Evaluation of therapeutic options for pancreatic pseudocysts.
A review of 81 patients with pancreatic pseudocyst was conducted to assess the value of different treatment modalities. Resection was associated with 18% mortality (two of 11 patients) and 36% morbidity. In three of nine patients undergoing external drainage a recurrent pseudocyst developed, and in one additional patient, a pancreatic fistula persisted. Internal drainage by cystogastrostomy (21 patients) resulted in 9.5% mortality and 9.5% morbidity, whereas cystojejunostomy (33 patients) was associated with a 6% mortality and 6% morbidity. Endoscopic drainage through the posterior wall of the stomach was unsuccessful in the two patients in which it was used. Internal drainage into the stomach, duodenum, or jejunum is a safe and effective approach for most pseudocysts. Persistent symptoms following surgical treatment were primarily related to failure to recognize multiple cysts and/or pancreatic duct obstruction and dilation characteristic or chronic pancreatitis.


Psychosocial Adjustment and Sibling Relationships in Siblings of Children with Autism Spectrum Disorder: Risk and Protective Factors.
This study compared sibling adjustment and relationships in siblings of children with Autism Spectrum Disorder (ASD-Sibs; n = 69) and siblings of children with typical development (TD-Sibs; n = 93). ASD-Sibs and TD-Sibs demonstrated similar emotional/behavioral adjustment. Older male ASD-Sibs were at increased risk for difficulties. Sibling relationships of ASD-Sibs involved less aggression, less involvement, and more avoidance than those of TD-Sibs. Partial support for a diathesis-stress conceptualization of sibling difficulties was found for ASD-Sibs. For TD-Sibs, broader autism phenotype (BAP) was related to psychosocial difficulties regardless of family stressors. For ASD-Sibs, BAP was related to difficulties only when family stressors were present. This suggests that having a sibling with ASD may be a protective factor that attenuates the negative impact of sibling BAP.


Microbial community dynamics responding to nutrient allocation associated with soybean cultivar 'Jake' ozone adaptation.
Tropospheric ozone (O3), a major air pollutant, leads to significant global yield loss in soybean [Glycine max (L.) Merr.]. Soybean cultivar 'Jake' shows O3 resilient traits in above-ground organs, but the root system remains sensitive to elevated O3 (eO3). Changing carbon (C) and nitrogen (N) resource composition during eO3 stress suggests that eO3 presumably alters belowground soil microbial communities and their driven nutrient transformation. Yet, the responses of belowground microbes to eO3 and their feedback on nutrient cycling in 'Jake' are unknown. In this study, we holistically investigated soil microbial communities associated with C and N dynamics and bacterial-fungal inter-kingdom networks in the rhizosphere and bulk soil at different developmental stages of 'Jake' grown under sub-ambient O3 [charcoal-filtered (CF) air, 12 h mean: 20 ppb] or eO3 (12 h mean: 87 ppb). The results demonstrated eO3 significantly decreased fungal diversity and complexity of microbial networks at different 'Jake' developmental stages, whereas bacterial diversity was more tolerant to eO3 in both bulk soil and rhizosphere. In the bulk soil, no O3-responsive microbial biomarkers were found to be associated with C and N content, implying eO3 may stimulate niche-based processes during 'Jake' growth. In contrast, this study identified O3-responsive microbial biomarkers that may contribute to the N acquisition (Chloroflexales) and C dynamics (Caldilineales, Thermomicrobiales, and Hypocreales) in the rhizosphere, which may support the O3 resilience of the 'Jake' cultivar. However, further investigation is required to confirm their specific contributions by determining changes in microbial gene expression. Overall, these findings conduce to an expanding knowledge base that O3 induces temporal and spatial changes in the effects of microbial and nutrient networks in the O3-tolerant agriculture ecosystems.


Balneotherapy for chronic venous insufficiency.
Chronic venous insufficiency (CVI) is a progressive and common disease that affects the superficial and deep venous systems of the lower limbs. CVI is characterised by valvular incompetence, reflux, venous obstruction, or a combination of these with consequent distal venous hypertension. Clinical manifestations of CVI include oedema, pain, skin changes, ulcerations and dilated skin veins in the lower limbs. It can result in a large financial burden on health systems. There is a wide variety of treatment options or therapies for CVI, ranging from surgery and medication to compression and physiotherapy. Balneotherapy (treatments involving water) is a relatively cheap option and potentially efficient way to deliver physical therapy for people with CVI. To assess the efficacy and safety of balneotherapy for the treatment of people with chronic venous insufficiency (CVI). The Cochrane Vascular Information Specialist searched the Cochrane Vascular Specialised Register, CENTRAL, MEDLINE, Embase, AMED and CINAHL databases, the World Health Organization International Clinical Trials Registry Platform and the Clinical Trials.gov trials register to August 2018. We searched the LILACS and IBECS databases. We also checked references, searched citations and contacted study authors to identify additional studies. We included randomised and quasi-randomised controlled trials comparing balneotherapy with no treatment or other types of treatment for CVI. We also included studies that used a combination of treatments. Two review authors independently reviewed studies retrieved by the search strategies. Both review authors independently assessed selected studies for complete analysis. We resolved conflicts through discussion. We attempted to contact trial authors for missing data, obtaining additional information. For binary outcomes (leg ulcer incidence and adverse events), we presented the results using odds ratio (OR) with 95% confidence intervals (CI). For continuous outcomes (disease severity, health-related quality of life (HRQoL), pain, oedema, skin pigmentation), we presented the results as a mean difference (MD) with 95% CI. We included seven randomised controlled trials with 891 participants (outpatients in secondary care). We found no quasi-randomised controlled trials. Six studies (836 participants) evaluated balneotherapy versus no treatment. One study evaluated balneotherapy versus a phlebotonic drug (melilotus officinalis) (55 participants). There was a lack of blinding of participants and investigators, imprecision and inconsistency, which downgraded the certainty of the evidence.For the balneotherapy versus no treatment comparison, there probably was no improvement in favour of balneotherapy in disease severity signs and symptom score as assessed using the Venous Clinical Severity Score (VCSS) (MD -1.66, 95% CI -4.14 to 0.83; 2 studies, 484 participants; moderate-certainty evidence). Balneotherapy probably resulted in a moderate improvement in HRQoL as assessed by the Chronic Venous Insufficiency Questionnaire 2 (CVIQ2) at three months (MD -9.38, 95% CI -18.18 to -0.57; 2 studies, 149 participants; moderate-certainty evidence), nine months (MD -10.46, 95% CI -11.81 to -9.11; 1 study; 55 participants; moderate-certainty evidence), and 12 months (MD -4.99, 95% CI -9.19 to -0.78; 2 studies, 455 participants; moderate-certainty evidence). There was no clear difference in HRQoL between balneotherapy and no treatment at six months (MD -1.64, 95% CI -9.18 to 5.89; 2 studies, 445 participants; moderate-certainty evidence). Balneotherapy probably slightly improved pain compared with no treatment (MD -1.23, 95% CI -1.33 to -1.13; 1 study; 390 participants; moderate-certainty evidence). There was no clear effect related to oedema between the two groups at 24 days (MD 43.28 mL, 95% CI -102.74 to 189.30; 2 studies, 153 participants; very-low certainty evidence). There probably was no improvement in favour of balneotherapy in the incidence of leg ulcers (OR 1.69, 95% CI 0.82 to 3.48; 2 studies, 449 participants; moderate-certainty evidence). There was probably a reduction in incidence of skin pigmentation changes in favour of balneotherapy at 12 months (pigmentation index: MD -3.59, 95% CI -4.02 to -3.16; 1 study; 59 participants; low-certainty evidence). The main complications reported included erysipelas (OR 2.58, 95% CI 0.65 to 10.22; 2 studies, 519 participants; moderate-certainty evidence), thromboembolic events (OR 0.35, 95% CI 0.09 to 1.42; 3 studies, 584 participants; moderate-certainty evidence) and palpitations (OR 0.33, 95% CI 0.01 to 8.52; 1 study; 59 participants; low-certainty evidence), with no clear evidence of an increase in reported adverse effects with balneotherapy. There were no serious adverse events reported in any of the studies.For the balneotherapy versus a phlebotonic drug (melilotus officinalis) comparison, we observed no clear difference in pain symptoms (OR 0.29, 95% CI 0.03 to 2.87; 1 study; 35 participants; very low-certainty evidence) and oedema (OR 0.21, 95% CI 0.02 to 2.27; 1 study; 35 participants; very low-certainty evidence). This single study did not report on the other outcomes of interest. We identified moderate- to low-certainty evidence that suggests that balneotherapy may result in a moderate improvement in pain, quality of life and skin pigmentation changes and has no clear effect on disease severity signs and symptoms score, adverse effects, leg ulcers and oedema when compared with no treatment. For future studies, measurements of outcomes such as disease severity sign and symptom score, quality of life, pain and oedema and choice of time points during follow-up must be standardised for adequate comparison between trials.


Preoperative and postoperative prediction of long-term meningioma outcomes.
Meningiomas are stratified according to tumor grade and extent of resection, often in isolation of other clinical variables. Here, we use machine learning (ML) to integrate demographic, clinical, radiographic and pathologic data to develop predictive models for meningioma outcomes. We developed a comprehensive database containing information from 235 patients who underwent surgery for 257 meningiomas at a single institution from 1990 to 2015. The median follow-up was 4.3 years, and resection specimens were re-evaluated according to current diagnostic criteria, revealing 128 WHO grade I, 104 grade II and 25 grade III meningiomas. A series of ML algorithms were trained and tuned by nested resampling to create models based on preoperative features, conventional postoperative features, or both. We compared different algorithms' accuracy as well as the unique insights they offered into the data. Machine learning models restricted to preoperative information, such as patient demographics and radiographic features, had similar accuracy for predicting local failure (AUC = 0.74) or overall survival (AUC = 0.68) as models based on meningioma grade and extent of resection (AUC = 0.73 and AUC = 0.72, respectively). Integrated models incorporating all available demographic, clinical, radiographic and pathologic data provided the most accurate estimates (AUC = 0.78 and AUC = 0.74, respectively). From these models, we developed decision trees and nomograms to estimate the risks of local failure or overall survival for meningioma patients. Clinical information has been historically underutilized in the prediction of meningioma outcomes. Predictive models trained on preoperative clinical data perform comparably to conventional models trained on meningioma grade and extent of resection. Combination of all available information can help stratify meningioma patients more accurately.


[Use of carnitine for stimulation of the growth and development of the mouse brain in undernourishment in early postnatal ontogeny].
The authors studied the possibilities of using carnitine for stimulation of the mouse brain development under malnutrition in the early postnatal ontogenesis. The results indicate that addition of carnitine to the diet during food rehabilitation stimulates the growth of the body and brain mass, increases the neocortex, hippocamp, cerebellum and the size of neuronal bodies in these structures, removes the effect of retarding the growth of neuronal processes and noticeably decreases the rate of dystrophic changes in the brain structures under study. Thus, the disturbances of the mouse brain development caused by malnutrition are completely eliminated after the animals are fed a full-value diet containing carnitine.


Direct cloning of a long restriction fragment aided with a jumping clone.
Long-range physical mapping with rare-cutting restriction enzymes (rare cutters) is an important step for structural analysis of complex genomes. Combination of two types of DNA clones bearing the rare-cutter sites, linking clones and jumping clones (Fig. 1a), facilitates the physical mapping [Poustka et al., Nature 325 (1987) 353-355]. A step followed by the physical mapping is the cloning of the large (rare-cutter-generated) restriction fragment of interest. For facilitating this step, we devised a method to directly clone a long restriction fragment without constructing the whole genomic DNA library using the jumping clone as starting material. The short DNA segments of a jumping clone, which are derived from the 5' and 3' terminal regions of the large restriction fragment, are inserted into the yeast artificial chromosome plasmid (pYAC) vector, and then converted into single strands with T7 gene 6-encoded 5'----3' exonuclease. The total genomic DNA digested with the restriction enzyme is also treated with the exonuclease to convert the terminal regions of the restriction fragments into single strands. In the resulting products, only the fragment corresponding to the jumping clone can form hybrids with the just-mentioned, single-stranded DNAs, which are connected to the pYAC, and only this fragment is cloned in yeast. We describe the protocol of this method with Escherichia coli DNA as a model experiment. Judging from the cloning efficiency, this method could be applied to cloning single-copy regions of the human genome, provided a jumping clone is available. The instability of inserts in the pYAC vector is also discussed.


Bacterial targets as potential indicators of diesel fuel toxicity in subantarctic soils.
Appropriate remediation targets or universal guidelines for polar regions do not currently exist, and a comprehensive understanding of the effects of diesel fuel on the natural microbial populations in polar and subpolar soils is lacking. Our aim was to investigate the response of the bacterial community to diesel fuel and to evaluate if these responses have the potential to be used as indicators of soil toxicity thresholds. We set up short- and long-exposure tests across a soil organic carbon gradient. Utilizing broad and targeted community indices, as well as functional genes involved in the nitrogen cycle, we investigated the bacterial community structure and its potential functioning in response to special Antarctic blend (SAB) diesel fuel. We found the primary effect of diesel fuel toxicity was a reduction in species richness, evenness, and phylogenetic diversity, with the resulting community heavily dominated by a few species, principally Pseudomonas. The decline in richness and phylogenetic diversity was linked to disruption of the nitrogen cycle, with species and functional genes involved in nitrification significantly reduced. Of the 11 targets we evaluated, we found the bacterial amoA gene indicative of potential ammonium oxidation, the most suitable indicator of toxicity. Dose-response modeling for this target generated an average effective concentration responsible for 20% change (EC20) of 155 mg kg(-1), which is consistent with previous Macquarie Island ecotoxicology assays. Unlike traditional single-species tolerance testing, bacterial targets allowed us to simultaneously evaluate more than 1,700 species from 39 phyla, inclusive of rare, sensitive, and functionally relevant portions of the community.


[The course of experimental staphylococcal sepsis in opposite mouse strains and first-generation hybrids].
The survival time and histological lesions of the kidneys, liver, heart, and lungs were studied in CBA/Sto, C3HA/Mv and F1 (C3HA/Mv x CBA/Sto) mice for 15 days after i.v. injections with S. aureus pathogenic strains CFU B-243 in doses of 10(9), 10(8) and 2.5 x 10(8) microbial cells. CBA/Sto mice were found relatively resistant and C3HA/Mv mice, susceptible to infection caused by different doses of S. aureus, this being associated with different morphological picture in the viscera. F1 hybrids were at least as susceptible to the infections as any of the parent strains, suggesting recessive inheritance of resistance to staphylococcal infection.


Predictive and Pharmacodynamic Biomarkers of Response to the Phosphatidylinositol 3-Kinase Inhibitor Taselisib in Breast Cancer Preclinical Models.
The PI3K signaling pathway serves as a central node in regulating cell survival, proliferation, and metabolism. PIK3CA, the gene encoding the PI3K catalytic subunit p110-alpha, is commonly altered in breast cancer resulting in the constitutive activation of the PI3K pathway. Using an unbiased cell line screening approach, we tested the sensitivity of breast cancer cell lines to taselisib, a potent PI3K inhibitor, and correlated sensitivity with key biomarkers (PIK3CA, HER2, PTEN, and ESR1). We further assessed how taselisib modulates downstream signaling in the different genomic backgrounds that occur within breast cancer. We found that sensitivity to taselisib correlated with the presence of PIK3CA mutations, but was independent of HER2 status. We further showed that HER2-amplified/PIK3CA wild-type cell lines are not as sensitive to taselisib when compared with HER2-amplified/PIK3CA-mutant cell lines. In a PIK3CA-mutant/PTEN null background, PI3K downstream signaling rebounded in the presence of taselisib correlating with decreased sensitivity at later time points. Finally, we observed that PIK3CA mutations cooccurred with mutations in the estrogen receptor (ER; ESR1) in metastatic tumors from patients with ER+ breast cancer. However, the cooccurrence of an ESR1 mutation with a PIK3CA mutation did not affect response to taselisib in a single agent setting or in combination with fulvestrant. In summary, these data suggest that development of taselisib in breast cancer should occur in a PIK3CA-mutant setting with cotreatments determined by the specific subtypes under investigation.


Intratumoral Iron Oxide Nanoparticle Hyperthermia and Radiation Cancer Treatment.
The potential synergism and benefit of combined hyperthermia and radiation for cancer treatment is well established, but has yet to be optimized clinically. Specifically, the delivery of heat via external arrays /applicators or interstitial antennas has not demonstrated the spatial precision or specificity necessary to achieve appropriate a highly positive therapeutic ratio. Recently, antibody directed and possibly even non-antibody directed iron oxide nanoparticle hyperthermia has shown significant promise as a tumor treatment modality. Our studies are designed to determine the effects (safety and efficacy) of iron oxide nanoparticle hyperthermia and external beam radiation in a murine breast cancer model. MTG-B murine breast cancer cells (1 × 106) were implanted subcutaneous in 7 week-old female C3H/HeJ mice and grown to a treatment size of 150 mm3 +/- 50 mm3. Tumors were then injected locally with iron oxide nanoparticles and heated via an alternating magnetic field (AMF) generator operated at approximately 160 kHz and 400 - 550 Oe. Tumor growth was monitored daily using standard 3-D caliper measurement technique and formula. specific Mouse tumors were heated using a cooled, 36 mm diameter square copper tube induction coil which provided optimal heating in a 1 cm wide region in the center of the coil. Double dextran coated 80 nm iron oxide nanoparticles (Triton Biosystems) were used in all studies. Intra-tumor, peri-tumor and rectal (core body) temperatures were continually measured throughout the treatment period. Preliminary in vivo nanoparticle-AMF hyperthermia (167 KHz and 400 or 550 Oe) studies demonstrated dose responsive cytotoxicity which enhanced the effects of external beam radiation. AMF associated eddy currents resulted in nonspecific temperature increases in exposed tissues which did not contain nanoparticles, however these effects were minor and not injurious to the mice. These studies also suggest that iron oxide nanoparticle hyperthermia is more effective than nonnanoparticle tumor heating techniques when similar thermal doses are applied. Initial electron and light microscopy studies of iron oxide nanoparticle and AMF exposed tumor cells show a rapid uptake of particles and acute cytotoxicity following AMF exposure.


Aggregation-induced emission spectral shift as a measure of local concentration of a pH-activatable rhodamine-based smart probe.
Generating activatable probes that report about molecular vicinity through contact-based mechanisms such as aggregation can be very convenient. Specifically, such probes change a particular spectral property only at the intended biologically relevant target. Xanthene derivatives, for example rhodamines, are able to form aggregates. It is typical to examine aggregation by absorption spectroscopy but for microscopy applications utilizing fluorescent probes it is very important to perform characterization by measuring fluorescence spectra. First we show that excitation spectra of aqueous solutions of rhodamine 6G can be very informative about the aggregation features. Next we establish the dependence of the fluorescence emission spectral maximum shift on the dimer concentration. The obtained information helped us confirm the possibility of aggregation of a recently designed and synthesized rhodamine 6G-based pH-activatable fluorescent probe and to study its pH and concentration dependence. The size of the aggregation-induced emission spectral shift at specific position on the sample can be measured by fluorescence microspectroscopy, which at particular pH allows estimation of the local concentration of the observed probe at microscopic level. Therefore, we show that besides aggregation-caused quenching and aggregation-induced emission also aggregation-induced emission spectral shift can be a useful photophysical phenomenon.


Evidence for individual discrimination and numerical assessment in collective antipredator behaviour in wild jackdaws (Corvus monedula).
Collective responses to threats occur throughout the animal kingdom but little is known about the cognitive processes underpinning them. Antipredator mobbing is one such response. Approaching a predator may be highly risky, but the individual risk declines and the likelihood of repelling the predator increases in larger mobbing groups. The ability to appraise the number of conspecifics involved in a mobbing event could therefore facilitate strategic decisions about whether to join. Mobs are commonly initiated by recruitment calls, which may provide valuable information to guide decision-making. We tested whether the number of wild jackdaws responding to recruitment calls was influenced by the number of callers. As predicted, playbacks simulating three or five callers tended to recruit more individuals than playbacks of one caller. Recruitment also substantially increased if recruits themselves produced calls. These results suggest that jackdaws use individual vocal discrimination to assess the number of conspecifics involved in initiating mobbing events, and use this information to guide their responses. Our results show support for the use of numerical assessment in antipredator mobbing responses and highlight the need for a greater understanding of the cognitive processes involved in collective behaviour.


Simultaneous radicular cyst and mucoepidermoid carcinoma in the maxilla: a diagnostic nightmare.
We present a case of a 20-year-old woman presenting initially with an asymptomatic palatal swelling. Radiographic examination showed a cyst at the right maxilla with bucco-lingual expansion and perforation of palatal bone. Incisional biopsy was carried out via a buccal approach and the result revealed a benign odontogenic cyst, in keeping with radicular cyst. The patient was then scheduled for cyst enucleation. During the procedure, it was found that the palatal lesion was unrelated to the maxillary cyst. Incisional biopsy of the palatal mass was carried out and revealed a low-grade mucoepidermoid carcinoma. The patient then had a partial maxillectomy with fibula flap reconstruction. There was no recurrence at postoperative 1 year follow-up and she was rehabilitated with dental implants.


Image-guided EBUS bronchoscopy system for lung-cancer staging.
The staging of the central-chest lymph nodes is a major step in the management of lung-cancer patients. For this purpose, the physician uses a device that integrates videobronchoscopy and an endobronchial ultrasound (EBUS) probe. To biopsy a lymph node, the physician first uses videobronchoscopy to navigate through the airways and then invokes EBUS to localize and biopsy the node. Unfortunately, this process proves difficult for many physicians, with the choice of biopsy site found by trial and error. We present a complete image-guided EBUS bronchoscopy system tailored to lymph-node staging. The system accepts a patient's 3D chest CT scan, an optional PET scan, and the EBUS bronchoscope's video sources as inputs. System workflow follows two phases: (1) procedure planning and (2) image-guided EBUS bronchoscopy. Procedure planning derives airway guidance routes that facilitate optimal EBUS scanning and nodal biopsy. During the live procedure, the system's graphical display suggests a series of device maneuvers to perform and provides multimodal visual cues for locating suitable biopsy sites. To this end, the system exploits data fusion to drive a multimodal virtual bronchoscope and other visualization tools that lead the physician through the process of device navigation and localization. A retrospective lung-cancer patient study and follow-on prospective patient study, performed within the standard clinical workflow, demonstrate the system's feasibility and functionality. For the prospective study, 60/60 selected lymph nodes (100%) were correctly localized using the system, and 30/33 biopsied nodes (91%) gave adequate tissue samples. Also, the mean procedure time including all user interactions was 6 min 43 s All of these measures improve upon benchmarks reported for other state-of-the-art systems and current practice. Overall, the system enabled safe, efficient EBUS-based localization and biopsy of lymph nodes.


Feasibility and first results of a group program to increase the frequency of cognitively stimulating leisure activities in people with mild cognitive impairment (AKTIVA-MCI).
AKTIVA-MCI is a program for patients with mild cognitive impairment (MCI) that aims to enhance participation in cognitively stimulating leisure activities. Participation in cognitively stimulating activities seems to be a potential strategy for people with MCI delaying cognitive decline for a while. In total, 35 MCI patients were enrolled in the pilot study of whom 29 completed the whole program (16 female, 71.1±7.5 years; Mini Mental Status Examination score: 28±2.2). Daily activity protocols were used to measure the frequency of participation in cognitively stimulating activities during the program (12 sessions). Additional standardized psychometric tests and questionnaires were used to assess cognition, mood, and subjective memory decline. Analyses of the daily activity protocols showed that during the intervention participants increased the frequency of several cognitively stimulating leisure activities. Comparison of pre-post data indicates no changes in cognitive status, mood, and subjective memory decline. These findings indicate that the program is suitable for patients with MCI.


Associations of Baseline and Longitudinal Serum Uromodulin With Kidney Failure and Mortality: Results From the African American Study of Kidney Disease and Hypertension (AASK) Trial.
Uromodulin (UMOD) is the most abundant protein found in urine and has emerged as a promising biomarker of tubule health. Circulating UMOD is also detectable, but at lower levels. We evaluated whether serum UMOD levels were associated with the risks of incident kidney failure with replacement therapy (KFRT) and mortality. Prospective cohort. Participants in AASK (the African American Study of Kidney Disease and Hypertension) with available stored serum samples from the 0-, 12-, and 24-month visits for biomarker measurement. Baseline log-transformed UMOD and change in UMOD over 2 years. KFRT and mortality. Cox proportional hazards and mixed-effects models. Among 500 participants with baseline serum UMOD levels (mean age, 54y; 37% female), 161 KFRT events occurred during a median of 8.5 years. After adjusting for baseline demographic factors, clinical factors, glomerular filtration rate, log-transformed urine protein-creatinine ratio, and randomized treatment groups, a 50% lower baseline UMOD level was independently associated with a 35% higher risk of KFRT (adjusted HR, 1.35; 95% CI, 1.07-1.70). For annual UMOD change, each 1-standard deviation lower change was associated with a 67% higher risk of KFRT (adjusted HR, 1.67; 95% CI, 1.41-1.99). Baseline UMOD and UMOD change were not associated with mortality. UMOD levels declined more steeply for metoprolol versus ramipril (P<0.001) as well as for intensive versus standard blood pressure goals (P = 0.002). Small sample size and limited generalizability. Lower UMOD levels at baseline and steeper declines in UMOD over time were associated with a higher risk of subsequent KFRT in a cohort of African American adults with chronic kidney disease and hypertension. Prior studies of uromodulin (UMOD), the most abundant protein in urine, and kidney disease have focused primarily on urinary UMOD levels. The present study evaluated associations of serum UMOD levels with the risks of kidney failure with replacement therapy (KFRT) and mortality in a cohort of African American adults with hypertension and chronic kidney disease. It found that participants with lower levels of UMOD at baseline were more likely to experience KFRT even after accounting for baseline kidney measures. Similarly, participants who experienced steeper annual declines in UMOD also had a heightened risk of kidney failure. Neither baseline nor annual change in UMOD was associated with mortality. Serum UMOD is a promising biomarker of kidney health.


Coxsackie adenovirus receptor and alpha nu beta3/alpha nu beta5 integrins in adenovirus gene transfer of rat cochlea.
This study was designed to determine whether Coxsackie adenovirus receptor (CAR) and alpha nu beta3/alpha nu beta5 integrin co-receptors are involved in adenovirus gene transfer in the rat cochlea. We find that CAR and integrin co-receptors are expressed in every cell subtype transduced by the adenoviral vector Ad5 DeltaE1-E3/cytomegalovirus/green fluorescent protein (GFP) on cochlear slices in vitro. The spiral ganglion neurons, which do not express CAR, were not transduced by the virus. Blocking these receptors by monoclonal antibodies decreased transgene expression, whereas disrupting tight junctions with ethylenediaminetetraacetic acid led to an increased transgene expression. However, sensory hair cells and strial cells also expressing CAR and alpha nu integrins were not transduced by the vector. GFP expression was also studied in vivo. Perilymphatic perfusion of adenovirus in vivo did not affect hearing and only cells lining the perilymphatic spaces were transduced. Endolymphatic perfusion resulted in low-frequency hearing loss and although some cells of the organ of Corti were efficiently transduced, the sensory and the strial cells were not. Transduced sensory and strial cells were occasionally observed in cochleas after single shot of adenovirus. Pretreatment with anti-CAR and anti-alpha nu antibodies decreases GFP expression in vivo, suggesting that the CAR/alpha nu integrin pathway is involved in adenovirus transduction in the cochlea.


First assessment at home of amyotrophic lateral sclerosis (ALS) patients by a nutrition network in the French region of Limousin.
Malnutrition is associated with poor survival among patients with amyotrophic lateral sclerosis (ALS). This study aimed to evaluate nutritional assessment by a network during first consultations in patients' homes. Patients identified by the regional ALS centre gave their informed consent. Assessment included functional, nutritional issues, evaluation of the need for help, whether personal or the use of aids, and noted any dietary supplementation and modification of the texture of food. Forty patients were seen a mean of 7.4 months after diagnosis; 52.5% had bulbar disease, 7.5% were malnourished; 29.4 ± 10.1 kcal/kg/day were consumed and protein intake was 1.3 ± 0.5 g/kg/day. Thirty-five percent of patients were anorexic, 43.8% reported taste disorders, and 70% had dysphagia, significantly associated with salivary stasis. Only 30% of dysphagic patients ate texture-modified food, and 90% of patients with problems drinking liquids did not use a thickener. In conclusion, assessment at home by a nutritional network can be conducted promptly. Malnutrition is rare in early disease, despite the fact that patients' diets are often low in energy and dysphagia is common. Unexpected taste disorders are detected. Dysphagia is very common but inadequately addressed. Consequently, home assessment by the network led several beneficial interventions.


Identification and characterization of the in vitro synthesized gene products of bacteriophage M13.
Bacteriophage M13 replicative form (RF) DNA was used to direct coupled transcription and translation in cell-free extracts prepared from Escherichia coli. By using RF DNA, isolated from cells infected with appropriate amber mutants of this phage, it has been possible to identify the products of genes I through IV. By using the same methods no gene-product relationship could be demonstrated for genes VI and VII. Coupled in vitro protein synthesis studies on RF-III DNA, a linear double-stranded DNA molecule, obtained after cleavage of either RF-I or RF-II DNA with the restriction endonuclease R.Hin11 from Haemophilus influenzae, indicated that the cleavage site for this enzyme is located in gene II. The in vitro products of both gene III and gene VIII are about 30 and six amino acids longer, respectively, than their native counterparts present within the virion. These results suggest that the latter proteins arise in vivo by cleavage of precursor molecules. Coupled transcription and translation studies on a DNA fragment which only contained the genetic information coding for gene IV protein, obtained after cleavage of RF DNA with the restriction endonuclease R.Hap11 from Haemophilus aphirophilus, indicated that a large number of the in vitro synthesized polypeptides are the result of premature chain termination.


Network propagation of rare variants in Alzheimer's disease reveals tissue-specific hub genes and communities.
State-of-the-art rare variant association testing methods aggregate the contribution of rare variants in biologically relevant genomic regions to boost statistical power. However, testing single genes separately does not consider the complex interaction landscape of genes, nor the downstream effects of non-synonymous variants on protein structure and function. Here we present the NETwork Propagation-based Assessment of Genetic Events (NETPAGE), an integrative approach aimed at investigating the biological pathways through which rare variation results in complex disease phenotypes. We applied NETPAGE to sporadic, late-onset Alzheimer's disease (AD), using whole-genome sequencing from the AD Neuroimaging Initiative (ADNI) cohort, as well as whole-exome sequencing from the AD Sequencing Project (ADSP). NETPAGE is based on network propagation, a framework that models information flow on a graph and simulates the percolation of genetic variation through tissue-specific gene interaction networks. The result of network propagation is a set of smoothed gene scores that can be tested for association with disease status through sparse regression. The application of NETPAGE to AD enabled the identification of a set of connected genes whose smoothed variation profile was robustly associated to case-control status, based on gene interactions in the hippocampus. Additionally, smoothed scores significantly correlated with risk of conversion to AD in Mild Cognitive Impairment (MCI) subjects. Lastly, we investigated tissue-specific transcriptional dysregulation of the core genes in two independent RNA-seq datasets, as well as significant enrichments in terms of gene sets with known connections to AD. We present a framework that enables enhanced genetic association testing for a wide range of traits, diseases, and sample sizes.


The role of surface polysaccharide in determining the resistance of Serratia marcescens to serum killing.
Two O14:H12 strains of Serratia marcescens with different sensitivities to killing by normal pooled human serum were investigated. Complement binding, studied by measuring hydrophobicity and using rocket immunoelectrophoresis with anti-human C3, showed the sensitive cells (S1220) rapidly bound and fixed complement whereas the resistant cells (4444-60) bound less C3b. The strains had identical membrane protein composition. Crossed immunoelectrophoresis suggested that in S1220 cells the polysaccharide material including LPS was less antigenic and present in smaller amounts than in 4444-60 cells. This was confirmed by examining extracted polysaccharide material chemically and by SDS-PAGE. The resistant strain had 33% more phenol-extractable polysaccharide material than the sensitive strain, possibly comprising LPS with longer O antigen chain lengths, or a microcapsule of O antigen polysaccharide. Extra polysaccharide material on the surface of the resistant strain prevents complement components binding and reaching the hydrophobic membrane where lytic lesions occur.


Atherosclerotic plaque characterization by 0.5-mm-slice multislice computed tomographic imaging.
It has been proposed that 0.5-mm-slice multislice computed tomography (MSCT) is a noninvasive tool for the detection of atherosclerotic plaque, but the validity of such an assessment has not been demonstrated by an invasive investigation. The present study was performed to compare the 0.5-mm-slice MSCT density of plaques with intravascular ultrasound (IVUS) findings. Atherosclerotic plaques were characterized in 37 consecutive patients undergoing percutaneous interventions. Based on the IVUS echogenecity, the plaques were classified as soft (n=18), fibrous (n=40) or calcified (n=40). In these 98 plaques, 0.5-mm-slice MSCT plaque density was calculated in 443 regions-of-interest, including 331 lesional foci and 112 luminal cross-sections, and represented as Hounsfield units (HU). MSCT density of the 3 types of plaque was 11+/-12 HU, 78+/-21 HU, and 516 +/-198 HU respectively. Computed tomography density of the (contrast-filled) lumen was 258+/-43 HU. There were statistically highly significant differences in the densitometric characteristics among the 4 groups (soft, fibrous, calcified plaque and lumen) by nonparametric Kruskal-Wallis test (p<0.0001). The IVUS-based coronary plaque configuration can be accurately identified by 0.5-mm slice MSCT. Noninvasive assessment of plaque characterization will ensure emphasis on the vessel wall beyond the vascular lumen.


Methods of induction of labor and women's experience: a population-based cohort study with mediation analyses.
Negative childbirth experience may affect mother wellbeing and health. However, it is rarely evaluated in studies comparing methods of induction of labor (IoL). To compare women's experience of IoL according to the method, considering the mediating role of interventions and complications of delivery. We used data from the MEDIP prospective population-based cohort, including all women with IoL during one month in seven French perinatal networks. The experience of IoL, assessed at 2 months postpartum, was first compared between cervical ripening and oxytocin, and secondarily between different cervical ripening methods. Mediation analyses were used to measure the direct and indirect effects of cervical ripening on maternal experience, through delivery with interventions or complications. The response rate was 47.8% (n = 1453/3042). Compared with oxytocin (n = 541), cervical ripening (n = 910) was associated less often with feelings that labor went 'as expected' (adjusted risk ratio for the direct effect 0.78, 95%CI [0.70-0.88]), length of labor was 'acceptable' (0.76[0.71-0.82]), 'vaginal discomfort' was absent (0.77[0.69-0.85]) and with lower global satisfaction (0.90[0.84-0.96]). Interventions and complications mediated between 6 and 35% of the total effect of cervical ripening on maternal experience. Compared to the dinoprostone insert, maternal experience was not significantly different with the other prostaglandins. The balloon catheter was associated with less pain. Cervical ripening was associated with a less positive experience of childbirth, whatever the method, only partly explained by interventions and complications of delivery. Counselling and support of women requiring cervical ripening might be enhanced to improve the experience of IoL.


Relative daughter cell volume and position of the division furrow in Tetrahymena.
The relative daughter cell volume (RDCV) values for Tetrahymena pyriformis were determined at division on live cells. It was found that the anterior cell is generally larger than the posterior cell, and that the RDCV values are distributed in groups 5-6% apart. The RDCV value was found to be independent of predivision cell volume, indicating that the mother cell is divided into proportional volumes. The cells seem, however, not to assess volume directly but rather a parameter related to the cell volume. Furthermore, the RDCV value was found to increase during cell division, so that the final value is not reached until actual separation of daughter cells. It is suggested that the division furrow is positioned so that the area of the cell surface lying between the old oral apparatus and the posterior pole of the cell is divided into equal parts. It is further suggested that several alternative values of the RDCV are possible, only one of which is expressed in each cell. The early division furrow is placed anteriorly to its final position, and its location is adjusted during cytokinesis.


Fusion injection of Rous Sarcoma virus proteins into Rous sarcoma virus-transformed, non-producing hamster cells causes release of infectious virus.
Purified virus proteins from transformation-defective (td) mutants of Rous sarcoma virus PrA or PrB were trapped in human erythrocyte ghosts which, after resealing, were fusion-injected into hamster RBH cells or rat TWERC cells. These cell lines are non-productively transformed by subgroup C Rous sarcoma virus. After fusion injection the hamster RBH cells released transforming subgroup C Rous sarcoma virus. No infectious virus could be rescued from rat TWERC cells. Since previous experiments have shown that fusion injection of the purified Rous sarcoma virus protein p15 into hamster RBH cells caused cleavage of the precursor protein pr76 to form the virus group-specific antigen (gag) but did not induce infectious virus, we conclude that in addition to p15 other virus proteins are required to induce virus rescue in hamster RBH cells.


Diverse chemotypes disrupt ion homeostasis in the Malaria parasite.
The antimalarial spiroindolones disrupt Plasmodium falciparum Na(+) regulation and induce an alkalinization of the parasite cytosol. It has been proposed that they do so by inhibiting PfATP4, a parasite plasma membrane P-type ATPase postulated to export Na(+) and import H(+) equivalents. Here, we screened the 400 antiplasmodial compounds of the open access 'Malaria Box' for their effects on parasite ion regulation. Twenty eight compounds affected parasite Na(+) and pH regulation in a manner consistent with PfATP4 inhibition. Six of these, with chemically diverse structures, were selected for further analysis. All six showed reduced antiplasmodial activity against spiroindolone-resistant parasites carrying mutations in pfatp4. We exposed parasites to incrementally increasing concentrations of two of the six compounds and in both cases obtained resistant parasites with mutations in pfatp4. The finding that diverse chemotypes have an apparently similar mechanism of action indicates that PfATP4 may be a significant Achilles' heel for the parasite.


Identification of cell wall deficient forms of M. avium subsp. paratuberculosis in paraffin embedded tissues from animals with Johne's disease by in situ hybridization.
M. avium subsp. paratuberculosis (M. paratuberculosis) is the causative agent of Johne's disease (JD) in ruminants leading to enormous economical losses in dairy and meat industries worldwide. During the subclinical stage of the disease, the infected animals are difficult if not impossible to detect by the available diagnostic tests including the PCR based ones. Although only considered an animal pathogen, cell wall deficient (CWD) forms of M. paratuberculosis have been isolated from patients with sarcoidosis and Crohn's disease (idiopathic diseases) in humans. Hence, the CWD form of this organism has been suspected to play a role in the pathogenesis of these diseases by persisting in the affected tissues and triggering a localized immune response and pathology. Differentiating between the CWD and acid-fast forms of this organism may lead to the determination of whether the CWD form is the pathogenic form in the subclinical cases of JD in animals and/or the etiologic agent for the above human diseases. To localize such organisms in tissue sections, CWD forms of mycobacteria were prepared in vitro and injected into beef cubes which were then formalin fixed and paraffin embedded. An in situ hybridization (ISH) technique, combined with the IS900 M. paratuberculosis-specific probe labeled with digoxigenin, was developed for the detection of nucleic acids specifically from the CWD forms but not their acid-fast forms in tissue sections. Specificity was confirmed by the negative finding with an irrelevant probe and with control tissue preparations containing CWD cells of related mycobacteria and unrelated organisms. This ISH procedure provides a way to distinguish between the acid-fast and CWD forms of M. paratuberculosis and to localize them in tissue sections. ISH may prove useful to evaluate the significance of CWD forms of M. paratuberculosis in the pathogenesis of JD, Crohn's disease and sarcoidosis.


Neuromuscular actuation of biohybrid motile bots.
The integration of muscle cells with soft robotics in recent years has led to the development of biohybrid machines capable of untethered locomotion. A major frontier that currently remains unexplored is neuronal actuation and control of such muscle-powered biohybrid machines. As a step toward this goal, we present here a biohybrid swimmer driven by on-board neuromuscular units. The body of the swimmer consists of a free-standing soft scaffold, skeletal muscle tissue, and optogenetic stem cell-derived neural cluster containing motor neurons. Myoblasts embedded in extracellular matrix self-organize into a muscle tissue guided by the geometry of the scaffold, and the resulting muscle tissue is cocultured in situ with a neural cluster. Motor neurons then extend neurites selectively toward the muscle and innervate it, developing functional neuromuscular units. Based on this initial construct, we computationally designed, optimized, and implemented light-sensitive flagellar swimmers actuated by these neuromuscular units. Cyclic muscle contractions, induced by neural stimulation, drive time-irreversible flagellar dynamics, thereby providing thrust for untethered forward locomotion of the swimmer. Overall, this work demonstrates an example of a biohybrid robot implementing neuromuscular actuation and illustrates a path toward the forward design and control of neuron-enabled biohybrid machines.


Abnormal brain development in newborns with congenital heart disease.
Congenital heart disease in newborns is associated with global impairment in development. We characterized brain metabolism and microstructure, as measures of brain maturation, in newborns with congenital heart disease before they underwent heart surgery. We studied 41 term newborns with congenital heart disease--29 who had transposition of the great arteries and 12 who had single-ventricle physiology--with the use of magnetic resonance imaging (MRI), magnetic resonance spectroscopy (MRS), and diffusion tensor imaging (DTI) before cardiac surgery. We calculated the ratio of N-acetylaspartate to choline (which increases with brain maturation), the ratio of lactate to choline (which decreases with maturation), average diffusivity (which decreases with maturation), and fractional anisotropy of white-matter tracts (which increases with maturation). We compared these findings with those in 16 control newborns of a similar gestational age. As compared with control newborns, those with congenital heart disease had a decrease of 10% in the ratio of N-acetylaspartate to choline (P=0.003), an increase of 28% in the ratio of lactate to choline (P=0.08), an increase of 4% in average diffusivity (P<0.001), and a decrease of 12% in white-matter fractional anisotropy (P<0.001). Preoperative brain injury, as seen on MRI, was not significantly associated with findings on MRS or DTI. White-matter injury was observed in 13 newborns with congenital heart disease (32%) and in no control newborns. Term newborns with congenital heart disease have widespread brain abnormalities before they undergo cardiac surgery. The imaging findings in such newborns are similar to those in premature newborns and may reflect abnormal brain development in utero.


Titanium dioxide nanoparticles detoxify pirimicarb under UV irradiation at ambient intensities.
Titanium dioxide nanoparticles (nTiO₂) form reactive oxygen species (ROS) under irradiation by ultraviolet light (UV). This known photocatalytic activity may finally affect the presence and toxicity of organic environmental chemicals, which have not yet been studied at ambient UV intensity. The authors used a three-factorial design to evaluate the interaction of the carbamate insecticide pirimicarb (initial nominal concentration, 20 µg/L), ambient UV irradiation (40 W/m² for 15 min), and nTiO₂(~100 nm; 2.0 mg/L). Pirimicarb, pirimicarb × UV, and pirimicarb × nTiO₂ treatments revealed a median immobilization of Daphnia magna after 72 h ranging between 70 and 80%. This effect seemed to be caused by the initial nominal pirimicarb concentration. However, UV irradiation before an exposure of daphnids in the presence of 2.0 mg nTiO₂/L reduced pirimicarb concentrations to values below the limit of quantification, likely because of the formation of ROS. This reduction was associated with an almost complete removal of toxicity for D. magna. Furthermore, during a second experiment, 0.2 mg nTiO₂/L in combination with 15 min UV irradiation reduced pirimicarb concentrations by approximately 30%. These results indicate a detoxification and therefore remediation potential of the combined application of nTiO₂ and UV irradiation at ambient levels. This potential has not been documented to date in surface waters, where nTiO₂ concentrations in the low to medium µg/L range may occur.


Outcome weighted ψ-learning for individualized treatment rules.
An individualized treatment rule is often employed to maximize a certain patient-specific clinical outcome based on his/her clinical or genomic characteristics as well as heterogeneous response to treatments. Although developing such a rule is conceptually important to personalized medicine, existing methods such as the partial least squares Qian and Murphy (2011) suffers from the difficulty of indirect maximization of a patient's clinical outcome, while the outcome weighted learning Y. Zhao, Zeng, Rush, and Kosorok (2012) is not robust against any perturbation of the outcome. In this article, we propose a weighted ψ-learning method to optimize an individualized treatment rule, which is robust against any data perturbation near the decision boundary by seeking the maximum separation. To solve nonconvex minimization, we employ a difference convex algorithm to relax the non-convex minimization iteratively based on a decomposition of the cost function into a difference of two convex functions. On this ground, we also introduce a variable selection method for further removing redundant variables for a higher performance. Finally, we illustrate the proposed method by simulations and a lung health study and demonstrate that it yields higher performances in terms of accuracy of prediction of individualized treatment.


An unusual mixed infection in a diabetic patient: Citrobacter freundii and Peptococcus magnus.
A case of an unusual mixed infection of the foot involving Citrobacter freundii and Peptococcus magnus is presented. Characteristics and treatment of infections caused by these organisms are reviewed. The possible effects of an infection of this type in a compromised patient are also presented.


Massive Submucosal Ganglia in Colonic Inertia.
- Colonic inertia is a debilitating form of primary chronic constipation with unknown etiology and diagnostic criteria, often requiring pancolectomy. We have occasionally observed massively enlarged submucosal ganglia containing at least 20 perikarya, in addition to previously described giant ganglia with greater than 8 perikarya, in cases of colonic inertia. These massively enlarged ganglia have yet to be formally recognized. - To determine whether such "massive submucosal ganglia," defined as ganglia harboring at least 20 perikarya, characterize colonic inertia. - We retrospectively reviewed specimens from colectomies of patients with colonic inertia and compared the prevalence of massive submucosal ganglia occurring in this setting to the prevalence of massive submucosal ganglia occurring in a set of control specimens from patients lacking chronic constipation. - Seven of 8 specimens affected by colonic inertia harbored 1 to 4 massive ganglia, for a total of 11 massive ganglia. One specimen lacked massive ganglia but had limited sampling and nearly massive ganglia. Massive ganglia occupied both superficial and deep submucosal plexus. The patient with 4 massive ganglia also had 1 mitotically active giant ganglion. Only 1 massive ganglion occupied the entire set of 10 specimens from patients lacking chronic constipation. - We performed the first, albeit distinctly small, study of massive submucosal ganglia and showed that massive ganglia may be linked to colonic inertia. Further, larger studies are necessary to determine whether massive ganglia are pathogenetic or secondary phenomena, and whether massive ganglia or mitotically active ganglia distinguish colonic inertia from other types of chronic constipation.


Integrating molecular networks with genetic variant interpretation for precision medicine.
More reliable and cheaper sequencing technologies have revealed the vast mutational landscapes characteristic of many phenotypes. The analysis of such genetic variants has led to successful identification of altered proteins underlying many Mendelian disorders. Nevertheless the simple one-variant one-phenotype model valid for many monogenic diseases does not capture the complexity of polygenic traits and disorders. Although experimental and computational approaches have improved detection of functionally deleterious variants and important interactions between gene products, the development of comprehensive models relating genotype and phenotypes remains a challenge in the field of genomic medicine. In this context, a new view of the pathologic state as significant perturbation of the network of interactions between biomolecules is crucial for the identification of biochemical pathways associated with complex phenotypes. Seminal studies in systems biology combined the analysis of genetic variation with protein-protein interaction networks to demonstrate that even as biological systems evolve to be robust to genetic variation, their topologies create disease vulnerabilities. More recent analyses model the impact of genetic variants as changes to the "wiring" of the interactome to better capture heterogeneity in genotype-phenotype relationships. These studies lay the foundation for using networks to predict variant effects at scale using machine-learning or algorithmic approaches. A wealth of databases and resources for the annotation of genotype-phenotype relationships have been developed to support developments in this area. This overview describes how study of the molecular interactome has generated insights linking the organization of biological systems to disease mechanism, and how this information can enable precision medicine. This article is categorized under: Translational, Genomic, and Systems Medicine > Translational Medicine Biological Mechanisms > Cell Signaling Models of Systems Properties and Processes > Mechanistic Models Analytical and Computational Methods > Computational Methods.


Surviving the storms: Emergency preparedness in Texas nursing facilities and assisted living facilities.
This study assesses the preparedness of long-term care facilities in Texas responding to Hurricanes Katrina and Rita. A 41-item questionnaire was mailed to facilities; the response rate was 42%. Among responding facilities, 4513 residents were evacuated, and 6% of respondents reported resident death. Financial losses were reported by 8% of nursing facilities and 45% of assisted living facilities due to transportation and staff overtime. Respondents indicated the need for improved disaster preparednesstraining, better coordination, and transportation. Changes in policy and practice will lead to better trained staff who will provide the care residents need for improved health outcomes during future public health disasters.


Dbl oncogene expression in MCF-10 A epithelial cells disrupts mammary acinar architecture, induces EMT and angiogenic factor secretion.
The proteins of the Dbl family are guanine nucleotide exchange factors (GEFs) of Rho GTPases and are known to be involved in cell growth regulation. Alterations of the normal function of these proteins lead to pathological processes such as developmental disorders, neoplastic transformation, and tumor metastasis. We have previously demonstrated that expression of Dbl oncogene in lens epithelial cells modulates genes encoding proteins involved in epithelial-mesenchymal-transition (EMT) and induces angiogenesis in the lens. Our present study was undertaken to investigate the role of Dbl oncogene in epithelial cells transformation, providing new insights into carcinoma progression.To assess how Dbl oncogene can modulate EMT, cell migration, morphogenesis, and expression of pro-apoptotic and angiogenic factors we utilized bi- and 3-dimensional cultures of MCF-10 A cells. We show that upon Dbl expression MCF-10 A cells undergo EMT. In addition, we found that Dbl overexpression sustains Cdc42 and Rac activation inducing morphological alterations, characterized by the presence of lamellipodia and conferring a high migratory capacity to the cells. Moreover, Dbl expressing MCF-10 A cells form altered 3D structures and can induce angiogenesis by producing proangiogenic factors such as CCL2. These results support a role for Dbl oncogene in epithelial cell differentiation and transformation and suggest the relevance of GEF deregulation in tumor onset and progression.


The learning environment in the obstetrics and gynecology clerkship: an exploratory study of students' perceptions before and after the clerkship.
For reasons that remain not entirely clear, Obstetrics and Gynecology (Ob/Gyn) clerkships often exhibit comparatively higher rates of medical student mistreatment. To explore perceptions of our local learning environment, focus groups were held with students yet to start (pre-students) and students having completed (post-students) their Ob/Gyn clerkship. Topics of discussion included learning expectations and experiences, perceptions of mistreatment, and suggestions for improving the learning environment and student treatment. Using a naturalistic approach, we conducted a conventional content analysis to identify emergent themes. Nine pre-students and nine post-students participated. While pre-students anticipated being actively engaged, they also expected - based on peer accounts - to be subject to an unwelcoming learning environment on the Ob/Gyn clerkship, despite working hard to become team members. Due to patient advocacy and protection concerns, post-students reported low levels of student involvement and, subsequently, an overall passive learning experience. Students from both groups offered valuable suggestions for improving the learning environment and student treatment. The sensitive nature of Ob/Gyn clinical encounters may lead to overprotective behaviors that contribute to students feeling mistreated and excluded from patient care and team membership. Students' experiences during Ob/Gyn clerkships could be improved by better balancing patient advocacy and student involvement. Practical implications to address these issues are offered.


[Effect of porphyrins and their derivatives on biosynthesis of vitamin B 12 and the development of Propionibacterium shermanii].
The effect of uroporphyrin, coproporphyrin and their cobalt-containing derivatives on the biosynthesis of vitamin B12 and development of propionibacterium shermanii was studied. The compounds under study stimulated the vitamin synthesis by growing cultures and resting suspensions of these bacteria. Cobalt porphyrins as the sole source of cobalt were used in the vitamin B12 biosynthesis. An addition of cobalt porphyrins to the growing culture of propionic bacteria increased in accumulation of their biomass. Possible mechanisms of porphyrin involvement in the biosynthesis of vitamin B12 and the specific role of cobalt porphyrins in the bacterial activity are discussed.


Metabolic depression: a chronic depressive subtype? Findings from the InCHIANTI study of older persons.
Several studies report a cross-sectional association between metabolic syndrome and depression. Possibly, metabolic syndrome promotes onset or chronicity of depression. However, such a longitudinal link has not yet been confirmed. This study examines whether metabolic syndrome or its components are associated with onset and chronicity of depression. Secondary analyses were performed on data from 823 participants (≥ 65 years of age) in the InCHIANTI study, a prospective, population-based cohort study of older persons. From 1998 to 2000, the study sample was randomly selected from the population registry of 2 sites in Italy using a multistage stratified sampling method. Baseline data collection consisted of a home interview and a medical evaluation at the study clinic. Follow-up for each participant occurred after 3 years and 6 years. Metabolic syndrome at baseline was defined as ≥ 3 of the following: abdominal obesity, high triglycerides, low high-density lipoprotein cholesterol, high blood pressure, and high fasting glucose. Depressive symptoms were assessed using the Center for Epidemiologic Studies Depression scale (CES-D) at baseline and after 3 and 6 years. Sample characteristics were compared between persons with and without depression at baseline using χ² and t statistics. Logistic regression analyses were conducted separately in persons with and without depression at baseline to test whether metabolic syndrome at baseline could predict onset and chronicity of depression at follow-up. At baseline, 235 persons had metabolic syndrome, and 168 were depressed (CES-D score ≥ 20). Among persons not depressed at baseline, 26.0% developed depression. Higher waist circumference increased the odds of depression onset (adjusted OR per SD increase = 1.28; 95% CI, 1.05-1.56), but there was no association between other metabolic syndrome components and onset of depression. Among persons depressed at baseline, depression had a chronic character in 69.0% of persons without and 88.5% of persons with metabolic syndrome. Metabolic syndrome was associated with an almost 3-fold increase in the odds of chronicity of depression (adjusted OR = 2.66; 95% CI, 1.01-7.00), with almost every metabolic syndrome component contributing to this association. In late life, waist circumference, but not metabolic syndrome, predicted onset of depression. Depressed persons with metabolic syndrome were more likely to have persistent or recurrent depression. The latter may suggest that depression with metabolic abnormalities, which could be labeled metabolic depression, identifies a chronic subtype of depression.


Multimerin-2 maintains vascular stability and permeability.
Multimerin-2 is an extracellular matrix glycoprotein and member of the elastin microfibril interface-located (EMILIN) family of proteins. Multimerin-2 is deposited along blood vessels and we previously demonstrated that it regulates the VEGFA/VEGFR2 signaling axis and angiogenesis. However, its role in modulating vascular homeostasis remains largely unexplored. Here we identified Multimerin-2 as a key molecule required to maintain vascular stability. RNAi knockdown of Multimerin-2 in endothelial cells led to cell-cell junctional instability and increased permeability. Mechanistically cell-cell junction dismantlement occurred through the phosphorylation of VEGFR2 at Tyr951, activation of Src and phosphorylation of VE-cadherin. To provide an in vivo validation for these in vitro effects, we generated Multimerin-2-/- (Mmrn2-/-) mice. Although Mmrn2-/- mice developed normally and displayed no gross abnormalities, endothelial cells displayed cell junctional defects associated with increased levels of VEGFR2 phospho-Tyr949 (the murine counterpart of human Tyr951), impaired pericyte recruitment and increased vascular leakage. Of note, tumor associated vessels were defective in Mmrn2-/- mice, with increased number of small and often collapsed vessels, concurrent with a significant depletion of pericytic coverage. Consequently, the Mmrn2-/- vessels were less perfused and leakier, leading to increased tumor hypoxia. Chemotherapy efficacy was markedly impaired in Mmrn2-/- mice and this was associated with poor drug delivery to the tumor xenografts. Collectively, our findings demonstrate that Multimerin-2 is required for proper vessel homeostasis and stabilization, and unveil the possibility to utilize expression levels of this glycoprotein in predicting chemotherapy efficacy.


A collaborative resource to build consensus for automated left ventricular segmentation of cardiac MR images.
A collaborative framework was initiated to establish a community resource of ground truth segmentations from cardiac MRI. Multi-site, multi-vendor cardiac MRI datasets comprising 95 patients (73 men, 22 women; mean age 62.73±11.24years) with coronary artery disease and prior myocardial infarction, were randomly selected from data made available by the Cardiac Atlas Project (Fonseca et al., 2011). Three semi- and two fully-automated raters segmented the left ventricular myocardium from short-axis cardiac MR images as part of a challenge introduced at the STACOM 2011 MICCAI workshop (Suinesiaputra et al., 2012). Consensus myocardium images were generated based on the Expectation-Maximization principle implemented by the STAPLE algorithm (Warfield et al., 2004). The mean sensitivity, specificity, positive predictive and negative predictive values ranged between 0.63 and 0.85, 0.60 and 0.98, 0.56 and 0.94, and 0.83 and 0.92, respectively, against the STAPLE consensus. Spatial and temporal agreement varied in different amounts for each rater. STAPLE produced high quality consensus images if the region of interest was limited to the area of discrepancy between raters. To maintain the quality of the consensus, an objective measure based on the candidate automated rater performance distribution is proposed. The consensus segmentation based on a combination of manual and automated raters were more consistent than any particular rater, even those with manual input. The consensus is expected to improve with the addition of new automated contributions. This resource is open for future contributions, and is available as a test bed for the evaluation of new segmentation algorithms, through the Cardiac Atlas Project (www.cardiacatlas.org).


Occupational farm work activities influence workers' indoor home microbiome.
Farm work entails a heterogeneous mixture of exposures that vary considerably across farms and farmers. Farm work is associated with various health outcomes, both adverse and beneficial. One mechanism by which farming exposures can impact health is through the microbiome, including the indoor home environment microbiome. It is unknown how individual occupational exposures shape the microbial composition in workers' homes. We investigated associations between farm work activities, including specific tasks and pesticide use, and the indoor microbiome in the homes of 468 male farmers. Participants were licensed pesticide applicators, mostly farmers, enrolled in the Agricultural Lung Health Study from 2008 to 2011. Vacuumed dust from participants' bedrooms underwent whole-genome shotgun sequencing for indoor microbiome assessment. Using questionnaire data, we evaluated 6 farm work tasks (processing of either hay, silage, animal feed, fertilizer, or soy/grains, and cleaning grain bins) and 19 pesticide ingredients currently used in the past year, plus 7 banned persistent pesticide ingredients ever used. All 6 work tasks were associated with increased microbial diversity levels, with a positive dose-response for the total number of tasks performed (P = 0.001). All tasks were associated with altered microbial compositions (weighted UniFrac P = 0.001) and with higher abundance of specific microbes, including soil-based commensal microbes such as Haloterrigena. Among the 19 pesticides, current use of glyphosate and past use of lindane were associated with increased microbial diversity (P = 0.02-0.04). Ten currently used pesticides and all 7 banned pesticides were associated with altered microbial composition (P = 0.001-0.04). Six pesticides were associated with differential abundance of certain microbes. Different farm activities and exposures can uniquely impact the dust microbiome inside homes. Our work suggests that changes to the home microbiome could serve as one pathway for how occupational exposures impact the health of workers and their cohabitating family members, offering possible future intervention targets.


Destabilization of DNA through interstrand crosslinking by UO22.
UO22+ was shown to form an interstrand crosslink between two different strands of a single DNA molecule. This crosslink hardly affected the hydrogen bonds between nucleobase pairs but destabilized the π-π stacking between the two nucleobases in the vicinity of UO22+-bound phosphate. Thereby, the fragility of the DNA backbone increased upon UO22+ binding.


[Laboratory performance of primary care units in tuberculosis diagnosis in Pelotas, Brazil].
In order to assess laboratory performance of Primary Care Units (UAP, Spanish acronym) in TB diagnosis though the sputum smear test in Pelotas, Brazil, a descriptive study was completed which included all sputum smear tests requested at eleven primary care units from 2007 to 2010. To assess laboratory performance, the number of sputum smear tests requested was quantified as well as the time between the request and the reading of the glass slides at the lab. 477 requests were registered at UAP. Of this total, only 51,8% were found in the laboratory's record book, the lowest performance was 24,2% and the highest 71,4%. Regarding the reading of glass slides, it was determined that 27,7% readings took 8 days or more. It is concluded that the primary care units in Pelotas have a low laboratory performance in TB diagnosis by sputum smear tests.


Protease inhibitor and triple-drug therapy: cellular immune parameters are not restored in pediatric AIDS patients after 6 months of treatment.
To assess whether treatment of HIV-positive children by antiretroviral drugs for a 6-month period would improve immune function significantly. Immunological assessment of 89 HIV-positive children who received protease inhibitor monotherapy for 12-16 weeks as part of phase I/II studies, followed by triple antiretroviral therapy for an additional 12 weeks, was conducted. Immunological parameters were assessed in vitro at four time points (at enrollment, at weeks 2-4, at weeks 12-16, and at weeks 24-28). Assessments included: cytokine production by monocytes, T-cell proliferation to mitogen or recall antigens (including an HIV antigen) and apoptotic cell death. Plasma levels of tumor necrosis factor (TNF)-alpha and soluble TNF receptor (sTNF-R) were also measured, in addition to CD4+ T-lymphocyte counts and viral load. In addition, limited analyses were performed on samples from 17 children after 120 weeks of therapy, including 104 weeks of triple therapy. At enrollment, the 89 children exhibited severe immune defects. Antiretroviral therapy raised CD4+ T-lymphocyte counts significantly and decreased viral loads. In contrast, the in vitro immune parameters studied were not improved, except for plasma levels of sTNF-RII which decreased in parallel with the decrease in viral load. In addition, there was a trend towards increased skin test reactivity for the ritonavir-treated children. No differences were seen in the immune parameters whether the patients were treated with mono- or triple therapy. Results obtained after 120 weeks of therapy demonstrated that defective interleukin-12 production was not restored by long-term therapy. After 6 months of therapy, with the exception of decreased sTNF-RII levels, and a trend towards increased skin test reactivity, restoration of several defective cellular immune responses did not occur despite significantly decreased viral loads and increased CD4+ T-lymphocyte counts.


Implementation of unit-based interventions to improve teamwork and patient safety on a medical service.
In a prior study involving 2 medical units, Structured Interdisciplinary Rounds (SIDRs) improved teamwork and reduced adverse events (AEs). SIDR was implemented on 5 additional units, and a pre- versus postintervention comparison was performed. SIDR combined a structured format for communication with daily interprofessional meetings. Teamwork was assessed using the Safety Attitudes Questionnaire (score range = 0-100), and AEs were identified using queries of information systems confirmed by 2 physician researchers. Paired analyses for 82 professionals completing surveys both pre and post implementation revealed improved teamwork (mean 76.8 ± 14.3 vs 80.5 ± 11.6; P = .02), which was driven mainly by nurses (76.4 ± 14.1 vs 80.8 ± 10.4; P = .009). The AE rate was similar across study periods (3.90 vs 4.07 per 100 patient days; adjusted IRR = 1.08; P = .60). SIDR improved teamwork yet did not reduce AEs. Higher baseline teamwork scores and lower AE rates than the prior study may reflect a positive cultural shift that began prior to the current study.


Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants.
The estimation of nuclear DNA content has been by far the most popular application of flow cytometry in plants. Because flow cytometry measures relative fluorescence intensities of nuclei stained by a DNA fluorochrome, ploidy determination, and estimation of the nuclear DNA content in absolute units both require comparison to a reference standard of known DNA content. This implies that the quality of the results obtained depends on the standard selection and use. Internal standardization, when the nuclei of an unknown sample and the reference standard are isolated, stained, and measured simultaneously, is mandatory for precise measurements. As DNA peaks representing G1 /G0 nuclei of the sample and standard appear on the same histogram of fluorescence intensity, the quotient of their position on the fluorescence intensity axis provides the quotient of DNA amounts. For the estimation of DNA amounts in absolute units, a number of well-established standards are now available to cover the range of known plant genome sizes. Since there are different standards in use, the standard and the genome size assigned to it has always to be reported. When none of the established standards fits, the introduction of a new standard species is needed. For this purpose, the regression line approach or simultaneous analysis of the candidate standard with several established standards should be prioritized. Moreover, the newly selected standard organism has to fulfill a number of requirements: it should be easy to identify and maintain, taxonomically unambiguous, globally available, with known genome size stability, lacking problematic metabolites, suitable for isolation of sufficient amounts of nuclei, and enabling measurements with low coefficients of variation of DNA peaks, hence suitable for the preparation of high quality samples.


Estimation of the proportion of bacterial nitrogen in canine feces using diaminopimelic acid as an internal bacterial marker.
A bacterial marker can be used to determine the portion of fecal N that is of bacterial origin, as well as the effect of dietary factors on the bacterial N in feces of the dog. Two experiments were conducted to determine the efficacy of diaminopimelic acid (DAPA) and purines as bacterial markers in dogs. In Exp. 1, five adult female dogs were fed the same commercial diet. In Exp. 2, 50 dogs were fed one of four test diets (as-fed basis): a prebiotic-free control or diets containing either 1% chicory, 1% mannanoligosaccharide (MOS), or 1% chicory plus 1% MOS. Fresh feces were collected in both experiments and used to isolate a bacteria-rich sample (BRS) by differential centrifugation. In Exp. 1, the BRS had a N:purine ratio of 0.66 and N:DAPA ratio of 18.9. The CV for the N:purine ratio (20.7%) was much higher than that for the N:DAPA ratio (6.1%), indicating that DAPA resulted in a less variable estimate of fecal bacterial N. Using either marker, approximately 50% of the fecal N was estimated to be of bacterial origin. In Exp. 2, the N:DAPA ratio of the BRS did not differ (P = 0.14) among treatments. The BRS from dogs fed prebiotic-containing diets had treatment averages for N:DAPA ratios ranging from 16.9 to 18.5, whereas BRS from dogs fed the control diet had a ratio of 15.9. Averaged across all dogs, approximately 46% of fecal N was of bacterial origin. When calculating fecal bacterial concentrations using the average N:DAPA ratio for all dogs, little difference existed in the estimation compared with using individual values. The value resulting from use of the average ratio was approximately 13% higher than when using the individual ratios for dogs fed the control diet, which was due to the lower N:DAPA ratio for dogs fed the control diet compared with dogs fed the other treatments. Based on the consistency of the N:DAPA ratio of the BRS, DAPA seems to be a suitable marker for estimation of bacterial N in the feces of dogs.


Demystifying the CT Radiation Dose Sheet.
The radiation dose sheet generated by the CT scanner is a form that displays important information about an examination. It functions as a road map for the examination, detailing what CT examinations were performed and what parameters were used to perform them. One essential element of the radiation dose sheet, the volume CT dose index (CTDIvol), is a commonly used radiation dose index that is displayed on most CT scanners. The CTDIvol is used for quality control and is helpful for comparing the radiation output among different protocols and different scanners. The dose-length product (DLP) is a radiation dose index that builds on the CTDIvol by incorporation of the scan length. The DLP is combined with a conversion coefficient and used to determine the effective dose from the CT examination. Determining the effective dose is a way to estimate the whole-body radiation dose, even if the CT examination is confined to a smaller part of the body. In addition to these values, other data about the study from the CT scanner manufacturer, including the tube voltage and tube current-time product, usually are displayed on CT scanners. These values are major determinants of the image quality and radiation dose. The radiation dose sheet is a useful tool for radiologists, technologists, and physicists, allowing them to comprehend the technical details of a CT examination. The authors describe the components of the radiation dose sheet, the relationships of these components with one another, and the contributions of these components to the radiation dose. ©RSNA, 2022.


Gender and being born overseas influences the amount of acute stroke therapy.
To identify patient factors that influence the amount, frequency and intensity of physical and occupational therapy received by patients each weekday within 14 days of stroke. Exploration of data from studies conducted to monitor activity and therapy. Stroke patients receiving active treatment (not for palliative care). Physiotherapists and occupational therapists. Therapy data for 274 patients from 7 hospitals were included. Patients received a median of 40.0 min of therapy (physiotherapy plus occupational therapy) per weekday. Multivariable regression found that women had 22% less total therapy per weekday (factor change 0.78, 95% confidence interval (CI): 0.66-0.9, p = 0.001) and a decreased likelihood of receiving two or more sessions of therapy per weekday. Those born overseas had 23% less high intensity therapy per weekday (Factor change 0.77, 95% CI: 0.71-0.84, p < 0.001). Those with more severe stroke had a greater likelihood of receiving two or more sessions of therapy per weekday (OR = 1.05, 95% CI: 1.02-1.09, p = 0.006) but increasing severity increased the odds of receiving no high intensity therapy by 7% (factor change 1.07, 95% CI: 1.02-1.11, p = 0.002). Age did not exhibit a significant association. There is some evidence that patient factors may influence the amount of therapy provided. Physiotherapists and occupational therapists should be aware of potential biases associated with therapy provision.


Long-term continuation of methotrexate therapy in giant cell arteritis patients in clinical practice.
To assess the long-term continuation of methotrexate (MTX) in a cohort of patients with giant cell arteritis (GCA) in daily clinical practice. Factors associated with its discontinuation rate were also investigated. A longitudinal study from 1991-2014, was performed. GCA patients with MTX and followed-up in a rheumatology outpatient clinic of Madrid during the study period were included. discontinuation of MTX due to: adverse drug reactions (ADR moderate and severe); inefficacy; sustained clinical response; patient decision. Covariables: sociodemographic, clinical and therapy. Incidence rates (IR) of MTX discontinuation per 100 patient-years with their 95% confidence interval (CI) were estimated using survival techniques. Factors associated to specific discontinuation causes were analysed using Cox models. We included 108 patients (244 patient-years). The IR was 37.2 [30.3-45.7]. The IR due to ADR, severe ADR, sustained clinical response and inefficacy was 20.8 [15.8-27.4]; 5.7 [3.3-9.6]; 8.2 [5.3-12.7] and 2.8 [1.3-6.0], respectively. Regarding multivariate analysis, younger patients, baseline cardiovascular disease, taking more glucocorticoids and lower initial doses of MTX were associated to a higher discontinuation rate due to inefficacy. Factors influencing the suspension due to ADRs were: older age, baseline. Chronic obstructive pulmonary disease, higher baseline erythrocyte sedimentation rate, several specific clinical patterns at diagnosis, and higher maximum dose of MTX during the follow up. In the final model for sustained clinical response older patients and more recurrences were independently associated to less discontinuation rate. We provide further data of the potential safety of long-term MTX in the management of GCA. We have also found several factors influencing the continuation of MTX.


Investigating dendrites and side reactions in sodium-oxygen batteries for improved cycle lives.
The development of sodium-oxygen batteries with high round-trip efficiencies is hindered by the short cycle lives. Sodium dendrite formation and oxygen crossover are identified as two major issues. By employing an ion selective membrane, the cycle life of sodium-oxygen batteries has been greatly improved.


Preparation and evaluation of microencapsulated fast melt tablets of ambroxol hydrochloride.
Natural resources in general and plant materials in particular are receiving more attention due to their safety as pharmaceutical excipients. Present work assessed the potential of a natural polysaccharide, pectin to mask the bitter taste of ambroxol hydrochloride, by microencapsulation technique, and its possibility to formulate as a fast disintegrating dosage form. Taste masking is an important developmental challenge in fast dissolving drug delivery system since it dissolves or disintegrates in the patient's mouth in close proximity to the taste buds. The prepared microspheres by emulsion solvent evaporation technique possessed good sphericity, smooth surface morphology, uniform and narrow size distribution (10-90 mum), when analyzed by scanning electron microscopy, laser diffraction and optical microscopy. Method of preparation has influenced the particle size and drug loading efficiency. Drug-polymer compatibility was confirmed by Fourier transform infrared spectroscopy and thin layer chromatography. DSC and X-ray diffraction studies revealed that the drug was dispersed inside the microspheres in the form of an insoluble matrix. The formation of microspheres was affected by glass transition temperature of the polymer, surfactant, type of plasticizers, volume of internal phase, stirrer speed etc. Fast dissolving tablets were prepared by the modification of melt granulation technique. The resulting granules were found to melt fast at body temperature, have smooth mouth feel and good physical stability. This study demonstrated that pectin could be a right choice in developing patient favored formulations for bitter drugs and can be utilized in fast disintegrating dosage forms as well.


Determination of Aluminum, Chromium, and Barium Concentrations in Infant Formula Marketed in Lebanon.
Infant formula is a major nutritional component for many infants and toddlers. However, the presence of contaminants, such as toxic metals, may pose increased health risks to infants. An investigation of the total concentrations of the metals aluminum (Al), barium (Ba), and chromium (Cr) in infant formulae marketed in Lebanon was performed. Powdered dairy and nondairy infant formula samples were collected from all commercially available brands (n = 39) in the Lebanese market on two production dates (78 samples in total) and analyzed for these three metals with inductively coupled plasma mass spectrometry. All brands contained detectable concentrations of Al and Ba, Cr was detected in 95% of brands. Mean (±standard deviation) concentrations of the metals were estimated as 1.54 ± 1.43 (Al), 0.256 ± 0.593 (Ba), and 0.168 ± 0.143 (Cr) (μg/g). The concentration ranges in the powdered formula were 0.080 to 7.93 (Al), 0.038 to 5.35 (Ba), and 0.041 to 0.348 (Cr) μg/g. A significant difference in the mean concentrations of Al, Ba, and Cr for the two production dates of a single brand was observed in 92, 59, and 83% of samples, respectively. The mean concentration of Al in the soy-based formula was significantly higher than that of aluminum in milk-based and corn-based formulas (P = 0.018). Cr concentrations in the continuation special formulations were significantly higher than those in the beginner formulations (P = 0.008). Our study provides the first publicly available information on metal contamination in infant formulas in Lebanon and reveals the need for frequent monitoring and surveillance of these products intended for infant consumption.


Factors influencing virus-host cell interplay.
The Conference on Retroviruses and Opportunistic Infections (CROI) provides an annual international forum for basic scientists and clinical and global health researchers to present and become current on the most recent advances in the field of HIV and AIDS research. The 18th conference contained a number of strong basic science sessions. HIV-1 infection of the cell is opposed by cellular factors that attack the viral replication cycle at various points. However, the virus has evolved defenses against these innate cellular antiviral proteins. CROI continues to be a strong forum for presentation of the most recent developments in this area of research. In addition, there were numerous presentations on cellular factors that regulate virus-host cell interplay as well as on research that is providing detailed insight into the mechanism of action of integrase inhibitors. Some presentations focused on approaches to studying and intervening with viral latency, particularly in primary cell models. Research on the use of zinc-finger nucleases to knock out CC chemokine receptor R5 expression in CD34+ stem cells also received a lot of interest. The hope is that these strategies will provide new therapeutic approaches to generate resistance to HIV-1 infection.


Patient and doctor perspectives on HIV screening in the emergency department: A prospective cross-sectional study.
The emergency department (ED) is mentioned specifically in the Swiss HIV testing recommendations as a site at which patients can benefit from expanded HIV testing to optimise early HIV diagnosis. At our centre, where local HIV seroprevalence is 0.2-0.4%, 1% of all patients presenting to the ED are tested for HIV. Barriers to HIV testing, from the patient and doctor perspective, and patient acceptability of rapid HIV testing were examined in this study. Between October 2014 and May 2015, 100 discrete patient-doctor encounter pairs undertook a survey in the ED of Lausanne University Hospital, Switzerland. Patients completed a questionnaire on HIV risk factors and were offered free rapid HIV testing (INSTI™). For every patient included, the treating doctor was asked if HIV testing had 1) been indicated according to the national testing recommendations, 2) mentioned, and 3) offered during the consultation. Of 100 patients, 30 had indications for HIV testing through risk factors or a suggestive presenting complaint (PC). Fifty patients accepted rapid testing; no test was reactive. Of 50 patients declining testing, 82% considered themselves not at risk or had recently tested negative and 16% wished to focus on their PC. ED doctors identified 20 patients with testing indications, mentioned testing to nine and offered testing to six. The main reason for doctors not mentioning or not offering testing was the wish to focus on the PC. Patients and doctors at our ED share the testing barrier of wishing to focus on the PC. Rapid HIV testing offered in parallel to the patient-doctor consultation increased the testing rate from 6% (offered by doctors) to 50%. Introducing this service would enable testing of patients not offered tests by their doctors and reduce missed opportunities for early HIV diagnosis.


[Role of measurement of nitric oxide in respiratory diseases].
Nitric oxide (NO) is a molecule with a homeostatic role in a number of physiological processes. Concerning respiratory diseases, exhaled nitric oxide (FE(NO)) is a marker of airway inflammation, and its measurement can be easily performed. To date, a large number of publications defined the performances of FE(NO). Studies on asthma attributed to FE(NO) an important role in diagnosis and especially in monitoring the effects of anti-inflammatory treatment, which label it as an "inflammometer" to be used as a guide in therapy algorithms. Less consistent results were thus far obtained in chronic obstructive lung disease, in which FE(NO) levels seem usually higher than in healthy subjects but lower than in asthma, unless an eosinophil inflammation is present, and in rhinosinusitis, where the levels of nasal NO (nNO) are low, probably because of a reduced NO flow into the nose due to mucosal swelling, while they increase after an effective treatment. Among other respiratory disorders, such as cystic fibrosis and primary ciliary dyskinesia, nNO levels are particularly low in the latter (possibly for the trapping and altered NO metabolism caused by dense secretions, and by decreased NO synthase activity) and suggest nNO as a good screening tool for such disease.


Mustard seed meal mixtures: management of Meloidogyne incognita on pepper and potential phytotoxicity.
Meals produced when oil is extracted from seeds in the Brassicaceae have been shown to suppress weeds and soilborne pathogens. These seed meals are commonly used individually as soil amendments; the goal of this research was to evaluate seed meal mixes of Brassica juncea (Bj) and Sinapis alba (Sa) against Meloidogyne incognita. Seed meals from Bj 'Pacific Gold' and Sa 'IdaGold' were tested alone and in combinations to determine rates and application times that would suppress M. incognita on pepper (Capsicum annuum) without phytotoxicity. Rates of soil application (% w/w) for the phytotoxicity study were: 0.5 Sa, 0.2 Bj, 0.25 Sa + 0.25 Bj, 0.375 Sa + 0.125 Bj, 0.125 Sa + 0.375 Bj, and 0, applied 0 - 5 weeks before transplant. Overall, 0.2% Bj was the least toxic meal to pepper seedlings. By comparison, 0.5% S. alba seed meal did not reduce lettuce (Lactuca sativa) seed germination at week 0, but all seed meal treatments containing B. juncea prevented or significantly reduced germination at week 0. The seed meals did not affect lettuce seed germination at weeks 1-5, but hypocotyl growth was reduced by all except 0.2% Bj at weeks 1, 4 and 5. Brassica juncea and Sa meals were tested for M. incognita suppression at 0.2, 0.15, 0.1 and 0.05%; mixtures were 0.1% Sa + 0.1% Bj, 0.15% Sa + 0.05% Bj, and 0.05% Sa + 0.15% Bj. All treatments were applied 2 weeks before transplant. The 0.2% Bj and 0.05% Sa + 0.15% Bj treatments overall had the longest shoots and highest fresh weights. Eggs per g root were lowest with 0.1 - 0.2% Bj amendments and the seed meal mixtures. The results indicate that Bj and some Bj + Sa mixtures can be applied close to transplant to suppress M. incognita populations on pepper; consequently, a seed meal mixture could be selected to provide activity against more than one pest or pathogen. For pepper, care should be taken in formulating mixtures so that Sa rates are low compared to Bj.


The interaction of ryanoids with individual ryanodine receptor channels.
Ryanodine binds with high affinity and specificity to a class of Ca(2+)-release channels known as ryanodine receptors (RyR). The interaction with RyR results in a dramatic alteration in function with open probability (Po) increasing markedly and rates of ion translocation modified. We have investigated the features of ryanodine that govern the interaction of the ligand with RyR and the mechanisms underlying the subsequent alterations in function by monitoring the effects of congeners and derivatives of ryanodine (ryanoids) on individual RyR2 channels. While the interaction of all tested ryanoids results in an increased Po, the amplitude of the modified conductance state depends upon the structure of the ryanoid. We propose that different rates of cation translocation observed in the various RyR-ryanoid complexes represent different conformations of the channel stabilized by specific conformers of the ligand. On the time scale of a single channel experiment ryanodine binds irreversibly to the channel. However, alterations in structure yield some ryanoids with dissociation rate constants orders of magnitude greater than ryanodine. The probability of occurrence of the RyR-ryanoid complex is sensitive to trans-membrane voltage, with the vast majority of the influence of potential arising from a voltage-driven alteration in the affinity of the ryanoid-binding site.


Involvement of the bed nucleus of the stria terminalis in initial conditioning and rapid reconditioning following extinction of contextual fear.
Although a great deal is known about neurobiological mechanisms of initial conditioning and extinction, relatively little is known about mechanisms involved in the return of behavior following extinction. In this article, we examine the effects of temporarily inactivating the bed nucleus of the stria terminalis (BNST) on initial conditioning and postextinction reconditioning. We investigate effects in unsignaled contextual fear conditioning, in which animals initially receive strong contextual conditioning, followed by three sessions of nonreinforced context exposure (extinction), and then receive a single context-shock reconditioning trial. In 2 experiments with male Long Evans rats, we evaluated the effects of delivery of a muscimol/baclofen cocktail to the BNST prior to initial conditioning or reconditioning. In Experiment 1, we found that a single context-shock pairing results in more freezing following extinction than when it is the initial conditioning trial. This rapid reconditioning effect was impaired by BNST inactivation. In Experiment 2, we found that BNST inactivation also causes a deficit in freezing after strong initial conditioning. These findings suggest that the BNST is involved in both initial conditioning and postextinction reconditioning. We discuss implications of these findings for current thinking about BNST function in learning and memory processes. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


[Metabolic syndroma long term results].
Several studies in obesity verify more effectiveness in weight-loss if diet and exercise were combined with behavioral modification. The main problem with all obesity treatments is a return to baseline weight after treatment ends. We have some evidence, that long-term behavioral methods may be successful to induce long-term weight-loss maintenance. Strategies for improving weight-loss maintenance in rehabilitation are discussed