specific.
We have investigated the role of p38MAPK in human airway smooth muscle (HASM) proliferation in response to thrombin and bFGF. The regulation of cyclin D1 mRNA, cyclin D1, cyclin E and p21Cip1 protein levels, and the extent of retinoblastoma protein (pRb) phosphorylation in response to activation of p38MAPK have also been examined. Two distinct inhibitors of p38MAPK, SB 203580 (10 microm) and SB 202190 (10 microm), prevented bFGF (0.3-3 nm)-stimulated cell proliferation, but had no effect on the response to thrombin (0.3-3 U ml(-1)). In cells incubated with thrombin or bFGF for 20 h, there was an increase in p38MAPK phosphorylation in response to bFGF, but not to thrombin. Thrombin and bFGF-stimulated increases in ERK phosphorylation and cyclin D1 mRNA and protein levels were not influenced by SB 203580 pre-treatment. Similarly, cyclin E and p21Cip1 protein levels, measured after 20 h incubation with mitogen, did not appear to be regulated by SB 203580 (10 microm). Although both thrombin and bFGF significantly increased levels of pRb phosphorylation, SB 203580 (10 microm) inhibited only bFGF-stimulated pRb phosphorylation. In addition, SB 203580 (10 microm) selectively inhibited bFGF-stimulated DNA synthesis, suggesting that the antimitogenic actions of SB 203580 on pRb phosphorylation cause cell cycle arrest at late G1 phase. In conclusion, these results indicate that p38MAPK is involved in bFGF-, but not in thrombin-stimulated HASM proliferation. The activation of the p38MAPK pathway by bFGF, but not by thrombin, regulates the phosphorylation of pRb without influencing cyclin D1 expression.


Effects of quinidine and diphenylhydantoin on membrane resistance in smooth muscle.
The effects of quinidine and diphenylhydantoin on the membrane electrical resistance of guinea-pig taenia coli have been studied by means of the double sucrose gap tachnique. At concentrations ranging from 2 to 7.10-minus 5 quinidine induces an evident dose-related increase of membrane resistance, as indicated by the increase of the electrotonic potential. These effects are unaffected by tetrodotoxin (10(-6)) and in the presence of various changes of ionic environment (replacement of Na by Li, increase of K or Ca or Mg concentrations). Only a concomitant rise of both K and Ca (or Mg) can prevent the effects of quinidine on membrane resistance. The rate of repetitive discharge under sustained electrical depolarization is not affected by quinidine, at concentrations increasing membrane resistance. The experiments with diphenylhydantoin showed that this drug is practically ineffective on membrane resistance, but induces a decrease of the rate of repetitive discharge under sustained depolarization.


Biomechanical Regulation of Endothelium-dependent Events Critical for Adaptive Remodeling.
Alterations in hemodynamic shear stress acting on the vascular endothelium are critical for adaptive arterial remodeling. The molecular mechanisms regulating this process, however, remain largely uncharacterized. Here, we sought to define the responses evoked in endothelial cells exposed to shear stress waveforms characteristic of coronary collateral vessels and the subsequent paracrine effects on smooth muscle cells. A lumped parameter model of the human coronary collateral circulation was used to simulate normal and adaptive remodeling coronary collateral shear stress waveforms. These waveforms were then applied to cultured human endothelial cells (EC), and the resulting differences in EC gene expression were assessed by genome-wide transcriptional profiling to identify genes distinctly regulated by collateral flow. Analysis of these transcriptional programs identified several genes to be differentially regulated by collateral flow, including genes important for endothelium-smooth muscle interactions. In particular, the transcription factor KLF2 was up-regulated by the adaptive remodeling coronary collateral waveform, and several of its downstream targets displayed the expected modulation, including the down-regulation of connective tissue growth factor. To assess the effect of endothelial KLF2 expression on smooth muscle cell migration, a three-dimensional microfluidic assay was developed. Using this three-dimensional system, we showed that KLF2-expressing EC co-cultured with SMC significantly reduce SMC migration compared with control EC and that this reduction can be rescued by the addition of exogenous connective tissue growth factor. Collectively, these results demonstrate that collateral flow evokes distinct EC gene expression profiles and functional phenotypes that subsequently influence vascular events important for adaptive remodeling.


Porous organic polymers for high-performance supercapacitors.
With the aim of addressing the global warming issue and fossil energy shortage, eco-friendly and sustainable renewable energy technologies are urgently needed. In comparison to energy conversion, studies on energy storage fall behind and remain largely to be explored. By storing energy from electrochemical processes at the electrode surface, supercapacitors (SCs) bridge the performance gap between electrostatic double-layer capacitors and batteries. Organic electrode materials have drawn extensive attention because of their special power density, good round trip efficiency and excellent cycle stability. Porous organic polymers (POPs) have drawn extensive attention as attractive electrode materials in SCs. In this review, we present and discuss recent advancements and design principles of POPs as efficient electrode materials for SCs from the perspectives of synthetic strategies and the structure-performance relationships of POPs. Finally, we put forward the outlook and prospects of POPs for SCs.