Mandibular nerve fascicular cross-face for sensitive recovery after mandibulectomy: a new technique.
Trigeminal nerve damage after mandibulectomy is a condition that may occur not infrequently when oncologic resections of the maxillo-facial district are performed. In the last decades microsurgery has made possible effective osteomuscular reconstructions using vascularised free flaps. Nevertheless, this procedure, if assuring satisfactory results, involved the sacrifice of the mandibular nerve. Nowadays, supramicrosurgical innovation has shifted the gold-standard to a higher level, allowing the complete sensitive recovery after mandibulectomy. The cross-face nerve transfer technique is an innovative procedure that provides excellent nerve regeneration in mini-invasive operations, avoiding visible scars and any deficiency in donor sites. We suggest that the cross-face nerve transfer is the surgical modality of choice to restore lower lip and chin sensibility after mandibulectomy.


A novel method for detergent concentration determination.
A fast and precise method for detergent concentration determination is presented. (Patent applications for the method described here have been submitted (EP05011904 and US60/702,261). Depending on the interest of the scientific community, the system will be commercialized. (For further information contact Hervé-W. Rémigy at the e-mail address below.) A small droplet of the detergent solution is deposited on a piece of Parafilm M and side views are recorded by two orthogonally arranged TV cameras. The droplet contours are then approximated by ellipses to determine the contact angles. Comparison of the observed contact angle values to calibrated standard curves of known detergent concentrations gives the concentration of the detergent assessed. A range of commonly used detergents was studied to demonstrate the reproducibility and precision of this simple method. As a first application, the detergent binding capacity of the Escherichia coli galactose/proton symporter (GalP) was assessed. Aggregation of GalP was observed when <260 +/- 5 dodecyl-beta,D-maltoside molecules were bound to one GalP molecule. These measurements document the efficacy of the drop-shape based detergent concentration determination described.


Steady-state cerebral autoregulation in older adults with amnestic mild cognitive impairment: linear mixed model analysis.
We examined whether the efficacy of steady-state cerebral autoregulation (CA) is reduced in older adults with amnestic mild cognitive impairment (aMCI), a prodromal stage of clinical Alzheimer disease (AD). Forty-two patients with aMCI and 24 cognitively normal older adults (NC) of similar age, sex, and education underwent stepwise decreases and increases in mean arterial pressure (MAP) induced by intravenous infusion of sodium nitroprusside and phenylephrine, respectively. Changes in cerebral blood flow (CBF) were measured repeatedly in the internal carotid and vertebral artery. Linear mixed modeling, including random effects of both individual intercept and regression slope, was used to quantify the MAP-CBF relationship accounting for nonindependent, repeated CBF measures. Changes in end-tidal CO2 (EtCO2) associated with changes in MAP were also included in the model to account for their effects on CBF. Marginal mean values of MAP were reduced by 13-14 mmHg during sodium nitroprusside and increased by 20-24 mmHg during phenylephrine infusion in both groups with similar doses of drug infusion. A steeper slope of changes in CBF in response to changes in MAP was observed in aMCI relative to NC, indicating reduced efficacy of CA (MAP × Group, P = 0.040). These findings suggest that cerebrovascular dysfunction may occur early in the development of AD.NEW & NOTEWORTHY Cerebral autoregulation is a fundamental regulatory mechanism to protect brain perfusion against changes in blood pressure that, if impaired, may contribute to the development of Alzheimer's disease. Using a linear mixed model, we demonstrated that the efficacy of cerebral autoregulation, assessed during stepwise changes in arterial pressure, was reduced in individuals with amnestic mild cognitive impairment, a prodromal stage of Alzheimer's disease. These findings support the hypothesis that cerebrovascular dysfunction may be an important underlying pathophysiological mechanism for the development of clinical Alzheimer's disease.


L-Arginine prevents air embolism-induced acute lung injury in rats.
Pulmonary air embolism, causing vessel obstruction and primary or secondary reactions of blood, can lead to acute lung injury. In addition, nitric oxide has been known to play a key role in various causes of lung injury. In this study we employed the isolated rat lung model to investigate the effects of l-arginine on air embolism-induced lung injury. Randomized, controlled study. Animal-care facility procedure room. Forty-two adult male Sprague-Dawley rats each weighing 250-350 g. Infusion of air at the rate of 0.25 mL/min for 1 min into the pulmonary artery in isolated and perfused rat lung resulted in pulmonary hypertension and lung edema. Air embolism elicited a significant increase in microvascular permeability as measured by the capillary filtration coefficient, lung weight gain, lung weight-to-body weight ratio, pulmonary arterial pressure, and protein concentration of bronchoalveolar lavage fluid. Pretreatment with L-arginine (4 mmol/L) significantly attenuated the acute lung injury induced by air embolism as shown by a significant decrease in all of the assessed variables but did not alter the pulmonary arterial pressure (p < .05). The protective effect of l-arginine was blocked when N(G)-nitro-L-arginine methyl ester (5 mmol/L) was added. Pretreatment with N(G)-nitro-L-arginine methyl ester exacerbated air embolism-induced lung injury. Our findings suggest that L-arginine can prevent air embolism-induced lung injury.


Epidemiology of clinical trials of medicines in respiratory diseases in Europe and Italy.
Clinical trials play a key role in advancing medical knowledge, improving patient care and promoting economic growth in Europe. We have assessed the clinical trial activity in any respiratory diseases in Europe, with a specific focus on Italy. Information from public sources (EFPIA, clinicaltrials.gov, clinicaltrialsregister. eu, AIFA) was used to describe clinical trial activity of in respiratory diseases in Europe and by country. In 2015, 3908 clinical trials were reported in Europe, 386 in respiratory diseases (9.9%). Germany was the first country both as absolute number (76 trials) and as percentage within country trials (14%), followed by Poland. Spain, Italy and France were the countries with the lowest number and percentage of trials in respiratory diseases. In 2013, the Italian Drug Agency reported 9 trials with respiratory compounds in Italy (2.1% of overall trials, 12ˆ position in the therapeutic area rank), 33% in phase 2 and 66% in phase 3. No phase 1 or phase 4 trials were reported for respiratory trials. Prevalence of respiratory trials by non-profit sponsors (28.3%) was below the average for the country (38.3%). Europe has a greater potential for clinical research on drugs for respiratory diseases, particularly in countries with less activity, such as Spain, France and Italy, that should identify and implement actions to increase attractiveness for clinical trials of drugs.


Experimental and Calculated Spectra of π-Stacked Mild Charge-Transfer Complexes: Jet-Cooled Perylene·(Tetrachloroethene)n, n = 1,2.
The S0 ↔ S1 spectra of the mild charge-transfer (CT) complexes perylene·tetrachloroethene (P·4ClE) and perylene·(tetrachloroethene)2 (P·(4ClE)2) are investigated by two-color resonant two-photon ionization (2C-R2PI) and dispersed fluorescence spectroscopy in supersonic jets. The S0 → S1 vibrationless transitions of P·4ClE and P·(4ClE)2 are shifted by δν = -451 and -858 cm(-1) relative to perylene, translating to excited-state dissociation energy increases of 5.4 and 10.3 kJ/mol, respectively. The red shift is ∼30% larger than that of perylene·trans-1,2-dichloroethene; therefore, the increase in chlorination increases the excited-state stabilization and CT character of the interaction, but the electronic excitation remains largely confined to the perylene moiety. The 2C-R2PI and fluorescence spectra of P·4ClE exhibit strong progressions in the perylene intramolecular twist (1au) vibration (42 cm(-1) in S0 and 55 cm(-1) in S1), signaling that perylene deforms along its twist coordinate upon electronic excitation. The intermolecular stretching (Tz) and internal rotation (Rc) vibrations are weak; therefore, the P·4ClE intermolecular potential energy surface (IPES) changes little during the S0 ↔ S1 transition. The minimum-energy structures and inter- and intramolecular vibrational frequencies of P·4ClE and P·(4ClE)2 are calculated with the dispersion-corrected density functional theory (DFT) methods B97-D3, ωB97X-D, M06, and M06-2X and the spin-consistent-scaled (SCS) variant of the approximate second-order coupled-cluster method, SCS-CC2. All methods predict the global minima to be π-stacked centered coplanar structures with the long axis of tetrachloroethene rotated by τ ≈ 60° relative to the perylene long axis. The calculated binding energies are in the range of -D0 = 28-35 kJ/mol. A second minimum is predicted with τ ≈ 25°, with ∼1 kJ/mol smaller binding energy. Although both monomers are achiral, both the P·4ClE and P·(4ClE)2 complexes are chiral. The best agreement for adiabatic excitation energies and vibrational frequencies is observed for the ωB97X-D and M06-2X DFT methods.


Intensification of the phenomenon of violence in the family environment of teenagers.
Childhood and adolescence are the periods of life when the experience of violence accumulates. As socially weaker individuals, children and teenagers are exposed to violence. The factors that increase the risk of child maltreatment include, above all, social and cultural factors and the stress that family suffer from. The literature on this subject distinguishes four categories of child maltreatment, namely: emotional, physical, negligence and sexual abuse. The survey involved 250 representatives of high school teenagers aged 15-20, including 145 girls and 105 boys. The research method was the survey estimating the Scale of Battered Child Syndrome (for teenagers and adults). The results show that a big group of teenagers admitted to having experienced at least one of four kinds of domestic violence. The group is not uniform, however, and the socio-cultural factors that affect the kind and intensification of the phenomenon of violence have been revealed. The most frequent reasons for using violence are: low level of education, unemployment of parents and material status connected with this fact, low frequency of attendance to religious services, alcohol abuse, and place of living. On account of the intensification of the phenomenon of violence in the domestic environment and both direct and distant consequences of the phenomenon in the form of mental and physical disorders of individuals as well as the dangers for the proper development of the society that result from it, there is a need to continue doing research on this phenomenon.


Failure in cervical total disc arthroplasty: single institution experience, systematic review of the literature, and proposal of the RUSH TDA failure classification system.
Cervical total disc arthroplasty (TDA) is an alternative procedure to anterior cervical discectomy and fusion that facilitates neural decompression while both preserving motion of the spinal unit and decreasing the risk for degenerative changes at adjacent segments. However, due to its more recent introduction in clinical practice and low complication rates, the modes by which TDA may fail remain to be described. This study sought to identify the modes and frequencies of cervical TDA failure in order to propose a novel classification system. Retrospective cohort and systematic review. Patients who underwent single or two-level TDA for cervical radiculopathy or myelopathy at a single institution and in the literature of medium and large prospective studies. Cervical TDA failure, defined as subsequent surgical intervention at the index segment. This study retrospectively reviewed patients who underwent single or two-level TDA for cervical radiculopathy or myelopathy at a single institution to identify the potential implant failure modes. A systematic review and meta-analysis of prospective data in the literature was then performed to further supplement failure mode identification and to describe the rates at which the various failure types occurred. Statistical analysis included between-group comparisons of Non-Failed and Failed patients and frequencies of each failure type among Failed patients. A retrospective review at our institution of 169 patients (201 levels) identified eight failures, for a failure rate of 4.7%. Additionally, seven patients were revised who had the primary surgery at an outside institution. The systematic review of 3976 patients (4525 levels) identified 165 (4.1%) additional failures. Using this data, six primary failure types were classified, with several subtypes. These include recurrent or persistent index-level stenosis (Type I); migration (Type II) presenting as gross extrusion (A) or endplate failure with subsidence/acute fracture (B); instability (Type III) due to mechanical loosening (A), septic loosening (B), or device fracture (C); device motion loss (Type IV) such as "locking" of the device in kyphosis; implantation error (Type V) due to malposition (A) or improper sizing (B); and wear (Type VI) either without osteolysis (A) or with wear-particle-induced osteolysis (B). Stenosis (Type I) was the most common mode of failure found both through retrospective review and in the literature. Cervical TDA fails through six primary mechanisms. While rates of certain failures requiring subsequent surgical intervention are low, it is possible that these complications may become more prevalent upon further longitudinal observation. Thus, future application and validation of this classification system is warranted to evaluate how failure frequencies change over time and with larger patient samples.


Descriptor-Free Deep Learning QSAR Model for the Fraction Unbound in Human Plasma.
Chemical-specific parameters are either measured in vitro or estimated using quantitative structure-activity relationship (QSAR) models. The existing body of QSAR work relies on extracting a set of descriptors or fingerprints, subset selection, and training a machine learning model. In this work, we used a state-of-the-art natural language processing model, Bidirectional Encoder Representations from Transformers, which allowed us to circumvent the need for calculation of these chemical descriptors. In this approach, simplified molecular-input line-entry system (SMILES) strings were embedded in a high-dimensional space using a two-stage training approach. The model was first pre-trained on a masked SMILES token task and then fine-tuned on a QSAR prediction task. The pre-training task learned meaningful high-dimensional embeddings based upon the relationships between the chemical tokens in the SMILES strings derived from the "in-stock" portion of the ZINC 15 dataset─a large dataset of commercially available chemicals. The fine-tuning task then perturbed the pre-trained embeddings to facilitate prediction of a specific QSAR endpoint of interest. The power of this model stems from the ability to reuse the pre-trained model for multiple different fine-tuning tasks, reducing the computational burden of developing multiple models for different endpoints. We used our framework to develop a predictive model for fraction unbound in human plasma (fu,p). This approach is flexible, requires minimum domain expertise, and can be generalized for other parameters of interest for rapid and accurate estimation of absorption, distribution, metabolism, excretion, and toxicity.


Cooperative effect between immunotherapy and antiangiogenic therapy leads to effective tumor rejection in tolerant Her-2/neu mice.
Immunotherapy is an attractive strategy for cancer treatment. However, self-tolerance is one of the major mechanisms that dampen immune responses against self-tumor antigens. We have demonstrated that Her-2/neu transgenic mice (neu mice) are tolerant to neu antigens and contain only a low avidity repertoire for neu. However, this repertoire has antitumor activity. Immunizations of neu mice are capable of activating the low-avidity T cells that, at best, retard the tumor growth. To increase the efficacy of the antitumor responses in neu mice, we hypothesized that immunotherapy in combination with antiangiogenic therapy would be a more efficient strategy for tumor eradication. The rationale for using this combination was that by decreasing the growth rate of the tumor with an antiangiogenic therapy, the low-avidity repertoire of neu mice stimulated by immunotherapeutic intervention would be more effective in destroying the slow growing tumor. To test this hypothesis, we stably expressed a soluble form of the Flt-1 vascular endothelial growth factor receptor (sFlt-1) on N202.1A cells, using a retrovirus vector. Expression of sFlt-1 on N202.1A (N202-Flt) cells significantly inhibited the tumor growth compared with N202.1A parental cells. In contrast to the application of immunotherapy alone or antiangiogenic therapy alone, which delayed the tumor growth, the combination of the two therapies provided complete inhibition of tumor growth in Her-2/neu mice. These results indicate that the use of tumor targeting with immunotherapy in simultaneous combination with antiangiogenic therapy provides a more efficient strategy for the treatment of solid tumors.


Molecular imaging of MMP activity discriminates unstable from stable plaque phenotypes in shear-stress induced murine atherosclerosis.
As atherosclerotic plaque ruptures are the primary cause of ischaemic events, their preventive identification by imaging remains a clinical challenge. Matrix metalloproteinases (MMP) are involved in plaque progression and destabilisation and are therefore promising targets to characterize rupture-prone unstable plaques. This study aims at evaluating MMP imaging to discriminate unstable from stable plaque phenotypes. ApoE deficient mice (ApoE-/-) on a high cholesterol diet underwent implantation of a tapered cuff around the right common carotid artery (CCA) inducing a highly inflamed atherosclerotic plaque upstream (US) and a more stable plaque phenotype downstream (DS) of the cuff. 8 weeks after surgery, the MMP inhibitor-based photoprobe Cy5.5-AF443 was administered i.v. 3h prior to in situ and ex vivo fluorescence reflectance imaging of the CCAs. Thereafter, CCAs were analysed regarding plaque size, presence of macrophages, and MMP-2 and MMP-9 concentrations by immunohistochemistry and ELISA. We found a significantly higher uptake of Cy5.5-AF443 in US as compared to DS plaques in situ (1.29 vs. 1.06 plaque-to-background ratio; p<0.001), which was confirmed by ex vivo measurements. Immunohistochemistry revealed a higher presence of macrophages, MMP-2 and MMP-9 in US compared to DS plaques. Accordingly, MMP-2 concentrations were significantly higher in US plaques (47.2±7.6 vs. 29.6±4.6 ng/mg; p<0.05). In the ApoE-/- cuff model MMP-2 and MMP-9 activities are significantly higher in upstream low shear stress-induced unstable atherosclerotic plaques as compared to downstream more stable plaque phenotypes. MMP inhibitor-based fluorescence molecular imaging allows visualization of these differences in shear stress-induced atherosclerosis.


Dynamic Electrical Source Imaging (DESI) of Seizures and Interictal Epileptic Discharges Without Ensemble Averaging.
We propose an algorithm for electrical source imaging of epileptic discharges that takes a data-driven approach to regularizing the dynamics of solutions. The method is based on linear system identification on short time segments, combined with a classical inverse solution approach. Whereas ensemble averaging of segments or epochs discards inter-segment variations by averaging across them, our approach explicitly models them. Indeed, it may even be possible to avoid the need for the time-consuming process of marking epochs containing discharges altogether. We demonstrate that this approach can produce both stable and accurate inverse solutions in experiments using simulated data and real data from epilepsy patients. In an illustrative example, we show that we are able to image propagation using this approach. We show that when applied to imaging seizure data, our approach reproducibly localized frequent seizure activity to within the margins of surgeries that led to patients' seizure freedom. The same approach could be used in the planning of epilepsy surgeries, as a way to localize potentially epileptogenic tissue that should be resected.


Donor funding priorities for communicable disease control in the developing world.
Prior research has considered donor funding for developing world health by recipient and donor country but not by disease. Examining funding by disease is critical since diseases may be in competition with one another for priority and donors may be making allocation decisions in ways that do not correspond to developing world need. In this study I calculate donor funding for 20 historically high-burden communicable diseases for the years 1996 to 2003 and examine factors that may explain variance in priority levels among diseases. I consider funding for developing world health from 42 major donors, classifying grants according to the communicable disease targeted. Data show that funding does not correspond closely with burden. Acute respiratory infections comprise more than a quarter of the burden among these diseases but receive less than 3% of direct aid. Malaria also stands out as a high-burden neglected disease. The evidence indicates that neither developing world need nor industrialized world interests explain all funding patterns, and that donors may be imitating one another in ways that do not take into account problems in the developing world. There is an urgent need for a major increase in funding for communicable disease control in the developing world, and for more balanced allocation of the resources already provided.


Draft genome sequence of Arthrospira platensis C1 (PCC9438).
Arthrospira platensis is a cyanobacterium that is extensively cultivated outdoors on a large commercial scale for consumption as a food for humans and animals. It can be grown in monoculture under highly alkaline conditions, making it attractive for industrial production. Here we describe the complete genome sequence of A. platensis C1 strain and its annotation. The A. platensis C1 genome contains 6,089,210 bp including 6,108 protein-coding genes and 45 RNA genes, and no plasmids. The genome information has been used for further comparative analysis, particularly of metabolic pathways, photosynthetic efficiency and barriers to gene transfer.


Metabolic syndrome and the development of chronic kidney disease among 118 924 non-diabetic Taiwanese in a retrospective cohort.
Metabolic syndrome (MetS) is a common risk factor for cardiovascular and chronic kidney disease (CKD) in Western populations; however, no prospective studies have examined MetS as a risk factor for CKD in Chinese adults. The incidence of CKD and the prospective link between MetS (defined by two criteria: modified Adult Treatment Panel III (ATP-III) and the International Diabetes Federation (IDF)) and CKD among 118,924 Taiwanese participants without baseline diabetes, aged 20-74 years with a mean 3.7 years follow up, was examined. CKD was measured by using estimated glomerular filtration rate or dipstick proteinuria (1+). The association between MetS or combination patterns of MetS abnormalities and CKD was evaluated using Cox models with adjustment for confounders. The incidence of CKD was 288/10,000 person-years (95% confidence interval (CI), 283-293). The findings showed that central obesity (OB), high blood pressure (BP) and high triglyceride were considered to be the major metabolic events in the study cohort. Incidences and hazard ratios (HR) on CKD had evidently increasing trends with the number of MetS components. The multivariable-adjusted HR for CKD associated with ATP-III-MetS was 1.30 (95% CI, 1.24-1.36). Equivalent HR for IDF-MetS were 1.37 (95% CI, 1.30-1.44). The associations were still observed when analyzing by stratifying incident diabetes and adjusting hypertension status. MetS induces an increased risk for CKD independent of baseline confounding factors and subsequent incident diabetes modified the associations lightly. The mechanism through which MetS may cause CKD in this population likely is the development of multiple metabolic pathogenic processes together.


Perfusion cells for studying regional variation in oral mucosal permeability in humans. 2. A specialized transport mechanism in D-glucose absorption exists in dorsum of tongue.
To clarify the site of d-glucose absorption in human oral cavity, newly designed perfusion cells were applied to five different sites in the human oral cavity, i.e., the dorsum of the tongue, the ventral surface of the tongue, the labial mucosa, the floor of the mouth, and the buccal mucosa. The solution of D-glucose was perfused for 1 h and the rate of absorption was calculated from the amount that disappeared from the perfusate. D-Glucose was absorbed rapidly from the dorsum of the tongue and the absorption was saturable. The saturable absorption was also observed in the ventral surface of the tongue, but not in the other three sites. The rate of D-glucose absorption in the dorsum and the ventral surface of the tongue was significantly larger than that of L-glucose, while in the other sites they were not significantly different. The presence of a specialized transport system for D-glucose absorption in the dorsum of human tongue was suggested.


The future of disasters studies: new disasters and the case of the Horn of Africa.
Past disasters have been well studied, but the challenge of using the findings to improve the management of future events remains a daunting task. This paper argues that there are new and complex disasters of which the state itself has become the main source, as reflected in the Horn of Africa. This region is characterised by increasing vulnerability owing to the alarming decline of democracy and the rise of authoritarianism. These new disasters are less researched because of the hazard of conducting fieldwork in such environments. However, there is mounting evidence to highlight the possibility of performing research in these settings, but not by employing traditional methods; rather, these tools may need to be customised for use. One key policy implication here is that donors may need to invest more money in analysing these new disasters and they may need to consider building and strengthening genuine partnerships between Northern and Southern research institutions.


Primary Peritonitis: An Index Case of Mycoplasma hominis Infection in a Healthy Female.
Primary peritonitis in healthy immunocompetent individuals is rare. Several case reports of Streptococcus species causing peritonitis have been described. Here, we present the first case of Mycoplasma hominis as the cause of primary peritonitis in a healthy woman. A 42-year-old female with history of uterine fibroids was admitted with abdominal pain and intraperitoneal fluid of unknown etiology. She was initially managed nonoperatively and empirically treated with broad spectrum antibiotics. Blood and urine cultures were unrevealing. Increasing abdominal pain and peritoneal fluid prompted diagnostic laparoscopy which revealed a dense fibrinous exudate covering the entire peritoneal cavity. Peritoneal fluid and biopsies were sent for cytology and culture. The peritoneal fluid was eventually sent for 16 s ribosomal analysis, which discovered Mycoplasma hominis RNA. Her antibiotics were narrowed, and she eventually made a full recovery. M. hominis is a rare source of systemic infection but has been known to colonize the urogenital tract and cause localized infections. This is the first presentation of M. hominis causing primary peritonitis in a healthy immunocompetent female. Multidisciplinary management of these patients is critical to achieve a timely diagnosis. Surgical exploration is often unavoidable to rule out secondary peritonitis.


Electromechanical properties of pressure-actuated poly(dimethylsiloxane) microfluidic push-down valves.
Pressure-actuated poly(dimethylsiloxane) (PDMS) valves have been characterized with respect to their electromechanical properties. Measurements of the valve opening and closing times, threshold pressures, and impedance spectra for closed valves can be used to assess the quality of the devices in general, determine their suitability for specialized applications, such as providing electrical isolated fluidic compartments for planar patch clamping, and specify ideal operating conditions. For our particular valve designs, we report valve opening times of the order of 10-100 micros, making them suitable for rapid buffer exchange applications. They can effectively provide reversible electrical isolation between adjacent fluidic compartments with typical resistances of 5 Gohms in the closed state, which meets the gigaohm requirement for patch clamping applications.


On-board recordings reveal no jamming avoidance in wild bats.
Animals often deal with situations in which vast sensory input is received simultaneously. They therefore must possess sophisticated mechanisms to select important input and ignore the rest. In bat echolocation, this problem is at its extreme. Echolocating bats emit sound signals and analyse the returning echoes to sense their environment. Bats from the same species use signals with similar frequencies. Nearby bats therefore face the difficulty of distinguishing their own echoes from the signals of other bats, a problem often referred to as jamming. Because bats commonly fly in large groups, jamming might simultaneously occur from numerous directions and at many frequencies. Jamming is a special case of the general phenomenon of sensory segregation. Another well-known example is the human problem of following conversation within a crowd. In both situations, a flood of auditory incoming signals must be parsed into important versus irrelevant information. Here, we present a novel method, fitting wild bats with a miniature microphone, which allows studying jamming from the bat's 'point of view'. Previous studies suggested that bats deal with jamming by shifting their echolocation frequency. On-board recordings suggest otherwise. Bats shifted their frequencies, but they did so because they were responding to the conspecifics as though they were nearby objects rather than avoiding being jammed by them. We show how bats could use alternative measures to deal with jamming instead of shifting their frequency. Despite its intuitive appeal, a spectral jamming avoidance response might not be the prime mechanism to avoid sensory interference from conspecifics.


Effect of lipoteichoic acid on thermotropic membrane properties.
Lipoteichoic acid, diglucosyldiacylglycerol, and phosphatidylglycerol isolated from Staphylococcus aureus were embedded in dipalmitoylglycerophosphoglycerol vesicles, and their thermotropic influence on this matrix was studied by differential scanning calorimetry. The natural fatty acids of phosphatidylglycerol effected peak broadening and a decrease in molar heat capacity. These effects were more pronounced with the glycolipid, which also increased the main transition temperature. With the lipoteichoic acid mixtures, two broad main transition peaks were observed, possibly due to different levels of lipoteichoic acid in vesicles. Both peaks showed a further upshift in transition temperatures and a pronounced decrease in molar heat capacity. Since the diacylglycerol moieties of all three amphiphiles were practically identical, the differences in the thermotropic effects have to be ascribed to the different structures of the head groups. Diglucosyldiacylglycerol is proposed to exert an additional effect by hydrogen bonding the hydroxyls of the sugar rings to their phospholipid neighbors. The stronger effect of lipoteichoic acid points to dynamic interactions of the long hydrophilic chain with the vesicle surface, which stabilize the membrane structure.


CRISPR/Cas9-mediated one step bi-allelic change of genomic DNA in iPSCs and human RPE cells in vitro with dual antibiotic selection.
CRISPR/Cas9 causes double-stranded DNA breaks that can undergo DNA repair either via non-homologous end joining (NHEJ) or, in the presence of a template, homology-directed repair (HDR). HDR is typically used to insert a specific genetic modification into the genome but has low efficiency compared to NHEJ, which is lowered even further when trying to create a homozygous change. In this study we devised a novel approach for homozygous single base editing based on utilising simultaneously two donor DNA templates cloned in plasmids with different antibiotic resistant genes. The donor templates were co-transfected alongside the CRISPR/Cas9 machinery into cells and a double antibiotic selection was optimised and allowed the isolation of viable desired clones. We applied the method for obtaining isogenic cells homozygous for variant B cystatin C, a recessive risk factor for age-related macular degeneration and Alzheimer's disease, in both induced Pluripotent Stem Cells (iPSCs) and a human RPE cell line. Bi-allelic gene edited clones were validated by sequencing, demonstrating that the double antibiotic templates approach worked efficiently for both iPSCs and human differentiated cells. We propose that this one step gene editing approach can be used to improve the specificity and frequency of introducing homozygous modifications in mammalian cells.


Long-term Cosmetic Alignment Following Surgery for Esotropia Versus Exotropia in Childhood: A Comparison Using Survival Curves.
Although long-term results following repair of esotropia and exotropia in childhood have been reported, the two entities have never been directly compared. Records of all children younger than 18 years who underwent horizontal strabismus surgery from 2000 through 2012 were reviewed. Children with structural eye pathology, severe vision loss, neurologic disorders, and incomitant or combined horizontal and vertical deviations were excluded. Failure was defined by a second horizontal surgery or a primary position deviation greater than 20 prism diopters (PD) at any time during follow-up (as long as 10 years). A threshold of 20 PD was chosen because families often consider deviations important enough to warrant a first or second surgery if it is larger but not if it is smaller. Of 317 children, 235 with esotropia had surgery at a mean age of 42 months and 82 with exotropia had surgery at a mean age of 60 months. Overall, surgery was successful in 78% of those with esotropia and 65% of those with exotropia. A second surgery was performed in 29 (12%) of those with esotropia and 15 (18%) of those with exotropia. The mean deviation at last follow-up, using absolute values, was 9 PD for esotropia and 10 PD for exotropia. Survival curve success was better for esotropia (P = .008). By survival curve analysis, success was more likely among children with esotropia. Many "failures" slipped beyond the 20 PD threshold only transiently and had deviations that were not noticeable to family or friends. Overall, children did reasonably well long-term following surgery for both esotropia and exotropia. Only 14% of the entire group required a second surgery during an average of 41 months of follow-up.


Examination of the hierarchical self-esteem model in adults with physical disability.
The primary purpose of this research was to examine the validity of the hierarchical physical self-perceptions model developed by Fox and Corbin (1989) within a population of physically disabled individuals. The secondary purpose was to compare the physical self-perceptions of those who were physically active and physically inactive. 97 adults with physical disabilities completed a self-esteem scale and the Physical Self-perception Profile, Japanese short version to assess General Physical Self-worth and its subdomains of Perceived Sports Competence, Physical Condition, Attractive Body, and Physical Strength. Path analysis showed adequate fit of the data to the hierarchical physical self-perception model. In general, physical self-perceptions were rated more favorably by physically active participants.


mtFociCounter for automated single-cell mitochondrial nucleoid quantification and reproducible foci analysis.
Mitochondrial DNA (mtDNA) encodes the core subunits for OXPHOS, essential in near-all eukaryotes. Packed into distinct foci (nucleoids) inside mitochondria, the number of mtDNA copies differs between cell-types and is affected in several human diseases. Currently, common protocols estimate per-cell mtDNA-molecule numbers by sequencing or qPCR from bulk samples. However, this does not allow insight into cell-to-cell heterogeneity and can mask phenotypical sub-populations. Here, we present mtFociCounter, a single-cell image analysis tool for reproducible quantification of nucleoids and other foci. mtFociCounter is a light-weight, open-source freeware and overcomes current limitations to reproducible single-cell analysis of mitochondrial foci. We demonstrate its use by analysing 2165 single fibroblasts, and observe a large cell-to-cell heterogeneity in nucleoid numbers. In addition, mtFociCounter quantifies mitochondrial content and our results show good correlation (R = 0.90) between nucleoid number and mitochondrial area, and we find nucleoid density is less variable than nucleoid numbers in wild-type cells. Finally, we demonstrate mtFociCounter readily detects differences in foci-numbers upon sample treatment, and applies to Mitochondrial RNA Granules and superresolution microscopy. mtFociCounter provides a versatile solution to reproducibly quantify cellular foci in single cells and our results highlight the importance of accounting for cell-to-cell variance and mitochondrial context in mitochondrial foci analysis.


Design of Next-Generation DGAT2 Inhibitor PF-07202954 with Longer Predicted Half-Life.
Diacylglycerol O-acyltransferase 2 (DGAT2) inhibitors have been shown to lower liver triglyceride content and are being explored clinically as a treatment for non-alcoholic steatohepatitis (NASH). This work details efforts to find an extended-half-life DGAT2 inhibitor. A basic moiety was added to a known inhibitor template, and the basicity and lipophilicity were fine-tuned by the addition of electrophilic fluorines. A weakly basic profile was required to find an appropriate balance of potency, clearance, and permeability. This work culminated in the discovery of PF-07202954 (12), a weakly basic DGAT2 inhibitor that has advanced to clinical studies. This molecule displays a higher volume of distribution and longer half-life in preclinical species, in keeping with its physicochemical profile, and lowers liver triglyceride content in a Western-diet-fed rat model.


Medicinal cannabis: Principal cannabinoids concentration and their stability evaluated by a high performance liquid chromatography coupled to diode array and quadrupole time of flight mass spectrometry method.
In the last few years, there has been a boost in the use of cannabis-based extracts for medicinal purposes, although their preparation procedure has not been standardized but rather decided by the individual pharmacists. The present work describes the development of a simple and rapid high performance liquid chromatography method with UV detection (HPLC-UV) for the qualitative and quantitative determination of the principal cannabinoids (CBD-A, CBD, CBN, THC and THC-A) that could be applied to all cannabis-based medicinal extracts (CMEs) and easily performed by a pharmacist. In order to evaluate the identity and purity of the analytes, a high-resolution mass spectrometry (HPLC-ESI-QTOF) analysis was also carried out. Full method validation has been performed in terms of specificity, selectivity, linearity, recovery, dilution integrity and thermal stability. Moreover, the influence of the solvent (ethyl alcohol and olive oil) was evaluated on cannabinoids degradation rate. An alternative extraction method has then been proposed in order to preserve cannabis monoterpene component in final CMEs.


Clinical efficacy and safety of sulbactam/ampicillin in patients suffering from chronic liver disease.
Sulbactam is a new beta-lactamase inhibitor with pharmacokinetic characteristics in humans similar to those of ampicillin. A total of 41 patients hospitalized in the Clinic of Infectious Diseases, University of Naples, for chronic liver diseases, were treated with sulbactam/ampicillin (ratio 1:2) for urinary, respiratory, biliary tract or soft tissue infections. Sulbactam/ampicillin was administered im or iv at a dosage of 3-9 g/day depending on the site and severity of the infection. All the patients treated with sulbactam/ampicillin had clinical signs and symptoms of infection, and all the organisms isolated were sensitive to sulbactam/ampicillin (MIC less than 16 mg/l). For both Gram-positive and Gram-negative bacteria the sulbactam/ampicillin MICs were much lower than the ampicillin MICs. In agreement with the favourable in-vitro results, we observed good therapeutic efficacy. 85% of the patients recovered or improved within a few days of therapy, with no clinical relapses, and in 81% of the infections the responsible bacteria were completely eradicated. We observed a low number of side effects (3/41 oral candidosis; 3/41 pain at the im injection site) and no change in the blood chemistry tests.


Moesin-ezrin-radixin-like protein (merlin) mediates protein interacting with the carboxyl terminus-1 (PICT-1)-induced growth inhibition of glioblastoma cells in the nucleus.
Moesin-ezrin-radixin-like protein (merlin) has long been considered a unique tumour suppressor that inhibits mitogenic signalling only at the membrane-cytoskeleton interface. However, the nucleocytoplasmic shuttling of merlin in a cell cycle-dependent manner has recently been observed, indicating that merlin may also exert its tumour-suppressive activity by interacting with specific nuclear protein partners. We have identified protein interacting with carboxyl terminus 1 (PICT-1) as a novel merlin-binding partner. Although the detailed mechanisms are not fully understood, several lines of evidence have previously implicated PICT-1 as a candidate tumour suppressor, including its phosphatase and tensin homolog deleted on chromosome 10 (PTEN)-dependent growth-suppression and cell-killing activities. We show here that PICT-1 is localised to the nucleolus, and Ser518-dephosphorylated merlin (the growth-inhibitory form of merlin) can interact with PICT-1 in the nucleolus. Ectopic expression of PICT-1, both in PTEN-positive HeLa cells and in PTEN-deficient U251 cells, effectively represses cyclin D1 expression, arrests the cell cycle at G0/G1, and promotes cell apoptosis. PICT-1 (1-356), a carboxyl-terminus truncated mutant that has lost the ability to bind merlin, has a markedly reduced inhibitory effect on the cell cycle and proliferation. Knockdown of merlin expression by siRNA attenuates the inhibitory effects induced by PICT-1 over-expression. We propose that merlin mediates PICT-1-induced growth inhibition by translocating to the nucleolus and binding PICT-1.


Functional and Evolutionary Implications of the Distribution of Phosphagens in Primitive-Type Spermatozoa.
External fertilization is considered to be the primitive condition in metazoans. The spermatozoa of such organisms typically display a common primitive-type morphology that is present in a range of phyla. These spermatozoa are extremely polarized cells in that the site of ATP synthesis (mitochondria in midpiece) is located at large diffusion distances from the ATP sink (dynein ATPases in the flagellum). Spermatozoa of polychaetes, sipunculids, echiuroids, echinoderms, and tunicates contain the phosphagen creatine phosphate or express the corresponding phosphagen kinase creatine kinase (or both), even when other phosphagens/phosphagen kinases are present in somatic tissues and eggs. The selective expression of the creatine kinase system in these spermatozoa may be related to potential advantages in the cellular transport of energy. To evaluate this possibility, we compared the efficacy of the major phosphagen systems for cellular transport of energy. We used a facilitated diffusion model for spatial ATP buffering, taking into account relative differences in diffusivity and thermodynamic poise. At low ratios of [total phosphagen pool]/ [total adenine nucleotide pool] (CG+P/CAd ratio), creatine phosphate carried a higher fraction of total high-energy phosphate (J) than the other phosphagens. However, J values for all phosphagens were greater than 0.9, and these differences disappeared as the CG+P/CAd ratio was increased. Thus, the functional benefit of using CP, rather than other phosphagens, in energy transport is quite limited. The creatine kinase system became associated with primitive-type spermatozoa early in metazoan evolution. This association is not necessarily related to inherent advantages of this phosphagen system for buffering of ATP, but may be linked to historical events in the evolution of the cell phenotype.


Effect of Printing Parameters on Tensile, Dynamic Mechanical, and Thermoelectric Properties of FDM 3D Printed CABS/ZnO Composites.
Fused deposition modelling (FDM) has been widely used in medical appliances, automobile, aircraft and aerospace, household appliances, toys, and many other fields. The ease of processing, low cost and high flexibility of FDM technique are strong advantages compared to other techniques for thermoelectric polymer composite fabrication. This research work focuses on the effect of two crucial printing parameters (infill density and printing pattern) on the tensile, dynamic mechanical, and thermoelectric properties of conductive acrylonitrile butadiene styrene/zinc oxide (CABS/ZnO composites fabricated by FDM technique. Results revealed significant improvement in tensile strength and Young's modulus, with a decrease in elongation at break with infill density. Improvement in dynamic storage modulus was observed when infill density changed from 50% to 100%. However, the loss modulus and damping factor reduced gradually. The increase of thermal conductivity was relatively smaller compared to the improvement of electrical conductivity and Seebeck coefficient, therefore, the calculated figure of merit (ZT) value increased with infill density. Line pattern performed better than rectilinear, especially in tensile properties and electrical conductivity. From the results obtained, FDM-fabricated CABS/ZnO showed much potential as a promising candidate for thermoelectric application.


Evaluation of viral genome assembly and diversity estimation in deep metagenomes.
Viruses have unique properties, small genome and regions of high similarity, whose effects on metagenomic assemblies have not been characterized so far. This study uses diverse in silico simulated viromes to evaluate how extensively genomes can be assembled using different sequencing platforms and assemblers. Further, it investigates the suitability of different methods to estimate viral diversity in metagenomes. We created in silico metagenomes mimicking various platforms at different sequencing depths. The CLC assembler revealed subpar compared to IDBA_UD and CAMERA , which are metagenomic-specific. Up to a saturation point, Illumina platforms proved more capable of reconstructing large portions of viral genomes compared to 454. Read length was an important factor for limiting chimericity, while scaffolding marginally improved contig length and accuracy. The genome length of the various viruses in the metagenomes did not significantly affect genome reconstruction, but the co-existence of highly similar genomes was detrimental. When evaluating diversity estimation tools, we found that PHACCS results were more accurate than those from CatchAll and clustering, which were both orders of magnitude above expected. Assemblers designed specifically for the analysis of metagenomes should be used to facilitate the creation of high-quality long contigs. Despite the high coverage possible, scientists should not expect to always obtain complete genomes, because their reconstruction may be hindered by co-existing species bearing highly similar genomic regions. Further development of metagenomics-oriented assemblers may help bypass these limitations in future studies. Meanwhile, the lack of fully reconstructed communities keeps methods to estimate viral diversity relevant. While none of the three methods tested had absolute precision, only PHACCS was deemed suitable for comparative studies.


Critical Overview on Pure Chitosan-based Scaffolds for Bone Tissue Engineering: Clinical insights in Dentistry.
Bone Tissue Engineering (BTE) is a field of regenerative medicine continuously improving, thanks to the development of new biomaterials used as grafts or scaffolds for repairing bone defects. In recent years, chitosan, a natural biopolymer extracted mainly from crustacean shells, has demonstrated unique and desirable characteristics for BTE applications, such as: biocompatibility, biodegradability, and osteoconductive behavior. Additionally, the presence of numerous active amine groups in its chemical structure allows it to be easily modified. Data suggest that chitosan scaffolds are highly biomimetic, and show an interesting bioactivity, and antibacterial behavior. We have demonstrated, in a critical overview, how chitosan-based scaffolds may hold great interest for BTE applications in medical and dental applications. Future research should be focused on the use of chitosan-scaffolds combined with other biomaterials or bioactive molecules, to increase their overall regenerative potential, also in critical-sized defects. In conclusion, chitosan can be considered a promising biomaterial in BTE and clinical dentistry.


Algorithm for mapping cutaneous tissue oxygen concentration using hyperspectral imaging.
The measurement of tissue oxygenation plays an important role in the diagnosis and therapeutic assessment of a large variety of diseases. Many different methods have been developed and are currently applied in clinical practice for the measurement of tissue oxygenation. Unfortunately, each of these methods has its own limitations. In this paper we proposed the use of hyperspectral imaging as new method for the assessment of the tissue oxygenation level. To extract this information from hyperspectral images a new algorithm for mapping cutaneous tissue oxygen concentration was developed. This algorithm takes into account and solves some problems related to setting and calculation of some parameters derived from hyperspectral images. The algorithm was tested with good results on synthetic images and then validated on the fingers of a hand with different blood irrigation states. The results obtained have proved the ability of hyperspectral imaging together with the developed algorithm to map the oxy- and deoxyhemoglobin distribution on the analyzed fingers. These are only preliminary results and other studies should be done before this approach to be used in the clinical setting for the diagnosis and monitoring of various diseases.


Arthroscopic-assisted focal resurfacing of the knee: surgical technique and preliminary results of 13 patients at 2 years follow-up.
The purpose of this study is to describe an arthroscopic-assisted surgical technique for focal resurfacing of medial tibio-femoral compartment and to present the preliminary clinical and radiographic results in a case series of 13 consecutive patients at a mean follow-up of 29 months. All patients were treated with the presented procedure for Ahlback grade 3 medial compartment osteoarthritis. Subjective evaluation was based on a visual analog scale for pain self-assessment. Objective clinical evaluation was based on Hospital for Special Surgery score. Range of motion was evaluated with a manual goniometer. Radiographic evaluation compared hip-knee-ankle angle pre- and post-operatively. Clinical and functional results were satisfactory. Hospital for Special Surgery score and visual analog scale for pain self-assessment showed significant improvements (P < 0.0001 and P = 0.0002, respectively). ROM and axial alignment were not significantly different respect to pre-operative values. Despite the small sample size and short follow-up, this study has shown that an arthroscopic-assisted focal resurfacing of the knee in selected cases can be a viable option for early onset degenerative joint disease, providing satisfactory pain relief and good functional results at 2 years follow-up. Retrospective case series, Level IV.


Risk factors of hepatitis B virus infection among blood donors in Duhok city, Kurdistan Region, Iraq.
Hepatitis B virus (HBV) infection is a public health problem. The lack of information about the seroprevalence and risk factors is an obstacle for preventive public health plans to reduce the burden of viral hepatitis. Therefore, this study was conducted in Iraq, where no studies had been performed to determine the prevalence and risk factors of HBV infection. Blood samples were collected form 438 blood donors attending blood bank in Duhok city. Serum samples were tested for HBV core-antibodies (HBcAb) and HBV surface-antigen (HBsAg) by ELISA. Various risk factors were recorded and multivariate analysis was performed. 5/438 (1.14%) of the subjects were HBsAg positive (HBsAg and HBcAb positive) and 36/438 (8.2%) were HBcAb positive. Hence, 41 cases were exposed to HBV and data analysis was based on that. Univariate analysis showed that there were significant associations between history of illegitimate sexual contact, history of alcohol or history of dental surgeries and HBV exposure (p<0.05 for all). Then, multivariate analysis was conducted to find HBV exposure predictive factors. It was found that history of dental surgery was a predictive factor for exposure to the virus (P=0.03, OR: 2.397). This study suggested that the history of dental surgery was predictive for HBV transmission in Duhok city. Further population-based study is needed to determine HBV risk factors in the society and public health plan based on that should be considered.


Mechanism and Experimental Verification of Luteolin for the Treatment of Osteoporosis Based on Network Pharmacology.
To explore the molecular mechanism of luteolin in the treatment of osteoporosis (OP) by network pharmacological prediction and experimentation. The target proteins of luteolin were obtained with the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). OP-related proteins were extracted from the Comparative Toxicogenomics Database (CTD) and GeneCards and DisGeNET databases. We imported the common protein targets of luteolin and OP into the STRING database to obtain the relationships between the targets. The common target proteins of luteolin and OP were assessed by KEGG and GO enrichment analyses with the DAVID database. Animal experiments were conducted to verify the effect of luteolin on bone mineral density in ovariectomised (OVX) rats. Finally, the effects of luteolin on key signalling pathways were verified by cell experiments in vitro. Forty-four targets of luteolin involved in the treatment of OP, including key target proteins such as TP53, AKT1, HSP90AA1, JUN, RELA, CASP3, and MAPK1, were screened. KEGG enrichment analysis found that luteolin inhibits OP by regulating the PI3K-Akt, TNF, oestrogen and p53 signalling pathways. The results of animal experiments showed that bone mass in the low-dose luteolin group (Luteolin-L group, 10 mg/kg), high-dose luteolin group (Luteolin-H group, 50 mg/kg) and positive drug group was significantly higher than that in the OVX group (P<0.05). Western blot (WB) analysis showed that the protein expression levels of Collagen I, Osteopontin and RUNX2 in bone marrow mesenchymal stem cells (BMSCs) cultured with 0.5, 1 and 5 μM luteolin for 48 h were significantly higher than those in the dimethyl sulfoxide (DMSO) group (P<0.05). In vitro cell experiments showed that the p-PI3K/PI3K and p-Akt/Akt expression ratios in BMSCs cultured with 0.5, 1 and 5 μM luteolin for 48 h were also significantly higher than those in the DMSO group (P<0.05). Luteolin has multitarget and multichannel effects in the treatment of OP. Luteolin could reduce bone loss in OVX rats, which may be due to its ability to promote the osteogenic differentiation of BMSCs by regulating the activity of the PI3K-Akt signalling pathway.


Retinal disease masquerading as spasmus nutans.
Two children with congenital stationary night blindness were originally diagnosed as having spasmus nutans. Both children had the typical features of spasmus nutans including asymmetric nystagmus, head shaking, and torticollis. The diagnosis of congenital stationary night blindness was established only after each child underwent electroretinography. The nystagmus associated with retinal disease can mimic many of the features of spasmus nutans. Children suspected of having spasmus nutans should undergo complete ophthalmologic examination and electroretinography if they are myopic.


Characterization of the microsomal epoxide hydrolase gene in patients with anticonvulsant adverse drug reactions.
Therapy with the aromatic anticonvulsants phenytoin, phenobarbital and carbamazepine has been associated with the occurrence of rare idiosyncratic hypersensitivity reactions. These drugs are thought to be activated to potentially reactive arene oxide (epoxide) metabolites by cytochrome P450-dependent monooxygenation, while liver microsomal epoxide hydrolase (mEH) plays a detoxifying role by converting such reactive intermediates to non-toxic dihydrodiols. Evidence from in vitro lymphocyte toxicity tests and enzyme inhibitor studies has suggested that an inherited defect in mEH function may be responsible for the enhanced drug toxicity observed in affected individuals. To test this hypothesis we designed methods to directly compare mEH gene structure in patients presenting with anticonvulsant adverse reactions and in control subjects in which no in vivo or in vitro toxicity to anticonvulsants could be demonstrated. Southern analysis of peripheral lymphocyte DNA using a full-length mEH cDNA as hybridization probe revealed no gross differences in mEH gene structure in selected patients when compared with DNA samples from unaffected control subjects. The human mEH gene was then cloned and characterized from a control individual. Nine exons were identified within a 22 kb region and sequences of selected regions, including all exons, were determined. Single strand conformation polymorphism (SSCP) analysis was performed on all exonic regions in genomic DNA from each of 26 subjects, including six unrelated patients with previous toxicity to anticonvulsant therapy and seven siblings (three of whom had displayed toxicity). Several distinct SSCP patterns could be observed among the subjects tested, each corresponding to a specific point mutation within one of the amplified fragments of the mEH gene. However, none of the SSCP patterns reflecting point mutations was correlated with the occurrence of anticonvulsant toxicity. From these observations we conclude that a genetic defect altering the structure and function of the mEH protein is unlikely to be responsible for predisposing patients to anticonvulsant adverse reactions.


Airway narrowing measured by spirometry and impulse oscillometry following room temperature and cold temperature exercise.
The efficacy of using impulse oscillometry (IOS) as an indirect measure of airflow obstruction compared to spirometry after exercise challenges in the evaluation of exercise-induced bronchoconstriction (EIB) has not been fully appreciated. The objective was to compare airway responses following room temperature and cold temperature exercise challenges, and to compare whether IOS variables relate to spirometry variables. Spirometry and IOS were performed at baseline and for 20 min after challenge at 5-min intervals. Two 6-min exercise challenges, inhaling either room temperature (22.0 degrees C) or cold temperature (- 1 degrees C) dry medical-grade bottled air. At least 48 h was observed between these randomly assigned challenges. Twenty-two physically active individuals (12 women and 10 men) with probable EIB. Subjects performed 6 min of stationary cycle ergometry while breathing either cold or room temperature medical-grade dry bottled air. Subjects were instructed to exercise at the highest intensity sustainable for the duration of the challenge. Heart rate and kilojoules of work performed were documented to verify exercise intensity. Strong correlations were observed within testing modalities for post-room temperature and post-cold temperature exercise spirometry and IOS values. Spirometry revealed no differences in postexercise peak falls in lung function between conditions; however, IOS identified significant differences in respiratory resistance (p < 0.05), with room temperature-inspired air being more potent than cold temperature-inspired air. Correlations were found between spirometric and IOS measures of change in airway function for both exercise challenges, indicating close equivalency of the methods. The challenges appeared to elicit the EIB response by a similar mechanism of water loss, and cold temperature did not have an additive effect. IOS detected a difference in degree of response between the temperatures, whereas spirometry indicated no difference, suggesting that IOS is a more sensitive measure of change in airway function.


Small cell carcinoma of the urinary bladder--a new case report.
Primary pure small cell carcinoma of the urinary bladder is an extremely rare and highly aggressive tumor with an average five-year survival rate of less than 10% as cited by multiple case reports. It accounts for about 0.5-1% of all bladder tumors. We present the case of a 44-years-old man, smoker (10 cigarettes/day) hospitalized in the Department of Urology, from the "Prof. dr. Th. Burghele" Hospital, Bucharest, for one month intermittent hematuria. Ultrasonography showed a sessile tumoral mass, sized 37/30mm. Transurethral resection of the tumor mass was performed and tissue fragments were sent to the pathologic lab to establish the histologic type, the degree of differentiation and invasion. Fragments of the tumor were fixed in 10% formaldehyde, paraffin embedded and processed as standard technique; the sections were stained with HE, VG and immunohistochemically with: CROMO, EMA, NSE, CD56, NK1, p53 and betaHCG. The microscopic examination reveled a tumor proliferation composed of two distinct components: extensive small cells areas and foci of typical low grade (G2) papillary urothelial carcinoma. The small cell are uniformly, round, with increased nucleo-cytoplasmic ratio, eosinophyl cytoplasm, hyperchromatic nuclei, finely granular chromatin and inconspicuous nucleoli. Immunohistochemical stains showed diffuse positive staining of the small cell component for CROMO, EMA, NSE, CD56, NK1 and urothelial carcinoma component stained focally for betaHCG. The rate of cell proliferation was increased (p53 - 80% positive reaction). Conclusions. A diagnosis of small cell carcinoma coexisting with low-grade urothelial carcinoma was established. Because of aggressive behavior and distinct treatment, the pathologist should watch out for the presence of small cell carcinoma component.


Laboratory studies with a new broad-spectrum penicillin, pirbenicillin.
Pirbenicillin {6-[d-2-phenyl-2(N-4-pyridylformimidoylaminoacetamido) -acetamido]-penicillanic acid} showed broad-spectrum antibacterial activity in vitro and also in the treatment of experimental infections after parenteral administration to mice. Against Pseudomonas aeruginosa, a three- to fourfold potency advantage over carbenicillin was seen both in vitro and in vivo. The in vitro antibacterial spectrum of pirbenicillin includes Escherichia coli, Serratia, Citrobacter, and Enterobacter isolates, against which it exhibited minimal inhibitory concentration values comparable to those of carbenicillin. However, mice infected with E. coli and Serratia were protected at doses of pirbenicillin that were two to four times lower than those required of carbenicillin. Pirbenicillin was more active than carbenicillin against gram-positive bacteria, especially Streptococcus faecalis. It was less active than carbenicillin against Proteus spp. and was inactive against ampicillin-resistant E. coli strains. Pirbenicillin was bactericidal at concentrations generally equal to or only two-fold higher than the minimal inhibitory concentration. With appropriately buffered media, pirbenicillin demonstrated eight- and fourfold better minimal bactericidal concentration values towards Pseudomonas isolates than those of carbenicillin and ticarcillin, respectively.


Natural Human Immunity Against Staphylococcal Protein A Relies on Effector Functions Triggered by IgG3.
Staphylococcal protein A (SpA) is a multifunctional, highly conserved virulence factor of Staphylococcus aureus. By binding the Fc portion of all human IgG subclasses apart from IgG3, SpA interferes with antibody and complement deposition on the bacterial surface, impairing staphylococcal clearance by phagocytosis. Because of its anti-opsonic properties, SpA is not investigated as a surface antigen to mediate bacterial phagocytosis. Herein we investigate human sera for the presence of SpA-opsonizing antibodies. The screening revealed that sera containing IgG3 against SpA were able to correctly opsonize the target and drive Fcγ receptor-mediated interactions and phagocytosis. We demonstrated that IgG3 Fc is significantly more efficient in inducing phagocytosis of SpA-expressing S. aureus as compared to IgG1 Fc in an assay resembling physiological conditions. Furthermore, we show that the capacity of SpA antibodies to induce phagocytosis depends on the specific epitope recognized by the IgGs on SpA molecules. Overall, our results suggest that anti-SpA IgG3 antibodies could favor the anti-staphylococcal response in humans, paving the way towards the identification of a correlate of protection against staphylococcal infections.


Dimensionality and Interface Engineering of 2D Homologous Perovskites for Boosted Charge-Carrier Transport and Photodetection Performances.
Two-dimensional (2D) homologous halide perovskite (HP) microcrystallines have emerged as a promising alternative light-sensitive material; however, the undesirable quantum confinement effect and severe interfacial charge-carrier scattering still hamper their applications in photodetectors (PDs). Here we propose a novel postsynthetic treatment to simultaneously solve both problems. 2D (OA)2FAn-1PbnBr3n+1 (OA and FA represent octadecylamine and formamidine) microplatelet film was immersed in solution containing FA+, leading to improvements in two aspects. First, the dimensionality of 2D HPs was increased through an exchange reaction between OA+ and FA+, which meliorates the quantum confinement effect and facilitates the separation of electrons and holes; second, the free-standing 2D HP microcrystallines were fused for promoted interdomain charge-carrier transport. The treated PDs achieved a 3600 and 4200% increase in external quantum yield and responsivity up to 7100% and 32 A/W, respectively, and the rise/decay time was shortened by two orders of magnitude to 0.25/1.45 ms.


Rapid Eye Movement (REM) sleep behavior disorder: a sleep disturbance affecting mainly older men.
Rapid Eye Movement (REM) sleep behavior disorder is characterized by the intermittent loss of REM-related muscle atonia and the appearance of elaborated motor behaviors (sometimes violent behavior) and vocalizations associated with dream mentation. Nine patients were diagnosed in our Sleep Disorders Unit with this syndrome during the period August 1997-April 2000. All were male, average age 67.9 +/- 6.9 years. The complaint of all our patients was the occurrence of violent or injurious sleep behavior mainly during the dream stage. Jumping or falling out of bed and slapping or beating their wives were more common. None had history or showed signs of dementia, Parkinson or other neurodegenerative diseases. A relative high amount of SWS (20.9%) was found. Seven showed an intermittent increase in chin EMG tonus while the other two had an almost continuous high chin EMG tonus during REM sleep. We did not observe any violent motor behavior during the polysomnographic recordings. Phasic activities during REM sleep were high but density quantification was not performed. Six patients had also Periodic Limb Movement (PLM) Disorders, four had also Obstructive Sleep Apnea (OSA) Syndrome. The treatment recommended to all patients was Clonazepam beginning with a 0.5-mg dose. Four patients reported a decrease or disappearance of sleep agitation and nightmares and were very happy with the treatment and without side effects. The others decided not to try Clonazepam or stopped after a few days of using it. RBD appears to be a sleep disturbance affecting mainly aged men. Its violent expression may frighten the patients and their bed-partners and may cause injury to both. In some cases this sleep disorder seems to be an early manifestation of a neurodegenerative disorder while in others it may represent only an idiopathic form. Clonazepam at lower doses is a good agent for the treatment of this condition.


Atomic force microscope mediated chromatography.
An atomic force microscope (AFM) is presented as an instrument for rapid, miniaturized chromatography. The AFM is used to inject a sample, provide shear driven liquid flow over a functionalized substrate, and detect separated components. The components are then analyzed with surface enhanced Raman spectroscopy using AFM deposition of gold nanoparticles on the separated bands. This AFM mediated chromatography (AFM-MC) is demonstrated using lipophilic dyes and normal phase chemistry. A significant reduction in both size and separation time scales is achieved with 25 μm length scale and 1 s separation times. AFM-MC has general applications to trace chemical analysis and microfluidics.


[Effects of methyl protodioscin on [Ca2+]i and ATPase activity in cardiomyocytes and analysis of mechanisms].
To study the effects of methyl protodioscin on the [Ca2+]i and the ATPase activity in cardiomyocytes, as well as their mechanisms. The cardiomyocytes were randomly divided into three groups, the control group treated with no serumal DMEM, the MPD group treated with MPD and the dilthiazem group treated with dilthiazem. Fluorospectrophotometer was used to determined the level of myocardial cell intracellular Ca2+ [Ca2+]i. In the experiment of ATPase activity on cellular membrane, the cardiomyocytes were randomly divided into two groups, the control group treated with no serumal DMEM, the MPD group treated with MPD. The activity of Na+-K+-ATPase,Ca2+-Mg2+-ATPase and Mg2+-ATP ATPase were determined. The quantitative analysis of SERCA2a mRNA expression was studied by RT-PCR that the groups and treatments in cardiomyocytes same as the experiment for ATPase activity assay. Under the quiescent condition, compared to the control group, the level of [Ca2+]i in cardiomyocytes of the MPD group and dilthiazem group was no different. After treatment with 40 mmol x L(-1) KCl, [Ca2+] was significantly lower in the MPD group and the dilthiazem group, and the intensity of peak value in time course of 60 s, the dilthiazem group and the MPD group also were lower than the control group (P < 0.001). Ca2+-Mg2+-ATPase and Na+-K+-ATPase in cultured rat were increased after treated with MPD compared to treatment with no serumal DMEM (P < 0.05, P < 0.01), but Mg2+-ATPase in these groups had no different. The expression of SERCA2a mRNA between the MPD group and the control group was no different. MPD could not up-regulated or down-regulated SERCA2a in endocytoplasmic reticulum. Methyl protodioscin could block the volt dependent form calcium channel in cellular membrane, and up-regulate the function of sodium pump and calcium pump, so that it could remain low calcium in the internal environment in cardiomyocytes.


Effects of alternative techniques of low-dose heparin administration on hematoma formation.
A small clinical trial was conducted to compare a modified subcutaneous injection technique with the standard subcutaneous method. The data in this study do not strongly support the use of either technique. However, the modified subcutaneous method may be preferable in that fewer injection-site hematomas formed. Further research with a larger population is indicated before either method can be recommended over the other.


Development of 30 novel polymorphic Expressed Sequence Tags (EST)-derived microsatellite markers for the Miiuy Croaker, Miichthys miiuy.
Expressed sequence tags (ESTs) can be used to identify microsatellite markers. We developed 30 polymorphic microsatellite markers from 5053 ESTs of the Miichthys miiuy. Out of 123 EST derived microsatellites for which PCR primers were designed, 30 loci were polymorphic in 30 individuals from a single natural population with 2-13 alleles per locus. The observed and expected heterozygosities were from 0.1024 to 0.7917 and from 0.2732 to 0.8845, respectively. Nine loci deviated from the Hardy-Weinberg equilibrium, and linkage disequilibrium was significant between 22 pairs of loci. These polymorphic microsatellite loci will be useful for genetic diversity analysis and molecule-assisted breeding for M. miiuy.


Purification and characterization of high Ca2+-requiring neutral proteases from porcine and bovine brains.
Porcine and bovine brain high Ca2+-requiring neutral proteases were purified to homogeneity by the same isolation procedures, and their properties were compared. A high degree of similarity existed between the two proteases. The purification procedures included ion-exchange chromatography on DEAE-cellulose, hydrophobic chromatography on phenyl-Sepharose CL-4B, second DEAE-cellulose chromatography, second phenyl-Sepharose CL-4B chromatography, and gel filtration on Ultrogel AcA 34. Both purified enzymes were composed of Mr 75,000 and 29,000 subunits, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Both enzymes required 250 microM Ca2+ for half-maximal activity and 700 microM Ca2+ for maximal activity. Sr2+ and Ba2+, but not Mg2+ or Mn2+, also activated both enzymes but not as effectively as Ca2+. Both enzymes displayed maximum activity at pH 7.5-8.0. Leupeptin, antipain, and trans-epoxysuccinyl-L-leucylagmatine inhibited both enzymes. Neurofilament triplet proteins and microtubule-associated proteins were extensively hydrolyzed by both proteases, but tubulin and actin were not hydrolyzed. The amino acid compositions of the two proteases were very similar. Antisera against bovine brain protease cross-reacted with porcine brain protease when examined by immunoelectrotransfer blot techniques.


Quantification of Hydrogen Bond Strength Based on Interaction Coordinates: A New Approach.
A new approach to quantify hydrogen bond strengths based on interaction coordinates (HBSBIC) is proposed and is very promising. In this research, it is assumed that the projected force field of the fictitious three atoms fragment (DHA) where D is the proton donor and A is the proton acceptor from the full molecular force field of the H-bonded complex characterizes the hydrogen bond. The "interaction coordinate (IC)" derived from the internal compliance matrix elements of this three-atom fragment measures how the DH covalent bond (its electron density) responds to constrained optimization when the HA hydrogen bond is stretched by a known amount (its electron density is perturbed by a specified amount). This response of the DH bond, based on how the IC depends on the electron density along the HA bond, is a measure of the hydrogen bond strength. The inter- and intramolecular hydrogen bond strengths for a variety of chemical and biological systems are reported. When defined and evaluated using the IC approach, the HBSBIC index leads to satisfactory results. Because this involves only a three-atom fragment for each hydrogen bond, the approach should open up new directions in the study of "appropriate small fragments" in large biomolecules.


Dually eligible for Medicare and Medicaid: two for one or double jeopardy?
This issue brief describes the characteristics of the population of individuals known as "dual eligibles," who are eligible for health insurance coverage through both Medicare and Medicaid. It also looks at the differences between "full Medicaid" and "supplemental Medicaid" dual eligibles and the ongoing challenges associated with enrollment and eligibility, integration and coordination, and managed care. The paper presents several examples of integrated care programs designed to better serve the dual-eligible population, including the Program of All-Inclusive Care for the Elderly, Evercare, social health maintenance organizations, and state/federal initiatives such as the Wisconsin Partnership Program, Texas STAR+PLUS, and others. Finally, it considers the implications for dual eligibles of the House and Senate Medicare prescription drug proposals.


Priorities in prevention research for eating disorders.
This article discusses potential areas for research in the prevention of eating disorders. Prevention research encompasses a large range of activities; reviews are available (Crisp 1988; Shisslak et al. 1987; USDHHS/McKnight 1996). This article focuses on school-based primary preventive interventions.


Familial cancer aggregation and the risk of lung cancer.
Around 90% of lung cancer worldwide is attributable to cigarette smoking, although less than 20% of cigarette smokers develop lung cancer. Other factors such as diet, chronic lung diseases, occupation and possibly environmental agents also contribute to this cancer. Genetic factors seem to play a role in lung cancer, but the precise characteristics influencing lung cancer susceptibility are not known, since genetic factors are easily obscured by the strong environmental determinants of lung cancer, particularly smoking. To estimate the effect that cancer occurrence among first-degree relatives has on the risk of lung cancer. Hospital-based case-control study. The metropolitan region of São Paulo, Brazil. 334 incident lung cancer cases and 578 controls matched by hospitals. By means of a structured questionnaire, cases and controls were interviewed about cancer occurrence in first-degree relatives, tobacco smoking, exposure to passive smoking, occupation, migration and socioeconomic status. Non-conditional logistic regression was used to calculate the risk of familial cancer aggregation, the effect of cancer in first-degree relatives and smoking in conjunction, and for controlling confounders. The adjusted odds ratio (OR) revealed a slight, but not statistically significant, excess risk of lung cancer for subjects with a history of lung cancer in relatives (OR 1.21; 95% confidence interval [CI] 0.50 - 2.92). The same was found among those with a history of other tobacco-related cancers in relatives (OR 1.36; 95% CI 0.87 - 2.14). A step gradient effect was observed regarding lung cancer risk, in accordance with increases in the number of pack-years of cigarette consumption. An interaction between familial cancer aggregation and tobacco smoking was detected. A mildly elevated risk of lung cancer among persons with a positive history of lung and other tobacco-related cancers was observed. The finding of an interaction between the variables of familial cancer aggregation and smoking suggests that familial cancer aggregation could be considered as a marker of susceptibility, increasing the risk of lung cancer among smokers. These results improve our knowledge of lung carcinogenesis and can guide future cancer genetic studies.


Scintillation index for the optical wave in the vertical oceanic link with anisotropic tilt angle.
The influence of the ocean depth and anisotropic tilt angle on vertical underwater wireless optical communication (UWOC) systems is considered in this study. We propose a power spectrum model of oceanic turbulence with an anisotropic tilt angle for the first time. Thereafter, the expression of the scintillation index is derived for a spherical wave propagating over anisotropic oceanic turbulence in the vertical link. In addition, considering the temperature and salinity, relevant data of the Atlantic and Pacific oceans at different depths are selected to study further the effect of ocean depth on the scintillation index. The results indicate that the scintillation index strongly depends on the ocean depth and anisotropic tilt angle. Moreover, the scintillation index is also related to other parameters, such as temperature and salinity, kinematic viscosity, the anisotropic factor, optical wavelength, and propagation distance. The presented results can be beneficial in designing optical wireless communication systems in the ocean environment.


Strategic planning. Part V of the dental management pyramid series.
Strategic planning is not something that should be taken lightly. It is the course by which you will map out your own personal future, as well as that of your practice. You cannot look ahead until you and your dental team are ready to accurately and responsibly assess the strengths, weaknesses, opportunities and threats facing your practice today. Putting in the work to do so will leave you and your practice in a better position to grow and prosper in the long run.


Interaction of the v-Rel oncoprotein with cellular transcription factor Sp1.
We previously showed that v-Rel, the oncoprotein of the avian retrovirus Rev-T, can increase expression from promoters containing binding sites for the cellular transcription factor Sp1 in chicken embryo fibroblasts (S. Sif, A.J. Capobianco, and T.D. Gilmore, Oncogene 8:2501-2509, 1993). In those experiments, v-Rel appeared to increase the transactivating function of Sp1; that is, v-Rel stimulated transactivation by a GAL4-Sp1 protein that lacked the Sp1 DNA-binding domain. We have now shown that in vitro-synthesized v-Rel and GAL4-Sp1 form a complex that can be immunoprecipitated with either anti-Sp1 or anti-v-Rel antiserum. We have also shown that a glutathione S-transferase (GST)-Sp1 fusion protein can specifically interact with in vitro-translated v-Rel and with in vivo-synthesized v-Rel from transformed chicken spleen cells. In addition, we have found that the abilities of wild-type and two mutant forms of v-Rel to increase transactivation by Sp1 in vivo correlate with their abilities to interact with Sp1 in vitro. The sequences important for the interaction of v-Rel with Sp1 in vitro have been mapped to the first 147 amino acids of v-Rel. Other Rel proteins, such as c-Rel, RelA, p52, and p50, were also able to form a complex with Sp1 in vitro. These results suggest that v-Rel increases expression from Sp1 site-containing promoters by functionally interacting with Sp1 and that cellular Rel proteins and Sp1 are likely to interact to influence transcription from natural promoters.


A measurement theory of illusory conjunctions.
Illusory conjunctions refer to the incorrect perceptual combination of correctly perceived features, such as color and shape. Research on the phenomenon has been hampered by the lack of a measurement theory that accounts for guessing features, as well as the incorrect combination of correctly perceived features. Recently, several investigators have suggested using multinomial models as a tool for measuring feature integration. The authors examined the adequacy of these models in 2 experiments by testing whether model parameters reflect changes in stimulus factors. In a third experiment, confidence ratings were used as a tool for testing the model. Multinomial models accurately reflected both variations in stimulus factors and observers' trial-by-trial confidence ratings.


The influence of the microbial quality of wastewater, lettuce cultivars and enumeration technique when estimating the microbial contamination of wastewater-irrigated lettuce.
This study investigated the volume of wastewater retained on the surface of three different varieties of lettuce, Iceberg, Cos, and Oak leaf, following submersion in wastewater of different microbial qualities (10, 102, 103, and 104 E. coli MPN/100 mL) as a surrogate method for estimation of contamination of spray-irrigated lettuce. Uniquely, Escherichia coli was enumerated, after submersion, on both the outer and inner leaves and in a composite sample of lettuce. E. coli were enumerated using two techniques. Firstly, from samples of leaves - the direct method. Secondly, using an indirect method, where the E. coli concentrations were estimated from the volume of wastewater retained by the lettuce and the E. coli concentration of the wastewater. The results showed that different varieties of lettuce retained significantly different volumes of wastewater (p < 0.01). No statistical differences (p > 0.01) were detected between E. coli counts obtained from different parts of lettuce, nor between the direct and indirect enumeration methods. Statistically significant linear relationships were derived relating the E. coli concentration of the wastewater in which the lettuces were submerged to the subsequent E. coli count on each variety the lettuce.


Achalasia: current therapeutic options.
Achalasia is a chronic incurable esophageal motility disorder characterized by impaired lower esophageal sphincter (LES) relaxation and loss of esophageal peristalsis. Although rare, it is currently the most common primary esophageal motility disorder, with an annual incidence of around 1.6 per 100,000 persons and prevalence of around 10.8/100,000 persons. Symptoms of achalasia include dysphagia to both solids and liquids, regurgitation, aspiration, chest pain and weight loss. As the underlying etiology of achalasia remains unclear, there is currently no curative treatment for achalasia. Management of achalasia mainly involves improving the esophageal outflow in order to provide symptomatic relief to patients. The most effective treatment options for achalasia include pneumatic dilation, Heller myotomy and peroral endoscopic myotomy (POEM), with the latter increasingly emerging as the treatment of choice for many patients. This review focusses on evidence for current and emerging treatment options for achalasia with a particular emphasis on POEM.


Glyoxalase I-type hemithioacetal isomerization reactivity of a mononuclear Ni(II) deprotonated amide complex.
The synthesis, characterization, and hemithioacetal isomerization reactivity of a mononuclear Ni(II) deprotonated amide complex, [(bppppa-)Ni]ClO4.CH3OH (1, bppppa- = monoanion of N,N-bis-[(6-phenyl-2-pyridyl)methyl]-N-[(6-pivaloylamido-2-pyridyl)methyl]amine), are reported. Complex 1 was characterized by X-ray crystallography, 1H NMR, UV-vis, FTIR, and elemental analysis. Treatment of 1 with an equimolar amount of the hemithioacetal PhC(O)CH(OH)SCD3 in dry acetonitrile results in the production of the thioester PhCH(OH)C(O)SCD3 in approximately 60% yield. This reaction is conveniently monitored via 2H NMR spectroscopy. A protonated analogue of 1, [(bppppa)Ni](ClO4)2 (2), is unreactive with the hemithioacetal, thus indicating the requirement of the anionic chelate ligand in 1 for hemithioacetal isomerization reactivity. Complex 1 is unreactive with the thioester product, PhCH(OH)C(O)SCD3, which indicates that the pKa value for the PhCH(OH)C(O)SCD3 proton of the thioester must be significantly higher than the pKa value of the C-H proton of the hemithioacetal (PhC(O)CH(OH)SCD3). Complex 1 is the first well-characterized Ni(II) coordination complex to exhibit reactivity relevant to Ni(II)-containing E. coli glyoxalase I. Treatment of NiBr2.2H2O with PhC(O)CH(OH)SCD3 in the presence of 1-methylpyrrolidine also yields thioester product, albeit the reaction is slower and involves the formation of multiple -SCD3 labeled species, as detected by 2H NMR spectroscopy. The results of this study provide the first insight into hemithioacetal isomerization promoted by a synthetic Ni(II) coordination complex versus a simple Ni(II) ion.


Pathophysiological Responses Associated with Exposure to TQ and EGCG Using CAOV-3 Ovarian Cancer Like Cells.
Ovarian cancer is the fifth most common cancer and the leading cause of mortality among the gynecologic cancers. Ovarian cancer is a very devastating disease it is rarely diagnosed in its early stages, and it is usually quite advanced by the time diagnosis is made leading to very poor outcomes. It is believed that in advanced stage ovarian cancer cells produce a protein that enables the tumor cells to proliferate and become resistance to conventional drug treatments. Development of new treatment options strongly relies on understanding the ovarian carcinoma ability to proliferate and to attack the cells using combination drug treatments that are synergistic. Recent studies in our laboratory have indicated that the exposure of Thymoquinone (TQ) and Epigallocatechin-3-gallate (EGCG) to adenocarcinoma of the prostate, colon, and pancreas. This study was executed to investigate the effectiveness of TQ and EGCG on reducing early stage ovarian cancer cells. A total of 72 wells were plated with (105) Caov-3 ovarian cancer cells according to standard lab protocols. Each group was subdivided into 4 groups. Group 1 served as control and groups 2, 3, and 4 were treated with TQ (16 µM), EGCG (3 µM), and TQ + EGCG, respectively. Biomarkers and morphological evaluations were performed following standard lab techniques. The results of the study revealed: (1) an increase in nitric oxide following administration of EGCG and the combination at 24 and 48 hours (p<0.05) and (2) no membrane or cellular damage to the cells at all phases. The results of this study suggest used in combination have shown to be more effective method for manipulating Caov-3 ovarian cancer cells than when used alone.


Glucose is a key metabolic regulator of osteoclasts; glucose stimulated increases in ATP/ADP ratio and calmodulin kinase II activity.
Glucose-stimulated increases in osteoclast activity are mediated, at least in part, by transcriptional regulation of H+-ATPase expression through a mechanism involving p38 mitogen-activated protein kinase. We hypothesized that early events in the glucose-dependent signaling pathway would be similar to those identified in other glucose-sensitive cells, such as islet beta-cells, including rapid changes in the cellular ATP/ADP ratio and mobilization of intracellular Ca2+. We demonstrate that glucose stimulates a prolonged 50% increase in the ATP/ADP ratio that was maximal 30 s after glucose concentrations were increased. Glucose stimulated a transient 30% increase in calcium/calmodulin-dependent kinase II (CaMK II) activity that was maximal 3 min after the glucose concentration was increased. CaMK II was activated maximally by 3 mmol D-glucose/L in 3-min assays. Activation of CaMK II in the presence of the nonmetabolizable glucose analog 2-deoxyglucose was 2-fold greater than with D-glucose but was unchanged by glucosamine. Pretreatment of osteoclasts with the intracellular Ca2+ chelator BAPTA-AM inhibited glucose transport by 75%. BAPTA-AM treatment also prevented glucose-dependent stimulation of CaMK II. The data indicate that osteoclasts utilize a glucose-sensing mechanism similar to that of beta-cells and that glucose-stimulated signaling in osteoclasts involves changes in the ATP/ADP ratio and mobilization of intracellular Ca2+, resulting in activation of CaMK II.


National survey of risk factors for non-communicable disease in Vietnam: prevalence estimates and an assessment of their validity.
To estimate the prevalence of non-communicable disease (NCD) risk factors at a provincial level in Vietnam, and to assess whether the summary estimates allow reliable inferences to be drawn regarding regional differences in risk factors and associations between them. Participants (n = 14706, 53.5 % females) aged 25-64 years were selected by multi-stage stratified cluster sampling from eight provinces each representing one of the eight geographical regions of Vietnam. Measurements were made using the World Health Organization STEPS protocols. Data were analysed using complex survey methods. Differences by sex in mean years of schooling (males 8.26 ± 0.20, females 7.00 ± 0.18), proportions of current smokers (males 57.70 %, females 1.73 %), and binge-drinkers (males 25.11 %, females 0.63 %), and regional differences in diet, reflected the geographical and socio-cultural characteristics of the country. Provinces with a higher proportion of urban population had greater mean levels of BMI (r = 0.82), and lesser proportions of active people (r = -0.89). The associations between the summary estimates were generally plausible (e.g. physical activity and BMI, r = -0.80) but overstated, and with some anomalous findings due to characterisation of smoking and hypertension by STEPS protocols. This report provides an extensive description of the sex-specific and regional distribution of NCD risk factors in Vietnam and an account of some health-related consequences of industrialisation in its early stages. The STEPS protocols can be utilized to provide aggregate data for valid between-population comparisons, but with important caveats identified.


E-Cigarette Use Among Never-Smoking California Students.
We determined the extent to which adolescents who have never used tobacco try e-cigarettes. Data on the prevalence and correlates of e-cigarette use among 482,179 California middle and high school students are from the 2013-2014 California Healthy Kids Survey. Overall, 24.4% had ever used e-cigarettes (13.4% have never used tobacco and 11.0% have used tobacco), and 12.9% were current e-cigarette users (5.9% have never used tobacco). Among those who have never used tobacco, males and older students were more likely to use e-cigarettes than females and younger students. Hispanics (odds ratio [OR] = 1.60; confidence interval [CI] = 1.53, 1.67) and those of other races (OR = 1.24; CI = 1.19, 1.29) were more likely than Whites to have ever used e-cigarettes, but only among those who had never used smokeless tobacco and never smoked a whole cigarette. E-cigarette use is very prevalent among California students who have never smoked tobacco, especially among Hispanic and other race students, males, and older students.


A customized retroviral vector confers marker gene expression in osteoclast lineage cells.
Osteoclasts play a seminal role in many skeletal diseases and therefore are candidates for cell-based gene delivery systems to treat disorders of bone. As an initial step toward developing osteoclast-mediated gene delivery systems, we have made and analyzed a customized Molony-Murine leukemia virus (MMLV)-based retroviral vector containing elements of the osteoclast-specific tartrate-resistant acid phosphatase (TRAP) gene. RAW 264.7 cells were transduced with the customized vector (E3) and differentiated along macrophage or osteoclast lineages. E3 contained a truncated form of the human nerve growth factor receptor (NGFR) as a reporter gene. NGFR expression increased with RANK-ligand (RANK-L) treatment but not with macrophage (gamma-IFN/LPS treatment) differentiation. Enhanced NGFR expression peaked 48 h after RANK-L treatment. Electrophoretic mobility shift assays (EMSA) analysis of the TRAP gene regulatory elements in E3 identified a single 27 bp DNA probe, which specifically bound protein from RANK-L-treated cells. DNA sequence revealed AP-1 binding sites, and analysis with mutant probes implied that the sites were functional. EMSA supershift analysis identified Fos protein interacting with the 27 bp probe. In summary, insertion of sequence -962 to -868 from the TRAP gene into the U3 region of the MMLV LTR confers RANK-L induced retroviral gene expression via Fos family protein interaction at AP-1 sites.


Subjective experience or objective process: understanding the gap between values and practice for involving patients in designing patient-centred care.
Patient-centred care and patient involvement are increasingly central concepts in health policy in the UK and elsewhere. However, there is little consensus regarding their definition or how to achieve "patient-centred" care in everyday practice or how to involve patients in service redesign initiatives. The purpose of this paper is to explore these issues from the perspective of key stakeholders within National Health Service (NHS) hospitals in the UK. Semi-structured interviews, covering a range of topics related to service redesign, were conducted with 77 key stakeholders across three NHS Trusts in the West Midlands. In total, 20 of these stakeholders were re-interviewed 18 months later. Data were managed and analysed using the Framework Method. While patient-centred care and patient involvement were regularly cited as important to the stakeholders, a gap persisted between values and reported practice. This gap is explained through close examination of the ways in which the concepts were used by stakeholders, and identifying the way in which they were adapted to fit other organizational priorities. The value placed on positive subjective experience changed to concerns about objective measurement of the patients as they move through the system. Increased awareness and reflection on the conceptual tensions between objective processes and subjective experiences could highlight reasons why patient-centred values fail to translate into improved practice. The paper describes and explains a previously unarticulated tension in health organisations between values and practice in patient centred care and patient involvement in service redesign.