Functional reconstitution of rhodopsin into tubular lipid bilayers supported by nanoporous media.
We report on a novel reconstitution method for G-protein-coupled receptors (GPCRs) that yields detergent-free, single, tubular membranes in porous anodic aluminum oxide (AAO) filters at concentrations sufficient for structural studies by solid-state NMR. The tubular membranes line the inner surface of pores that traverse the filters, permitting easy removal of detergents during sample preparation as well as delivery of ligands for functional studies. Reconstitution of bovine rhodopsin into AAO filters did not interfere with rhodopsin function. Photoactivation of rhodopsin in AAO pores, monitored by UV-vis spectrophotometry, was indistinguishable from rhodopsin in unsupported unilamellar liposomes. The rhodopsin in AAO pores is G-protein binding competent as shown by a [35S]GTPgammaS binding assay. The lipid-rhodopsin interaction was investigated by 2H NMR on sn-1- or sn-2-chain perdeuterated 1-stearoyl-2-docosahexaenoyl-sn-glycero-3-phospholine as a matrix lipid. Rhodopsin incorporation increased mosaic spread of bilayer orientations and contributed to spectral density of motions with correlation times in the range of nano- to microseconds, detected as a significant reduction in spin-spin relaxation times. The change in lipid chain order parameters due to interaction with rhodopsin was insignificant.


Bivariate Genomic Footprinting Detects Changes in Transcription Factor Activity.
In response to activating signals, transcription factors (TFs) bind DNA and regulate gene expression. TF binding can be measured by protection of the bound sequence from DNase digestion (i.e., footprint). Here, we report that 80% of TF binding motifs do not show a measurable footprint, partly because of a variable cleavage pattern within the motif sequence. To more faithfully portray the effect of TFs on chromatin, we developed an algorithm that captures two TF-dependent effects on chromatin accessibility: footprinting and motif-flanking accessibility. The algorithm, termed bivariate genomic footprinting (BaGFoot), efficiently detects TF activity. BaGFoot is robust to different accessibility assays (DNase-seq, ATAC-seq), all examined peak-calling programs, and a variety of cut bias correction approaches. BaGFoot reliably predicts TF binding and provides valuable information regarding the TFs affecting chromatin accessibility in various biological systems and following various biological events, including in cases where an absolute footprint cannot be determined.


Comparison of diagnostic tests for hymenoptera sting allergy.
The authors examined correlations among individual Hymenoptera venom skin tests, venom radioallergosorbent tests (RAST), venom-induced leukocyte histamine release (LHR) assays and individual Hymenoptera whole body extract (WBE) skin tests in 37 patients with histories of systemic reactions to Hymenoptera stings. Significant positive correlations were seen between the venom skin test results and results from either the venom RAST or the LHR assay for most venoms. There was a relatively high frequency of positive WBE skin tests in association with other negative tests. The ultimate diagnostic test for stinging insect sensitivity is a deliberate sting challenge; in lieu of such a sting the clinical history and the venom skin test appear to provide the best estimate of clinical Hymenoptera sting sensitivity.


Weight Loss Outcomes and Lifestyle Patterns Following Sleeve Gastrectomy: an 8-Year Retrospective Study of 212 Patients.
Sleeve gastrectomy (SG) is an effective treatment for extreme obesity; however, long-term weight loss outcomes remain largely understudied. We aimed to examine the long-term weight changes following SG and patient characteristics and lifestyle patterns related to weight loss outcomes. Data from medical records of patients operated in a tertiary university medical center between 2008 and 2014 were reviewed, along with information derived from a telephone questionnaire. Information included the following: current medical status, medications, side effects, behavior, lifestyle habits, and weight changes. A total of 212 patients (69.3% females) were included, with a median follow-up duration of 8 years post-SG. Mean age and baseline body mass index (BMI) of participants were 39.7 ± 12.0 years and 42.2 ± 4.9 kg/m2, respectively. Mean BMI, percentage excess weight loss, and percentage total body weight loss were 33.1 ± 6.1, 55.5 ± 27.5%, and 21.7 ± 10.7%, respectively. Higher baseline BMI was found to be the strongest independent predictor for insufficient weight loss (OR = 0.90, P = 0.001, 95% CI 0.85, 0.96). Sweetened beverage intake, usage of psychiatric medications, higher initial BMI, and lower age were significant predictors for increased weight gain from nadir weight (P < 0.0001, P = 0.005, P = 0.035, and P < 0.0001, respectively). SG patients were found to maintain a notable long-term weight loss. Nevertheless, weight regain and insufficient weight loss were prevalent in the long-term post-operative period, and were related to certain lifestyle patterns. Clinical practice should emphasize the relationship between post-operative weight loss outcomes and specific behaviors. Efforts should be made to educate patients on the need for lifelong follow-up and support.


Cell-free DNA mutations as biomarkers in breast cancer patients receiving tamoxifen.
The aim was to identify mutations in serum cell-free DNA (cfDNA) associated with disease progression on tamoxifen treatment in metastatic breast cancer (MBC). Sera available at start of therapy, during therapy and at disease progression were selected from 10 estrogen receptor (ER)-positive breast cancer patients. DNA from primary tumor and normal tissue and cfDNA from minute amounts of sera were analyzed by targeted next generation sequencing (NGS) of 45 genes (1,242 exons). At disease progression, stop-gain single nucleotide variants (SNVs) for CREBBP (1 patient) and SMAD4 (1 patient) and non-synonymous SNVs for AKAP9 (1 patient), PIK3CA (2 patients) and TP53 (2 patients) were found. Mutations in CREBBP and SMAD4 have only been occasionally reported in breast cancer. All mutations, except for AKAP9, were also present in the primary tumor but not detected in all blood specimens preceding progression. More sensitive detection by deeper re-sequencing and digital PCR confirmed the occurrence of circulating tumor DNA (ctDNA) and these biomarkers in blood specimens.


Liblomycin-mediated DNA cleavage in human head and neck squamous carcinoma cells and purified DNA.
Liblomycin (LBM), a novel bleomycin analogue, and bleomycin A2 (BLM A2) were compared with respect to their relative potential to inhibit growth in a human head and neck squamous carcinoma cell line and to produce DNA damage within cellular DNA and nuclei DNA and against isolated naked DNA. Against the BLM-sensitive cell line 183A, the concentration of LBM that inhibits cell growth by 50% was 1.1 microM for a 30-min drug exposure, while it was 23 microM for BLM A2. Drug-mediated DNA double-strand cleavage within cells was compared with the relative ability of these drugs to produce DNA cleavage in isolated 183A cell nuclei. Though 30-min exposures of cells to equimolar concentrations of both drugs resulted in 4-fold greater cellular DNA damage by LBM than BLM A2, the two drugs were nearly equipotent in producing DNA injury within isolated nuclei. Against Simian virus 40 DNA, however, LBM was 10-fold less effective than BLM A2 in producing Forms II and III DNA from Form I DNA. Radioactivity from either [3H]BLM A2 or 125I-LBM found associated with cells after a 30-min incubation period was also assessed in the 183A cell line. The exposure of cells to radiolabeled drug (1 microM) resulted in a 71-fold greater amount of cell-associated radioactivity for LBM than for BLM A2. The relative abilities of the 183A cell line to partially reseal LBM- or BLM A2-mediated DNA double-strand breaks were also assessed. No preferential repair of overall drug-mediated DNA injury, however, was observed. Finally, drug-mediated specific cleavage sites on pBR322 DNA were determined. At doses that gave the same extent of DNA cleavage, both BLM A2 and LBM gave similar patterns of strand scission, although minor differences were observed. Taken together, these data demonstrate that the greater efficacy of LBM against the BLM-sensitive head and neck squamous cell line is due mainly to LBM's greater association with cells over a defined time period, even though the DNA cleaving ability of LBM is relatively lower than that of BLM A2.


Sodium alginate-magnesium aluminum silicate composite gels: characterization of flow behavior, microviscosity, and drug diffusivity.
The aims of the present study were to characterize the flow behavior and thixotropic properties of sodium alginate-magnesium aluminum silicate (SA-MAS) composite gels with various ratios of SA and MAS, and to investigate the drug diffusivity and microviscosity of the composite gels. Moreover, interaction of SA and MAS in the form of dry composite was examined by using Fourier Transform Infrared (FTIR), and a possible structure model of SA-MAS composite gel was illustrated. Incorporating MAS into the SA gels provided higher viscosity and changed the flow behavior from Newtonian to pseudoplastic with thixotropy. This was due to the formation of electrostatic force and intermolecular hydrogen bonding between SA and MAS, leading to a denser matrix structure of the composite gels. Increasing the content of MAS decreased the drug diffusivity but increased the microviscosity of the composite gels. The denser matrix structure of the composite gels had a higher tortuosity, resulting in slower drug diffusion through water-filled channels in the gels. This finding suggested that incorporating MAS into the SA gels could improve the flow behavior and sustain drug release from the gels because of the formation of a matrix structure between SA and MAS in the gels.


A descriptive analysis of seizure events among adults who participated in a computer-based assessment.
The purpose of this study was to document seizure events associated with the use of a computer-based assessment and to describe the contextual factors surrounding these seizure episodes. Study participants were adults with epilepsy who were enrolled at research sites in Atlanta and Boston. Subjects were asked to complete a computer-based assessment at 3 time points. Fourteen seizure events were documented; they occurred during 1.6% of all completed assessments (896) and affected 4.4% of the participants (320). The mean age of participants who experienced seizure events was 41.4 years; about 70% were female, and 70% were white. A variety of possible precipitating factors for seizure events included hunger, fatigue, stress, and medication changes. Participants indicated computer use could have triggered their seizures in 2 instances. These findings suggest use of computer-based assessments may pose minimal risks for adults with epilepsy, particularly those without a history of photosensitivity epilepsy.


Birt-Hogg-Dubé Syndrome - report of two cases with two new mutations.
Birt-Hogg-Dubé syndrome (BHDS) is a rare autosomal dominant genodermatosis characterized by cutaneous fibrofolliculomas and/or trichodiscomas, lung cysts, spontaneous pneumothorax and renal tumors. However, its clinical expression is highly variable. This syndrome is caused by germline mutations in the folliculin gene (FLCN) on chromosome 17p11.2. Two men, 60 and 39-year-old, presented with a several year history of asymptomatic whitish papules scattered over the face and neck. Skin biopsies revealed fibrofolliculomas. The clinical diagnosis of BHDS was corroborated by identification of new heterozygotic mutations in FLCN gene, in exon 6 (C.573_574delinsT) and in exon 9 (c.1015C>T), respectively. Computed tomography scan of the thorax and abdomen showed pulmonary cysts with no suspicious kidneys lesions, and, in the case of the second patient, a mass in left adrenal gland. Laparoscopic left adrenalectomy was performed and histopathological examination was compatible with a malignant perivascular epithelioid cell tumor. The presence of multiple fibrofolliculomas should raise the suspicion of BHDS. Patients with this syndrome, regardless of the detected mutation, should be carefully monitored to ensure that potentially serious disease-related conditions can be detected early.


Relative bioavailability of L-ascorbyl-2-polyphosphate in broiler chickens.
Broiler chick diets and drinking water were supplemented with two sources of vitamin C: crystalline L-ascorbic acid (AsA) or L-ascorbyl-2-polyphosphate (APP) to provide 0, 25, 50, 100, 200, 400, 800, 1,600, and 3,200 ppm (mg/kg) AsA. The bioavailability of APP relative to AsA, as estimated by the change in plasma AsA concentration, was evaluated during 24-h periods of supplementation. When provided in the feed, no differences in dietary AsA content were attributed to vitamin source. In contrast, APP administration at 25 and 50 ppm, resulted in higher (P < .001) AsA values in drinking water when compared with AsA supplementation. Plasma AsA values were elevated (P < .05) above baseline when either AsA or APP were supplemented in the feed or water at a level of 400 ppm or greater. Plasma AsA concentrations, following supplementation of the diets, were higher (P < .05) in AsA-treated (800 ppm) chicks when compared with APP-supplemented chicks. During water supplementation, AsA (800 ppm) and APP (3,200 ppm) administration resulted in higher plasma AsA values when compared with their alternate vitamin source. At all other levels of water supplementation, no differences in plasma AsA were associated with vitamin source. The absence of a consistent difference in plasma AsA, relative to vitamin source, suggests that the isolated differences observed may be due to chance. It was concluded that APP was of similar bioavailability to that of AsA, as estimated by the ability to elevate plasma AsA concentrations in broiler chicks.


First report of Schmallenberg virus infection in cattle and midges in Poland.
Two outbreaks of Schmallenberg virus (SBV) infection that coincided with the introduction of two bulls imported from France into two herds located in West Pomerania and Silesia provinces in Poland are described in detail. The first SBV real-time RT-PCR-positive result was obtained during routine testing of one of the imported bulls. The second bull and the affected farms were tracked by further investigation. Transmission of SBV into Polish cattle herds where the bulls were imported was confirmed by viral RNA detection in real-time RT-PCR, virus isolation followed by immunoperoxidase (IPX) staining and seroconversion. SBV RNA was detected also in Culicoides obsoletus pools caught in a trap located 5 km from one of the outbreaks. Testing nearly 900 samples collected prior to the two outbreaks from the same areas or provinces neighbouring with Germany where SBV cases had previously been detected gave negative results for the presence of SBV or specific antibodies. These cases are the first ones detected in cattle in Poland and provide evidence of recent transmission of the pathogen into the country and involvement of midge vectors.


Long-term excitability and fine tuning of nerve pedicles reinnervating strap muscles in the dog.
Contraction of paralyzed striated muscles has been restored by stimulating reinnervating pedicles with currents of low intensity. In order to allow clinical application, stable, long-term excitability must emulate the parameters necessary for the stimulation of normal motor nerves. In 6 dogs, the ansa hypoglossi nerve was implanted into the contralateral denervated sternohyoid muscle and surrounded with a bipolar cuff electrode. Three of the reinnervating pedicles were chronically paced with a Medtronic Itrel II Multiprogrammable Pulse Generator (0.5 V, 0.2 second on [30 pulses per second, 0.21-millisecond pulse width], 2.9 seconds off). At reexploration after 8 months (6 months for 1 dog), frank contraction confirmed by electromyography tracings occurred in all animals with currents in the range of 0.1 to 0.5 mA. Muscle force was further manipulated by selective release of blocking currents (600 Hz, 1.7 to 0.4 mA) superimposed over regular stimulation (50 Hz, 0.3 to 1.7 mA). Nerve and muscle vitality were histologically confirmed. Long-term, low-intensity conduction capabilities, fine tuning, good tolerance of implanted electrodes, and lack of fatigue suggest that reinnervating pedicles may be successfully used for pacing when clinically indicated.


Dynamical disease: Challenges for nonlinear dynamics and medicine.
Dynamical disease refers to illnesses that are associated with striking changes in the dynamics of some bodily function. There is a large literature in mathematics and physics which proposes mathematical models for the physiological systems and carries out analyses of the properties of these models using nonlinear dynamics concepts involving analyses of the stability and bifurcations of attractors. This paper discusses how these concepts can be applied to medicine.


Early estimates of seasonal influenza vaccine effectiveness in Europe: results from the I-MOVE multicentre case-control study, 2012/13.
We conducted a test-negative case-control study based in five European sentinel surveillance networks. The early 2012/13 adjusted influenza vaccine effectiveness was 78.2% (95% CI: 18.0 to 94.2) against influenza B, 62.1% (95% CI: -22.9 to 88.3%) against A(H1) pdm09, 41.9 (95% CI: -67.1 to 79.8) against A(H3N2) and 50.4% (95% CI: -20.7 to 79.6) against all influenza types in the target groups for vaccination. Efforts to improve influenza vaccines should continue to better protect those at risk of severe illness or complications.


Muscle Tone Assessment under General Anesthesia for Sjögren-Larsson Syndrome and Spasticity.
Study of muscle tone in individuals with severe spasticity (Modified Asworth Scale - MAS:3) under general anesthesia can confirm or rule out the eventual necessity of the impending spasticity relieving ablative neurosurgery by observing the hypertonia reduction and passive range of motion expansion. Therefore, what we measure under muscle relaxants is practically a fixed deformity. The study was performed on a girl with Sjögren-Larsson syndrome, presenting with icthyosis and spastic diplegia. Proposed intervention was Dorsal Rhizotomy. Under general anesthesia, with and without muscle relaxants, hypertonia was significantly reduced (MAS:1), but the angle of motion did not increase much. We decided not to perform such a neurosurgical procedure. In ambiguous situations, the proposed study can help in decision-making for spasticity treatment.


The role of robotic gait training coupled with virtual reality in boosting the rehabilitative outcomes in patients with multiple sclerosis.
Motor impairment is the most common symptom in multiple sclerosis (MS). Thus, a variety of new rehabilitative strategies, including robotic gait training, have been implemented, showing their effectiveness. The aim of our study was to investigate whether an intensive robotic gait training, preceding a traditional rehabilitative treatment, could be useful in improving and potentiating motor performance in MS patients. Forty-five patients, who fulfilled the inclusion criteria, were enrolled in this study and randomized into either the control group (CG) or the experimental group (EG). A complete clinical evaluation, including the Expanded Disability Severity Scale, the Functional Independence Measure, the Hamilton Rating Scale for Depression, the time up and go test (TUG), and the Tinetti balance scale, was performed at baseline (T0), after 6 week (T1), at the end of rehabilitative training (T2), and 1 month later (T3). A significant improvement was observed in the EG for all the outcome measures, whereas the CG showed an improvement only in TUG. In contrast, from T1 to T2, only CG significantly improved in all outcomes, whereas the EG had an improvement only regarding TUG. From T2 to T3, no significant differences in Functional Independence Measure scores emerged for both the groups, but a significant worsening in Tinetti balance scale and TUG was observed for the CG and in TUG for the EG. Our study provides evidence that robotic rehabilitationn coupled with two-dimensional virtual reality may be a valuable tool in promoting functional recovery in patients with MS.


Evaluation of the Effect of Comprehensive Nursing Interventions on Plaque Control in Patients with Periodontal Disease in the Context of Artificial Intelligence.
Plaque is a bacterial biofilm that adheres to each other and exists on the tooth surface, and new plaque can continuously reform after removing it from the tooth surface. The pathogenesis of periodontal disease is related to the bacteria, the host and the environment, with the bacteria and bacterial products in plaque being the main initiators of periodontal disease. The effective control of plaque is an effective method for the treatment and prevention of periodontal disease and is often underappreciated in clinical practice. For the traditional diagnostic method through experience and visual observation, it may lead to misdiagnosis and underdiagnosis. In order to accurately diagnose plaque disease, this study designed a convolutional neural network-based oral dental disease diagnosis system for oral care interventions to improve oral health awareness. Thus motivate patients to implement proper oral health care measures, and continuously and lifelong insist on thorough daily plaque removal to improve patients' physical health and quality of life in periodontal disease patients.


Estimation of Chest Wall Attenuation of 123I Emissions in Substernal Goiter: A Phantom Study.
In patients with substernal goiter, the generally accepted theory is that thyroid uptake measurements with iodine isotopes are underestimated because of attenuation by the chest wall. The extent of this underestimation is not well known. In this study, we calculated the attenuation of 123I emissions using a cadaver chest wall with a thyroid probe to better understand the potential severity of this underestimation. Methods: A 11.1-MBq capsule of 123I was measured using a thyroid probe directly in a standard neck phantom and behind a cadaver chest wall that included the soft tissues and bony structures (sternum). Results: The calculated attenuation of the iodine capsule was 18% for the neck phantom and 35% for the cadaver chest wall. Conclusion: Thyroid uptake in cases of substernal goiter may be underestimated by standard techniques using a neck phantom. The composition of the chest wall can vary greatly, and the substernal extent of the goiter would be difficult to calculate with high accuracy on a routine basis. Comparison between the cadaveric specimen and the phantom does give us a rough estimation of the differences in attenuation. Our findings suggest that attenuation by the chest wall can be substantial. Knowledge of the extent of the substernal component of the thyroid gland may be useful if the uptake measurement is used to calculate doses for treating hyperthyroidism in patients with substernal goiter.


Detecting Cochlear Synaptopathy Through Curvature Quantification of the Auditory Brainstem Response.
The sound-evoked electrical compound potential known as auditory brainstem response (ABR) represents the firing of a heterogenous population of auditory neurons in response to sound stimuli, and is often used for clinical diagnosis based on wave amplitude and latency. However, recent ABR applications to detect human cochlear synaptopathy have led to inconsistent results, mainly due to the high variability of ABR wave-1 amplitude. Here, rather than focusing on the amplitude of ABR wave 1, we evaluated the use of ABR wave curvature to detect cochlear synaptic loss. We first compared four curvature quantification methods using simulated ABR waves, and identified that the cubic spline method using five data points produced the most accurate quantification. We next evaluated this quantification method with ABR data from an established mouse model with cochlear synaptopathy. The data clearly demonstrated that curvature measurement is more sensitive and consistent in identifying cochlear synaptic loss in mice compared to the amplitude and latency measurements. We further tested this curvature method in a different mouse model presenting with otitis media. The change in curvature profile due to middle ear infection in otitis media is different from the profile of mice with cochlear synaptopathy. Thus, our study suggests that curvature quantification can be used to address the current ABR variability issue, and may lead to additional applications in the clinic diagnosis of hearing disorders.


Tissue hypersensitivity to dental materials.
Five cases of tissue hypersensitivity to Scutan and two to Impregum are reported. The reaction to Scutan was a burning sensation in the mucosa in contact with the material, followed by swelling and erythematous ulcers producing heavy discomfort. All symptoms gradually disappeared after removal of the material. To Impregum the reaction was basically the same, only less vigorous since contact with the material is of limited duration. All the cases were women in the menopause. Attention is drawn to the possibility of hormonal imbalance being a factor of importance in the development of hypersensitivity reactions to materials of this sort.


The analysis of corneal asphericity (Q value) and its related factors of 1,683 Chinese eyes older than 30 years.
To determine corneal Q value and its related factors in Chinese subjects older than 30 years. Cross sectional study. 1,683 participants (1,683 eyes) from the Handan Eye Study were involved, including 955 female and 728 male with average age of 53.64 years old (range from 30 to 107 years). The corneal Q values of anterior and posterior surfaces were measured at 3.0, 5.0 and 7.0mm aperture diameters using Bausch & Lomb Orbscan IIz (software version 3.12). Age, gender and refractive power were recorded. The average Q values of the anterior surface at 3.0, 5.0 and 7.0mm aperture diameters were -0.28±0.18, -0.28±0.18, and -0.29±0.18, respectively. The average Q value of the anterior surface at the 5.0mm aperture diameter was negatively correlated with age (B = -0.003, p<0.01) and the refractive power (B = -0.013, p = 0.016). The average Q values of the posterior surface at 3.0, 5.0, and 7.0mm were -0.26±0.216, -0.26±0.214, and -0.26±0.215, respectively. The average Q value of the posterior surface at the 5.0mm aperture diameter was positively correlated with age (B = 0.002, p = 0.036) and the refractive power (B = 0.016, p = 0.043). The corneal Q value of the elderly Chinese subjects is different from that of previously reported European and American subjects, and the Q value appears to be correlated with age and refractive power.


A new approach for atraumatic implant explantation and immediate implant installation.
This study reports a new approach for implant extraction in a convenient, fast, and atraumatic way and allowing a new implant placement at same surgical time. The Biotechnology Institute extraction kit consists of a wrench that allows a 200 N-cm countertorque force, an internal connection extractor, an extractor for external connection, and a set of ratchet handle extension pieces. In this study, a total of 91 implants installed in 42 patients were explanted. Extraction torques ranged from 80 to 200 N-cm. In those cases in which implant removal torques exceeded 200 N-cm, a 2-3-mm depth incision was done using a set of atraumatic trephines to avoid too high countertorque forces. The possibility of extracting osseointegrated implants opens new doors in oral implantology. The extraction kit allows dentists to re-treat cases and improve the final results, achieving the best outcomes for patients.


Acquired prion disease: iatrogenic CJD, variant CJD, kuru.
Human prion diseases can be classified as sporadic, hereditary or acquired. The cause of sporadic Creutzfeldt-Jakob disease (CJD) is unknown, hereditary cases are associated with mutations of the prion protein gene (PRNP) and acquired forms are caused by the transmission of infection from human to human or, as a zoonosis, from cattle to human. Although acquired forms of human prion disease are rare, the transmission of a fatal and untreatable neurological disorder has had major implications for public health and public policy.


Primary and Secondary Stability of Implants with Hydrophilic Surfaces in the Posterior Maxilla: A Split-Mouth Randomized Controlled Clinical Trial.
The aim of this split-mouth randomized clinical trial was to evaluate the primary and secondary stability of implants with hydrophilic surfaces in comparison to implants with conventional surfaces in the posterior region of the maxilla. Twenty patients with a bilateral edentulous ridge in the posterior area of the maxilla randomly received implants with two types of surfaces: (1) implants with the surface modified by double acid-etching and sandblasting (DAS, n = 20); and (2) implants with the surface modified by double acid-etching and sandblasting, stored in 0.9% saline solution to confer highly hydrophilic properties (DAS-H, n = 20) on the surface. The implants presented the same macrostructure with a hybrid design. The resonance frequency analysis was performed in order to obtain the implant stability quotient (ISQ) using Osstell. The ISQ analyses were performed just after placement of the implant (primary stability) and at 28, 40, and 90 days after the surgical procedure (secondary stability). There were no differences between the DAS and DAS-H surfaces in the primary stability or during the conversion of the primary to the secondary stability; however, there was a reduction in the stability of the implants at 28 days, which increased significantly at 40 and 90 days in both surfaces. It can be concluded that the surface wettability of implants with a hybrid macrostructure did not increase the primary and secondary implant stability in the posterior region of the maxilla.


Involvement of the p38/MK2 Pathway in MCLR Hepatotoxicity Revealed through MAPK Pharmacological Inhibition and Phosphoproteomics in HepaRG Cells.
Microcystin-leucine arginine (MCLR) is one of the most common and toxic microcystin variants, a class of cyanotoxins produced by cyanobacteria. A major molecular mechanism for MCLR-elicited liver toxicity involves the dysregulation of protein phosphorylation through protein phosphatase (PP) inhibition and mitogen-activated protein kinase (MAPK) modulation. In this study, specific pharmacological MAPK inhibitors were used in HepaRG cells to examine the pathways associated with MCLR cytotoxicity. SB203580 (SB), a p38 inhibitor, rescued HepaRG cell viability, whereas treatment with SP600125 (JNK inhibitor), MK2206 (AKT inhibitor), or N-acetylcysteine (reactive oxygen species scavenger) did not. Phosphoproteomic analysis revealed that phosphosites-which were altered by the addition of SB compared to MCLR treatment alone-included proteins involved in RNA processing, cytoskeletal stability, DNA damage response, protein degradation, and cell death. A closer analysis of specific proteins in some of these pathways indicated that SB reversed the MCLR-mediated phosphorylation of the necroptosis-associated proteins, the mixed lineage kinase domain-like protein (MLKL), receptor-interacting serine/threonine kinase 1 (RIP1), DNA damage response proteins, ataxia telangiectasia and Rad3-related kinase (ATR), and checkpoint kinase 1 (CHK1). Overall, these data implicate p38/MK2, DNA damage, and necroptosis in MCLR-mediated hepatotoxicity, and suggest these pathways may be targets for prevention prior to, or treatment after, MCLR toxicity.


Photodynamic activity of monocationic and non-charged methoxyphenylporphyrin derivatives in homogeneous and biological media.
A novel 5-[4-(trimethylammonium)phenyl]-10,15,20-tris(2,4,6-trimethoxyphenyl)porphyrin iodide (2) has been synthesized. A positive charge was incorporated at a peripheral position to increase the amphiphilic character of the structure. The photodynamic effect of the cationic porphyrin 2 was compared with that of non-charged 5-(4-aminophenyl)-10,15,20-tris(2,4,6-trimethoxyphenyl)porphyrin (1), both in a homogeneous medium bearing photooxidizable substrates and in vitro on the Hep-2 human larynx carcinoma cell line. Absorption and fluorescence spectroscopic studies in different media show that 2 is essentially unaggregated in solution, and also in human cells. The singlet molecular oxygen, O2(1delta(g)), production was evaluated using 9,10-dimethylanthracene in N,N-dimethylformamide, yielding phi(delta) values of approximately 0.66 for both porphyrins. The addition of beta-carotene suppresses the O2(1delta(g))-mediated photooxidation. L-Tryptophan and guanosine 5'-monophosphate were used as biological substrate models. Porphyrin 2 sensitizes the decomposition of both compounds faster than does 1. In the biological medium, no dark cytotoxicity was observed, even though a high porphyrin concentration (10 microM) and a long incubation time (24 h) were employed. Cell treatments were performed with 5 microM of porphyrin for 24 h. Under these conditions, the uptake of porphyrin 2 into Hep-2 was about 3 times higher than that of 1. Cell survival after irradiation with visible light was dependent upon both the light exposure level and intracellular sensitizer concentration. Thus, a higher photocytotoxic effect was found for porphyrin 2 in comparison to 1. These results show that the amphiphilic monocationic porphyrin 2 could be a promising model for phototherapeutic agents with potential applications in tumor cell inactivation by photodynamic therapy.


"Perhaps something of beauty can grow:" experiences of care farming for grief.
Care farming has been used to alleviate distress and increase wellbeing in various populations. This study provides an overview of how bereaved adults (N = 115) experienced a grief-specific care farm through a content analysis of open-ended survey questions. The care farm's nature spaces and interactions with animals emerged as important components of the experience, interacting with grief-related activities and experiences. Together, the spaces and species of the care farm provided a supportive context for integrating grief, processing emotions, and receiving compassionate support. Some participants also experienced changes in how they viewed their grief and improvements in interpersonal relationships.


A strabismus susceptibility locus on chromosome 7p.
Strabismus has been known to have a significant genetic component, but the mode of inheritance and the identity of the relevant genes have been enigmatic. This paper reports linkage analysis of nonsyndromic strabismus. The principal results of this study are: (i) the demonstrated feasibility of identifying and recruiting large families in which multiple members have (or had) strabismus; (ii) the linkage in one large family of a presumptive strabismus susceptibility locus to 7p22.1 with a multipoint logarithm of odds score of 4.51 under a model of recessive inheritance; and (iii) the failure to observe significant linkage to 7p in six other multiplex families, consistent with genetic heterogeneity among families. These findings suggest that it will be possible to localize and ultimately identify strabismus susceptibility genes by linkage analysis and mutation screening of candidate genes.


Constipation is casting a shadow over everyday life - a systematic review on older people's experience of living with constipation.
To explore and summarise best evidence of how constipation affects the daily living of older people from their own perspective. Furthermore, to assess how interventions aimed at treating constipation in older people affect patient-reported outcome such as quality of life. Constipation is a common and overlooked problem with an impact on everyday life, especially among older people. Older people seem to have individual preconceptions on constipation which can influence the strategies used to prevent and treat constipation. A systematic review, integrating findings from both qualitative and quantitative studies. Systematic searches were carried out in PubMed, CINAHL, PsycINFO and EMBASE on the 31st of July 2014. A search strategy was constructed with key concepts identified using PICO to identify quantitative studies and PIC(o) to identify qualitative studies. Search terms included constipation, elderly, aged, elderly people, aged people, quality of life, patient experience, patient perspective, meaning, emotion, psychological. Reference lists were searched manually. A total of nine studies were included in the review, five quantitative and four qualitative. Three main themes crystallised from the results of the included studies: bodily experiences, everyday life shadowed by constipation and adverse psychological effects. Constipation among older people was connected to subjective and comprehensive experiences. It had a negative impact on physical and mental well-being as well as the social life of older people. The review also showed that older people had individual and personal strategies, based on their own beliefs. Healthcare professionals need to be aware of the experiences of living with constipation as well as the range of strategies used by patients to prevent and treat constipation. The patient perspective on constipation needs to be integrated in the strategies and actions carried out by healthcare professionals.


Enhanced biological responses of a hydroxyapatite/TiO2 hybrid structure when surface electric charge is controlled using radiofrequency sputtering.
We investigated whether surface roughness and control of surface electric charge of a hydroxyapatite (HA)/titanium oxide (TiO2) hybrid coating could enhance biological responses associated with bone formation. After acid etching, a titanium surface was modified with HA and TiO2 by the dual sputtering deposition technique using radiofrequency sputtering. These surfaces were analyzed for surface roughness and surface electric charge intensity. Rat bone marrow-derived osteoblast-like cells were cultured on HA/TiO2 hybrid surfaces with different electric charges. The attachment and spreading behavior of these cells were significantly increased on the hybrid surface (p<0.05). In vivo experiment, the strength of bone-titanium implant integration with a hybrid surface was 3 times that of a control (p<0.05). The dual sputtering deposition technique created a HA/TiO2 hybrid structure. Our results show that the surface electric charge on a titanium surface is an important factor for enhancing biological responses.


[C-reactive protein as a tool in the diagnosis of anastomotic leak after colorectal surgery].
Anastomotic leak (AL) is one of the most severe complications after colorectal surgery. AL is often diagnosed late when the patient is presenting with septic symptoms. C-reactive protein (CRP) as a tool in the diagnosis of AL has been investigated in several studies. CRP is proven to have a high negative predictive value which allows safe discharge of patients with low CRP measurements on day 3 or 4 after surgery. CRP can assist in the diagnosis of AL but cannot be used as a single parameter.


Transport of hemolysin by Escherichia coli.
The hemolytic phenotype in Escherichia coli is determined by four genes. Two (hlyC and hlyA) determine the synthesis of a hemolytically active protein which is transported across the cytoplasmic membrane. The other two genes (hlyBa and hlyBb) encode two proteins which are located in the outer membrane and seem to form a specific transport system for hemolysin across the outer membrane. The primary product of gene hlyA is a protein (protein A) of 106,000 daltons which is nonhemolytic and which is not transported. No signal peptide can be recognized at its N-terminus. In the presence of the hlyC gene product (protein C), the 106,000-dalton protein is processed to the major proteolytic product of 58,000 daltons, which is hemolytically active and is transported across the cytoplasmic membrane. Several other proteolytic fragments of the 106,000-dalton protein are also generated. During the transport of the 58,000-dalton fragment (and possible other proteolytic fragments of hlyA gene product), the C protein remains in the cytoplasm. In the absence of hlyBa and hlyBb the entire hemolytic activity (mainly associated with the 58,000-dalton protein) is located in the periplasm: Studies on the location of hemolysin in hlyBa and hlyBb mutants suggest that the gene product of hlyBa (protein Ba) binds hemolysin and leads it through the outer membrane whereas the gene product of hlyBb (protein Bb) releases hemolysin from the outer membrane. This transport system is specific for E coli hemolysin. Other periplasmic enzymes of E coli and heterologous hemolysin (cereolysin) are not transported.


Occurrence and larval movement of Diatraea saccharalis (Lepidoptera: Crambidae) in seed mixes of non-Bt and Bt pyramid corn.
Larval movement of target pest populations among Bt and non-Bt plants is a major concern in the use of a seed mixture refuge strategy for Bt resistance management. In this study, occurrence and larval movement of the sugarcane borer, Diatraea saccharalis (F.), were evaluated in four planting patterns of non-Bt and Bt plants containing Genuity® SmartStax(TM) traits in 2009-2011. The four planting patterns were: (1) a pure stand of 27 Bt plants; (2) one non-Bt plant in the center, surrounded by 26 Bt plants; (3) a pure stand of 27 non-Bt plants; (4) one Bt plant in the center, surrounded by 26 non-Bt plants. Studies were conducted under four conditions: (1) open field with natural infestation; (2) greenhouse with artificial infestations; open field with artificial infestations (3) on the center plants only and (4) on every plant. The major objective of this study was to determine whether refuge plants in a seed mixture strategy could provide a comparable refuge population of D. saccharalis to a 'structured refuge' planting. Larvae of D. saccharalis showed the ability to move from infested plants to at least four plants away, as well as to adjacent rows, but the majority remained within the infested row. However, the number of larvae found on the non-Bt plants in the mixture plantings was not significantly reduced compared with the pure stand of non-Bt corn. The results of this study show that refuge plants in a seed mixture may be able to provide a comparable refuge population of D. saccharalis to a structured refuge planting.


AMBRA1 interplay with cullin E3 ubiquitin ligases regulates autophagy dynamics.
Autophagy maintains cellular homeostasis by degrading harmful or unnecessary intracellular components. How the autophagy response is induced rapidly and transiently remains largely unknown. We report that the E3 ubiquitin ligases Cullin-5 and Cullin-4 regulate the onset and termination of autophagy, respectively, by dynamically interacting with AMBRA1, a regulator of autophagy. Under normal conditions, Cullin-4 binding to AMBRA1 limits its protein abundance. Autophagy stimuli promote AMBRA1 stabilization by causing ULK1-dependent Cullin-4 release. Notably, Cullin-4/AMBRA1 dissociation is transient, and the re-established interaction triggers AMBRA1 degradation, terminating the autophagy response. Moreover, Cullin-4 inhibits the interaction between AMBRA1 and another Cullin E3 ligase. Indeed, upon Cullin-4 dissociation, AMBRA1 binds and inhibits Cullin-5, thus promoting the accumulation of the mTOR inhibitor DEPTOR. Through DEPTOR stabilization, AMBRA1 establishes a feedback loop that ensures the rapid onset of autophagy by enhancing mTOR inactivation. Our findings show that Cullin-mediated degradation of autophagy regulators temporally controls the autophagy response.


Hypoxia differentially enhances the effects of transforming growth factor-beta isoforms on the synthesis and secretion of glycosaminoglycans by human lung fibroblasts.
Interstitial lung diseases associated with hypoxia, such as lung fibrosis, are characterized by enhanced production of transforming growth factor-beta (TGF-beta) and increased deposition of extracellular matrix (ECM) molecules, including glycosaminoglycans (GAGs). In this study, we investigated the effect of hypoxia (3% O(2)) on TGF-beta-induced GAG synthesis by primary human pulmonary fibroblasts, established from lung biopsies. Total GAG synthesis was assessed by the incorporation of [(3)H]glucosamine into GAGs associated with the cell layer (cells and ECM) or secreted in the medium. GAGs were isolated and purified by gel filtration, fractionated by electrophoresis on cellulose acetate membranes, and characterized using GAG-degrading enzymes. GAG molecules identified in the cell layer and the medium were: hyaluronic acid, and chondroitin, dermatan, and heparan sulfates. All TGF-beta isoforms time dependently induced [(3)H]glucosamine incorporation into GAGs of the cell layer or the medium. Characterization of individual GAG molecules indicated that this was attributed to dermatan and heparan sulfates in the cell layer and to hyaluronic acid and chondroitin and dermatan sulfates in the medium. Hypoxia enhanced the effect of all TGF-beta isoforms, particularly that of TGF-beta3, on the secretion of hyaluronic acid and chondroitin and dermatan sulfates. In the cell layer, hypoxia stimulated only the effect of TGF-beta2-induced [(3)H]glucosamine incorporation into GAGs. Our data indicate that hypoxia differentially enhances the effect of TGF-beta isoforms on the secretion and deposition of GAGs and may hasten ECM remodeling associated with the pathogenesis of lung fibrosis.


[Evoked potentials extraction based on cross-talk resistant adaptive noise cancellation].
As Evoked Potentials are much lower in amplitude with respect to the on-going EEC, many trigger-related signals are needed for common averaging technique to enable the extraction of single-trail evoked potentials (EP). How to acquire EP through fewer evocations is an important research project. This paper proposes a cross-talk resistant adaptive noise cancellation method to extract EP. Together with the use of filtering technique and the common averaging technique, the present method needs much less evocations to acquire EP signals. According to the simulating experiment, it needs only several evocations or even only one evocation to get EP signals in good quality.


Beam based alignment and its relevance in Indus-2.
Initially in the Indus-2 storage ring, the closed orbit distortion (COD) could be best corrected to 1.3 mm rms in the horizontal and 0.43 mm rms in the vertical plane. The strength of the corrector magnets required high values for COD correction. This revealed that offsets in COD readout by the beam position monitors (BPMs) played a role in not achieving a rms COD lower than the above value. Thus, the offset between the electrical center of BPMs and the magnetic center of the nearest quadrupole magnet could be estimated using the beam based alignment (BBA) method. It prefers that the quadrupole magnet is able to be controlled individually and active shunt power supply (ASPS) system was designed for this purpose that works efficiently. This paper describes the methodology of BBA, topology of ASPS and its performance, and COD minimization using the measured BPM offsets. After BBA, the COD could be reduced to 0.45 mm rms and 0.2 mm rms in horizontal and vertical planes, respectively.


Synthetic, structural and theoretical studies of amidinate and guanidinate stabilised germanium(I) dimers.
The neutral germanium(i) dimers, [{Ge(Piso)}(2)] and [{Ge(Giso)}(2)], Piso = [(ArN)(2)CBu(t)](-), Giso = [(ArN)(2)CNPr(i)(2)](-), Ar = C(6)H(3)Pr(i)(2)-2,6, which are stabilised by bulky amidinate and guanidinate ligands respectively, have been prepared by reduction of the corresponding germanium(ii) chlorides, [Ge(Piso)Cl] and [Ge(Giso)Cl]; theoretical studies suggest that the Ge-Ge bonds of [{Ge(Piso)}(2)] and [{Ge(Giso)}(2)] are associated with their HOMOs, whilst their LUMOs have substantial Ge-Ge pi-bonding character.


A nutritional supplement containing lactoferrin stimulates the immune system, extends lifespan, and reduces amyloid β peptide toxicity in Caenorhabditis elegans.
Lactoferrin is a highly multifunctional glycoprotein involved in many physiological functions, including regulation of iron absorption and immune responses. Moreover, there is increasing evidence for neuroprotective effects of lactoferrin. We used Caenorhabditis elegans as a model to test the protective effects, both on phenotype and transcriptome, of a nutraceutical product based on lactoferrin liposomes. In a dose-dependent manner, the lactoferrin-based product protected against acute oxidative stress and extended lifespan of C. elegans N2. Furthermore, Paralysis of the transgenic C. elegans strain CL4176, caused by Aβ1-42 aggregates, was clearly ameliorated by treatment. Transcriptome analysis in treated nematodes indicated immune system stimulation, together with enhancement of processes involved in the oxidative stress response. The lactoferrin-based product also improved the protein homeostasis processes, cellular adhesion processes, and neurogenesis in the nematode. In summary, the tested product exerts protection against aging and neurodegeneration, modulating processes involved in oxidative stress response, protein homeostasis, synaptic function, and xenobiotic metabolism. This lactoferrin-based product is also able to stimulate the immune system, as well as improving reproductive status and energy metabolism. These findings suggest that oral supplementation with this lactoferrin-based product could improve the immune system and antioxidant capacity. Further studies to understand the molecular mechanisms related with neuronal function would be of interest.


Diagnosis of acute appendicitis in two databases. Evaluation of different neighborhoods with an LVQ neural network.
The use of an artificial neural network system was studied in the diagnosis of acute abdominal pain, especially acute appendicitis, with patients from Finland and Germany. Separate Learning Vector Quantization (LVQ) neural networks were trained with a training set from each database and also with a combined database. Each neural network was evaluated separately with a test set of cases from each database. With the combined database different neighborhood methods were compared to find the optimal choice for this decision-making problem. The acute appendicitis cases of the Finnish test data set were classified well with all the networks, but the cases of the German test set were difficult to classify for the Finnish network. The use of larger neighborhoods increased the sensitivity of the classification by nearly 10%. The differences in the results of the Finnish and German databases suggest that there are differences in the data collection or patient populations between centers. Therefore, care must be taken when using decision-support systems which have been developed in other centers. Neural networks offer a method to evaluate differences between databases. With the use of larger neighborhoods, the effects of the differences on the accuracy of the classification can be partly diminished.


Patient safety culture lives in departments and wards: multilevel partitioning of variance in patient safety culture.
Aim of study was to document 1) that patient safety culture scores vary considerably by hospital department and ward, and 2) that much of the variation is across the lowest level organizational units: the wards. Setting of study: 500-bed Norwegian university hospital, September-December 2006. Data collected from 1400 staff by (the Norwegian version of) the generic version of the Safety Attitudes Questionnaire (SAQ Short Form 2006). Multilevel analysis by MLwiN version 1.10. Considerable parts of the score variations were at the ward and department levels. More organization level variation was seen at the ward level than at the department level. Patient safety culture improvement efforts should not be limited to all-hospital interventions or interventions aimed at entire departments, but include involvement at the ward level, selectively aimed at low-scoring wards. Patient safety culture should be studied as closely to the patient as possible. There may be such a thing as "hospital safety culture" and the variance across hospital departments indicates the existence of department safety cultures. However, neglecting the study of patient safety culture at the ward level will mask important local variations. Safety culture research and improvement should not stop at the lowest formal level of the hospital (wards, out-patient clinics, ERs), but proceed to collect and analyze data on the micro-units within them.


Predicting Lexical Relations between Biomedical Terms: towards a Multilingual Morphosemantics-based System.
This paper addresses the issue of how semantic information can be automatically assigned to compound terms, i.e. both a definition and a set of semantic relations. This issue is particularly crucial when elaborating multilingual databases and when developing cross-language information retrieval systems. The paper shows how morpho-semantics can contribute in the constitution of multilingual lexical networks in biomedical corpora. It presents a system capable of labelling terms with morphologically related words, i.e. providing them with a definition, and grouping them according to synonymy, hyponymy and proximity relations. The approach requires the interaction of three techniques: (1) a la morphosemantic parser, (2) a multilingual table defining basic relations between word roots, and (3) a set of language-independant rules to draw up the list of related terms. This approach has been fully implemented for French, on an about 29,000 terms biomedical lexicon, resulting to more than 3,000 lexical families.


A Population Health Perspective on Racism and Racial Microaggressions.
Racism, one of the social determinants of health, often goes unnoticed by those less affected by its pernicious effects. The lived social experience of race has been linked to significant physical and mental health disparities. Individually or together, we know that racism and discrimination are associated with poorer health of persons from racial minority groups as evidenced in higher rates of mortality, earlier onset of disease, greater severity and progression of disease and higher levels of comorbidity and impairment. These disparities are persistent over time and, although may lessen in degree, are evident at every level of income and education. This article provides a glimpse of the impact of racism on individuals and groups, with a focus on microaggressions as a subtle but pervasive form of racism, and how it is an underlying causative factor for health disparities.


Cell transformation and tumor induction by Abelson murine leukemia virus in the absence of helper virus.
We investigated the role of the Moloney helper virus, Moloney murine leukemia virus (Mo-MuLV), in cell transformation and tumor induction by the defective Abelson murine leukemia virus (Ab-MuLV). A molecular clone of Ab-MuLV (P160 strain) was transfected into the psi 2 packaging cell line, and helper virus-free Ab-MuLV (psi 2) was harvested from the supernatant medium. Ab-MuLV (psi 2) was as efficient as helper virus-containing Ab-MuLV (Mo-MuLV) in the transformation of primary bone marrow cells in vitro. Inoculation of weanling BALB/c mice with Ab-MuLV (psi 2) induced nonthymic pre-B-cell lymphomas with high efficiency and short latency (28 days). Adult BALB/c mice were less sensitive to tumor induction by a factor of 100. Ab-MuLV (psi 2) did not induce tumors in weanling C57BL/6 mice, unlike Ab-MuLV (Mo-MuLV). Examination of the proviral integration pattern in Ab-MuLV (psi 2)-induced tumor cell DNA revealed that each of the tumors contained a single integrated provirus. Immunoprecipitation of viral-encoded proteins in helper virus-free tumor cell lines detected the P160 Ab-MuLV-transforming protein; however, no trace of the gag, pol, and env helper virus-encoded proteins was found. Our results indicate that integration and expression of a single Ab-MuLV genome is sufficient for efficient transformation of primary bone marrow cells by Ab-MuLV in vitro and tumor induction in susceptible mice. However, the helper virus may contribute to tumor induction in weanling resistant mice.


Clinical- and cost-effectiveness of telemedicine in type 2 diabetes mellitus: a systematic review and meta-analysis.
Emerging telemedicine programs offer potential low-cost solutions to the management of chronic disease. We sought to evaluate the clinical effectiveness and cost effectiveness of telemedicine approaches on glycemic control in patients with type 2 diabetes mellitus. Using terms related to type 2 diabetes and telemedicine, MEDLINE, Cochrane, EMBASE, and CINAHL Plus were searched to identify relevant studies published through February 28, 2014. Data from identified clinical trials were pooled according to telemedicine approach, and evaluated using conventional meta-analytical methods. We identified 47 articles, from 35 randomized controlled trials, reporting quantitative outcomes for hemoglobin A1c (HbA1c). Twelve of the 35 studies provided intervention via telephone, either in the form of a call or a text message; 19 studies tested internet-based programs, employing video-conferencing and/or informational websites; and four studies used interventions involving electronically transmitted recommendations made by clinicians in response to internet-based reporting by patients. Overall, pooled results from these studies revealed a small, but statistically significant, decrease in HbA1c following intervention, compared to conventional treatment (pooled difference in means=-0.37, 95% CI=-0.49 to -0.25, Z=-6.08, P<0.001). Only two of the 35 studies included assessment of cost-effectiveness. These studies were disparate, both in terms of overall expense and relative cost-effectiveness. Optimization of telemedicine approaches could potentially allow for more effective self-management of disease in type 2 diabetes patients, though evidence to-date is unconvincing. Furthermore, significant publication bias was detected, suggesting that the literature should be interpreted cautiously.


Acidified glycerol lysis test in various haemolytic anaemias.
Acidified glycerol lysis test (AGLT) was found to be an appropriate technique for detection of sphaerocytes in blood particularly in hereditary sphaerocytosis. The incubation of blood for 5 hours at 37 degrees C prior to measurement exerts positive effect on the reliability of the test and may further suggest the type of haemolytic disorder.


Organ-sparing treatment of advanced bladder cancer: a 10-year experience.
Radical cystectomy is considered as standard therapy for muscle-invasive bladder cancer. We present 10-year results of bladder-sparing treatment by conservative surgery and radiotherapy +/- chemotherapy. From 1982 through 1991, 245 consecutive patients, mean age 66 years, with invasive bladder cancer (T2-3 or poor prognostic T1, no distant metastases) entered a prospective protocol with the objective of bladder preservation. Treatment consisted of transurethral resection (complete, if possible) and definitive radiotherapy with 56 Gy maximum dose (50.4 Gy minimum target dose) in 28 fractions. Since 1985, 139 patients received a simultaneous chemotherapy on 5 days in the first and fifth treatment week with either 25 mg/m2 cisplatin daily (79 patients) or 65 mg/m2 carboplatin (60 patients). Cystectomy was performed as salvage treatment for residual or recurrent invasive disease. The median follow-up at the date of analysis (12-31-92) was 5.9 years. The overall survival was 47% after 5 years and 26% after 10 years. The 5-year survival according to the initial T-category was 60% for T1 (44 patients), 64% for T2 (47 patients), 43% for T3 (127 patients), and 16% for T4 (23 patients). The most important single prognostic factor was the amount of residual tumor after TUR (5-year survival 80% after R0, 53% after R1, and 31% after R2 resection, p < 0.01). Chemotherapy increased the rate of complete remission, but had no impact on 5-year survival (52% vs. 50%). Fifty-three salvage cystectomies were performed, all without severe complications, and 192 patients (79%) maintained a normal functioning bladder. The bladder preservation rate in 5-year survivors was 83%. Organ-sparing treatment of advanced bladder cancer by transurethral surgery and definitive radiotherapy or radiochemotherapy is feasible and effective. The survival in this series is as good as in any comparable cystectomy series. Eighty-three percent of long-term survivors maintained their functioning bladders.


Safe and efficient transdermal delivery of desmopressin acetate by iontophoresis in rats.
Iontophoretic delivery of desmopressin acetate (DDAVP) was assessed for delivery efficiency and drug stability, both in vitro and in vivo. The effect of current intensity and duration of current application on the decomposition of DDAVP was investigated in vitro. It was shown that when a current of 0.1 mA was applied for 5 min, the decomposition of DDAVP was negligible. In vivo experiments under the same conditions showed that the antidiuretic response to DDAVP persisted for about 6 h. Furthermore, when this iontophoresis was repeated 3 times at intervals of 4 h, the antidiuretic response persisted for about 11 h. These results suggest that repeated short-term iontophoresis is a safe and effective technique for transdermal delivery of DDAVP.


Electric and mass transport of a carbon nanotube encapsulating a copper nano-rod studied by in-situ transmission electron microscopy.
Electric properties and current-induced structural changes of carbon nanotubes (CNTs) encapsulating copper nano-rods were studied by in-situ transmission electron microscopy (TEM). The diameter and the length of a copper filled CNT were 18 nm and 256 nm, respectively. The thickness of the graphite layer was about 1 nm. The bias voltage was applied between the two ends of the CNT inside the TEM, and the current as well as TEM images were recorded simultaneously. At a bias voltage of 1.4 V, the current increased to 10 microA, corresponding to a current density of 4.0 x 10(6) A/cm2, and at the same time the nano-rods inside the CNT started to move to an end of the CNT. After the movement of the nano-rods, an empty CNT was left. Resistivities of the CNT and the copper nano-rod were measured to be 3.0 x 10(-5) ohm m and 1.2 x 10(-4) ohm m, respectively.


Elastic force restricts growth of the murine utricle.
Dysfunctions of hearing and balance are often irreversible in mammals owing to the inability of cells in the inner ear to proliferate and replace lost sensory receptors. To determine the molecular basis of this deficiency we have investigated the dynamics of growth and cellular proliferation in a murine vestibular organ, the utricle. Based on this analysis, we have created a theoretical model that captures the key features of the organ's morphogenesis. Our experimental data and model demonstrate that an elastic force opposes growth of the utricular sensory epithelium during development, confines cellular proliferation to the organ's periphery, and eventually arrests its growth. We find that an increase in cellular density and the subsequent degradation of the transcriptional cofactor Yap underlie this process. A reduction in mechanical constraints results in accumulation and nuclear translocation of Yap, which triggers proliferation and restores the utricle's growth; interfering with Yap's activity reverses this effect.


An archaeal CRISPR type III-B system exhibiting distinctive RNA targeting features and mediating dual RNA and DNA interference.
CRISPR-Cas systems provide a small RNA-based mechanism to defend against invasive genetic elements in archaea and bacteria. To investigate the in vivo mechanism of RNA interference by two type III-B systems (Cmr-α and Cmr-β) in Sulfolobus islandicus, a genetic assay was developed using plasmids carrying an artificial mini-CRISPR (AC) locus with a single spacer. After pAC plasmids were introduced into different strains, Northern analyses confirmed that mature crRNAs were produced from the plasmid-borne CRISPR loci, which then guided gene silencing to target gene expression. Spacer mutagenesis identified a trinucleotide sequence in the 3'-region of crRNA that was crucial for RNA interference. Studying mutants lacking Cmr-α or Cmr-β system showed that each Cmr complex exhibited RNA interference. Strikingly, these analyses further revealed that the two Cmr systems displayed distinctive interference features. Whereas Cmr-β complexes targeted transcripts and could be recycled in RNA cleavage, Cmr-α complexes probably targeted nascent RNA transcripts and remained associated with the substrate. Moreover, Cmr-β exhibited much stronger RNA cleavage activity than Cmr-α. Since we previously showed that S. islandicus Cmr-α mediated transcription-dependent DNA interference, the Cmr-α constitutes the first CRISPR system exhibiting dual targeting of RNA and DNA.


Kidney ATPase response in seawater-transferred rainbow trout (Oncorhynchus mykiss). Effect of salinity and fish size.
Two sizes of domesticated rainbow trout (Oncorhynchus mykiss, 40 +/- 0.67 and 180 +/- 3.9 g) were directly transferred to brackish water (9 ppt) and seawater (28 ppt). Kidney Na(+)-K(+)-ATPase and Mg(2+)-ATPase activities were measured in fresh water, and after long-term seawater adaptation (up to 21 days). Renal Na(+)-K(+)-ATPase activity increased after saltwater loading in small trout, while large trout displayed an unmodified ATPase activity. The smallest trout showed a low but progressive increase in renal Mg(2+)-ATPase activity after the transfer to both salinities. However, ATPase activity remained unchanged or significantly decreased in large trout after the transfer to seawater or brackish water, respectively.


The limits of chimpanzee-human comparisons for understanding human cognition.
Evolutionary questions require specialized approaches, part of which are comparisons between close relatives. However, to understand the origins of human tool behavior, comparisons with solely chimpanzees are insufficient, lacking the power to identify derived traits. Moreover, tool use is unlikely a unitary phenomenon. Large-scale comparative analyses provide an alternative and suggest that tool use co-evolves with a suite of cognitive traits.


Recurrent lipomatous tumor of hypopharynx: case report and literature review.
Liposarcoma is one of the most common soft-tissue sarcomas in adults, but head and neck are rarely involved, especially regions of the larynx and hypopharynx. According to Enzinger and Weiss, liposarcoma can be divided into 5 subtypes: well-differentiated, myxoid, round cell, pleomorphic and dedifferentiated. We present an unusual case of well-differentiated liposarcoma of the hypopharynx in a patient with previous three procedures of endoscopic removal of hypopharyngeal tumor classified as benign lipoma. Well-differentiated liposarcoma is a tumor of low-grade malignancy, which frequently recurs locally, but does not metastasize. Wide tumor resection with free margins is mandatory. Immunohistochemistry is a useful diagnostic tool. We also discuss recently published literature on this unusual presentation of well-differentiated liposarcoma.


Clinical significance of first trimester umbilical cord cysts.
A cystic mass of the umbilical cord was identified by transvaginal sonography in 10 first trimester pregnancies at a mean gestational age of 8 weeks 4 days (range, 8 weeks 1 day to 9 weeks 3 days) and at a mean crown-rump length of 20.5 mm (range, 15 to 25 mm). The cyst was solitary in all cases, the mean diameter was 4.6 mm (range, 3 to 6 mm), and the location was closer to the fetal insertion in two cases, in the middle of the cord in seven cases, and closer to the placental insertion in one case. Gestational sac and yolk sac diameters as well as the fetal heart rate were within normal ranges for gestational age in all cases. Information on detailed second trimester scans was available in nine cases, demonstrating complete resolution of the cyst and normal fetal anatomic survey in each case. These nine pregnancies were followed to delivery, and normal healthy infants were delivered at term in all cases. This series suggests that the incidental detection of umbilical cord cysts in early pregnancy is not associated with an adverse pregnancy outcome.


Synthesis, antibacterial and antifungal activity of some new pyrazoline and pyrazole derivatives.
A series of 2-pyrazolines 5-9 have been synthesized from α,β-unsaturated ketones 2-4. New 2-pyrazoline derivatives 13-15 bearing benzenesulfonamide moieties were then synthesized by condensing the appropriate chalcones 2-4 with 4-hydrazinyl benzenesulfonamide hydrochloride. Ethyl [1,2,4] triazolo[3,4-c][1,2,4]triazino[5,6-b]-5H-indole-5-ethanoate (26) and 1-(5H-[1,2,4]triazino[5,6-b] indol-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (32) were synthesized from 3-hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24). On the other hand ethyl[1,2,4]triazolo[3,4-c][1,2,4]triazino[5,6-b]-5,10-dihydroquinoxaline- 5-ethanoate (27) and 1-(5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl)-3-methyl-1H-pyrazol-5(4H)-one (33) were synthesized from 3-hydrazinyl-5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline (25) by reaction with diethyl malonate or ethyl acetoacetate, respectively. Condensation of 6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indole-2-carbaldehyde (1') with compound 24 or 25 afforded the corresponding Schiff's bases 36 and 37, respectively. Reaction of the Schiff's base 37 with benzoyl hydrazine or acetic anhydride afforded benzohydrazide derivative 39 and the cyclized compound 40, respectively. Furthermore, the pyrazole derivatives 42-44 were synthesized by cyclization of hydrazine derivative 25 with the prepared chalcones 2-4. All the newly synthesized compounds have been characterized on the basis of IR and 1H-NMR spectral data as well as physical data. Antimicrobial activity against the organisms E. coli ATCC8739 and P. aeruginosa ATCC 9027 as examples of Gram-negative bacteria, S. aureus ATCC 6583P as an example of Gram-positive bacteria and C. albicans ATCC 2091 as an example of a yeast-like fungus have been studied using the Nutrient Agar (NA) and Sabouraud Dextrose Agar (SDA) diffusion methods. The best performance was found for the compounds 16, 17, 19 and 20.


Engineering the Self-Assembly Induced Emission of Cu Nanoclusters by Au(I) Doping.
Aggregation-induced emission (AIE) and self-assembly induced emission (SAIE) effects have been employed to tune the emission properties of metal nanoclusters (NCs). However, it is still not possible to further enhance the photoluminescence quantum yields (PLQYs) and control the emission colors of the NCs using AIE and SAIE. On the basis of our recent work studying the contribution of Cu(I) defects in the SAIE of Cu NCs, in this article, Au(I) was doped into Cu NC self-assembled nanosheets (NSASs) to construct a more stable Au(I)-centered state. As a result, the PLQYs, emission stability, and tunability of emission colors of the Cu NSASs were significantly improved. Detailed studies reveal that the doped Au(I) induces a Au(I)-Cu(I) metallophilic interaction, which leads to a ligand-to-Cu-Au charge transfer, which facilitates the relaxation of excited electrons via a radiative pathway, thereby enhancing the emission intensity. The charge transfer from Cu to Au lowers the energy, thus leading to the red-shift of PL emission. Au(I) is likely doped into the Cu NSASs rather than in individual NCs, because 0.3% Au doping is enough to alter the emission properties. By mixing Au(I)-doped Cu NSASs with different emission colors (due to different Au doping percentages) as color conversion materials on commercially available 365 nm GaN chips, a white light-emitting diode prototype is fabricated.


Oxozinc carboxylate complexes: a new synthetic approach and the carboxylate ligand effect on the noncovalent-interactions-driven self-assembly.
An atom-efficient and mild synthesis of a series of oxozinc carboxylates [Zn(4)(μ(4)-O)(O(2)CR)(6)] [where R = Ph (2a), p-PhC(6)H(4) (2b), p-MeC(6)H(4) (2c), and p-MeSC(6)H(4) (2d)] from well-defined alkylzinc precursors and H(2)O is described. The molecular and crystal structures of the resulting complexes have been determined by single-crystal X-ray diffraction. A closer examination of their crystal structure provides a direct picture of the effect of the nature of substituents on the molecular self-assembly of the octahedral oxozinc through noncovalent interactions. It was revealed that these discrete oxozinc clusters can form diverse types of noncovalent assemblies ranging from structures representing zeolitic topologies in the case of 2a to soft porous materials with gated voids or open channels for the remaining molecular clusters.


Heterogeneous Molecular Catalysis of Electrochemical Reactions: Volcano Plots and Catalytic Tafel Plots.
We analyze here, in the framework of heterogeneous molecular catalysis, the reasons for the occurrence or nonoccurrence of volcanoes upon plotting the kinetics of the catalytic reaction versus the stabilization free energy of the primary intermediate of the catalytic process. As in the case of homogeneous molecular catalysis or catalysis by surface-active metallic sites, a strong motivation of such studies relates to modern energy challenges, particularly those involving small molecules, such as water, hydrogen, oxygen, proton, and carbon dioxide. This motivation is particularly pertinent for what concerns heterogeneous molecular catalysis, since it is commonly preferred to homogeneous molecular catalysis by the same molecules if only for chemical separation purposes and electrolytic cell architecture. As with the two other catalysis modes, the main drawback of the volcano plot approach is the basic assumption that the kinetic responses depend on a single descriptor, viz., the stabilization free energy of the primary intermediate. More comprehensive approaches, investigating the responses to the maximal number of experimental factors, and conveniently expressed as catalytic Tafel plots, should clearly be preferred. This is more so in the case of heterogeneous molecular catalysis in that additional transport factors in the supporting film may additionally affect the current-potential responses. This is attested by the noteworthy presence of maxima in catalytic Tafel plots as well as their dependence upon the cyclic voltammetric scan rate.


Product-enhanced reverse transcriptase assay for replication-competent retrovirus and lentivirus detection.
The product-enhanced reverse transcriptase (PERT) assay has been used to detect reverse transcriptase (RT) activity associated with retroviruses. Although the PERT assay has been proposed as a method for detection of replication-competent retrovirus (RCR) and lentivirus (RCL), it has not been rigorously compared with existing methods for RCR and RCL detection. We have assessed the PERT assay for detection of RCL and RCR that may contaminate lentiviral and retroviral vectors and compared it with published methods for RCL (p24gag ELISA/gag PCR) and RCR (S+/L-) detection. Our results suggest that the PERT assay is as sensitive as p24gag ELISA and gag PCR for detection of replication-competent HIV-1 in an RCL detection assay. Comparison of detection of replication-competent retroviruses, GALV and RD114, by extended S+/L- and PERT assays indicates that both assays can detect 1 IU of each virus. Our findings suggest that the PERT assay can be used for RCL and RCR testing of a variety of retroviral vectors regardless of the structure, sequence, and envelope of the vectors.


Severe Disease Activity May Predispose Patients to Post-colectomy Duodenitis Associated with Ulcerative Colitis.
Objective Diffuse mucosal inflammation in the duodenum, distinct from peptic ulcer disease, has been repeatedly reported in patients with ulcerative colitis (UC). The pathogenesis of this complication remains uncertain; however, colectomy for medically refractory UC appears to trigger duodenitis. Cases in which colectomy was performed for UC were analyzed to characterize UC-related duodenitis after colectomy. Methods A retrospective case-control study of UC-related duodenitis that developed after colectomy in medically refractory UC between January 2011 and June 2020 was conducted. UC-related duodenitis was diagnosed based on typical clinical, endoscopic, and histological findings, and no duodenitis was endoscopically defined by the normal duodenal mucosa. Clinical and laboratory data, disease severity, and medications used were collected and compared between the UC-related and non-duodenitis cases. Results Ten UC-related duodenitis and 35 non-duodenitis cases were identified among 45 patients with UC who underwent esophagogastroduodenoscopy after colectomy. Disease severity, defined by the C-reactive protein level and partial Mayo score prior to colectomy, was significantly higher in duodenitis patients than in non-duodenitis patients. In comparison to non-duodenitis patients, duodenitis patients more frequently received rescue therapies with calcineurin inhibitors or anti-TNF-α agents at the time of colectomy (100% vs. 65.7%). Conclusion Patients with UC with higher disease activity, especially those who require rescue therapies with calcineurin inhibitors and anti-TNF-α agents, may be prone to developing UC-related duodenitis after colectomy.


Genome-wide redistribution of MeCP2 in dorsal root ganglia after peripheral nerve injury.
Methyl-CpG-binding protein 2 (MeCP2), a protein with affinity for methylated cytosines, is crucial for neuronal development and function. MeCP2 regulates gene expression through activation, repression and chromatin remodeling. Mutations in MeCP2 cause Rett syndrome, and these patients display impaired nociception. We observed an increase in MeCP2 expression in mouse dorsal root ganglia (DRG) after peripheral nerve injury. The functional implication of increased MeCP2 is largely unknown. To identify regions of the genome bound by MeCP2 in the DRG and the changes induced by nerve injury, a chromatin immunoprecipitation of MeCP2 followed by sequencing (ChIP-seq) was performed 4 weeks after spared nerve injury (SNI). While the number of binding sites across the genome remained similar in the SNI model and sham control, SNI induced the redistribution of MeCP2 to transcriptionally relevant regions. To determine how differential binding of MeCP2 can affect gene expression in the DRG, we investigated mmu-miR-126, a microRNA locus that had enriched MeCP2 binding in the SNI model. Enriched MeCP2 binding to miR-126 locus after nerve injury repressed miR-126 expression, and this was not mediated by alterations in methylation pattern at the miR-126 locus. Downregulation of miR-126 resulted in the upregulation of its two target genes Dnmt1 and Vegfa in Neuro 2A cells and in SNI model compared to control. These target genes were significantly downregulated in Mecp2-null mice compared to wild-type littermates, indicating a regulatory role for MeCP2 in activating Dnmt1 and Vegfa expression. Intrathecal delivery of miR-126 was not sufficient to reverse nerve injury-induced mechanical and thermal hypersensitivity, but decreased Dnmt1 and Vegfa expression in the DRG. Our study shows a regulatory role for MeCP2 in that changes in global redistribution can result in direct and indirect modulation of gene expression in the DRG. Alterations in genome-wide binding of MeCP2 therefore provide a molecular basis for a better understanding of epigenetic regulation-induced molecular changes underlying nerve injury.


DeepSeg: deep neural network framework for automatic brain tumor segmentation using magnetic resonance FLAIR images.
Gliomas are the most common and aggressive type of brain tumors due to their infiltrative nature and rapid progression. The process of distinguishing tumor boundaries from healthy cells is still a challenging task in the clinical routine. Fluid-attenuated inversion recovery (FLAIR) MRI modality can provide the physician with information about tumor infiltration. Therefore, this paper proposes a new generic deep learning architecture, namely DeepSeg, for fully automated detection and segmentation of the brain lesion using FLAIR MRI data. The developed DeepSeg is a modular decoupling framework. It consists of two connected core parts based on an encoding and decoding relationship. The encoder part is a convolutional neural network (CNN) responsible for spatial information extraction. The resulting semantic map is inserted into the decoder part to get the full-resolution probability map. Based on modified U-Net architecture, different CNN models such as residual neural network (ResNet), dense convolutional network (DenseNet), and NASNet have been utilized in this study. The proposed deep learning architectures have been successfully tested and evaluated on-line based on MRI datasets of brain tumor segmentation (BraTS 2019) challenge, including s336 cases as training data and 125 cases for validation data. The dice and Hausdorff distance scores of obtained segmentation results are about 0.81 to 0.84 and 9.8 to 19.7 correspondingly. This study showed successful feasibility and comparative performance of applying different deep learning models in a new DeepSeg framework for automated brain tumor segmentation in FLAIR MR images. The proposed DeepSeg is open source and freely available at https://github.com/razeineldin/DeepSeg/.


Changes in Metabolic Parameters and Body Weight in Patients With Prediabetes Treated With Adjunctive Brexpiprazole for Major Depressive Disorder: Pooled Analysis of Short- and Long-Term Clinical Studies.
Objective: Certain atypical antipsychotics, while efficacious as adjunctive treatments in major depressive disorder (MDD), are associated with metabolic adverse effects and weight gain. This analysis determined the effect of adjunctive brexpiprazole on metabolic parameters and body weight in adults with MDD and prediabetes (ie, at risk of developing diabetes) based on pooled data from 3 short-term studies and 1 long-term study. Methods: The short-term studies were 6-week, randomized, double-blind, placebo-controlled studies of adjunctive oral brexpiprazole 1-3 mg/d in outpatients with MDD (DSM-IV-TR criteria) and inadequate response to antidepressant treatment, conducted between June 2011 and May 2016. The long-term study was a 26- to 52-week, open-label extension study conducted between October 2011 and May 2017. Prediabetes was defined based on fasting serum glucose and glycated hemoglobin (HbA1c) levels. Shifts in diabetes status and shifts/changes in fasting metabolic parameters and body weight were determined. Results: Most patients receiving adjunctive brexpiprazole maintained their baseline diabetes status in the short term (568/751; 75.6%) and long term (1,919/2,746; 69.9%). The incidence of categorical shifts in fasting metabolic parameters generally did not differ between treatment groups or between prediabetes and non-diabetes subgroups. Mean changes from baseline in metabolic parameters were small in the short term (all < 5 mg/dL) and long term (all < 6 mg/dL, except < 20 mg/dL for triglycerides). Moderate weight gain was observed in the short term (1.5 kg) and long term (3.4-4.1 kg). Conclusions: Adjunctive brexpiprazole had a limited impact on the metabolic profile of patients with MDD, regardless of diabetes status (prediabetes/non-diabetes). Trial Registration: Data used in this post hoc analysis came from studies with ClinicalTrials.gov identifiers NCT01360645, NCT01360632, NCT02196506, and NCT01360866.


68 Ga-somatostatin analogue PET-CT: Analysis of costs and benefits in a public hospital setting.
Between 2009 and 2012, 68 Ga-somatostatin analogue PET-CT progressively replaced 111 In-octreotide scintigraphy for imaging neuroendocrine tumours in WA public hospitals due to published literature demonstrating improved diagnostic accuracy and increased availability. Despite significantly improved sensitivity and specificity, 68 Ga-somatostatin analogue PET is currently unfunded in Australia. This study sought to undertake cost analysis of the two modalities in a public hospital setting and to compare them with regard to patient factors such as imaging time and radiation dose. This analysis was based on retrospective clinical data from 95 111 In-octreotide scintigraphies performed in 2007 and 2008 at Sir Charles Gairdner (SCGH) and Royal Perth (RPH) hospitals and 219 68 Ga-somatostatin analogue PET-CT studies performed in 2013 at SCGH. Whole body effective radiation dose was derived from the radiopharmaceutical and low-dose CT scan. The cost analysis included radiopharmaceutical and imaging costs. The median imaging time for an 111 In-octreotide scintigraphy was 152 min at SCGH, 100 min at RPH and 20 min for a 68 Ga-somatostatin analogue PET-CT scan. The mean effective radiation dose for 111 In-octreotide scintigraphy was 18.1 mSv at SCGH and 13.8 mSv at RPH. The effective dose for 68 Ga-somatostatin analogue PET-CT was 8.7-10.8 mSv. The average cost of 68 Ga-somatostatin analogue PET-CT was four times less than 111 In-octreotide scintigraphy. 68 Ga-somatostatin analogue PET-CT is not only more accurate than 111 In-octreotide scintigraphy, this study has also shown that it is significantly less expensive, delivers a lower radiation dose to patients and requires less imaging time for patients and staff. 68 Ga-somatostatin PET-CT provides an important combination of both reduced cost and improved clinical care for patients.


Effect of carbohydrate position on lysosomal transport of procathepsin L.
To study the role of carbohydrate in lysosomal protein transport, we engineered two novel glycosylation signals (Asn-X-Ser/Thr) into the cDNA of human procathepsin L, a lysosomal acid protease. We constructed six mutant cDNAs encoding glycosylation signals at mutant sites Asn-138, Asn-175, or both sites together, in the presence or absence of the wild-type Asn-204 site. We stably transfected wild-type and mutant cDNAs into NIH3T3 mouse fibroblasts and then used species-specific antibodies to determine the glycosylation status, phosphorylation, localization, and transport kinetics of recombinant human procathepsin L containing one, two, or three glycosylation sites. Both novel glycosylation sites were capable of being glycosylated, although Asn-175 was utilized only 30-50% of the time. Like the wild-type glycosylation at Asn-204, carbohydrates at Asn-138 and Asn-175 were completely sensitive to endoglycosidase H, and they were phosphorylated. Mutant proteins containing two carbohydrates were capable of being delivered to lysosomes, but there was not a consistent relationship between the efficiency of lysosomal delivery and carbohydrate content of the protein. Pulse-chase labeling revealed a unique biosynthetic pattern for proteins carrying the Asn-175 glycosylation sequence. Whereas wild-type procathepsin L and mutants bearing carbohydrate at Asn-138 appeared in lysosomes by about 60 min, proteins with carbohydrate at Asn-175 were processed to a lysosome-like polypeptide within 15 min. Temperature shift, brefeldin A, and NH4Cl experiments suggested that the rapid processing did not occur in the endoplasmic reticulum and that Asn-175 mutants could interact with the mannose 6-phosphate receptor. Taken together, our results are consistent with the interpretation that Asn-175 carbohydrate confers rapid transport to lysosomes. We may have identified a recognition domain in procathepsin L that is important for its interactions with the cellular transport machinery.


Detection of interstitial lung abnormalities on picture archive and communication system video monitors.
The purpose of this study was to compare the detection of interstitial lung abnormalities on video display workstation monitors between radiologists experienced with video image interpretation and radiologists who lack this experience. Twenty-four patients with interstitial lung abnormalities documented by high-resolution computed tomography (HRCT) and lung biopsy, and 26 control patients with no history of pulmonary disease or a normal HRCT and normal chest radiographs were studied. Images were acquired using storage phosphor digital radiography and displayed on 1,640 x 2,048 pixel resolution video monitors. Five board-certified radiologists evaluated the images in a blinded and randomized manner by using a six-point presence of abnormality grading scale. Three radiologists were from 1 to 4 years out of residency and considered to be experienced workstation monitor readers with between 1 to 3 years of video monitor image interpretation. For the inexperienced readers, one radiologist had no prior experience with reading images from a video monitor and was direct out of residency, and the other radiologist had less than 4 months of intermittent exposure and was 1 year out of residency. Sensitivity and specificity were determined for individual readers. Positive predictive values, negative predictive values, accuracy, and receiver-operating curves were also generated. A comparison was made between experienced and inexperienced readers. For readers experienced with video monitor image interpretation, the sensitivity ranged from 87.5% to 92%, specificity from 69% to 92%, positive predictive value (PPV) from 73% to 87.5%, negative predictive value (NPV) from 87% to 90%, and accuracy from 80% to 88%. For inexperienced readers, these values were sensitivity 58%, specificity 50% to 65%, PPV 52% to 61%, NPV 56.5% to 63%, and accuracy 54% to 62%. Comparing image interpretation between experienced and inexperienced readers, there were statistically significant differences for sensitivity (P < .01), specificity (P < .01), PPV (P < .05), NPV (P < .05), accuracy (P < .05), and area under the receiver operator curve (Az) (P < .01). Within the respective experienced and inexperienced groups, no statistical significant differences were present. Our results show that digitally acquired chest radiographs displayed on high-resolution workstation monitors are adequate for the detection of interstitial lung abnormalities when the images are interpreted by radiologists experienced with video image interpretation. Radiologists inexperienced with video monitor image interpretation, however, cannot reliably interpret images for the detection of interstitial lung abnormalities.


Discovery of Novel α-Aminophosphonates with Hydrazone as Potential Antiviral Agents Combined With Active Fragment and Molecular Docking.
A series of novel α-aminophosphonate derivatives containing hydrazone were designed and synthesized based on active fragments. Bioassay results demonstrated that title compounds possessed good activities against tobacco mosaic virus. Among them, compounds 6a, 6g, 6i, and 6j were equivalent to the commercial antiviral agents like dufulin. On structure optimization-based molecular docking, compound 6k was synthesized and displayed excellent activity with values of 65.1% curative activity, 74.3% protective activity, and 94.3% inactivation activity, which were significantly superior to the commercial antiviral agents dufulin and ningnanmycin. Therefore, this study indicated that new lead compounds could be developed by adopting a joint strategy with active fragments and molecular docking.


Long-term physiologic changes of intraocular pressure: a 10-year longitudinal analysis in young and middle-aged Japanese men.
Although the intraocular pressure (IOP) is the most important factor related to the onset and progression of glaucoma, there is little evidence on long-term aging effects on IOP. This article examines the changes of IOP over 10 years in normal eyes to assess physiologic changes related to aging. Population-based long-term longitudinal study. STUDY POPULATION AND OBSERVATION PROCEDURE: Two thousand nine hundred eighty-seven Japanese male aircraft crew members underwent IOP measurement by Goldmann apparatus and received physical and complete ophthalmologic examinations every year for 10 years. A total of 2330 healthy persons (21-49 years; mean age, 35.9+/-6.8 [standard deviation]) who had no history of treatment for ophthalmic diseases, current illnesses, and no missing data for 10 consecutive years were analyzed. For analysis of the longitudinal index (trend), the linear regression coefficients for 11 points of measurement were determined. Ophthalmologic and physiologic factors affecting the 10-year mean values and a trend of IOP were examined by multivariable linear regression analysis. Intraocular pressure tended to decrease with age in all age groups. The mean linear regression coefficient (right eye/left eye) = -0.076/-0.060 (95% confidence interval [CI]: (-0.094 to -0.057)/(-0.078 to -0.041), -0.073/-0.060 [95% CI: (-0.084 to -0.062)/(-0.071 to -0.049)], and -0.060/-0.050 [95%CI: (-0.075 to -0.046)/(-0.064 to -0.036)] (mmHg/year) in the 20s, 30s, and 40s, respectively). In investigating correlations between the 10-year mean values of IOP and factors examined in this study, multivariate analysis showed a significantly inverse correlation with spherical power (partial regression coefficient [B] = -0.155/-0.144) and positive correlation with esophoria (B = 0.536/0.521), systolic blood pressure (B = 0.021/0.022), heart rate (B = 0.024/0.024), and hematocrit (B = 0.041/0.043) with IOP. The trend of IOP was significantly positively associated with the trends of systemic factors: body mass index (BMI) (B = 0.117/0.119), blood pressure (systolic) (B = 0.020/0.020), and hematocrit (B = 0.057/0.045), but not with any ophthalmologic factor. The long-term observation clearly demonstrated that, in normal eyes, the IOP decreased with aging. The IOP value was negatively associated with spherical power and positively with esophoria, blood pressure, heart rate, and hematocrit. The change of IOP could be affected mainly by the change of body mas index, blood pressure, and hematocrit.


The Application of Bio-orthogonality for In Vivo Animal Imaging.
The application of bio-orthogonality has greatly facilitated numerous aspects of biological studies in recent years. In particular, bio-orthogonal chemistry has transformed biological research, including in vitro conjugate chemistry, target identification, and biomedical imaging. In this review, we highlighted examples of bio-orthogonal in vivo imaging published in recent years. We grouped the references into two major categories: bio-orthogonal chemistry-related imaging and in vivo imaging with bio-orthogonal nonconjugated pairing. Lastly, we discussed the challenges and opportunities of bio-orthogonality for in vivo imaging.


Adjuvant long-term immunochemotherapy for squamous cell carcinoma of the thoracic esophagus.
Between 1965 and 1978, 131 patients with primary squamous cell carcinoma of the thoracic esophagus underwent subtotal esophagectomy. Since 1972 we hae used active therapy involving pre- and postoperative irradiation and postoperative long-term cancer immunochemotherapy. When the surgical results of the active therapy and the historical control group, in which postoperative radiotherapy was used primarily, were compared, active therapy appeared to be high effective. The one, two, and three-year survival rates were 54.7, 36.3, and 27.1 per cent, respectively, in the former, and 32, 14, and 12 12 per cent, respectively, in the latter group. With respect to noncurative cases, the one-year survival rate was 42.2 per cent in the active therapy group, and 20 per cent in the control group.


Transformation of an Online Multidisciplinary Course into a Live Interprofessional Experience.
Objective. To design, implement, and assess an interprofessional education (IPE) course in the first professional year of students enrolled in eight different health professions programs. Design. An interprofessional faculty committee created a 1-credit hour required IPE course to not only teach students about the roles and responsibilities of each discipline and how they may contribute to an interprofessional team, but to also improve collaboration and team-based communication skills among health care professions students. Students were placed in interprofessional groups and met weekly to participate in didactic lectures, discussion sessions, and a standardized patient encounter. Assessment. Seven hundred and eighty-three health professions students were enrolled in the course, of which 130 students completed questionnaires at all three time points. Students were neutral about the course and found it moderately valuable (Mean 6.23 [on a scale from 1 to 10], interesting (Mean 5.61), and enjoyable (Mean 5.57). Written feedback from the course indicated that the majority of students enjoyed the standardized patient encounter and thought the course provided a valuable opportunity to interact with other students in other health professions programs. Conclusion. This required course served as an introductory interprofessional approach in preparing health professions students to learn from each other about their various roles and responsibilities and how each can contribute to the health care team.


Neuronal function of the mRNA decapping complex determines survival of Caenorhabditis elegans at high temperature through temporal regulation of heterochronic gene expression.
In response to adverse environmental cues, Caenorhabditis elegans larvae can temporarily arrest development at the second moult and form dauers, a diapause stage that allows for long-term survival. This process is largely regulated by certain evolutionarily conserved signal transduction pathways, but it is also affected by miRNA-mediated post-transcriptional control of gene expression. The 5'-3' mRNA decay mechanism contributes to miRNA-mediated silencing of target mRNAs in many organisms but how it affects developmental decisions during normal or stress conditions is largely unknown. Here, we show that loss of the mRNA decapping complex activity acting in the 5'-3' mRNA decay pathway inhibits dauer formation at the stressful high temperature of 27.5°C, and instead promotes early developmental arrest. Our genetic data suggest that this arrest phenotype correlates with dysregulation of heterochronic gene expression and an aberrant stabilization of lin-14 mRNA at early larval stages. Restoration of neuronal dcap-1 activity was sufficient to rescue growth phenotypes of dcap-1 mutants at both high and normal temperatures, implying the involvement of common developmental timing mechanisms. Our work unveils the crucial role of 5'-3' mRNA degradation in proper regulation of heterochronic gene expression programmes, which proved to be essential for survival under stressful conditions.