Intranuclear distribution of poly(A) RNA determined by electron microscope in situ hybridization.
We used a biotinylated poly(dT) probe to localize poly(A) RNA in HeLa cells at optical and electron microscope levels. We established that the fluorescent speckled staining pattern corresponds at the ultrastructural level to the labeling of perichromatin fibrils, at least part of the population of perichromatin granules, and clusters of interchromatin granules. Coiled bodies and the interchromatin granule-associated zones, a recently described subcompartment containing U1 but not U2 snRNA, were not labeled. The density of the labeling of interchromatin granule clusters exceeded by three to five times that of the surrounding extranucleolar area. These results are discussed in relation to the role of perichromatin fibrils in splicing of pre-mRNA and to the possible involvement of interchromatin granules in the assembly of mature spliceosomes as well as in sorting and/or coordination of RNA molecules to be transported to the cytoplasm.


Assessment of Pansharpening Methods Applied to WorldView-2 Imagery Fusion.
Since WorldView-2 (WV-2) images are widely used in various fields, there is a high demand for the use of high-quality pansharpened WV-2 images for different application purposes. With respect to the novelty of the WV-2 multispectral (MS) and panchromatic (PAN) bands, the performances of eight state-of-art pan-sharpening methods for WV-2 imagery including six datasets from three WV-2 scenes were assessed in this study using both quality indices and information indices, along with visual inspection. The normalized difference vegetation index, normalized difference water index, and morphological building index, which are widely used in applications related to land cover classification, the extraction of vegetation areas, buildings, and water bodies, were employed in this work to evaluate the performance of different pansharpening methods in terms of information presentation ability. The experimental results show that the Haze- and Ratio-based, adaptive Gram-Schmidt, Generalized Laplacian pyramids (GLP) methods using enhanced spectral distortion minimal model and enhanced context-based decision model methods are good choices for producing fused WV-2 images used for image interpretation and the extraction of urban buildings. The two GLP-based methods are better choices than the other methods, if the fused images will be used for applications related to vegetation and water-bodies.


Numerical evaluation of the Luneburg integral and ray tracing.
The Luneburg integral has many applications in optics and optoelectronics, among which is determination of the refractive-index profile of a Luneburg lens with a full or nonfull aperture. Consequently, computationally efficient and accurate methods for evaluating this integral represent an important challenge. An alternative approach to numerical evaluation of the Luneburg integral that is five times faster than existing methods is described. Several improvements in the ray-tracing procedure in gradient-index media are also presented. A combination of these methods increases the speed of ray tracing through the generalized Luneburg lens by as many as 2 orders of magnitude compared with earlier algorithms. The precision of our method can be easily controlled.


Randomised clinical trial: faecal microbiota transplantation by colonoscopy plus vancomycin for the treatment of severe refractory Clostridium difficile infection-single versus multiple infusions.
Faecal microbiota transplantation (FMT) is a highly effective treatment against recurrent Clostridium difficile infection. Far less evidence exists on the efficacy of FMT in treating severe Clostridium difficile infection refractory to antibiotics. To compare the efficacy of two FMT-based protocols associated with vancomycin in curing subjects with severe Clostridium difficile infection refractory to antibiotics. Subjects with severe Clostridium difficile infection refractory to antibiotics were randomly assigned to one of the two following treatment arms: (1) FMT-S, including a single faecal infusion via colonoscopy followed by a 14-day vancomycin course, (2) FMT-M, including multiple faecal infusions plus a 14-day vancomycin course. In the FMT-M group, all subjects received at least two infusions, while those with pseudomembranous colitis underwent further infusions until the disappearance of pseudomembranes. The primary outcome was the cure of refractory severe Clostridium difficile infection. Fifty six subjects, 28 in each treatment arm, were enrolled. Twenty one patients in the FMT-S group and 28 patients in the FMT-M group were cured (75% vs 100%, respectively, both in per protocol and intention-to-treat analyses; P = 0.01). No serious adverse events associated with any of the two treatment protocols were observed. A pseudomembrane-driven FMT protocol consisting of multiple faecal infusions and concomitant vancomycin was significantly more effective than a single faecal transplant followed by vancomycin in curing severe Clostridium difficile infection refractory to antibiotics. Clinical-Trials.gov registration number: NCT03427229.


Unexpected relationship between tympanometry and mortality in children with nontraumatic coma.
We sought to further examine the relationship between tympanometry and mortality after noting an unexpected association on assessment of baseline data of a study whose primary aim was to investigate the utility of noninvasive tympanic membrane displacement measurement for monitoring intracranial pressure in childhood coma. We recruited children who presented with acute nontraumatic coma to the high-dependency unit of Kilifi District Hospital on the rural coast of Kenya. We excluded children with sickle cell disease, epilepsy, and neurodevelopmental delay. We performed tympanometry on the right ear before tympanic membrane displacement analyzer measurements. All children were managed according to standard World Health Organization guidelines. We recruited 72 children with a median age of 3.2 years (interquartile range [IQR]: 2.0-4.3 years); 31 (43%) were female. Thirty-eight (53%) had cerebral malaria, 8 (11%) acute bacterial meningitis, 4 (6%) sepsis, and 22 (30%) encephalopathy of unknown etiology. Twenty (28%) children died. Tympanometry was normal in 25 (35%) children. Adjusting for diagnosis and clinical features of increased intracranial pressure, both associated with death on univariable analysis, children with abnormal tympanometry had greater odds of dying than did those with normal tympanometry (adjusted odds ratio: 17.0; 95% confidence interval: 1.9-152.4; P = .01). Children who died had a lower compliance (0.29 mL; IQR: 0.09-0.33 mL) compared with those who survived (0.48 mL; IQR: 0.29-0.70 mL) (P < .01). Abnormal tympanometry appears to be significantly associated with death in children with acute nontraumatic coma. This finding needs to be explored further through a prospective study that incorporates imaging and intensive physiologic monitoring.


[The treatment of skin ulcers in patients with systemic sclerosis].
Systemic Sclerosis (Ssc) is a complex disease of the connective tissue, characterized by progressive thickening and fibrosis of the skin and the internal organs and by diffused damage of the microvascular system. The fibrosis ones of the skin associated to the characteristic vascular alterations lead to the genesis of ulcers, more or less extended, often multiple, peripheral localization, chronic course, painful, able to influence patient's quality of life. Indeed, immunity reactivity, the thinning and the loss of elasticity of the skin, the peripheral neurological damage and the eventual drug assumption that can reduce regenerative/reparative abilities, can easily make an ulcer chronic and become infected complicating still more the patient disease, rendering more difficult the cure often, ulcer evolves to gangrene, and in some cases, in amputation too. For all these reasons, we have begun to study ulcers therapy (local and systemic), considering this activity it leave integrating of the charitable distance of the sclerodermic patient, putting to point on strategy both diagnostic and therapeutic, but above all with the primary scope, if possible, is to prevent ulcers, in contrary case, to alleviate the pain and to render the quality of the life of the patient better.


Differentiation-promoting activity of pomegranate (Punica granatum) fruit extracts in HL-60 human promyelocytic leukemia cells.
Differentiation refers to the ability of cancer cells to revert to their normal counterparts, and its induction represents an important noncytotoxic therapy for leukemia, and also breast, prostate, and other solid malignancies. Flavonoids are a group of differentiation-inducing chemicals with a potentially lower toxicology profile than retinoids. Flavonoid-rich polyphenol fractions from the pomegranate (Punica granatum) fruit exert anti-proliferative, anti-invasive, anti-eicosanoid, and pro-apoptotic actions in breast and prostate cancer cells and anti-angiogenic activities in vitro and in vivo. Here we tested flavonoid-rich fractions from fresh (J) and fermented (W) pomegranate juice and from an aqueous extraction of pomegranate pericarps (P) as potential differentiation-promoting agents of human HL-60 promyelocytic leukemia cells. Four assays were used to assess differentiation: nitro blue tetrazolium reducing activity, nonspecific esterase activity, specific esterase activity, and phagocytic activity. In addition, the effect of these extracts on HL-60 proliferation was evaluated. Extracts W and P were strong promoters of differentiation in all settings, with extract J showing only a relatively mild differentiation-promoting effect. The extracts had proportional inhibitory effects on HL-60 cell proliferation. The results highlight an important, previously unknown, mechanism of the cancer preventive and suppressive potential of pomegranate fermented juice and pericarp extracts.


Inflammatory IL-15 is required for optimal memory T cell responses.
Due to their ability to rapidly proliferate and produce effector cytokines, memory CD8+ T cells increase protection following reexposure to a pathogen. However, low inflammatory immunizations do not provide memory CD8+ T cells with a proliferation advantage over naive CD8+ T cells, suggesting that cell-extrinsic factors enhance memory CD8+ T cell proliferation in vivo. Herein, we demonstrate that inflammatory signals are critical for the rapid proliferation of memory CD8+ T cells following infection. Using murine models of viral infection and antigen exposure, we found that type I IFN-driven expression of IL-15 in response to viral infection prepares memory CD8+ T cells for rapid division independently of antigen reexposure by transiently inducing cell-cycle progression via a pathway dependent on mTOR complex-1 (mTORC1). Moreover, exposure to IL-15 allowed more rapid division of memory CD8+ T cells following antigen encounter and enhanced their protective capacity against viral infection. Together, these data reveal that inflammatory IL-15 promotes optimal responses by memory CD8+ T cells.


Ly-6C is a marker of memory CD8+ T cells.
This study examined long-term phenotypic and functional effects of TCR ligation in vivo. Flow cytometric analysis of T cells from mice treated with anti-CD3 revealed an increase in CD44 expression in both the CD4+ and CD8+ populations. The phenotypic changes were a result of TCR engagement, because treatment with staphylococcal enterotoxin B (SEB) resulted in a preferential increase in CD44 expression on the SEB-reactive V beta 8 T cells. In addition, the percentage of cells expressing Ly-6C increased among the CD8+ subset after anti-CD3 treatment and in the V beta 8+ CD8+ subset after treatment with SEB. Finally, the TCR transgenic (Tg) mouse strain 2C was used to confirm that the phenotypic changes can be induced by exposure to a physiologic ligand (H-2Ld). Before treatment, nearly all of the Tg+CD8+ cells were CD44low/Ly-6C-. Tg+ peritoneal exudate T cells isolated from mice challenged with P815 cells (H-2Ld) up-regulated Ly-6C and secreted higher levels of IFN-gamma on a per Tg+ CD8+ T cell basis after treatment. Taken together, these data indicate that in vivo TCR/CD3 engagement results in phenotypic and functional changes in T cells. Furthermore, Ly-6C expression correlates with an increase in IFN-gamma production after antigenic stimulation of CD8+ T cells, suggesting that it is a "memory" marker that correlates with Ag-specific functional changes in CD8+ T cells.


Observing the Transformation of Bodily Self-consciousness in the Squeeze-machine Experiment.
How is our bodily self-consciousness acquired, and how does it affect cognition? To investigate this, we conducted an experiment using the squeeze machine, a device that provides pressure along the length of the user's whole body. The squeeze machine is used to help autistic people relax. The inventor of the machine, Dr. Temple Grandin says that the squeeze machine, beyond bringing her relaxation, enables her to feel empathy for others. This claim is of considerable interest and raises the following two points; first, the problem of empathy in autism is an important issue and Squeeze Machine could be effective. Second, it suggests that the physical action of Squeeze Machine could provide an insight of mind-body problem to us. Here, we hypothesize that the squeeze machine focuses conscious to direct the bodily self, transforming bodily self-consciousness itself. Such intentionality could bring empathy to others. In this study, we tested whether bodily self-consciousness would be transformed through the squeeze-machine experience. In the first part of the protocol, we simplified the original design of Grandin's squeeze machine but ensured that it retained its relaxing effect. In the second part of the protocol, we adopted a preestablished method of peri-personal space (PPS) measurement to estimate changes in extended bodily space. The results showed that the boundaries of PPS that appeared in the control experiment disappeared during the use of the squeeze machine. Indeed, collected subjective reports suggest that bodily self-consciousness continually drifted between the point of action of the external force (squeeze-machine pressure) and of an internal force (balance among body parts that are usually not consciously controlled), leading to the abandonment of the immobilization of individual PPS.


Interactions between Hyaluronan and Its Receptors (CD44, RHAMM) Regulate the Activities of Inflammation and Cancer.
The glycosaminoglycan hyaluronan (HA), a major component of extracellular matrices, and cell surface receptors of HA have been proposed to have pivotal roles in cell proliferation, migration, and invasion, which are necessary for inflammation and cancer progression. CD44 and receptor for HA-mediated motility (RHAMM) are the two main HA-receptors whose biological functions in human and murine inflammations and tumor cells have been investigated comprehensively. HA was initially considered to be only an inert component of connective tissues, but is now known as a "dynamic" molecule with a constant turnover in many tissues through rapid metabolism that involves HA molecules of various sizes: high molecular weight HA (HMW HA), low molecular weight HA, and oligosaccharides. The intracellular signaling pathways initiated by HA interactions with CD44 and RHAMM that lead to inflammatory and tumorigenic responses are complex. Interestingly, these molecules have dual functions in inflammations and tumorigenesis. For example, the presence of CD44 is involved in initiation of arthritis, while the absence of CD44 by genetic deletion in an arthritis mouse model increases rather than decreases disease severity. Similar dual functions of CD44 exist in initiation and progression of cancer. RHAMM overexpression is most commonly linked to cancer progression, whereas loss of RHAMM is associated with malignant peripheral nerve sheath tumor growth. HA may similarly perform dual functions. An abundance of HMW HA can promote malignant cell proliferation and development of cancer, whereas antagonists to HA-CD44 signaling inhibit tumor cell growth in vitro and in vivo by interfering with HMW HA-CD44 interaction. This review describes the roles of HA interactions with CD44 and RHAMM in inflammatory responses and tumor development/progression, and how therapeutic strategies that block these key inflammatory/tumorigenic processes may be developed in rodent and human diseases.


Collaboration between primary care and psychiatric services: does it help family physicians?
To compare family physicians' reports of their experiences managing patients with psychiatric disorders in settings with and without access to collaborative mental health services. Survey using a questionnaire adapted from a similar study in Australia. Family physicians were asked about their knowledge, skills, and degree of comfort in managing the following psychiatric disorders derived from the primary care version of the 10th edition of the International Classification of Diseases: psychosis, depression, anxiety, childhood disorders, and stress-related disorders. We also compared the 2 groups of physicians regarding their satisfaction with mental health services in general. The Capital District Health Authority (CDHA) in Nova Scotia. All family physicians practising in the CDHA. Self-reported knowledge, skills, and degree of comfort in managing psychiatric problems; satisfaction with mental health services, adjusted for family physicians' demographics; and stated interest in mental health. We received 101 responses (37 from physicians with access to collaborative care and 64 from physicians without access) from 7 communities in the CDHA. Family physicians who had access to collaborative care reported significantly greater knowledge in the areas of psychosis, alcohol or substance use, and childhood behavioural problems; and better skills in managing psychosis, alcohol or substance use, childhood depression or anxiety, childhood behavioural disorders, and relationship problems. Their comfort levels in managing relationship problems and childhood behavioural disorders were also significantly higher. Family physicians with access to collaborative care were significantly more satisfied with mental health services, over and above shared care. All these differences remained significant after controlling for sex, level of interest in mental health, and years in practice. Family physicians with access to collaborative care reported greater knowledge, better skills, and more comfort in managing psychiatric disorders and greater satisfaction with mental health services. Further work is needed to establish why this is so and to determine any effect on patient outcomes, such as symptoms, quality of life, and psychosocial functioning.


Structure of a type IV secretion system.
Bacterial type IV secretion systems translocate virulence factors into eukaryotic cells, distribute genetic material between bacteria and have shown potential as a tool for the genetic modification of human cells. Given the complex choreography of the substrate through the secretion apparatus, the molecular mechanism of the type IV secretion system has proved difficult to dissect in the absence of structural data for the entire machinery. Here we use electron microscopy to reconstruct the type IV secretion system encoded by the Escherichia coli R388 conjugative plasmid. We show that eight proteins assemble in an intricate stoichiometric relationship to form an approximately 3 megadalton nanomachine that spans the entire cell envelope. The structure comprises an outer membrane-associated core complex connected by a central stalk to a substantial inner membrane complex that is dominated by a battery of 12 VirB4 ATPase subunits organized as side-by-side hexameric barrels. Our results show a secretion system with markedly different architecture, and consequently mechanism, to other known bacterial secretion systems.


Monoclonal antibodies specific for peptide epitopes of the epidermal growth factor receptor's extracellular domain.
The ErbB tyrosine kinase receptor family plays an important role in normal cellular growth and differentiation. In addition, ErbB receptor family members are commonly amplified and overexpressed in various human neoplasms and tumor-derived cell lines, where it is believed that increased signalling as a result of receptor overexpression may play an important role in oncogenesis. Consequently, ErbB receptor family members are being investigated rigorously as potential biomarkers of cancer and as therapeutic targets in malignant tissues. Numerous studies now demonstrate the existence of "soluble" ErbB (sErbB) analogs in normal and cancerous tissues. These sErbB proteins embody the extracellular domain (ECD) of the receptor only; they are generated by either proteolytic cleavage or from truncated, alternatively spliced mRNA transcripts. Recently, we have identified an alternate transcript of the human c-erbB1 (Epidermal Growth Factor Receptor) proto-oncogene from placenta that encodes a sErbB1 protein of 60-kDa. This protein, p60 sErbB1, is glycosylated and secreted when expressed in transfected tissue culture cells in vitro. Although "soluble" receptor analogs may play important physiological roles in intercellular communication, tissue morphogenesis, tissue regeneration and repair, and embryogenesis by inhibiting or stimulating specific mitogenic and pattern forming signals, their mechanism of action has not been thoroughly elucidated. To further characterize sErbB1 expression in human tissues and cell lines and to better understand their role in carcinogenesis and normal development, we have generated monoclonal antibodies (MAbs) toward specific peptide epitopes of ErbB1 extracellular subdomains III and IV. These antibody reagents are described here and should be useful experimental, preparative, analytical, diagnostic, and therapeutic reagents for the study of sErbB1 molecules in normal development and cancer.


The trafficking protein SYP121 of Arabidopsis connects programmed stomatal closure and K⁺ channel activity with vegetative growth.
The vesicle-trafficking protein SYP121 (SYR1/PEN1) was originally identified in association with ion channel control at the plasma membrane of stomatal guard cells, although stomata of the Arabidopsis syp121 loss-of-function mutant close normally in ABA and high Ca²⁺. We have now uncovered a set of stomatal phenotypes in the syp121 mutant that reduce CO₂ assimilation, slow vegetative growth and increase water use efficiency in the whole plant, conditional upon high light intensities and low relative humidity. Stomatal opening and the rise in stomatal transpiration of the mutant was delayed in the light and following Ca²⁺-evoked closure, consistent with a constitutive form of so-called programmed stomatal closure. Delayed reopening was observed in the syp121, but not in the syp122 mutant lacking the homologous gene product; the delay was rescued by complementation with wild-type SYP121 and was phenocopied in wild-type plants in the presence of the vesicle-trafficking inhibitor Brefeldin A. K⁺ channel current that normally mediates K⁺ uptake for stomatal opening was suppressed in the syp121 mutant and, following closure, its recovery was slowed compared to guard cells of wild-type plants. Evoked stomatal closure was accompanied by internalisation of GFP-tagged KAT1 K⁺ channels in both wild-type and syp121 mutant guard cells, but their subsequently recycling was slowed in the mutant. Our findings indicate that SYP121 facilitates stomatal reopening and they suggest that K⁺ channel traffic and recycling to the plasma membrane underpins the stress memory phenomenon of programmed closure in stomata. Additionally, they underline the significance of vesicle traffic for whole-plant water use and biomass production, tying SYP121 function to guard cell membrane transport and stomatal control.


Thermal decomposition of ammonium hexachloroosmate.
Structural changes of (NH4)2[OsCl6] occurring during thermal decomposition in a reduction atmosphere have been studied in situ using combined energy-dispersive X-ray absorption spectroscopy (ED-XAFS) and powder X-ray diffraction (PXRD). According to PXRD, (NH4)2[OsCl6] transforms directly to metallic Os without the formation of any crystalline intermediates but through a plateau where no reactions occur. XANES and EXAFS data by means of Multivariate Curve Resolution (MCR) analysis show that thermal decomposition occurs with the formation of an amorphous intermediate {OsCl4}x with a possible polymeric structure. Being revealed for the first time the intermediate was subjected to determine the local atomic structure around osmium. The thermal decomposition of hexachloroosmate is much more complex and occurs within a minimum two-step process, which has never been observed before.


Effector CD8+ T-cell Engraftment and Antitumor Immunity in Lymphodepleted Hosts Is IL7Rα Dependent.
Adoptive cellular therapy, in which activated tumor-reactive T cells are transferred into lymphodepleted recipients, is a promising cancer treatment option. Activation of T cells decreases IL7 responsiveness; therefore, IL15 is generally considered the main driver of effector T-cell responses in this setting. However, we found in lymphodepleted mice that CD8(+) T cells activated with IL12 showed enhanced engraftment that was initially dependent on host IL7, but not IL15. Mechanistically, enhanced IL7 responsiveness was conferred by elevated IL7Rα expression, which was critical for antitumor immunity. Elevated IL7Rα expression was achievable without IL12, as polyclonal CD8(+) T cells activated with high T-cell receptor (TCR) stimulation depended on T-cell IL7Rα expression and host IL7 for maximal engraftment. Finally, IL12 conditioning during the activation of human CD8(+) T cells, including TCR-modified T cells generated using a clinically relevant protocol, led to enhanced IL7Rα expression. Our results demonstrate the importance of the donor IL7Rα/host IL7 axis for effector CD8(+) T-cell engraftment and suggest novel strategies to improve adoptive cellular therapy as a cancer treatment.


Analyzing Spinal Shape Changes During Posture Training Using a Wearable Device.
Lower back pain is one of the most prevalent diseases in Western societies. A large percentage of European and American populations suffer from back pain at some point in their lives. One successful approach to address lower back pain is postural training, which can be supported by wearable devices, providing real-time feedback about the user's posture. In this work, we analyze the changes in posture induced by postural training. To this end, we compare snapshots before and after training, as measured by the Gokhale SpineTracker™. Considering pairs of before and after snapshots in different positions (standing, sitting, and bending), we introduce a feature space, that allows for unsupervised clustering. We show that resulting clusters represent certain groups of postural changes, which are meaningful to professional posture trainers.


Obesity-related metabolite profiles of black women spanning the epidemiologic transition.
In developed countries, specific metabolites have been associated with obesity and metabolic diseases, e.g. type 2 diabetes. It is unknown whether a similar profile persists across populations of African-origin, at increased risk for obesity and related diseases. In a cross-sectional study of normal-weight and obese black women (33.3 ± 6.3 years) from the US (N = 69, 65 % obese), South Africa (SA, N = 97, 49 % obese) and Ghana (N = 82, 33 % obese) serum metabolite profiles were characterized via gas chromatography-time of flight/mass spectrometry. In US and SA women, BMI correlated with branched-chain and aromatic amino acids, as well as dopamine and aminoadipic acid. The relationship between BMI and lipid metabolites differed by site; BMI correlated positively with palmitoleic acid (16:1) in the US; negatively with stearic acid (18:0) in SA, and positively with arachidonic acid (20:4) in Ghana. BMI was also positively associated with sugar-related metabolites in the US; i.e. uric acid, and mannitol, and with glucosamine, glucoronic acid and mannitol in SA. While we identified a common amino acid metabolite profile associated with obesity in black women from the US and SA, we also found site-specific obesity-related metabolites suggesting that the local environment is a key moderator of obesity.


Cell-Based Therapies for Joint Disease in Veterinary Medicine: What We Have Learned and What We Need to Know.
Biological cell-based therapies for the treatment of joint disease in veterinary patients include autologous-conditioned serum, platelet-rich plasma, and expanded or non-expanded mesenchymal stem cell products. This narrative review outlines the processing and known mechanism of action of these therapies and reviews current preclinical and clinical efficacy in joint disease in the context of the processing type and study design. The significance of variation for biological activity and consequently regulatory approval is also discussed. There is significant variation in study outcomes for canine and equine cell-based products derived from whole blood or stem cell sources such as adipose and bone marrow. Variation can be attributed to altering bio-composition due to factors including preparation technique and source. In addition, study design factors like selection of cases with early vs. late stage osteoarthritis (OA), or with intra-articular soft tissue injury, influence outcome variation. In this under-regulated field, variation raises concerns for product safety, consistency, and efficacy. Cell-based therapies used for OA meet the Food and Drug Administration's (FDA's) definition of a drug; however, researchers must consider their approach to veterinary cell-based research to meet future regulatory demands. This review explains the USA's FDA guidelines as an example pathway for cell-based therapies to demonstrate safety, effectiveness, and manufacturing consistency. An understanding of the variation in production consistency, effectiveness, and regulatory concerns is essential for practitioners and researchers to determine what products are indicated for the treatment of joint disease and tactics to improve the quality of future research.


Tissue-targeted therapy of autoimmune diabetes using dendritic cells transduced to express IL-4 in NOD mice.
A deficit in IL-4 production has been previously reported in both diabetic human patients and non-obese diabetic (NOD) mice. In addition, re-introducing IL-4 into NOD mice systemically, or as a transgene, led to a beneficial outcome in most studies. Here, we show that prediabetic, 12-week old female NOD mice have a deficit in IL-4 expression in the pancreatic lymph nodes (PLN) compared to age-matched diabetes-resistant NOD.B10 mice. By bioluminescence imaging, we demonstrated that the PLN was preferentially targeted by bone marrow-derived dendritic cells (DCs) following intravenous (IV) administration. Following IV injection of DCs transduced to express IL-4 (DC/IL-4) into 12-week old NOD mice, it was possible to significantly delay or prevent the onset of hyperglycemia. We then focused on the PLN to monitor, by microarray analysis, changes in gene expression induced by DC/IL-4 and observed a rapid normalization of the expression of many genes, that were otherwise under-expressed compared to NOD.B10 PLN. The protective effect of DC/IL-4 required both MHC and IL-4 expression by the DCs. Thus, adoptive cellular therapy, using DCs modified to express IL-4, offers an effective, tissue-targeted cellular therapy to prevent diabetes in NOD mice at an advanced stage of pre-diabetes, and may offer a safe approach to consider for treatment of high risk human pre-diabetic patients.


Is this pill an antibiotic or a painkiller? Improving the identification of oral antibiotics for better use.
In this Viewpoint, we discuss how the identification of oral antibiotics and their distinction from other commonly used medicines can be challenging for consumers, suppliers, and health-care professionals. There is a large variation in the names that people use to refer to antibiotics and these often relate to their physical appearance, although antibiotics come in many different physical presentations. We also reflect on how the physical appearance of medicine influences health care and public health by affecting communication between patients and health-care professionals, dispensing , medicine use, and the public understanding of health campaigns. Furthermore, we report expert and stakeholder consultations on improving the identification of oral antibiotics and discuss next steps towards a new identification system for antibiotics. We propose to use the physical appearance as a tool to support and nudge awareness about antibiotics and their responsible use.


Increase in p53 protein levels by presenilin 1 gene mutations and its inhibition by secretase inhibitors.
Presenilin 1 (PS1) gene mutations are the major causes of early-onset familial Alzheimer's disease and are known to increase amyloid-beta42 (Abeta42) production as well as to promote apoptosis. We have recently reported that intracellular Abeta42 activates p53 mRNA expression and promotes p53-dependent apoptosis. Here, we examined the p53 mRNA and protein levels in cells transfected with wild-type and I143T/G384A mutant PS1 genes. Although the baseline p53 mRNA levels remained unaltered, the p53 protein levels were significantly elevated in mutant PS1-transfected cells. Treatments with apoptosis-inducing agents induced significant elevation of the p53 protein but not p53 mRNA levels in mutant PS1-transfected cells. Treatment with a beta-secretase inhibitor and gamma-secretase inhibitor decreased the intracellular Abeta levels in amyloid-beta protein precursor (AbetaPP) and PS1-double transfected cells, and restrained upregulation of the p53 protein levels in the mutant PS1-transfected cells. Also, we found that proteasome activity was decreased in mutant PS1-transfected cells compared to wild-type PS1-transfected cells. Proteasome activity was further decreased in AbetaPP/PS1-double transfected cells. Taken together, p53-dependent apoptosis upregulated by the I143T/G384A mutant PS1 gene may be associated, at least in part, with intracellular Abeta and proteasome impairment.


Large-amplitude plasma wave generation with a high-intensity short-pulse beat wave.
A short-pulse laser beat wave scheme for advanced particle accelerator applications is examined. A short, intense (3-ps, >10(18)-W cm(-2)) two-frequency laser pulse is produced by use of a modified chirped-pulse amplification scheme and is shown to produce relativistic plasma waves during interactions with low-density plasmas. The generation of plasma waves was observed by measurement of forward Raman scattering. Resonance was found to occur at an electron density many times that expected, owing to ponderomotive displacement of plasma within the focal region.


Dynamic modulation of subthalamic nucleus activity facilitates adaptive behavior.
Adapting actions to changing goals and environments is central to intelligent behavior. There is evidence that the basal ganglia play a crucial role in reinforcing or adapting actions depending on their outcome. However, the corresponding electrophysiological correlates in the basal ganglia and the extent to which these causally contribute to action adaptation in humans is unclear. Here, we recorded electrophysiological activity and applied bursts of electrical stimulation to the subthalamic nucleus, a core area of the basal ganglia, in 16 patients with Parkinson's disease (PD) on medication using temporarily externalized deep brain stimulation (DBS) electrodes. Patients as well as 16 age- and gender-matched healthy participants attempted to produce forces as close as possible to a target force to collect a maximum number of points. The target force changed over trials without being explicitly shown on the screen so that participants had to infer target force based on the feedback they received after each movement. Patients and healthy participants were able to adapt their force according to the feedback they received (P < 0.001). At the neural level, decreases in subthalamic beta (13 to 30 Hz) activity reflected poorer outcomes and stronger action adaptation in 2 distinct time windows (Pcluster-corrected < 0.05). Stimulation of the subthalamic nucleus reduced beta activity and led to stronger action adaptation if applied within the time windows when subthalamic activity reflected action outcomes and adaptation (Pcluster-corrected < 0.05). The more the stimulation volume was connected to motor cortex, the stronger was this behavioral effect (Pcorrected = 0.037). These results suggest that dynamic modulation of the subthalamic nucleus and interconnected cortical areas facilitates adaptive behavior.


α8-integrins are required for hippocampal long-term potentiation but not for hippocampal-dependent learning.
Integrins are heterodimeric transmembrane cell adhesion receptors that are essential for a wide range of biological functions via cell-matrix and cell-cell interactions. Recent studies have provided evidence that some of the subunits in the integrin family are involved in synaptic and behavioral plasticity. To further understand the role of integrins in the mammalian central nervous system, we generated a postnatal forebrain and excitatory neuron-specific knockout of alpha8-integrin in the mouse. Behavioral studies showed that the mutant mice are normal in multiple hippocampal-dependent learning tasks, including a T-maze, non-match-to-place working memory task for which other integrin subunits like alpha3- and beta1-integrin are required. In contrast, mice mutant for alpha8-integrin exhibited a specific impairment of long-term potentiation (LTP) at Schaffer collateral-CA1 synapses, whereas basal synaptic transmission, paired-pulse facilitation and long-term depression (LTD) remained unaffected. Because LTP is also impaired in the absence of alpha3-integrin, our results indicate that multiple integrin molecules are required for the normal expression of LTP, and different integrins display distinct roles in behavioral and neurophysiological processes like synaptic plasticity.


Provider Beliefs and Practices About Assessing Sexual Orientation in Two Veterans Health Affairs Hospitals.
Despite known health disparities for lesbian, gay, and bisexual (LGB) individuals, research in the civilian sector has documented low rates of patient disclosure and provider assessment of this domain. Very little is known about Veterans Health Affairs (VHA) providers' care of LGB veterans, a population that has been relatively invisible until recently because of the vestiges of the Department of Defense policy of "Don't Ask, Don't Tell." This study examined the attitudes, beliefs, and clinical practices of VHA healthcare providers regarding sexual minority veterans. Physical and mental health VHA healthcare providers (n=202) from two southern VHA hospitals completed an anonymous self-report questionnaire. Measures included comfort in providing care to LGB veterans, factors affecting decisions about assessing sexual orientation with veterans, and attitudes toward sexual minority individuals. Although approximately half of VHA providers thought that sexual orientation should be routinely discussed, the providers rarely assessed this issue with their patients. Over half of providers believed that veterans would disclose their sexual orientation if it was important to them, and almost half of providers believed sexual orientation is not relevant to healthcare. Many VHA providers may be unaware of the unique health disparities experienced by LGB individuals. Culturally appropriate care cannot be provided to LGB veterans unless providers explicitly assess sexual orientation in healthcare visits. Central to this assessment is providing patients with a clear rationale for the purpose of the assessment and related documentation. Staff training is needed to address providers' beliefs and reservations about discussing sexual orientation that emerged in this investigation.


Application of mass spectrometry-based MHC immunopeptidome profiling in neoantigen identification for tumor immunotherapy.
One of the challenges for cancer vaccine and adoptive T-cell-based immunotherapy is to identify the major histocompatibility complex (MHC)-associated non-self neoantigens recognized by T cells. T cell epitope in silico prediction algorithms have been widely used for neoantigen prediction; nonetheless, this platform lacks the experimental evidence of directly identification of the presented epitopes on cell surface. Currently, mass spectrometry (MS)-based proteomics is an advanced analytical technology for large-scale peptide sequencing, which has become a powerful tool for directly profiling the immunopeptidome presented by MHC molecules. Integrating with next-generation sequencing, proteogenomic analysis provides the "gold standard" for neoantigen identification at protein level. This method discovers the tumor-specific neoantigens derived from somatic mutations, proteasome splicing, noncoding RNA, and post-translational modified antigens. Herein, we review basis of antigen processing and presentation, tumor antigen classification, existing approaches for neoantigen discovery, quantitative proteomics, epitope prediction programs, and advantages and drawbacks of proteomics workflow for MHC immunopeptidome profiling. Furthermore, we summarize 40 recently published reports addressing the fundamental theory, breakthrough and most advanced updates for the mass spectrometry-based neoantigen discovery for cancer immunotherapy.


Automatic determination of diltiazem and desacetyldiltiazem in human plasma using liquid-solid extraction on disposable cartridges coupled to HPLC--Part II: Optimization of liquid-solid extraction.
An automatic liquid-solid extraction (LSE) procedure to be coupled to HPLC for the determination of diltiazem and desacetyldiltiazem in plasma has been developed. The LSE operations are performed on disposable extraction cartridges (DECs) by means of a sample processor equipped with a robotic arm holding a needle through which the different liquids are dispensed. The operating parameters of LSE have been optimized with respect to recovery, detectability and reproducibility by using, whenever possible, aqueous solutions of the analytes. Different kinds of DECs have been tested. For the compounds studied, DECs filled with 50 mg of cyanopropyl silica have been selected. The influence of the pH of the buffer used in the washing step has been studied, leading finally to the selection of the same phosphate buffer (pH 7.4) as in the HPLC mobile phase. The minimum volume of methanol which still gives a nearly complete elution of the analytes from the extraction cartridges has been determined. Under these conditions, a high sensitivity can be obtained without an evaporation step. Moreover, the volume of buffer to be added to the methanolic eluate before injection into the HPLC system has been optimized in such a way that a focusing effect is obtained at the top of the analytical column while the dilution of the extract is minimized.


The CopenHeartSF trial--comprehensive sexual rehabilitation programme for male patients with implantable cardioverter defibrillator or ischaemic heart disease and impaired sexual function: protocol of a randomised clinical trial.
Sexuality is an important part of people's physical and mental health. Patients with heart disease often suffer from sexual dysfunction. Sexual dysfunction has a negative impact on quality of life and well-being in persons with heart disease, and sexual dysfunction is associated with anxiety and depression. Treatment and care possibilities seem to be lacking. Studies indicate that non-pharmacological interventions such as exercise training and psychoeducation possess the potential of reducing sexual dysfunction in patients with heart disease. The CopenHeartSF trial will investigate the effect of a comprehensive sexual rehabilitation programme versus usual care. CopenHeartSF is an investigator-initiated randomised clinical superiority trial with blinded outcome assessment, with 1:1 central randomisation to sexual rehabilitation plus usual care versus usual care alone. Based on sample size calculations, 154 male patients with impaired sexual function due to implantable cardioverter defibrillator or ischaemic heart disease will be included from two university hospitals in Denmark. All patients receive usual care and patients allocated to the experimental intervention group follow a 12-week sexual rehabilitation programme consisting of an individualised exercise programme and psychoeducative consultation with a specially trained nurse. The primary outcome is sexual function measured by the International Index of Erectile Function. The secondary outcome measure is psychosocial adjustment to illness by the Psychosocial Adjustment to Illness Scale, sexual domain. A number of explorative analyses will also be conducted. CopenHeartSF is approved by the regional ethics committee (no H-4-2012-168) and the Danish Data Protection Agency (no 2007-58-0015) and is performed in accordance with good clinical practice and the Declaration of Helsinki in its latest form. Clinicaltrials.gov identifier: NCT01796353.


Comparison of oral aspirin versus topical applied methyl salicylate for platelet inhibition.
Oral acetylsalicylic acid (aspirin) is the primary antiplatelet therapy in the treatment of acute myocardial infarction and acute coronary syndrome. Methyl salicylate (MS; oil of wintergreen) is compounded into many over-the-counter antiinflammatory muscle preparations and has been shown to inhibit platelet aggregation locally and to be absorbed systemically. To assess the ability of topically applied MS to inhibit systemic platelet aggregation for patients who are unable to tolerate oral drug therapy. A randomized, prospective, blinded, crossover study was conducted in 9 healthy men, aged 30-46 years. All subjects ingested 162 mg of aspirin or applied 5 g of 30% MS preparation to their anterior thighs. There was a minimum 2-week washout period between study arms. Blood and urine were collected at baseline and at 6 hours. An aggregometer measured platelet aggregation over time against 5 standard concentrations of epinephrine, and a mean area under the curve (AUC) was calculated. Urinary metabolites of thromboxane B(2) were measured by a standard enzyme immunoassay. Differences in and between groups at baseline and 6 hours were tested by the Wilcoxon signed-rank test. Baseline platelet aggregation did not differ significantly between the 2 arms of the study (median AUC [% aggregation(*)min]; binominal confidence intervals): aspirin 183; 139 to 292 versus MS 197; 118 to 445 (p = 0.51). Both aspirin and MS produced statistically significant platelet inhibition; aspirin decreased the AUC from 183; 139 to 292 to 85; 48 to 128 (p = 0.008) and MS decreased the AUC from 197; 118 to 445 to 112; 88 to 306 (p = 0.011). No significant difference was detected between baseline and 6-hour thromboxane levels for either aspirin (p = 0.779) or MS (p = 0.327). Topical MS and oral aspirin both significantly decrease platelet aggregation in healthy human volunteers.


Integration of the Intrinsic Surgical Risk With Patient Comorbidities and Severity of Congenital Cardiac Disease Does Not Improve Risk Stratification in Children Undergoing Noncardiac Surgery.
The objective of this study is to estimate the surgical risk of noncardiac procedures on the incidence of 30-day mortality in children with congenital heart disease. Children with congenital heart disease undergoing noncardiac surgery from 2012 to 2016 and included in the American College of Surgeons National Surgical Quality Improvement Program (ACS NSQIP) Pediatric database were included in the derivation cohort, while the 2017 database was used as a validation cohort. Intrinsic surgical risk quartiles were built utilizing 30-day mortality rates for each Current Procedural Terminology code and relative value units to create 2 groups defined as low surgical risk (quartiles 1-3) and high surgical risk procedures (quartile 4). We used multivariable logistic regression to determine the predictors for 30-day mortality including patient comorbidities and intrinsic surgical risk. A partially external validation of the model was performed using the 2017 version of the database. We included 37,658 children with congenital heart disease undergoing noncardiac surgery with an incidence of overall 30-day mortality of 1.7% in the derivation cohort and 1.5% in the validation cohort (n = 13,129). Intrinsic surgical risk of procedures represented by Current Procedural Terminology procedural codes and relative value units risk quartiles was significantly associated with 30-day mortality (unadjusted P < .001). Predicted probability of 30-day mortality ranges from 0.2% (95% confidence interval [CI], 0.2-0.2) with no comorbidities to 39.6% (95% CI, 23.2-56.0) when all comorbidities were present among high surgical risk procedures and from 0.3% (95% CI, 0.3-0.3) to 54.8% (95% CI, 39.4-70.1) among low surgical risk procedures. An excellent discrimination was reported for the multivariable model with area under the curve (AUC) of 0.86 (95% CI, 0.85-0.88). High surgical risk was not associated with increased odds of 30-day mortality after adjustment for all other predictors (adjusted odds ratio [OR]: 0.75, 95% CI, 0.62-0.91). We also estimated the discriminative ability of a model that does not include the surgical risk (0.86 [95% CI, 0.84-0.88], with P value for the direct comparison of the AUC of the 2 models = 0.831). The multivariable model obtained from an external validation cohort reported an optimism corrected AUC of 0.88 (95% CI, 0.85-0.91). Our study demonstrates that integration of intrinsic surgical risk to comorbidities and severity of cardiac disease does not improve prediction of 30-day mortality in children undergoing noncardiac surgery. In children with congenital heart disease, patient comorbidities, and severity of the cardiac lesion are the predominant predictors of 30-day mortality.


Unsupervised clustering of temporal patterns in high-dimensional neuronal ensembles using a novel dissimilarity measure.
Temporally ordered multi-neuron patterns likely encode information in the brain. We introduce an unsupervised method, SPOTDisClust (Spike Pattern Optimal Transport Dissimilarity Clustering), for their detection from high-dimensional neural ensembles. SPOTDisClust measures similarity between two ensemble spike patterns by determining the minimum transport cost of transforming their corresponding normalized cross-correlation matrices into each other (SPOTDis). Then, it performs density-based clustering based on the resulting inter-pattern dissimilarity matrix. SPOTDisClust does not require binning and can detect complex patterns (beyond sequential activation) even when high levels of out-of-pattern "noise" spiking are present. Our method handles efficiently the additional information from increasingly large neuronal ensembles and can detect a number of patterns that far exceeds the number of recorded neurons. In an application to neural ensemble data from macaque monkey V1 cortex, SPOTDisClust can identify different moving stimulus directions on the sole basis of temporal spiking patterns.


Standardization of the twinning rate.
There is a great interest in comparing twinning rates. These comparisons can be performed between different time periods for a specific population, between different regions within the same country, and between different populations. However, there are several factors (maternal age, parity, urbanization, etc.) that influence the twinning rate. The most dominant one is maternal age, and because the age distribution of the mother varies, it is necessary to standardize the data to make these comparisons. If we want to compare the twinning rates in different countries, we have to face the problem that the composition of the data from different countries may differ to a great extent. The applicable method is determined by the data of the lowest quality. Often the available data do not allow the traditional (direct and/or indirect) methods of standardization. Under such circumstances other methods have to be used. Earlier, Fellman and Eriksson (1987) proposed and successfully applied a new method. In this article we discuss the standardization problem in more detail. We suggest different methods and apply them to different data on twinning from Australia, Finland, and Baden-Württemberg (West Germany). The new standardization methods give standardized twinning rates similar to the rates obtained by traditional methods. It is noted that, irrespective of standardization method, changes in maternal age alone cannot explain temporal or regional variations in the twinning rate. Other factors that may raise or lower the twinning rate are decreasing parity, sociodemographic changes with increased communication, which causes the breakup of isolates, and deteriorating physical condition of mothers as a result of increased industrialization and urbanization.


Observations of severe and lethal coalitionary attacks in wild mountain gorillas.
In humans and chimpanzees, most intraspecific killing occurs during coalitionary intergroup conflict. In the closely related genus Gorilla, such behavior has not been described. We report three cases of multi-male, multi-female wild mountain gorilla (G. beringei) groups attacking extra-group males. The behavior was strikingly similar to reports in chimpanzees, but was never observed in gorillas until after a demographic transition left ~25% of the population living in large social groups with multiple (3+) males. Resource competition is generally considered a motivator of great apes' (including humans) violent intergroup conflict, but mountain gorillas are non-territorial herbivores with low feeding competition. While adult male gorillas have a defensible resource (i.e. females) and nursing/pregnant females are likely motivated to drive off potentially infanticidal intruders, the participation of others (e.g. juveniles, sub-adults, cycling females) is harder to explain. We speculate that the potential for severe group disruption when current alpha males are severely injured or killed may provide sufficient motivation when the costs to participants are low. These observations suggest that the gorilla population's recent increase in multi-male groups facilitated the emergence of such behavior, and indicates social structure is a key predictor of coalitionary aggression even in the absence of meaningful resource stress.


FLUORIDE: A REVIEW OF USE AND EFFECTS ON HEALTH.
Appropriate oral health care is fundamental for any individual's health. Dental caries is still one of the major public health problems. The most effective way of caries prevention is the use of fluoride. The aim of our research was to review the literature about fluoride toxicity and to inform physicians, dentists and public health specialists whether fluoride use is expedient and safe. Data we used in our review were systematically searched and collected from web pages and documents published from different international institutions. Fluoride occurs naturally in our environment but we consume it in small amounts. Exposure can occur through dietary intake, respiration and fluoride supplements. The most important factor for fluoride presence in alimentation is fluoridated water. Methods, which led to greater fluoride exposure and lowered caries prevalence, are considered to be one of the greatest accomplishments in the 20th century`s public dental health. During pregnancy, the placenta acts as a barrier. The fluoride, therefore, crosses the placenta in low concentrations. Fluoride can be transmitted through the plasma into the mother's milk; however, the concentration is low. The most important action of fluoride is topical, when it is present in the saliva in the appropriate concentration. The most important effect of fluoride on caries incidence is through its role in the process of remineralization and demineralization of tooth enamel. Acute toxicity can occur after ingesting one or more doses of fluoride over a short time period which then leads to poisoning. Today, poisoning is mainly due to unsupervised ingestion of products for dental and oral hygiene and over-fluoridated water. Even though fluoride can be toxic in extremely high concentrations, it`s topical use is safe. The European Academy of Paediatric Dentistry (EAPD) recommends a preventive topical use of fluoride supplements because of their cariostatic effect.


Parallel gene analysis with allele-specific padlock probes and tag microarrays.
Parallel, highly specific analysis methods are required to take advantage of the extensive information about DNA sequence variation and of expressed sequences. We present a scalable laboratory technique suitable to analyze numerous target sequences in multiplexed assays. Sets of padlock probes were applied to analyze single nucleotide variation directly in total genomic DNA or cDNA for parallel genotyping or gene expression analysis. All reacted probes were then co-amplified and identified by hybridization to a standard tag oligonucleotide array. The technique was illustrated by analyzing normal and pathogenic variation within the Wilson disease-related ATP7B gene, both at the level of DNA and RNA, using allele-specific padlock probes.


Theory of multiplate resonant reflectors.
The standard equations of thin film optics are applied to multiplate resonant reflectors of the kind used with ruby lasers. The numerical results show considerable disagreement with predictions based on approximate theory. A design is proposed that gives high reflectivity and is insensitive to thickness errors.


Replication of an Egfr-wing shape association in a wild-caught cohort of Drosophila melanogaster.
Linkage disequilibrium mapping has been used extensively in medical and evolutionary genetics to map causal polymorphisms within genes associated with disease status or phenotypic variation for a trait. However, the initial findings of most nonhuman studies have not been replicated in subsequent studies, due in part to false positives, as well as additional factors that can render true positives unreplicable. These factors may be more severe when the initial study is performed using an experimental population of organisms reared under controlled lab conditions. We demonstrate that despite considerable phenotypic differences for wing shape between a lab-reared experimental population and a wild-caught cohort of Drosophila melanogaster, an association between a putative regulatory polymorphism in Egfr and wing shape can be replicated. These results are discussed both within the framework of future association-mapping studies and within the context of the evolutionary dynamics of alleles in populations.


Teaching professionalism: a survey of physical therapy educators.
This study examined the opinions of physical therapy faculty relative to teaching and fostering professionalism in entry-level physical therapy education. A paper-and-pencil survey was mailed to a random sample of 318 physical therapy educators across the United States. Of the 318 surveys mailed, 166 participants (response rate, 52%) completed and returned the survey. Descriptive analysis revealed that 98% of the physical therapy educators view professionalism as an important component of a physical therapy curriculum. Eighty-nine percent of the respondents expressed concern about the professional behaviors of one or more of their entry-level students; however, based on the frequency of negative behaviors, these concerns appear to be isolated to a small percentage of the students. The most frequent negative behaviors observed among entry-level physical therapy students included tardiness and lack of personal responsibility. Respondents identified clinical reasoning, integrity, and honesty as the three most important professional skills for a physical therapist. The three most common teaching methods used to foster professionalism included generic abilities, small group discussion, and related reading assignments. Professional socialization is clearly a concern among physical therapy faculty for a few entry-level physical therapy students; however, based on the results of this investigation, it appears that most entry-level physical therapy students are making a smooth transition to professional socialization. The information obtained from this investigation may be useful to allied health professionals and educators to help promote professionalism among entry-level students enrolled in professional programs.


Leg Amputations Among Texans Remote From Experienced Surgical Care.
Surgical outcomes may differ between low-volume and experienced hospitals. We sought to identify characteristics of remote patients-those living more than 50 miles from an experienced center-who underwent leg amputations for peripheral artery disease (PAD) and foot complications at low-volume and experienced hospitals and identify regions of Texas where such patients live. Publicly available Texas hospitalization data from 2004 through 2009 were used to identify patients with PAD who underwent leg amputation for foot complications, including foot ulcers, foot infections, and gangrene. Geocoding was used to further identify a subset of remote patients and to estimate distances from zip code of residence to hospital in which care was received. Among all leg amputations, 850 (18.6%) were performed on patients classified as remote, and 3723 (81.4%) were performed on patients classified as nonremote. Compared with nonremote patients, remote patients were more often categorized as white and more frequently received Medicare and/or Medicaid. Of the subset of remote patients, those at low-volume hospitals were older, were less often categorized as Hispanic, more often had Medicaid coverage, were also more frequently admitted through the emergency department, and often had a foot infection compared with those at experienced centers. Geospatial analysis identified five concentrated geographic areas of remote patients who live more than 50 miles from an experienced center. These findings suggest travel distance may at least influence, if not constrain, the choice of hospital for patients with PAD and foot complications. Efforts to decrease leg amputations among remote patients should be focused on five specific geographic areas of Texas.


Characterization of neuromas in peripheral nerves and their effects on heterotopic bone formation.
The formation of neuromas involves expansion of the cellular components of peripheral nerves. The onset of these disorganized tumors involves activation of sensory nerves and neuroinflammation. Particularly problematic in neuroma is arborization of axons leading to extreme, neuropathic pain. The most common sites for neuroma are the ends of transected nerves following injury; however, this rodent model does not reliably result in neuroma formation. In this study, we established a rodent model of neuroma in which the sciatic nerve was loosely ligated with two chromic gut sutures. This model formed neuromas reliably (∼95%), presumably through activation of the neural inflammatory cascade. Resulting neuromas had a disorganized structure and a significant number of replicating cells. Quantification of changes in perineurial and Schwann cells showed a significant increase in these populations. Immunohistochemical analysis showed the presence of β-tubulin 3 in the rapidly expanding nerve and a decrease in neurofilament heavy chain compared to the normal nerve, suggesting the axons forming a disorganized structure. Measurement of the permeability of the blood-nerve barrier shows that it opened almost immediately and remained open as long as 10 days. Studies using an antagonist of the β3-adrenergic receptor (L-748,337) or cromolyn showed a significant reduction in tumor size and cell expansion as determined by flow cytometry, with an improvement in the animal's gait detected using a Catwalk system. Previous studies in our laboratory have shown that heterotopic ossification is also a result of the activation of neuroinflammation. Since heterotopic ossification and neuroma often occur together in amputees, they were induced in the same limbs of the study animals. More heterotopic bone was formed in animals with neuromas as compared to those without. These data collectively suggest that perturbation of early neuroinflammation with compounds such as L-748,337 and cromolyn may reduce formation of neuromas.


Sodium pump activity and calcium relaxation in vascular smooth muscle of deoxycorticosterone acetate-salt rats.
The Na+-K+ pump activity was determined in femoral arterial smooth muscle from deoxycorticosterone acetate (DOCA)-salt hypertensive rats using potassium relaxation and ouabain-sensitive 86Rb uptake as indices. The membrane-stabilizing effect of calcium and its relation to Na+-K+ pump activity also were examined. Femoral arteries from DOCA-salt rats exhibited a greater relaxation in response to potassium addition after contraction with norepinephrine in a low potassium (0.6 mM) Krebs solution. The concentration of potassium required to produce a 50% relaxation was significantly less in DOCA-salt rats. Ouabain-sensitive 86Rb uptake was significantly greater at 3, 10, and 20 minutes of 86Rb incubation in femoral arteries from DOCA-salt rats. Linear regression analysis revealed a significant correlation between the uptake of 86Rb and time of incubation in both control and DOCA-salt rats. A significant difference in the slopes of the regression lines showed that the rate of uptake was greater in DOCA-salt rats. No difference was observed in ouabain-insensitive 86Rb uptake. A dose-dependent relaxation in response to increasing concentrations of calcium following contraction to norepinephrine was observed in femoral arteries from control and DOCA-salt rats. The relaxation was directly dependent on the level of extracellular potassium and was blocked by ouabain. Femoral arteries from DOCA-salt rats relaxed to a significantly greater extent in response to calcium at each level of potassium when compared with controls. These results provide further evidence for an increase in Na+-K+ pump activity in vascular smooth muscle from DOCA-salt hypertensive rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Cathelicidin attenuates hyperoxia-induced kidney injury in newborn rats.
Aim: Supplemental oxygen is often used to treat neonates with respiratory disorders. Human and animal studies have demonstrated that neonatal hyperoxia increases oxidative stress and induces damage and collagen deposition in kidney during the perinatal period. Cathelicidin LL-37 is one important group of human antimicrobial peptides which exhibits antioxidant activity and its overexpression resists hyperoxia-induced oxidative stress. This study was designed to evaluate the protective effects of cathelicidin in hyperoxia-induced kidney injury in newborn rats. Methods: Sprague-Dawley rat pups were reared in either room air (RA) or hyperoxia (85% O2) and were randomly treated with low-dose (4 mg/kg) and high-dose (8 mg/kg) cathelicidin in normal saline (NS) administered intraperitoneally on postnatal days 1-6. The following six groups were obtained: RA + NS, RA + low-dose cathelicidin, RA + high-dose cathelicidin, O2 + NS, O2 + low-dose cathelicidin, and O2 + high-dose cathelicidin. Kidneys were taken for Western blot and histological analyses on postnatal day 7. Results: The hyperoxia-reared rats exhibited significantly lower body weights and anti-inflammatory M2 macrophages, but the kidney injury scores, oxidative stress marker 8-hydroxy-2'-deoxyguanosine (8-OHdG)-positive cells, pro-inflammatory M1 macrophages, collagen deposition, and NF-κB expression were higher than did the RA-reared rats. Conclusions: Cathelicidin treatment attenuated kidney injury as evidenced by lower kidney injury scores, 8-OHdG-positive cells, collagen deposition, and reversion of hyperoxia-induced M1/M2 macrophage polarization. The role of Cathelicidin in ameliorates kidney injury of the hyperoxia newborn rats was accompanied by decreased NF-κB expression, which probably through the modulating NF-κB activity in the kidney.


Concentrations and solubility of selected trace metals in leaf and bagged black teas commercialized in Poland.
The objective of this study was to determine the concentrations of heavy metals in bagged and leaf black teas of the same brand and evaluate the percentage transfer of metals to tea infusion to assess the consumer exposure. Ten leaf black teas and 10 bagged black teas of the same brand available in Poland were analyzed for Zn, Mn, Cd, Pb, Ni, Co, Cr, Al, and Fe concentrations both in dry material and their infusion. The bagged teas contained higher amounts of Pb, Mn, Fe, Ni, Al, and Cr compared with leaf teas of the same brand, whereas the infusions of bagged tea contained higher levels of Mn, Ni, Al, and Cr compared with leaf tea infusions. Generally, the most abundant trace metals in both types of tea were Al and Mn. There was a wide variation in percentage transfer of elements from the dry tea materials to the infusions. The solubility of Ni and Mn was the highest, whereas Fe was insoluble and only a small portion of this metal content may leach into infusion. With respect to the acceptable daily intake of metals, the infusions of both bagged and leaf teas analyzed were found to be safe for human consumption.


Public Views on Models for Accessing Genomic and Health Data for Research: Mixed Methods Study.
The literature abounds with increasing numbers of research studies using genomic data in combination with health data (eg, health records and phenotypic and lifestyle data), with great potential for large-scale research and precision medicine. However, concerns have been raised about social acceptability and risks posed for individuals and their kin. Although there has been public engagement on various aspects of this topic, there is a lack of information about public views on data access models. This study aimed to address the lack of information on the social acceptability of access models for reusing genomic data collected for research in conjunction with health data. Models considered were open web-based access, released externally to researchers, and access within a data safe haven. Views were ascertained using a series of 8 public workshops (N=116). The workshops included an explanation of benefits and risks in using genomic data with health data, a facilitated discussion, and an exit questionnaire. The resulting quantitative data were analyzed using descriptive and inferential statistics, and the qualitative data were analyzed for emerging themes. Respondents placed a high value on the reuse of genomic data but raised concerns including data misuse, information governance, and discrimination. They showed a preference for giving consent and use of data within a safe haven over external release or open access. Perceived risks with open access included data being used by unscrupulous parties, with external release included data security, and with safe havens included the need for robust safeguards. This is the first known study exploring public views of access models for reusing anonymized genomic and health data in research. It indicated that people are generally amenable but prefer data safe havens because of perceived sensitivities. We recommend that public views be incorporated into guidance on models for the reuse of genomic and health data.


Modulation of N400 in chronic non-fluent aphasia using low frequency Repetitive Transcranial Magnetic Stimulation (rTMS).
Low frequency Repetitive Transcranial Magnetic Stimulation (rTMS) has previously been applied to language homologues in non-fluent populations of persons with aphasia yielding significant improvements in behavioral language function up to 43 months post stimulation. The present study aimed to investigate the electrophysiological correlates associated with the application of rTMS through measurement of the semantic based N400 Event-related brain potentials (ERP) component. Low frequency (1 Hz) rTMS was applied to the anterior portion of the homologue to Broca's area (pars triangularis), for 20 min per day for 10 days, using a stereotactic neuronavigational system. Twelve non-fluent persons with aphasia, 2-6 years post stroke were stimulated. Six participants were randomly assigned to receive real stimulation and six participants were randomly assigned to receive a blind sham control condition. ERP measures were recorded at baseline, 1 week and 2 months subsequent to stimulation. The findings demonstrate treatment related changes observed in the stimulation group when compared to the placebo control group at 2 months post stimulation indicating neuromodulation of N400 as a result of rTMS. No treatment related changes were identified in the stimulation group, when compared to the sham group from baseline to 1 week post stimulation. The electrophysiological results represent the capacity of rTMS to modulate neural language networks and measures of lexical-semantic function in participants with non-fluent aphasia and suggest that time may be an important factor in brain reorganization subsequent to rTMS.


The Economics of Human Development and Social Mobility.
This paper distills and extends recent research on the economics of human development and social mobility. It summarizes the evidence from diverse literatures on the importance of early life conditions in shaping multiple life skills and the evidence on critical and sensitive investment periods for shaping different skills. It presents economic models that rationalize the evidence and unify the treatment effect and family influence literatures. The evidence on the empirical and policy importance of credit constraints in forming skills is examined. There is little support for the claim that untargeted income transfer policies to poor families significantly boost child outcomes. Mentoring, parenting, and attachment are essential features of successful families and interventions to shape skills at all stages of childhood. The next wave of family studies will better capture the active role of the emerging autonomous child in learning and responding to the actions of parents, mentors and teachers.


Interventions for health workforce retention in rural and remote areas: a systematic review.
Attracting and retaining sufficient health workers to provide adequate services for residents of rural and remote areas has global significance. High income countries (HICs) face challenges in staffing rural areas, which are often perceived by health workers as less attractive workplaces. The objective of this review was to examine the quantifiable associations between interventions to retain health workers in rural and remote areas of HICs, and workforce retention. The review considers studies of rural or remote health workers in HICs where participants have experienced interventions, support measures or incentive programs intended to increase retention. Experimental, quasi-experimental and observational study designs including cohort, case-control, cross-sectional and case series studies published since 2010 were eligible for inclusion. The Joanna Briggs Institute methodology for reviews of risk and aetiology was used. Databases searched included MEDLINE (OVID), CINAHL (EBSCO), Embase, Web of Science and Informit. Of 2649 identified articles, 34 were included, with a total of 58,188 participants. All study designs were observational, limiting certainty of findings. Evidence relating to the retention of non-medical health professionals was scant. There is growing evidence that preferential selection of students who grew up in a rural area is associated with increased rural retention. Undertaking substantial lengths of rural training during basic university training or during post-graduate training were each associated with higher rural retention, as was supporting existing rural health professionals to extend their skills or upgrade their qualifications. Regulatory interventions requiring return-of-service (ROS) in a rural area in exchange for visa waivers, access to professional licenses or provider numbers were associated with comparatively low rural retention, especially once the ROS period was complete. Rural retention was higher if ROS was in exchange for loan repayments. Educational interventions such as preferential selection of rural students and distributed training in rural areas are associated with increased rural retention of health professionals. Strongly coercive interventions are associated with comparatively lower rural retention than interventions that involve less coercion. Policy makers seeking rural retention in the medium and longer term would be prudent to strengthen rural training pathways and limit the use of strongly coercive interventions.


MRI in patients with otovestibular complaints of unknown origin.
The place of MRI in the diagnostic work-up of patients with bilateral perceptive hearing loss, tinnitus and vertigo is under discussion. The purpose of this study is to investigate the role of MRI in patients with otovestibular and cranial nerve complaints of unknown aetiology. After thorough otologic examination, 430 patients were consecutively referred for an MR examination of the cerebellopontine angle. The detection rate for essential lesions was 4.9%. Two groups of retrocochlear lesions were frequently observed: central WMLs/atrophy and neurovascular conflict affecting a cranial nerve. MR imaging of the cerebellopontine angle, fossa posterior and petrous bones makes it possible to observe abnormalities of the vestibulocochlear nerve and inner ear. Additional T2-weighted FSE images of the whole brain make it possible to evaluate the occurrence of early central lesions. This imaging protocol can diagnose essential lesions relating directly to the complaint in 4.9% of the patients with hearing loss, subjective tinnitus or vertigo. We frequently observed two groups of lesions of uncertain significance in our study population. WMLs are present in 50% of patients with a mean age of 59 years. In the younger subpopulation aged under 51 years the prevalence of WMLs is 24%. It remains unclear whether these lesions can be accounted for by the diversity of symptoms with which the patients presented. In addition, we found a high number of neurovascular conflicts involving different cranial nerves.


Particle imaging velocimetry evaluation of intracranial stents in sidewall aneurysm: hemodynamic transition related to the stent design.
We investigated the flow modifications induced by a large panel of commercial-off-the-shelf (COTS) intracranial stents in an idealized sidewall intracranial aneurysm (IA). Flow velocities in IA silicone model were assessed with and without stent implantation using particle imaging velocimetry (PIV). The use of the recently developed multi-time-lag method has allowed for uniform and precise measurements of both high and low velocities at IA neck and dome, respectively. Flow modification analysis of both regular (RSs) and flow diverter stents (FDSs) was subsequently correlated with relevant geometrical stent parameters. Flow reduction was found to be highly sensitive to stent porosity variations for regular stents RSs and moderately sensitive for FDSs. Consequently, two distinct IA flow change trends, with velocity reductions up to 50% and 90%, were identified for high-porosity RS and low-porosity FDS, respectively. The intermediate porosity (88%) regular braided stent provided the limit at which the transition in flow change trend occurred with a flow reduction of 84%. This transition occurred with decreasing stent porosity, as the driving force in IA neck changed from shear stress to differential pressure. Therefore, these results suggest that stents with intermediate porosities could possibly provide similar flow change patterns to FDS, favourable to curative thrombogenesis in IAs.


Ensemble Docking Coupled to Linear Interaction Energy Calculations for Identification of Coronavirus Main Protease (3CLpro) Non-Covalent Small-Molecule Inhibitors.
SARS-CoV-2, or severe acute respiratory syndrome coronavirus 2, represents a new strain of Coronaviridae. In the closing 2019 to early 2020 months, the virus caused a global pandemic of COVID-19 disease. We performed a virtual screening study in order to identify potential inhibitors of the SARS-CoV-2 main viral protease (3CLpro or Mpro). For this purpose, we developed a novel approach using ensemble docking high-throughput virtual screening directly coupled with subsequent Linear Interaction Energy (LIE) calculations to maximize the conformational space sampling and to assess the binding affinity of identified inhibitors. A large database of small commercial compounds was prepared, and top-scoring hits were identified with two compounds singled out, namely 1-[(R)-2-(1,3-benzimidazol-2-yl)-1-pyrrolidinyl]-2-(4-methyl-1,4-diazepan-1-yl)-1-ethanone and [({(S)-1-[(1H-indol-2-yl)methyl]-3-pyrrolidinyl}methyl)amino](5-methyl-2H-pyrazol-3-yl)formaldehyde. Moreover, we obtained a favorable binding free energy of the identified compounds, and using contact analysis we confirmed their stable binding modes in the 3CLpro active site. These compounds will facilitate further 3CLpro inhibitor design.


Predictors of sun protection behaviors and severe sunburn in an international online study.
The incidence of melanoma continues to increase in many countries, and primary prevention of melanoma includes avoidance of sunburn as well as adequate sun protection behavior. The aim of this study was to examine the prevalence of self-reported sun protection behaviors and sunburn in users of the Internet, and to identify the demographic, clinical, and attitudinal/motivational correlates of sun protection behaviors. Self-report data were gathered on behalf of the GenoMEL consortium using an online survey available in 10 different languages, and 8,178 individuals successfully completed at least 80% of survey items, with 73% of respondents from Europe, 12% from Australia, 7% from the United States, 2% from Israel, and 6% from other countries. Half of all respondents and 27% of those with a previous melanoma reported at least one severe sunburn during the previous 12 months. The strongest factors associated with sun protection behavior were perceived barriers to protection (beta = -0.44/beta = -0.37), and respondents who reported a positive attitude toward suntans were less likely to protect (beta = -0.16/beta = -0.14). Reported use of protective clothing and shade, as well as avoidance of midday sun exposure, were more strongly related to reduced risk of sunburn than sunscreen use. Despite widespread dissemination of public health messages about the importance of sun protection, a substantial proportion of this international sample, including respondents with a previous melanoma, reported inadequate sun protection behaviors resulting in severe sunburn. Future strategies to decrease sunburn should target the practical, social, and psychological barriers associated with nonuptake of sun protection.


Fenugreek Seed Powder Nullified Aluminium Chloride Induced Memory Loss, Biochemical Changes, Aβ Burden and Apoptosis via Regulating Akt/GSK3β Signaling Pathway.
Alzheimer's disease (AD) is the most common form of dementia that mainly affects the cognitive functions of the aged populations. Trigonella foenum-graecum (L.) (fenugreek), a traditionally well utilized medicinal plant ubiquitously used as one of the main food additive worldwide, is known to have numerous beneficial health effects. Fenugreek seed extract could be able to inhibit the activity of acetylcholinesterase (AChE), a key enzyme involved in the pathogenesis of AD, and further shown to have anti-parkinsonic effect. The present study was aimed to explore the neuroprotective effect of fenugreek seed powder (FSP) against aluminium chloride (AlCl3) induced experimental AD model. Administration of germinated FSP (2.5, 5 and 10% mixed with ground standard rat feed) protected AlCl3 induced memory and learning impairments, Al overload, AChE hyperactivity, amyloid β (Aβ) burden and apoptosis via activating Akt/GSK3β pathway. Our present data could confirm the neuroprotective effect of fenugreek seeds. Further these results could lead a possible therapeutics for the management of neurodegenerative diseases including AD in future.


Hair cycle stage of the mouse vibrissa follicle determines subsequent fiber growth and follicle behavior in vitro.
The establishment of culture models representative of all aspects of in vivo hair follicle behavior is an important goal for theoretic and analytic studies. Rodent vibrissa follicles have regular, predictable, and relatively short growth cycles. In this investigation, we took advantage of these properties; we classified mouse vibrissa follicles according to different phases in the hair cycle and then compared fiber growth and morphologic changes in culture. Follicles isolated in the early phase of the growth cycle produced fine growing fibers with an average growth that exceeded 3 mm over 15 d. Even when hair growth had slowed or halted subsequently, histology showed that these follicles retained an anagen-like morphology. By contrast, follicles isolated toward the end of the growing cycle produced thicker fibers for much shorter periods, after which growth ceased and the fibers lifted up from the base of the follicle. Internally, these specimens resembled their telogen counterparts in situ. Follicles isolated in mid-growth demonstrated intermediate fiber growth characteristics. In organ culture, mouse vibrissa follicles therefore closely reflect their in vivo origin in growth characteristics and cycle timing. These data provide new opportunities for studying hair growth cycle mechanisms in vitro, but present a caveat for quantitative studies because there may be a greater growth cycle-related variation than has previously been assumed.


Risk of ischemic stroke with tamoxifen treatment for breast cancer: a meta-analysis.
To assess the overall risk of stroke, specifically ischemic stroke, associated with tamoxifen use by performing a meta-analysis of data reported in breast cancer trials. Tamoxifen increases the risk of venous thromboembolism in women with breast cancer, but its relationship to stroke risk is uncertain. A systematic review of randomized controlled trials of tamoxifen for breast cancer management and prevention published since 1980 was performed using MEDLINE. The summary odds ratio (OR) and 95% CI were calculated using the Mantel-Haenszel method, followed by a statistical test for heterogeneity. Nine trials met the inclusion criteria, and six trials specified ischemic stroke outcomes. The Mantel-Haenszel summary OR was 1.82 (95% CI, 1.41 to 2.36) for ischemic stroke and 1.40 (1.14 to 1.72) for any stroke. The chi2 heterogeneity test was 6.0 (p > 0.1) for ischemic stroke and 16.1 (p < 0.05) for any stroke. The random-effects summary OR of Der Simonian and Laird for any stroke was 1.29 (0.92 to 1.81). During a mean follow-up period of 4.9 years, the frequency of ischemic stroke was 0.71% with tamoxifen vs 0.39% for controls (absolute increased risk, 0.32%; number needed to harm [NNH], 313). Women with breast cancer who were treated with tamoxifen had an 82% increased risk of ischemic stroke and a 29% increased risk of any stroke, but the absolute risk is small. Further studies assessing prespecified cerebrovascular outcomes are ongoing and will further clarify the risk of stroke associated with tamoxifen use.


Severe coronary artery disease after radiation therapy of the chest and mediastinum: clinical presentation and treatment.
To define the clinical and angiographic features and the therapeutic problems in patients with coronary artery disease after therapeutic irradiation of the chest. An observational retrospective study. The cardiac catheterisation laboratory, university medical school. 15 subjects (8 men and 7 women, aged 25-56 years, mean 44) examined in the cardiac catheterisation laboratory, who had significant coronary artery disease years after having radiation treatment to the chest and anterior mediastinum. In the early stages of the study angiography was performed because of typical symptoms of ischaemic heart disease. Later on it was performed because of a high index of suspicion in people with signs of extensive radiation heart damage. Clinical and electrocardiographic evidence of ischaemic heart disease; echocardiographic signs of pericardial, myocardial or valvar involvement; angiographic evidence of coronary arterial stenosis, with special attention to the ostia; haemodynamic and angiographic signs of pericardial, myocardial, and valvar disease. Survival and symptomatic and functional status were ascertained after medical or surgical treatment. The patients were relatively young and had no risk factors. Seven patients had no signs or symptoms of ischaemic heart disease. Ten patients had ostial stenosis, which was associated with extensive involvement of other cardiac structures in nine of them. Seven required surgical treatment for coronary artery disease. Two died, one at surgery and the other one six months later. Five patients had complications associated with irradiation. Coronary arterial disease can be reasonably ascribed to the effects of chest irradiation when the patients are young and free from risk factors, especially if the obstructions are ostial and there is important damage to other cardiac structures. In patients with damage to other cardiac structures angina and infarction are often absent and coronary angiography seems to be mandatory. Patients often require surgical treatment and postoperative complications are common.


Linear latent force models using Gaussian processes.
Purely data-driven approaches for machine learning present difficulties when data are scarce relative to the complexity of the model or when the model is forced to extrapolate. On the other hand, purely mechanistic approaches need to identify and specify all the interactions in the problem at hand (which may not be feasible) and still leave the issue of how to parameterize the system. In this paper, we present a hybrid approach using Gaussian processes and differential equations to combine data-driven modeling with a physical model of the system. We show how different, physically inspired, kernel functions can be developed through sensible, simple, mechanistic assumptions about the underlying system. The versatility of our approach is illustrated with three case studies from motion capture, computational biology, and geostatistics.


Photometric stereo via expectation maximization.
This paper presents a robust and automatic approach to photometric stereo, where the two main components, namely surface normals and visible surfaces, are respectively optimized by Expectation Maximization (EM). A dense set of input images is conveniently captured using a digital video camera while a handheld spotlight is being moved around the target object and a small mirror sphere. In our approach, the inherently complex optimization problem is simplified into a two-step optimization, where EM is employed in each step: 1) Using the dense input, the weight or importance of each observation is alternately optimized with the normal and albedo at each pixel and 2) using the optimized normals and employing the Markov Random Fields (MRFs), surface integrabilities and discontinuities are alternately optimized in visible surface reconstruction. Our mathematical derivation gives simple updating rules for the EM algorithms, leading to a stable, practical, and parameter-free implementation that is very robust even in the presence of complex geometry, shadows, highlight, and transparency. We present high-quality results on normal and visible surface reconstruction, where fine geometric details are automatically recovered by our method.


Human papillomavirus-associated adenocarcinoma of the base of the tongue.
Human papillomavirus (HPV) is a major cause of oropharyngeal squamous cell carcinoma with characteristic clinical and pathologic features relative to their non-HPV-associated counterparts. Here we describe 2 cases of HPV-associated adenocarcinoma of the oropharynx. Both cases arose at the base of the tongue, and neither had the histologic or immunohistochemical features of a primary salivary gland tumor or metastasis from another location. One patient had metastases to neck lymph nodes and the lungs and died of disease 37 months after diagnosis. Evidence for an HPV association consisted of strong diffuse expression of p16, polymerase chain reaction-based detection of HPV16 DNA sequences, and localization of HPV by in situ hybridization within tumor cells of both primary and metastatic lesions. These results further expand the spectrum of HPV-associated head and neck malignancy. This rare entity should be distinguished from primary salivary gland adenocarcinoma and may be a candidate for HPV-specific targeted therapies.


Structural insights into the binding of vascular endothelial growth factor-B by VEGFR-1(D2): recognition and specificity.
The formation of blood vessels (angiogenesis) is a highly orchestrated sequence of events involving crucial receptor-ligand interactions. Angiogenesis is critical for physiological processes such as development, wound healing, reproduction, tissue regeneration, and remodeling. It also plays a major role in sustaining tumor progression and chronic inflammation. Vascular endothelial growth factor (VEGF)-B, a member of the VEGF family of angiogenic growth factors, effects blood vessel formation by binding to a tyrosine kinase receptor, VEGFR-1. There is growing evidence of the important role played by VEGF-B in physiological and pathological vasculogenesis. Development of VEGF-B antagonists, which inhibit the interaction of this molecule with its cognate receptor, would be important for the treatment of pathologies associated specifically with this growth factor. In this study, we present the crystal structure of the complex of VEGF-B with domain 2 of VEGFR-1 at 2.7 A resolution. Our analysis reveals that each molecule of the ligand engages two receptor molecules using two symmetrical binding sites. Based on these interactions, we identify the receptor-binding determinants on VEGF-B and shed light on the differences in specificity towards VEGFR-1 among the different VEGF homologs.


Er,Cr:YSGG 2780 nm Laser Treatment of Lip Melanin Hyperpigmentation.
In the current research, Er,Cr:YSGG laser been used in ablative mode to peel the pigmentation, and many sessions were done to completely remove the melanin pigmentations. 85 patients were selected and Er,Cr : YSGG laser with 2780 nm wavelength in ablative mode (iPlus, Biolase, USA) was used to treat patients with dark lip melanin pigmentation, using gold handpiece and MZ 10 tip (diameter of 1 mm), and the parameters were set as follows: energy density 28.7 J/cm2, frequency 30 Hz, water cooling 100%, and air cooling 60%. The vast majority of the patients (82.4%) had mild adverse effects after the lip depigmentation and 74.1% of patients reported complete improvement of the lip with a high satisfaction rate (84.7%) during the follow-up period with a low level of relapse. Er,Cr:YSGG 2670 nm is an effective tool to achieve excellent esthetic results in the treatment of lip melanin pigmentation; it is very well tolerated by patients with minimal adverse effects.


The value of periprostatic fascia thickness and fascia preservation as prognostic factors of erectile function after nerve-sparing robot-assisted radical prostatectomy.
To determine the correlation of preoperative fascia thickness (FT) and intraoperative fascia preservation (FP) with erectile function (EF) after nerve-sparing robot-assisted radical prostatectomy (RARP). Our analysis included 106 patients, with localized prostate cancer and no erectile dysfunction (ED) before RARP, assessed with preoperative 3 Tesla (3 T) multiparametric magnetic resonance imaging (MRI). FP score was defined as the extent of FP from the base to the apex of the prostate, quantitatively assessed by the surgeon. Median fascia thickness (MFT) per patient was defined as the sum of the median FT of 12 MRI regions. Preserved MFT (pMFT) was the sum of the saved MFT. The percentage of pFMT (ppMFT) was also calculated. Fascia surface (FS) was measured on MRI and it was combined with FP score resulting in preserved FS (pFS) and percentage of pFS (ppFS). FP score, pMFT, ppMFT, pFS and ppFS were significantly lower (p < 0.0001) in patients with ED. In the multivariate regression analysis, lower FP score [odds ratio (OR) 0.721, p = 0.03] and lower ppMFT (OR 0.001, p = 0.027) were independent predictors of ED. ROC analysis showed the highest area under the curve for ppMFT (0.787) and FP score (0.767) followed by pMFT (0.755) and ppFS (0.743). MRI-determined periprostatic FT combined with intraoperative FP score are correlated to postprostatectomy EF. Based on the hypothesis that a thicker fascia forms a protective layer for the nerves, we recommend assessing FT preoperatively to counsel men for the odds of preserving EF after RARP.