Erythropoietin promotes the growth of tumors lacking its receptor and decreases survival of tumor-bearing mice by enhancing angiogenesis.
Erythropoietin (Epo), a known hematopoietic growth factor, has been reported to promote tumor growth and angiogenesis in Epo receptor (EpoR)-positive tumors, but its effects on EpoR-negative tumors have not been clearly shown. Here, we show that Epo accelerates the growth of EpoR-negative tumors by promoting tumor angiogenesis. Mice were inoculated with Lewis lung carcinoma cells and treated with Epo. Erythropoietin accelerated tumor growth and increased intratumoral microvessel density, although it did not accelerate Lewis lung carcinoma cell tumor proliferation in vitro. To observe the direct effect of Epo on endothelial cells, we examined human dermal microvascular endothelial cells (HMVECs) that expressed EpoR. Erythropoietin induced the proliferation of HMVECs and protected them from H2O2-induced cell death. Erythropoietin activated the extracellular signal-regulated kinase signaling pathway and up-regulated the expression of the downstream antiapoptotic protein Bcl-xL in HMVECs. Moreover, in both the absence and presence of tumors, in vivo treatment of mice with Epo increased circulating endothelial progenitor cells. To investigate the role of Epo in a primary tumor model, we inoculated the chemical carcinogen methylcholanthrene (MCA) subcutaneously into mice at two doses, a high or a low dose, which induced fibrosarcoma, and treated them with Epo. Erythropoietin promoted tumor growth after MCA inoculation at both doses and decreased the overall survival of the mice inoculated with the high-dose MCA. However, Epo did not increase the incidence of fibrosarcoma at either dose. Lewis lung carcinoma cells and MCA-induced fibrosarcomas did not express EpoR. These results suggest that Epo accelerates the growth of tumors that lack EpoR expression by promoting tumor angiogenesis.


Age differences in exposure and reactions to interpersonal tensions: a daily diary study.
This study examines age differences in exposure and reactivity to interpersonal tensions. The data are from the National Study of Daily Experiences in which participants ages 25 to 74 (N = 666) completed phone interviews wherein they described interpersonal tensions and rated the stressfulness of the tensions each evening for 8 days. Coders rated descriptions for types of behavioral reactions. Multilevel models revealed older adults reported fewer interpersonal tensions, were more likely to report tensions with spouses, were less likely to report tensions with children, experienced less stress, and were less likely to argue and more likely to do nothing in response to tensions than were younger adults. Age differences in emotional and behavioral reactions did not appear to be due to variations in exposure to tensions. The discussion centers on why older people may be better able to regulate their reactions to problems than younger people.


[Evaluation of the influenza vaccine effectiveness among children aged 6 to 72 months based on the test-negative case control study design].
Objective: To assess the effectiveness of seasonal influenza vaccine among children aged 6 to 72 months. Methods: The test-negative case control study was conducted based on available surveillance data which was from China Influenza Surveillance Information system (CSIS). From October 2016 to April 2017 and from October 2017 to April 2018,1 161 cases aged 6-72 months with influenza-like illness in Yongkang and Yiwu city, were selected as the study subjects, and the cases with influenza test-positive were selected as the case group (403 cases). Test-negative subjects were selected as control group (758 cases). The etiology and immunization data of the subjects were obtained from CSIS and Immune Information and Management System (IIMS) respectively. Vaccine effectiveness was estimated using multivariate logistic regression model,and the mixed effects of non-randomized control in TNCC study were equalized by using the propensity score (PS) method in the statistical analysis. Results: The age of the subjects was (2.44±1.60) years,and there were 681 boys (58.66%). The age of case group was (2.62±1.58) years, and there were 246 boys (61.04%). The case group was including of 237 cases (58.81%) of influenza A (H3N2), 92 cases (22.83%) of influenza A (H1N1) pmd09, 62 cases (15.38%) of influenza B(Victoria) lineage, 11 cases (2.73%) of influenza B (Yamagata) lineage and one case (0.25%) co-infection of influenza [A(H(3)N(2))+B (Victoria)]. The mean age of the control group was (2.35±1.61) years,and there were 435 boys (57.39%). Overall vaccine effectiveness (VE) against all type influenza for two seasons combined was 58% (95%CI: 31%-74%). An analysis by age groups showed 68% (95%CI:41%-82%) of the VE estimate among children aged 36-72 months while it was 28%(95%CI:-80%-71%)of the VE estimate among children aged 6-35 months. The VE estimate value was 54% (95%CI:16%-75%) against all type influenza and 65% (95%CI:24%-83%) against influenza A (H(3)N(2)) during the 2016-2017 season. During the 2017-2018 season, the VE estimate value was 69% (95%CI:18%-88%) against all type influenza. Conclusion: Influenza vaccine is effective in preventing influenza virus infection during the flu season,especially the effect among children aged 36-72 months is higher compared to that among children aged 6-35 months.


Pathology of a live attenuated anti-schistosome vaccine in mice.
Tissue responses of mice to intramuscular injection of 50 kR 60Co-attenuated schistosomula of Schistosoma mansoni were studied. Controls included injection of unattenuated schistosomula, medium alone, antigen-coated beads, and alum-adsorbed tetanus/diphtheria toxoids. Primary reactions to tissue-confined deposits of injected schistosomula, whether attenuated or not, were relatively intense and prolonged. Parasite attrition proceeded steadily, with most destroyed by the 7th day; however, a few intact organisms persisted up to 4 weeks. Cryopreservation did not alter the course of parasite attrition nor host reaction. Irradiated larvae were not found in lymph nodes, lungs, or liver. Neutrophils dominated the early reactions and were gradually replaced by mononuclear phagocytes, lymphoid cells, and eosinophils. Fibroblast proliferation and muscle regeneration began by day 3; reaction size and intensity peaked by day 7. From weeks 1-4, inflammatory infiltrates and regenerative proliferation underwent gradual involution, and injection sites were healed with no scarring by the end of 4-5 weeks. Mice primed by infection or by prior injection showed an accelerated course of inflammation, enhanced tissue eosinophilia, and more rapid healing. An unwanted, but prominent, feature of schistosomular vaccine reactions in mice was tracking of the inflammatory infiltrate along connective tissue septal and nerve sheaths, the latter raising the question of the pain potential of the vaccine. To conclude, in mice, attenuated schistosomular vaccines cause relatively marked local inflammatory responses but no systemic lesions at all, and their injection sites heal without permanent damage.


Increased endothelin-1 responsiveness in human coronary artery smooth muscle cells exposed to 1,25-dihydroxyvitamin D(3).
Low blood concentrations of 25-hydroxyvitamin D(3) are associated with increased mortality, while some studies suggest improved cardiovascular outcomes with vitamin D(3) supplementation in chronic kidney disease. However, the physiological effects of vitamin D(3) on the cardiovascular system remain poorly understood making it difficult to determine whether vitamin D(3) supplementation might provide cardiovascular benefit or even cause harm. Thus here we investigated the effects of chronic 1,25-dihydroxyvitamin D(3) treatment on intracellular signaling in human coronary artery smooth muscle cells (HCASMCs) and found that 1,25-dihydroxyvitamin D(3) significantly potentiated endothelin (ET-1) signaling. Specifically, 1,25-dihydroxyvitamin D(3) (24-h pretreatment) caused a more than threefold enhancement in both ET-1-induced intracellular calcium mobilization and extracellular signal-regulated kinase (ERK) activation. This 1,25-dihydroxyvitamin D(3)-elicited signaling enhancement was not observed for either vasopressin or carbachol. With the use of endothelin receptor (ETR) isoform-selective antagonists, ETRA was found to be primarily responsible for the 1,25-dihydroxyvitamin D(3)-induced ET-1 responsiveness and yet ETRA mRNA expression and protein abundance were unaltered following 1,25-dihydroxyvitamin D(3) treatment. While there was an increase in ETRB mRNA expression in response to 1,25-dihydroxyvitamin D(3), the protein abundance of ETRB was again unchanged. Finally, ETRA/ETRB heterodimerization was not detected in HCASMCs in either the absence or presence of 1,25-dihydroxyvitamin D(3). Together, these data show for the first time that 1,25-dihydroxyvitamin D(3) enhances endothelin responsiveness in HCASMCs and that the effect is mediated through ETRA.


The biology of photodynamic therapy.
The subcellular, cellular and tissue/tumour interactions with non-toxic photosensitizing chemicals plus non-thermal visible light (photodynamic therapy (PDT) are reviewed. The extent to which endothelium/vasculature is the primary target is discussed, and the biochemical opportunities for manipulating outcome highlighted. The nature of tumour destruction by PDT lends itself to imaging outcome by MRI and PET.


Expression of tdTomato and luciferase in a murine lung cancer alters the growth and immune microenvironment of the tumor.
Imaging techniques based on fluorescence and bioluminescence have been important tools in visualizing tumor progression and studying the effect of drugs and immunotherapies on tumor immune microenvironment in animal models of cancer. However, transgenic expression of foreign proteins may induce immune responses in immunocompetent syngeneic tumor transplant models and augment the efficacy of experimental drugs. In this study, we show that the growth rate of Lewis lung carcinoma (LL/2) tumors was reduced after transduction of tdTomato and luciferase (tdTomato/Luc) compared to the parental cell line. tdTomato/Luc expression by LL/2 cells altered the tumor microenvironment by increasing tumor-infiltrating lymphocytes (TILs) while inhibiting tumor-induced myeloid-derived suppressor cells (MDSCs). Interestingly, tdTomato/Luc expression did not alter the response of LL/2 tumors to anti-PD-1 and anti-CTLA-4 antibodies. These results suggest that the use of tdTomato/Luc-transduced cancer cells to conduct studies in immune competent mice may lead to cell-extrinsic tdTomato/Luc-induced alterations in tumor growth and tumor immune microenvironment that need to be taken into consideration when evaluating the efficacy of anti-cancer drugs and vaccines in immunocompetent animal models.


Dietary fibers affect viscosity of solutions and simulated human gastric and small intestinal digesta.
Two experiments were conducted to determine the viscosities of both soluble and insoluble dietary fibers. In Expt. 1, corn bran, defatted rice bran, guar gum, gum xanthan, oat bran, psyllium, soy hulls, stabilized rice bran, wheat bran, wood cellulose, and 2 methylcellulose controls (Ticacel 42, Ticacel 43) were hydrated in water overnight at 0.5, 1, 1.5, or 2% concentrations. In Expt. 2, guar gum, oat bran, psyllium, rice bran, wheat bran, and wood cellulose were subjected to a 2-stage in vitro gastric and small intestinal digestion simulation model. Viscosity was measured every 2 and 3 h during gastric and small intestinal simulation, respectively. Viscosities in both experiments were measured at multiple shear rates. Viscosities of all fiber solutions were concentration- and shear rate-dependent. Rice brans, soy hulls, and wood cellulose had the lowest viscosities, whereas guar gum, psyllium, and xanthan gum had the highest viscosities, regardless of concentration. During gastric simulation, viscosity was higher (P < 0.05) at 4 h than at 0 h for guar gum, psyllium, rice bran, and wheat bran. During small intestinal simulation, viscosities were higher (P < 0.05) between 3 and 9 h compared with 18 h for guar gum, oat bran, and rice bran. Guar gum, psyllium, and oat bran exhibited viscous characteristics throughout small intestinal simulation, indicating potential for these fibers to elicit blood glucose and lipid attenuation. Wheat and rice brans and wood cellulose did not exhibit viscous characteristics throughout small intestinal digestion; thus, they may be beneficial for laxation.


Clinical significance of TT virus infection in patients with chronic hepatitis C.
The TT virus (TTV) is a novel DNA virus that has recently been identified. The clinical significance of TTV infection in patients with chronic hepatitis C has not been determined. The aim of this study was to determine the prevalence and possible role of TTV in a well characterized population with chronic hepatitis C infection. Ninety patients with chronic HCV and known time of HCV acquisition were selected from approximately 250 patients followed at our institution. Characteristics including age, sex, histology, and length of disease were recorded. Direct sequencing of the NS5 region was used for HCV genotyping. TTV DNA detection was based on PCR. TTV infection was present in 24 of 90 (27%) HCV patients. Patients were divided into four groups based on stage of disease; chronic hepatitis (CH, 29 patients), compensated cirrhosis (CC, 17 patients), decompensated cirrhosis (DC, 28 patients), and hepatocellular carcinoma (HCC, 16 patients). TTV was present in 2/29 (7%), 2/17 (12%), 11/28 (39%), and 9/16 (56%) in those with CAH, CC, DC, and HCC respectively. TTV was significantly more prevalent among those with advanced disease (DC and HCC) compared to those with stable disease (CH and CC; p = 0.001). Mean age, sex, and the time from exposure to HCV to development of complications were similar in TTV-positive and -negative patients. TTV infection was more common in patients infected with HCV genotype 1b. Univariate analysis showed that length of HCV infection, HCV genotype 1b, and TTV infection were important in predicting the stage of liver disease in HCV patients. However, after adjusting for length of HCV infection, TTV but not HCV genotype was important in predicting the stage of liver disease. We conclude that 1) TTV infection is common in patients with chronic HCV; 2) TTV infection is more prevalent among patients with advanced HCV-associated liver disease (DC and HCC) than in those with stable disease (CH and CC); and 3) TTV infection is more common in patients with HCV genotype 1b but is independent from genotype in predicting the stage of HCV-associated liver disease.


Acceleration of Monte Carlo-based scatter compensation for cardiac SPECT.
Single proton emission computed tomography (SPECT) images are degraded by photon scatter making scatter compensation essential for accurate reconstruction. Reconstruction-based scatter compensation with Monte Carlo (MC) modelling of scatter shows promise for accurate scatter correction, but it is normally hampered by long computation times. The aim of this work was to accelerate the MC-based scatter compensation using coarse grid and intermittent scatter modelling. The acceleration methods were compared to un-accelerated implementation using MC-simulated projection data of the mathematical cardiac torso (MCAT) phantom modelling (99m)Tc uptake and clinical myocardial perfusion studies. The results showed that when combined the acceleration methods reduced the reconstruction time for 10 ordered subset expectation maximization (OS-EM) iterations from 56 to 11 min without a significant reduction in image quality indicating that the coarse grid and intermittent scatter modelling are suitable for MC-based scatter compensation in cardiac SPECT.


Evaluating an undergraduate nursing student telehealth placement for community-dwelling frail older people during the COVID-19 pandemic.
To evaluate a rapid response student telehealth placement experience implementing interRAI assessments of community-dwelling frail older people during the COVID-19 pandemic. To identify lessons to inform future telehealth clinical placements. New Zealand undertakes assessment of older people with disabilities using the interRAI contact assessment tool for less complex conditions and home care assessment tool for complex needs. New Zealand entered lockdown in March 2020 in response to COVID-19. New Zealand's most vulnerable community members required urgent needs assessment. A clinical placement whereby 3rd year undergraduate nursing students trained by interRAI-NZ educators worked remotely from home delivering telehealth assessment for 'at risk' older people across the Waikato District, New Zealand. This represented the first telehealth experience within an undergraduate nursing program approved by the New Zealand Nursing Council. A case study evaluation utilising mixed method questionnaire and qualitative techniques within an interpretive paradigm. 19 third year students in the fifth semester of a Bachelor of Nursing program and 5 nursing staff members engaged in delivery of the initiative completed pre- and post-placement short answer questionnaires. Reflective diaries were maintained by students on placement. Post-placement interviews and focus group discussions provided in-depth data. COREQ guidelines informed analysis and reporting. Student and tutor responses showed consistent themes: tackling COVID-19; implementation requirements; nursing competencies; provider relationships; and community insights. These provide insight and highlight lessons learnt from this initiative. Student confidence in therapeutic engagement and clinical assessment and interest in aged care was increased, confirming the viability and importance of this inaugural telehealth student placement initiative. Graduate work readiness is enhanced through telehealth placement experience and interRAI assessor training. These are recommended as core components of future nursing education programs.


An Impermeant Ganetespib Analog Inhibits Extracellular Hsp90-Mediated Cancer Cell Migration that Involves Lysyl Oxidase 2-like Protein.
Extracellular Hsp90 (eHsp90) activates a number of client proteins outside of cancer cells required for migration and invasion. Therefore, eHsp90 may serve as a novel target for anti-metastatic drugs as its inhibition using impermeant Hsp90 inhibitors would not affect the numerous vital intracellular Hsp90 functions in normal cells. While some eHsp90 clients are known, it is important to establish other proteins that act outside the cell to validate eHsp90 as a drug target to limit cancer spread. Using mass spectrometry we identified two precursor proteins Galectin 3 binding protein (G3BP) and Lysyl oxidase 2-like protein (LOXL2) that associate with eHsp90 in MDA-MB231 breast cancer cell conditioned media and confirmed that LOXL2 binds to eHsp90 in immunoprecipitates. We introduce a novel impermeant Hsp90 inhibitor STA-12-7191 derived from ganetespib and show that it is markedly less toxic to cells and can inhibit cancer cell migration in a dose dependent manner. We used STA-12-7191 to test if LOXL2 and G3BP are potential eHsp90 clients. We showed that while LOXL2 can increase wound healing and compensate for STA-12-7191-mediated inhibition of wound closure, addition of G3BP had no affect on this assay. These findings support of role for LOXL2 in eHsp90 stimulated cancer cell migration and provide preliminary evidence for the use of STA-12-7191 to inhibit eHsp90 to limit cancer invasion.


Ultrasonic extraction followed by a novel filtration and clean-up device for screening of some polyphenols in tobaccos.
A simple and efficient HPLC method using a diode array detector (DAD) was developed for simultaneous determination of six polyphenols in tobaccos. Ultrasonic extraction was employed at 25 degrees C to extract the polyphenols present in tobaccos into anhydrous methanol. A novel filtration device linked to a clean-up cartridge was designed for simultaneous extract filtration and clean-up. Optimized HPLC-DAD analysis, with multi-wavelength detection, was used for determination of the polyphenols. Because the content of some of the polyphenols is too low to be quantified directly, a concentration step was necessary. Anhydrous methanol was employed for extraction of the polyphenols because of its high extraction efficiency and its low boiling point in the concentration step. Using the proposed method, six polyphenols were quantified in tobaccos (Nicotina tobaccum L.).


Thermoresponsive N,N-dialkylacrylamide copolymer blends as DNA sieving matrices with a thermally tunable mesh size.
In an earlier study we showed that a blend of thermoresponsive and nonthermoresponsive hydroxyalkylcelluloses could be used to create a thermally tunable polymer network for double-stranded (ds) DNA separation. Here, we show the generality of this approach using a family of polymers suited to a wider range of DNA separations: a blended mixture of N,N-dialkylacrylamide copolymers with different thermoresponsive behaviors. A mixture of 47% w/w N,N-diethylacrylamide (DEA)/53% w/w N,N-dimethylacrylamide (DMA) (DEA47; thermoresponsive, transition temperature = 55 degrees C in water) and 30% w/w DEA/70% w/w DMA (DEA30; nonthermoresponsive, transition temperature > 85 degrees C in water) copolymers in the ratio of 1:5 w/w DEA47:DEA30 was used to separate a dsDNA restriction digest (PhiX174-HaeIII). We investigated the effects of changing mesh size on dsDNA separation, as controlled by temperature. We observed good DNA separation performance with the copolymer blend at temperatures ranging from 25 degrees C to 48 degrees C. The separation selectivity was evaluated quantitatively for certain DNA fragment pairs as a function of temperature. The results were compared with those obtained with a control matrix consisting only of the nonthermoresponsive DEA30. Different DNA fragment pairs of various sizes show distinct temperature-dependent selectivities. Over the same temperature range, no significant temperature dependence of selectivity is observed for these DNA fragment pairs in the nonthermoresponsive control matrix. Overall, the results show similar trends in the temperature dependency of separation selectivity to what was previously observed in hydroxyalkylcellulose blends, for the same DNA fragment pairs. Finally, we showed that a ramped temperature scheme enables improved separation in the blended copolymer matrix for both small and large DNA fragments, simultaneously in a single capillary electrophoresis (CE) run.


Raloxifene affects brain activation patterns in postmenopausal women during visual encoding.
Recent brain imaging studies have shown that estrogens alter brain activation patterns upon working memory tasks in postmenopausal women. Estrogens, however, have many systemic side effects. We investigated the effect of the Selective estrogen receptor modulator (SERM) raloxifene on brain activation patterns during a memory task in postmenopausal women with functional magnetic resonance imaging (fMRI). Twenty postmenopausal and right handed women (mean age 65.7 years; SD 3.0) were included in this double blind, placebo controlled and randomized study. Whole brain fMRI was performed before and after three months of daily treatment with raloxifene 60 mg or placebo. Each scanning session consisted of a visual encoding task, a recognition test and a simple photic simulation test. Data analyses was performed with SPM99b software. Specific regions of interest for the tasks were defined based in previous experiments. Visual encoding activated the ventral route, posterior medial temporal lobe and frontal cortex in both groups. Treatment interactions for raloxifene compared to placebo were a decrease in activation in the left parahippocampal gyrus and left lingual gyrus, an increase in activation in the right superior frontal gyrus. The mean recognition test and the simple photic stimulation test showed no treatment interactions. Our results show that raloxifene affects brain activation patterns upon visual encoding in postmenopausal women.


The use of molecular technology to investigate trypanosome infections in tsetse flies at Liwonde Wild Life Reserve.
Trypanosomes are protozoan flagellates that cause human African trypanosomiasis (HAT) and African animal trypanosomiasis (AAT). HAT is caused by Trypanosoma brucei rhodesiense in East and Central Africa and T.b. gambiense in West Africa, whereas AAT is caused by a number of trypanosome species, including T. brucei brucei, T. evansi, T. vivax, T. congolense, T. godfreyi and T. simiae. The aim of this study was to establish if tsetse flies at Liwonde Wild Life Reserve (LWLR) are infected with these trypanosomes and thus pose a risk to both humans and animals within and surrounding the LWLR. A total of 150 tsetse flies were caught. Of these, 82 remained alive after capture and were dissected such that the mid-gut could be examined microscopically for trypanosomes. DNA extractions were performed from both mid-guts and the 68 dead flies using a Qiagen Kit. Amplification techniques involved the Internal Transcriber Spacer 1 (ITS 1) conventional polymerase chain reaction (PCR) with primers designed to identify trypanosome species, and Repetitive Insertion Mobile Element - Loop Mediated Isothermal Amplification (RIME LAMP), a sequence specific to T. brucei. Analysis showed that 79/82 (96.3%) of the mid-guts examined microscopically were positive for trypanosomes and that 75/150 (50%) of the DNA extracts (from the mid-gut, and tsetse fly carcasses) were positive for T. brucei, as determined by the RIME LAMP method. ITS1 PCR further showed that 87/150 (58.0%) flies were positive for trypanosomes, of which 56/87 (64.4%) were T. brucei, 9/87 (10.3%) were T. vivax; 7/87 (8.1%) were T. simiae; 6/87 (6.9%) were T. congolense, and 6/87 (6.9%) were T. godfreyi. Ten samples had a mixture of infections. Our analysis demonstrated a mixture of infections from trypanosome species in tsetse flies at LWLR, and that T. brucei, the species that causes HAT, was the most common. Our study successfully used molecular techniques to demonstrate the presence of T. b. rhodesiense at LWLR, a species that causes HAT in both East and Central Africa.


Tetrasubstituted phenanthrolines as highly potent, water-soluble, and selective G-quadruplex ligands.
Small molecules capable of stabilizing the G-quadruplex (G4) structure are of interest for the development of improved anticancer drugs. Novel 4,7-diamino-substituted 1,10-phenanthroline-2,9-dicarboxamides that represent hybrid structures of known phenanthroline-based ligands have been designed. An efficient synthetic route to the compounds has been developed and their interactions with various G4 sequences have been evaluated by Förster resonance energy transfer (FRET) melting assays, fluorescent intercalator displacement (FID), electrospray ionization mass spectrometry (ESI-MS), and circular dichroism (CD) spectroscopy. The preferred compounds have high aqueous solubility and are strong and potent G4 binders with a high selectivity over duplex DNA; thus, they represent a significant improvement over the lead compounds. Two of the compounds are inhibitors of HeLa and HT1080 cell proliferation.


The effects of medical group practice organizational factors on physicians' use of resources.
Few studies have systematically examined the influence of physician, patient, and practice characteristics on physician-directed use of resources within the overall environment of medical group practices and none have included the practice culture in the analysis. This study analyzes the effects of the structure and culture of medical group practices on the amount of resources used to manage uncomplicated hypertension episodes of care for enrollees in a Minneapolis/St. Paul HMO during 1990. Three findings emerged from this study: (1) resource use for a well-defined episode of care varies much more than one would expect in this highly competitive managed care environment; (2) the culture of the group practice appears to be more important than organizational structure in determining resource use for the treatment of hypertension; and (3) together the culture and structural variables only explain 8 percent of the variance in resource use. The study indicated that medical group practice organizations have less influence on physicians' practice styles than expected. The group practices studied are all located in a highly competitive managed care environment and these conditions should be causing them to create more standardized practice styles among their physicians. However, wide variations in individual physician practice styles account for most of the differences observed. Either much of the unexplained variance in resource use for this episode of care results from unobserved patient and illness characteristics, or managed healthcare is not yet causing medical group practices in Minnesota to challenge physicians' individualistic practice styles.


Multi-host model-based identification of Armillifer agkistrodontis (Pentastomida), a new zoonotic parasite from China.
Pentastomiasis is a rare parasitic infection of humans. Pentastomids are dioecious obligate parasites requiring multiple hosts to complete their lifecycle. Despite their worm-like appearance, they are commonly placed into a separate sub-class of the subphylum Crustacea, phylum Arthropoda. However, their systematic position is not uncontested and historically, they have been considered as a separate phylum. An appraisal of Armillifer agkistrodontis was performed in terms of morphology and genetic identification after its lifecycle had been established in a multi-host model, i.e., mice and rats as intermediate hosts, and snakes (Agkistrodon acutus and Python molurus) as definitive hosts. Different stages of the parasite, including eggs, larvae and adults, were isolated and examined morphologically using light and electron microscopes. Phylogenetic and cluster analysis were also undertaken, focusing on the 18S rRNA and the Cox1 gene. The time for lifecycle completion was about 14 months, including 4 months for the development of eggs to infectious larvae in the intermediate host and 10 months for infectious larvae to mature in the final host. The main morphological difference between A. armillatus and Linguatula serrata is the number of abdominal annuli. Based on the 18S rRNA sequence, the shortest hereditary distance was found between A. agkistrodontis and Raillietiella spp. The highest degree of homology in the Cox 1 nucleic acid sequences and predicted amino acid sequences was found between A. agkistrodontis and A. armillatus. This is the first time that a multi-host model of the entire lifecycle of A. agkistrodontis has been established. Morphologic and genetic analyses supported the notion that pentastomids should be placed into the phylum Arthropoda.


Intervention strategies for laryngotracheitis: impact of extended downtime and enhanced biosecurity auditing.
An outbreak of vaccinal infectious laryngotracheitis (LT) began in 2005 involving 57 ranches of two broiler companies in California. Standard biosecurity, and cleaning and disinfection programs along with vaccination, did not stop the outbreak. Due to the close proximity and number of birds in the same geographic area, the decision was made by both companies to attempt a joint regional and zonal depopulation strategy. The strategy involved extended downtime between flock placements on ranches located within close proximity to one another. This extended downtime on each ranch ranged from 30 to 91 days. An extensive biosecurity audit, with more than 70 items, was implemented. Briefly, this included heating all houses to 37 C for 100 hr, removing the litter, cleaning and disinfecting everything on the ranches, then again heating the houses to 37 C for 100 hr. Used litter was spread on crops away from poultry, or was sent to a litter processor for pasteurization. Extensive surveillance for LT at 28, 35, and 42 days of age was performed on the initial flocks, which had been placed immediately after the extended downtime. Since completion of this plan in early 2008, LT was diagnosed on only two of the previously 57 affected ranches. Those two ranches, and those within close proximity, went through the extended downtime program and biosecurity audit a second time. Currently, both companies are free of LT. This program lends credence to the importance of cooperation between companies to consider all the ranches within close proximity as the population at risk. In the control of LT in broilers, the program also highlights the necessity for extended downtime and enhanced biosecurity auditing of all flocks.


Balancing hope with reality: how neonatal nurses manage the uncertainty of caring for extremely premature babies.
This article aims to explore the ways in which neonatal nurses manage the uncertainty associated with the treatment and outcomes of extremely premature babies. Current literature suggests that survival rates of extremely premature babies have increased; however, the incidence of long-term problems has not decreased among survivors. The outcomes can often not be predicted; therefore, there is much uncertainty associated with survival and outcomes. Neonatal nurses care for babies and families during these times of uncertainty. This article will focus on how neonatal nurses manage and survive the challenges associated with uncertainty when caring for extremely premature babies. Qualitative. This article used a series of interviews in a qualitative study informed by phenomenological insights. The analysis of the interview data involved the discovery of thematic statements and the analysis of the emerging themes. Three themes captured the experience of working with uncertainty from the perspective of the neonatal nurses: 'Everything is fine, then they crash': When the honeymoon is over; '"I don't know which one is going to be fine": it's like a lottery' and 'Balancing hope with reality'. Uncertainty had both positive and negative aspects, because while ever there was uncertainty, there was room for hope. While initial uncertainty of the baby's prognosis and outcome gave the nurses hope, certainty of diagnosis and poor outcome could take that hope away. Increasing survival of extremely premature babies will see neonatal nurses caring for more babies ≤24 weeks gestation. Prematurity has risks associated with life-sustaining treatments. Uncertainty is a reality of life for these babies, their families and the nursing and medical teams who care for them. It is important to recognise the challenges associated with uncertainty.


Adherence to Oral Nutrition Supplementation in Gastrointestinal Cancer Patients: A Systematic Review of the Literature.
The prevalence of malnutrition is high in gastrointestinal (GI) cancer patients. The use of oral nutrition supplementation (ONS) as part of patients' nutritional therapy seems to be effective in the improvement of nutritional status. Nevertheless, oncology patients, experience several symptoms that negatively affect their compliance with ONS products. Τhe aim of this systematic review is to examine the factors affecting compliance with ONS in patients who underwent GI cancer surgery and/or adjuvant treatments. A systematic search was conducted to identify studies published until June 2023 that assessed compliance to ONS in GI cancer patients. Eleven studies fulfilled the eligibility criteria and were included in the analysis. Postoperative compliance with ONS among GI cancer surgery patients ranged between 26.2% and 71.1%, whereas in GI cancer patients receiving chemotherapy the average reported rate was 90.2%. The main reasons for noncompliance were the presence of GI symptoms, such as early satiety, bloating, and diarrhea after ONS consumption, as well as taste alterations that result in aversion to the provided ONS. Frequent monitoring of these patients is crucial in order to record adverse effects, identify patients that are in need of personalized guidance at an early stage and motivate them to follow their ONS plan.


Death and Dialysis After Transcatheter Aortic Valve Replacement: An Analysis of the STS/ACC TVT Registry.
The authors sought to elucidate the true incidence of renal replacement therapy (RRT) after transcatheter aortic valve replacement (TAVR). There is a wide discrepancy in the reported rate of RRT after TAVR (1.4% to 40%). The true incidence of RRT after TAVR is unknown. The STS/ACC TVT (Society of Thoracic Surgeons/American College of Cardiology Transcatheter Valve Therapy) registry was linked to the Centers for Medicare & Medicaid database to identify all patients that underwent TAVR from November 2011 through September 2015 and their outcomes. The authors compared rates of death, new RRT, and a composite of both as a function of pre-procedure glomerular filtration rate (GFR), both in stages of chronic kidney disease (CKD), as well as on a continuous scale. Pre-procedure GFR is associated with the risk of death and new RRT after TAVR when GFR is <60 ml/min/m2, and increases significantly when GFR falls below 30 ml/min/m2. Incremental increases in GFR of 5 ml/min/m2 were statistically significant (unadjusted hazard ratio: 0.71; p < 0.001) at 30 days, and continued to be significant at 1 year when pre-procedure GFR was <60 ml/min/m2. One in 3 CKD stage 4 patients will be dead within 1 year, with 14.6% (roughly 1 in 6) requiring dialysis. In CKD stage 5, more than one-third of patients will require RRT within 30 days; nearly two-thirds will require RRT at 1 year. In both unadjusted and adjusted analysis, pre-procedural GFR was associated with the outcomes of death and new RRT. Increasing CKD stage leads to an increased risk of death and/or RRT. Continuous analysis showed significant differences in outcomes in all levels of CKD when GFR was <60 ml/min/m2. Pre-procedure GFR should be considered when selecting CKD patients for TAVR.


Hemodynamics of distal revascularization-interval ligation.
Distal revascularization-interval ligation (DRIL) empirically corrects steal after arteriovenous fistula (AVF) creation in most cases, but because there is no topologic alteration in anatomy, it is unclear as to why it is effective. To explore this issue, nine symptomatic patients underwent intravascular pressure and flow measurements before and after DRIL following upper arm autologous AVFs. Mean pre-DRIL systolic pressure (mmHg; mean +/- SD) in the proximal brachial artery (PROX) was 102 +/- 17, while that at the AV anastomosis (AV ANAST) was 47 +/- 38 (p < 0.0006). Flow (mL/min) distal to AV ANAST was retrograde with the fistula open (-21 +/- 64) but became antegrade (58 +/- 29; p < 0.03) with occlusion of the fistula. Following DRIL, pressures at both PROX and AV ANAST sites did not change (104 +/- 24 and 51 +/- 43, respectively). However, pressure at the point at which the blood flow split to supply the hand or the fistula, now PROX, increased from 47 +/- 38 (pre-DRIL AV ANAST) to 104 +/- 24 (p < 0.0001). Pressure in the brachial artery distal to the ligature increased to 104 +/- 27 (p < 0.0001), flow at this point (to the hand) became antegrade (51 +/- 39; p < 0.03), and occlusion of the fistula did not significantly change pressure at this site. We hypothesize that improvement in hand perfusion following DRIL is due to a higher pressure at the point at which the blood flow splits to supply both hand and fistula (pre-DRIL: AV ANAST; post-DRIL: PROX), allowing antegrade flow down the new bypass to the lower pressure forearm. This increased pressure must be due to the increased resistance of the fistula created by interposing the arterial segment between the original AV ANAST and new PROX ANAST. As such, DRIL is schematically equivalent to banding, but resistance is increased in a fashion that is physiologically and empirically acceptable.


Integrated community models: A framework combining multispecies data sources to estimate the status, trends and dynamics of biodiversity.
Data deficiencies among rare or cryptic species preclude assessment of community-level processes using many existing approaches, limiting our understanding of the trends and stressors for large numbers of species. Yet evaluating the dynamics of whole communities, not just common or charismatic species, is critical to understanding and the responses of biodiversity to ongoing environmental pressures. A recent surge in both public science and government-funded data collection efforts has led to a wealth of biodiversity data. However, these data collection programmes use a wide range of sampling protocols (from unstructured, opportunistic observations of wildlife to well-structured, design-based programmes) and record information at a variety of spatiotemporal scales. As a result, available biodiversity data vary substantially in quantity and information content, which must be carefully reconciled for meaningful ecological analysis. Hierarchical modelling, including single-species integrated models and hierarchical community models, has improved our ability to assess and predict biodiversity trends and processes. Here, we highlight the emerging 'integrated community modelling' framework that combines both data integration and community modelling to improve inferences on species- and community-level dynamics. We illustrate the framework with a series of worked examples. Our three case studies demonstrate how integrated community models can be used to extend the geographic scope when evaluating species distributions and community-level richness patterns; discern population and community trends over time; and estimate demographic rates and population growth for communities of sympatric species. We implemented these worked examples using multiple software methods through the R platform via packages with formula-based interfaces and through development of custom code in JAGS, NIMBLE and Stan. Integrated community models provide an exciting approach to model biological and observational processes for multiple species using multiple data types and sources simultaneously, thus accounting for uncertainty and sampling error within a unified framework. By leveraging the combined benefits of both data integration and community modelling, integrated community models can produce valuable information about both common and rare species as well as community-level dynamics, allowing for holistic evaluation of the effects of global change on biodiversity.


[Modern concept of peripheral erectile mechanisms].
The article describes the peripheral mechanisms of erection and control over them. This knowledge has been acquired as a result of the recent development of pharmacological research designed to study the regulation of erectile smooth muscle tone. Smooth muscle fibres of the corpora cavernosa and arteries supplying the penis relax in response to a reduction of intracellular calcium. This relaxation allows both an increase of the blood flow to the penis and opening of sinusoid spaces. Cyclic nucleotides, cGMP and cAMP, are intracellular messengers of the mediators acting on smooth muscle fibres and regulating these intracellular calcium movements. Nitric oxide (NO) increases the intracellular cGMP concentration that triggers relaxation. Other proerectile mediators, such as vasoactive intestinal polypeptide, prostaglandin E1, are of the secondary importance. In contrast, neurotransmitters of the sympathetic nervous system (norepinephrine), neuropeptide Y, and endothelin induce contraction of cavernous smooth muscle fibres, thereby opposing erection. Oxygenation of the cavernous tissue is also an important factor in the regulation of local mechanisms of erection. Regulation of calcium sensitivity as well as functioning of intracellulare contact--gap junction are of certain interest. A better understanding of the peripheral pharmacology of erection opens the way to new pathophysiological and therapeutic prospects in the broad context of erectile dysfunction.


Beyond behavior modification: Benefits of social-emotional/self-regulation training for preschoolers with behavior problems.
The current study evaluated the initial efficacy of three intervention programs aimed at improving school readiness in preschool children with externalizing behavior problems (EBP). Participants for this study included 45 preschool children (76% boys; Mage=5.16years; 84% Hispanic/Latino background) with at-risk or clinically elevated levels of EBP. During the summer between preschool and kindergarten, children were randomized to receive three newly developed intervention packages. The first and most cost effective intervention package was an 8-week School Readiness Parenting Program (SRPP). Families randomized into the second and third intervention packages received not only the weekly SRPP, but children also attended two different versions of an intensive kindergarten summer readiness class (M-F, 8a.m.-5p.m.) that was part of an 8-week summer treatment program for pre-kindergarteners (STP-PreK). One version included the standard behavioral modification system and academic curriculum (STP-PreK) while the other additionally contained social-emotional and self-regulation training (STP-PreK Enhanced). Baseline, post-intervention, and 6-month follow-up data were collected on children's school readiness outcomes including parent, teacher, and objective assessment measures. Analyses using linear mixed models indicated that children's behavioral functioning significantly improved across all groups in a similar magnitude. Children in the STP-PreK Enhanced group, however, experienced greater growth across time in academic achievement, emotion knowledge, emotion regulation, and executive functioning compared to children in the other groups. These findings suggest that while parent training is sufficient to address children's behavioral difficulties, an intensive summer program that goes beyond behavioral modification and academic preparation by targeting socio-emotional and self-regulation skills can have incremental benefits across multiple aspects of school readiness.


Personality and gender differences in global perspective.
Men's and women's personalities appear to differ in several respects. Social role theories of development assume gender differences result primarily from perceived gender roles, gender socialization and sociostructural power differentials. As a consequence, social role theorists expect gender differences in personality to be smaller in cultures with more gender egalitarianism. Several large cross-cultural studies have generated sufficient data for evaluating these global personality predictions. Empirically, evidence suggests gender differences in most aspects of personality-Big Five traits, Dark Triad traits, self-esteem, subjective well-being, depression and values-are conspicuously larger in cultures with more egalitarian gender roles, gender socialization and sociopolitical gender equity. Similar patterns are evident when examining objectively measured attributes such as tested cognitive abilities and physical traits such as height and blood pressure. Social role theory appears inadequate for explaining some of the observed cultural variations in men's and women's personalities. Evolutionary theories regarding ecologically-evoked gender differences are described that may prove more useful in explaining global variation in human personality.


Hierarchical Fully Convolutional Network for Joint Atrophy Localization and Alzheimer's Disease Diagnosis Using Structural MRI.
Structural magnetic resonance imaging (sMRI) has been widely used for computer-aided diagnosis of neurodegenerative disorders, e.g., Alzheimer's disease (AD), due to its sensitivity to morphological changes caused by brain atrophy. Recently, a few deep learning methods (e.g., convolutional neural networks, CNNs) have been proposed to learn task-oriented features from sMRI for AD diagnosis, and achieved superior performance than the conventional learning-based methods using hand-crafted features. However, these existing CNN-based methods still require the pre-determination of informative locations in sMRI. That is, the stage of discriminative atrophy localization is isolated to the latter stages of feature extraction and classifier construction. In this paper, we propose a hierarchical fully convolutional network (H-FCN) to automatically identify discriminative local patches and regions in the whole brain sMRI, upon which multi-scale feature representations are then jointly learned and fused to construct hierarchical classification models for AD diagnosis. Our proposed H-FCN method was evaluated on a large cohort of subjects from two independent datasets (i.e., ADNI-1 and ADNI-2), demonstrating good performance on joint discriminative atrophy localization and brain disease diagnosis.


Immersive virtual reality and psychological well-being in adult chronic physical illness: systematic review.
Individuals with chronic physical illness are at increased risk of negative psychological sequelae. Immersive virtual reality (VR) is an emerging treatment that might reduce these negative effects and increase quality of life in individuals with chronic physical illness. To systematically review literature examining the use of immersive VR in adult populations with chronic physical illness to understand: (1) how immersive VR is used to improve psychological well-being of adults with chronic physical illness (2) what effect this immersive VR has on the psychological well-being of adults with chronic physical illness. Systematic literature review and meta-analysis. Searches of Ovid Medline/PubMed, PsycINFO, Embase, Web of Science and Scopus between July 1993 and March 2023 inclusive. 12 811 texts were identified; 31 met the inclusion criteria. Relaxing and engaging immersive VR interventions were shown to be acceptable and feasible among adults with cancer, dementia, cardiovascular disease, kidney disease and multiple sclerosis. Many of the studies reviewed were feasibility or pilot studies and so the evidence about effectiveness is more limited. The evidence, mostly from studies of people with cancer, suggests that immersive VR can have a positive effects on anticipatory anxiety symptoms and pain. Environment-based and game-based relaxing immersive VR offer novel interventions, with beneficial effects among people with cancer and, potentially, beneficial effects in those with other long-term physical illness.


Adoptive transfer of islet antigen-autoreactive T cell clones to transgenic NOD.Ea(d)mice induces diabetes indicating a lack of I-E mediated protection against activated effector T cells.
Transgenic insertion of the MHC class II Ea(d)gene in NOD mice restores I-E expression and prevents T-cell-mediated autoimmune diabetes (IDDM). The specific molecular and cellular mechanisms responsible for the diabetes resistance of transgenic NOD.Ea(d)mice remain unclear. We adoptively transferred islet antigen-specific T cell clones into NOD and transgenic NOD.Ea(d)mice to evaluate the level of protection provided by I-E expression against activated effector T cells. We have found that neither neonatal or 3-5-week-old I-E-expressing NOD.Ea(d)mice can completely inhibit the diabetogenic activities of activated islet antigen-specific T cell clones. These data indicate that Ealpha protein expression in NOD antigen presenting cells (APC) does not reduce islet autoantigen presentation in the context of I-A(g7)below the threshold required for stimulation of effector/memory diabetogenic T cells. Our results suggest that the mechanism of Ealpha protein-mediated diabetes resistance in NOD mice may be "antigen ignorance," in which the quantity of islet autoantigens presented in the context of I-A(g7)by APC is reduced below the threshold required to activate nai;ve islet antigen-specific T cells.


Studies of folic acid compounds in nature. IV. Folic acid compounds in soybeans and cow milk.
Folate compounds in soybean and cow's mild were identified by Sephadex chromatography and differential microbiological assaymsoybean contained mainly monoglutamates (ca. 52%), some diglutamates (ca. 16%), and polyglutamates (pentaglutamates representing the major portion). 5-CHO-H4PteGlun(1-6) constituted 65-70% of total folate activity; Cow's milk contained monoglutamates (60%) and polyglutamates (ranging from di- to hepta-conjugates). In contrast to soybean, 90-95% of milk folate was in the 5-CH3h4pteGlun(1-7) form.


Languages, DBMSs, and expert systems. Software for nurse decision making.
This article sheds some light on the task of selecting software for decision making by nurse administrators. The basics of software are described, defining and distinguishing software, firmware, programs, languages, and systems (including database management and expert systems). A discussion of evolving trends and a short discussion of how to go about selecting software systems to meet your needs are included.


Validation approach for a fast and simple targeted screening method for 75 antibiotics in meat and aquaculture products using LC-MS/MS.
An approach is described to validate a fast and simple targeted screening method for antibiotic analysis in meat and aquaculture products by LC-MS/MS. The strategy of validation was applied for a panel of 75 antibiotics belonging to different families, i.e., penicillins, cephalosporins, sulfonamides, macrolides, quinolones and phenicols. The samples were extracted once with acetonitrile, concentrated by evaporation and injected into the LC-MS/MS system. The approach chosen for the validation was based on the Community Reference Laboratory (CRL) guidelines for the validation of screening qualitative methods. The aim of the validation was to prove sufficient sensitivity of the method to detect all the targeted antibiotics at the level of interest, generally the maximum residue limit (MRL). A robustness study was also performed to test the influence of different factors. The validation showed that the method is valid to detect and identify 73 antibiotics of the 75 antibiotics studied in meat and aquaculture products at the validation levels.


The effect of sulphurous water in patients with osteoarthritis of hand. Double-blind, randomized, controlled follow-up study.
The aim of the study was to demonstrate the effectiveness of sulphurous water in patients with osteoarthritis of the hand. Forty-seven patients with osteoarthritis of the hand were enrolled into the double-blind, randomized, controlled study, satisfying ACR criteria. One group of the patients (n = 24) received balneotherapy, bathing in sulphurous thermal water for 20 min per occasion, 15 times in all during a period of 3 weeks. The control group (n = 21) had a bath exclusively in warm tap water. Assessments were carried out in both groups on four occasions: at the beginning and at the end of the treatment, and 3 and 6 months after the beginning of the treatment. The parameters studied were the following: pain in the hand, morning stiffness in the joints, grip strength of both hands, and Health Assessment Questionnaire Disability Index (HAQ) and AUSCAN Hand Osteoarthritis Index and EuroQol quality of life questionnaire. At the end of treatment, the improvement was more pronounced in the patient group treated with the sulphurous water. After 3 months, significant improvement could be detected in all parameters, except the morning stiffness and EQ5D. After 6 months, the values of pain, HAQ and AUSCAN continued to be significantly better in comparison with the baseline values. The improvement in quality of life was significant only at the end of the treatment, 6 months later not any longer. The difference between the two groups was significant after 3 months in point of pain and EQVAS. Balneotherapy and within this the sulphurous spa water alone may be effective for the attenuation of pain in patients with hand osteoarthrosis.


Sources of career disadvantage in nursing. A study of NHS Wales.
Despite the numerical predominance of women in nursing there is a marked concentration of women, especially those working part-time, in the lower echelons of the profession. The paper presents survey data and interview material from a study of qualified nurses in NHS Wales. By controlling for differences in education and experience in nursing work, it was found that comparable groups of female nurses received unequal employment opportunities. Women with dependent children were primarily located in the lower nurse grades irrespective of their qualifications and experience. Much of this was associated with inflexible working practices, and the low status of part-time work. Occupational downgrading for female returners was also a significant barrier to career advancement. However, these problems were not recognised by management. Management failed to evaluate the mechanics of their human resource policies in terms which matter to many nurses, in particular with regard to the management of diversity.


Melanoma brain metastases treated with stereotactic radiosurgery and concurrent pembrolizumab display marked regression; efficacy and safety of combined treatment.
Brain metastases are common in patients with metastatic melanoma. With increasing numbers of melanoma patients on anti-PD-1 therapy, we sought to evaluate the safety and initial response of brain metastases treated with concurrent pembrolizumab and radiation therapy. From an institutional database, we retrospectively identified patients with melanoma brain metastases treated with radiation therapy (RT) who received concurrent pembrolizumab. Concurrent treatment was defined as RT during pembrolizumab administration period and up to 4 months after most recent pembrolizumab treatment. Response was categorized by change in maximum diameter on first scheduled follow-up MRI. Lesion and patient specific outcomes including response, lesion control, brain control and overall survival were recorded and descriptively compared to contemporary treatments with RT and concurrent ipilimumab or RT without immunotherapy. From January 2014 through December 2015, we identified 21 patients who received concurrent radiation therapy and pembrolizumab for brain metastases or resection cavities that had at least one scheduled follow-up MRI. Eleven underwent stereotactic radiosurgery (SRS), 7 received hypofractionated radiation and 3 had whole brain treatment (WBRT). All treatments were well tolerated with no observed Grade 4 or 5 toxicities; Grade 3 edema and confusion occurred in 1 patient treated with WBRT after prior SRS. For metastases treated with SRS, at first scheduled follow-up MRI (median 57 days post SRS), 70% (16/23) exhibited complete (CR, n = 8) or partial response (PR, n = 8). The intracranial response rates (CR/PR) for patients treated with SRS and concurrent ipilimumab and SRS without concurrent immunotherapy was 32% and 22%, respectively. Concurrent pembrolizumab with brain RT appears safe in patients with metastatic melanoma, and SRS in particular is effective in markedly reducing the size of brain metastases at the time of first follow-up MRI. These results compare favorably to SRS in combination with ipilimumab and SRS without concurrent immunotherapy.


Role of the integrin-associated protein CD9 in binding between sperm ADAM 2 and the egg integrin alpha6beta1: implications for murine fertilization.
CD9 is a tetraspan protein that associates with several beta1 integrins, including alpha6beta1. Because alpha6beta1 is present on murine eggs and interacts with the sperm-surface glycoprotein ADAM 2 (fertilin beta), we first asked whether CD9 is present on murine eggs and whether it functions in sperm-egg binding and fusion. CD9 is present on the plasma membrane of oocytes in the ovary as well as on eggs isolated from the oviduct. The anti-CD9 mAb, JF9, potently inhibits sperm-egg binding and fusion in vitro in a dose-dependent manner. JF9 also disrupts binding of fluorescent beads coated with native fertilin or a recombinant fertilin beta disintegrin domain. (Both ligands bind to the egg via alpha6beta1.) Immunohistochemistry showed that CD9 is undetectable in the uterine epithelium, appears basolaterally and as prominent apical patches on the epithelium in the region between the uterus and the oviduct, and then persists apically in the oviduct. The integrin alpha6A subunit is found in similar apical patches in the region between the uterus and oviduct, but is confined to the basal aspect of the epithelium in the uterus and oviduct. Hence, alpha6A and CD9 both are expressed on the apical epithelial surface at the uterine-oviduct junction. These findings correlate with the observation that fertilin beta "knockout" sperm traverse the uterus but do not progress into the oviduct, contributing to the infertility of fertilin beta(-/-) male mice. Our results suggest that high-avidity binding between fertilin beta (ADAM 2) and alpha6beta1 requires cooperation between alpha6beta1 and CD9. Such cooperation may assist sperm passage into the oviduct as well as sperm-egg interactions.


[Treatment of Goldmann-Favre syndrome with cyclosporin A and bromocriptine].
Up to now several reports have described a disturbance of humoral and cellular immune response in patients with tapetoretinal degeneration, but a pathogenetic relation of the described immunological changes to the retinal disease was not established. Therefore immunosuppressive treatment has not been advocated for degenerative vitreoretinal disease. We now report about the natural course of Goldmann-Favre's vitreo-retinal degeneration and the results of a therapeutical trial with cyclosporin A and bromocriptine in two patients. Without therapy we saw a slowly progressive flat central retinoschisis with a serous macular exsudation in fluorescein-angiography. Additionally we found peripheral vitreoretinal changes, mild cellular vitreal infiltration, extinguished electro-retinogram and disturbed dark adaptation. Visual acuity and visual field were quite stabile. Under therapy with cyclosporin A we found a regression of the macular edema and flattening of the retinoschisis. One of the two cases showed a good improvement of visual acuity, in the other case a clear subjective improvement did not correlate with an increase in visual acuity. An additional therapy with bromocriptine did not bring further success, but we were able to reduce the cyclosporin A to a blood concentration of 70-100 ng per ml.


Endoscopic submucosal dissection for early gastric cancer: It is time to consider the quality of its outcomes.
Endoscopic resection, particularly endoscopic submucosal dissection (ESD), is widely used as a standard treatment modality for early gastric cancer (EGC) when the risk of lymph node metastasis is negligible. Compared with surgical gastrectomy, ESD is a minimally invasive procedure with additional advantages, such as preservation of the entire stomach and maintenance of the patient's quality of life. However, not all patients achieve curative resection after ESD of EGC. Several patients require surgical gastrectomy after ESD to achieve a curative treatment. Additional surgery after ESD, owing to non-curative resection, places considerable emotional and financial burdens on patients. Recently, as the number of endoscopists performing ESD has increased, the rate of non-curative resection after ESD has increased correspondingly. In order to decrease the non-curative resection rate, as well as determine the ideal rate of non-curative resection after ESD, it is time to consider quality indicators for the outcomes of ESD for EGC.


Background and stimulus-induced patterns of high metabolic activity in the visual cortex (area 17) of the squirrel and macaque monkey.
We have used 2-deoxy-D-[14C]glucose (2-DG) autoradiography and cytochrome oxidase histochemistry to examine background and stimulus-induced patterns of metabolic activity in monkey striate cortex. In squirrel monkeys (Saimiri sciureus) that binocularly or monocularly viewed diffuse white light or binocularly viewed bars of many orientations and spatial frequencies, 2-DG consumption was not uniform across the cortex but consisted of regularly spaced radial zones of high uptake. The zones extended through all laminae except IVc beta and, when viewed tangentially, formed separate patches 500 microns apart. The cytochrome oxidase stain in these animals also revealed patches of high metabolism which coincided with the 2-DG patches. Squirrel monkeys binocularly viewing vertical stripes showed parallel bands of increased 2-DG uptake in the cortex, while the cytochrome label in these animals remained patchy. When monkeys were kept in the dark during 2-DG exposure, 2-DG-labeled patches were not seen but cytochrome oxidase-positive patches remained. In macaque (Macaca nemestrina) monkeys, binocular stimulation with many orientations and spatial frequencies produced radial zones of high 2-DG uptake in layers I to IVa and VI. When viewed tangentially, these zones formed a dots-in-rows pattern with a spacing of 350 X 500 microns; cytochrome oxidase staining produced an identical pattern. Macaca differed from Saimiri in that monocular stimulation labeled alternate rows. These results indicate that there are radial zones of high background metabolism across squirrel and macaque monkey striate cortex. In Saimiri these zones do not appear to be related to an eye dominance system, while in Macaca they do. The presence of these zones of high metabolism may complicate the interpretation of 2-DG autoradiographs that result from specific visual stimuli.


Preoperative Assessment of Coronary Arteries by Cardiac Computed Tomography in Patients with Lung Cancer.
To retrospectively evaluate characteristics of coronary artery disease (CAD) in patients undergoing lung resection for cancer by using dual-source computed tomography (DSCT), thus increasing our understanding of perioperative evaluation. DSCT coronary angiography was performed for perioperative evaluation in 145 patients with non-small cell lung cancer (NSCLC) due to suspected CAD. The imaging characteristics of CAD including extent, distribution and types of plaques, and luminal stenosis and congenial anomalous including origin of coronary arteries and myocardial bridge (MB) were evaluated and compared by sex, smoking behavior, and pathological types. In total, coronary artery plaques were observed in 130 (89.7%) patients. A total of 205 coronary vessels (1.7 ± 1.3 per patient) and 278 segments (2.4 ± 2.3 per patient) were found to have plaques, respectively. The most frequently involved segment were the left anterior descending (LAD) artery (45.4%) and its proximal segment (24.1%) (p < 0.05). Mild narrowing (61.7%) and calcified plaques (52.9%) were most found (all p < 0.001). Types of plaques, luminal narrowing and extent were not significantly different by sex, smoking behavior, or pathological type. Anomalous origination of the right coronary ostium from left sinus was observed in 0.7% of patients. MB was observed in 16.6% of patients. Dual-source CT is a useful preoperative noninvasive test for CAD in patients planning to undergo lung resection for cancer.


High-dose combination alkylating agents with bone marrow support as initial treatment for metastatic breast cancer.
To evaluate the effect of high-dose chemotherapy in the treatment of metastatic breast cancer, we performed a phase II trial of a single treatment with high-dose cyclophosphamide (5,625 mg/m2), cisplatin (165 mg/m2), and carmustine (600 mg/m2), or melphalan (40 mg/m2) and bone marrow support as the initial chemotherapy for metastatic breast cancer. Twenty-two premenopausal patients with estrogen receptor negative, measurable metastatic disease were treated. Twelve of 22 patients (54%) obtained a complete response at a median 18 days. The overall response rate is 73% (complete and partial response). Median duration of response in the patients achieving complete response was 9.0 months with a median duration of survival for complete responders that is currently undefined. Relapse occurred predominantly at sites of pretreatment bulk disease or within areas of previous radiation therapy. Toxicity was frequent and five patients died of therapy-related complications. The results indicate that a single treatment with intensive combination alkylating agents with bone marrow support can produce more rapid and frequent complete responses than conventional chemotherapy when used as initial chemotherapy for metastatic breast cancer, although median disease-free and overall survival is not improved. Three patients (14%) remain in unmaintained remission beyond 16 months.


Clinical and penile Doppler outcomes using a modified, tourniquet free, Nesbit plication for severe Peyronie's disease.
Penile curvature (PC) can be surgically corrected by plication techniques or Nesbit corporoplasty. These shortening techniques can be complicated by post-operative: penile shortening, recurrent PC, palpable suture knots and erectile dysfunction. Furthermore, Nesbit procedures require the use of a penile tourniquet to avoid intraoperative bleeding. This observational study aims to assess the results of Nesbit modified corporoplasty, avoiding intraoperative use of tourniquet without risk of bleeding. The objective is to reduce penile ischemic anatomical and functional damages such as long-term erectile dysfunction. Between January 2010 and March 2019, a total of 64 patients with congenital penile curvature (CPC) and Peyronie's disease (PD) underwent surgical correction with a Nesbit modified technique first time described by Rolle et al., with minimal technical differences. The operation notes were retrospectively reviewed. In particular, we evaluated pre- and post-operative erectile functions using IIEF-5 score, penile Doppler ultrasonography and overall patient satisfaction. During operations, no intraoperative bleeding was noted, and no short-term complications such as hematomas or neurovascular bundle lesions were reported. At 6 months, no palpable subcutaneous indurations and no sensory change were detected. Post-operative penile shortening was reported in 38 (59.4%) patients (mean 0.83±0.79 cm), but it did not influence the high overall satisfaction rate of 91.4%. Only 2 patients reported a slightly partial recurrence of curvature (<15%) with no need for a redo surgery. Mean IIEF-5 score increased from 17.1±5.2 to 20.8±3.9 at 6 months and 21.8±3.4 at 12 months (P<0.001 in both cases). Mean PSV also significantly increased at the end of follow-up (28.5±6.1 at baseline vs. 31.0±7.1 at 12 months, P=0.03). Considering the optimal results in terms of erectile functions increasing and absence of PC recurrence (>15°), we think that Nesbit modified corporoplasty without tourniquet application during reconstruction is a safe and effective surgical procedure for all kind of shortening corporoplasty to reduce the time of penile ischemia, preventing even serious consequences for the normal physiology of erection.


Receptive fields of second-order neurons in the olfactory bulb of the hamster.
Electrical stimulation of nerve fibers emerging from different positions of the olfactory epithelium was used to determine the receptive fields for 52 olfactory bulb neurons in the hamster. The responses of olfactory bulb neurons were recorded extracellularly with metal-filled micropipettes. Suprathreshold stimuli (500 microA) were applied to each of eight standard epithelial positions spaced approximately 250 microns apart, and the position, or positions, which, when stimulated, produced a response in the bulb were taken as an index of the neuron's receptive field. The results indicate that most bulb neurons have very localized receptive fields limited to only one or two stimulating positions. Furthermore, there was a statistically significant correlation between the location of a neuron's receptive field in the olfactory epithelium and the recording depth of the neuron in the olfactory bulb (Spearman rank correlation coefficient, rs, 0.67, P < 0.001). These findings demonstrate that in the mammalian olfactory system there exists a topographical projection of input from localized regions in the epithelium onto the second-order neurons in the olfactory bulb.


Wearable Ambulatory Accelerometer System for Estimating Arterial Stiffness: A Pilot Study.
Given the gap between the crucial role of measuring arterial stiffness in cardiovascular disease prevention and the lack of a technology for frequent/continuous measurement to assess it without an operator, we have developed a wearable accelerometer-based system. It estimates local stiffness metrics (Ep, β, and AC) by employing a one-point patient-specific calibration on the features of acceleration plethysmogram (APG) signal. An in-vivo study on 12 subjects was conducted (a) to select suitable ones from the host features on which the calibration could be applied and (b) to assess the feasibility of reliably estimating the stiffness metrics post-exercise when calibrated prior. The acquired APG signals were found to be reliable (SNR > 38 dB) and repeatable (CoV < 10 %). By examining a correlation matrix, it was found that (a-b)/(a"-b") is a potential feature of consideration for calibration against the stiffness. Due to exercise intervention, the local stiffness metrics have physiologically perturbed by a significant amount (p < 0.05), as observed from the reference measurements. Estimated Ep was found to have statistically significant and strong correlation (r = 0.761, p < 0.05) with actual Ep value, whereas statistically significant and moderate correlation were found with estimated β (r = 0.682, p < 0.05) and estimated AC (r = 0.615, p < 0.05) with their respective actual measures. The system demonstrated its ability to estimate post-exercise stiffness metrics using the baseline calibration, even when subject to significant physiological changes.Clinical Relevance- This study reveals the potential of the developed wearable system to be used for continuous stiffness estimation even in the presence of hemodynamic perturbations.


Bacterial Lysis through Interference with Peptidoglycan Synthesis Increases Biofilm Formation by Nontypeable Haemophilus influenzae.
Nontypeable Haemophilus influenzae (NTHi) is an opportunistic pathogen that mainly causes otitis media in children and community-acquired pneumonia or exacerbations of chronic obstructive pulmonary disease in adults. A large variety of studies suggest that biofilm formation by NTHi may be an important step in the pathogenesis of this bacterium. However, the underlying mechanisms involved in this process are poorly elucidated. In this study, we used a transposon mutant library to identify bacterial genes involved in biofilm formation. The growth and biofilm formation of 4,172 transposon mutants were determined, and the involvement of the identified genes in biofilm formation was validated in in vitro experiments. Here, we present experimental data showing that increased bacterial lysis, through interference with peptidoglycan synthesis, results in elevated levels of extracellular DNA, which increased biofilm formation. Interestingly, similar results were obtained with subinhibitory concentrations of β-lactam antibiotics, known to interfere with peptidoglycan synthesis, but such an effect does not appear with other classes of antibiotics. These results indicate that treatment with β-lactam antibiotics, especially for β-lactam-resistant NTHi isolates, might increase resistance to antibiotics by increasing biofilm formation. IMPORTANCE Most, if not all, bacteria form a biofilm, a multicellular structure that protects them from antimicrobial actions of the host immune system and affords resistance to antibiotics. The latter is especially disturbing with the increase in multiresistant bacterial clones worldwide. Bacterial biofilm formation is a multistep process that starts with surface adhesion, after which attached bacteria divide and give rise to biomass. The actual steps required for Haemophilus influenzae biofilm formation are largely not known. We show that interference with peptidoglycan biosynthesis increases biofilm formation because of the release of bacterial genomic DNA. Subinhibitory concentrations of β-lactam antibiotics, which are often prescribed to treat H. influenzae infections, increase biofilm formation through a similar mechanism. Therefore, when β-lactam antibiotics do not reach their MIC in vivo, they might not only drive selection for β-lactam-resistant clones but also increase biofilm formation and resistance to other antimicrobial compounds.


In vitro characterization of mutant yeast RNA polymerase II with reduced binding for elongation factor TFIIS.
We have reported previously the isolation and genetic characterization of mutations in the gene encoding the largest subunit of yeast RNA polymerase II (RNAPII), which lead to 6-azauracil (6AU)-sensitive growth. It was suggested that these mutations affect the functional interaction between RNAPII and transcription-elongation factor TFIIS because the 6AU-sensitive phenotype of the mutant strains was similar to that of a strain defective in the production of TFIIS and can be suppressed by increasing the dosage of the yeast TFIIS-encoding gene, PPR2, RNAPIIs were purified and characterized from two independent 6AU-sensitive yeast mutants and from wild-type (wt) cells. In vitro, in the absence of TFIIS, the purified wt polymerase and the two mutant polymerases showed similar specific activity in polymerization, readthrough at intrinsic transcriptional arrest sites and nascent RNA cleavage. In contrast to the wt polymerase, both mutant polymerases were not stimulated by the addition of a 3-fold molar excess of TFIIS in assays of promoter-independent transcription, readthrough or cleavage. However, stimulation of the ability of the mutant RNAPIIs to cleave nascent RNA and to read through intrinsic arrest sites was observed at TFIIS:RNAPII molar ratios greater than 600:1. Consistent with these findings, the binding affinity of the mutant polymerases for TFIIS was found to be reduced by more than 50-fold compared with that of the wt enzyme. These studies demonstrate that TFIIS has an important role in the regulation of transcription by yeast RNAPII and identify a possible binding site for TFIIS on RNAPII.


[Fabry disease--a diagnostic and therapeutic challenge].
Fabry disease, unlike most other metabolic diseases, is prone to familiar and regional clustering because of X-linked inheritance and normal fertility in affected men. Affected individuals can be offered intravenous enzyme replacement therapy. In two counties in western Norway there are 41 affected individuals, giving a minimum prevalence of the disease of 1 in 17,000. The patient material is briefly presented. We discuss some of the administrative and financial challenges that this type of diseases present in our public health system. The great variability in disease expression and severity within and between families delays the diagnosis and necessitates a broad clinical follow up of affected persons. A false diagnosis is often made (e.g. MS, irritable colon, a psychiatric disorder, idiopathic hypertrophic cardiomyopathy, or kidney failure). The follow up regime is briefly described, with special emphasis on the practical and financial implications of enzyme replacement therapy in Norway.


Sequence changes in six variants of rice tungro bacilliform virus and their phylogenetic relationships.
The DNA of three biological variants, G1, Ic and G2, which originated from the same greenhouse isolate of rice tungro bacilliform virus (RTBV) at the International Rice Research Institute (IRRI), was cloned and sequenced. Comparison of the sequences revealed small differences in genome sizes. The variants were between 95 and 99% identical at the nucleotide and amino acid levels. Alignment of the three genome sequences with those of three published RTBV sequences (Phi-1, Phi-2 and Phi-3) revealed numerous nucleotide substitutions and some insertions and deletions. The published RTBV sequences originated from the same greenhouse isolate at IRRI 20, 11 and 9 years ago. All open reading frames (ORFs) and known functional domains were conserved across the six variants. The cysteine-rich region of ORF3 showed the greatest variation. When the six DNA sequences from IRRI were compared with that of an isolate from Malaysia (Serdang), similar changes were observed in the cysteine-rich region in addition to other nucleotide substitutions and deletions across the genome. The aligned nucleotide sequences of the IRRI variants and Serdang were used to analyse phylogenetic relationships by the bootstrapped parsimony, distance and maximum-likelihood methods. The isolates clustered in three groups: Serdang alone; Ic and G1; and Phi-1, Phi-2, Phi-3 and G2. The distribution of phylogenetically informative residues in the IRRI sequences shared with the Serdang sequence and the differing tree topologies for segments of the genome suggested that recombination, as well as substitutions and insertions or deletions, has played a role in the evolution of RTBV variants. The significance and implications of these evolutionary forces are discussed in comparison with badnaviruses and caulimoviruses.


Overlapping generations can promote altruistic behavior.
We use an inclusive fitness model to study the evolution of altruism in a patch-structured population in which there is positive probability of breeder survival from one generation to the next. We find first that breeder survival promotes altruism and second that there is a marked difference between benefits of fecundity and benefits of survival. Under the first altruism is more strongly favored, and under the second altruism is less strongly favored than in a randomly mixing population.


Altered adrenergic control in hypercholesterolemia.
Vascular reactivity is altered by hypercholesterolemia and atherosclerosis. Animal studies have shown that serotonergic mechanisms of vasoconstriction are enhanced, while the changes with adrenergic mechanisms are still equivocal. Serum lipids are increased with beta-blocker and diuretic therapy for hypertension. Thus, the increase in serum lipids may be responsible for both structural and functional abnormalities of the vascular system in hypertensive patients. In contrast, recent reports have suggested that alpha 1-adrenergic receptor inhibition, useful in hypertension therapy, may exert a beneficial effect on plasma lipids. We have also collected preliminary data in our laboratory examining the effects of alpha 1-adrenergic receptor inhibition with prazosin on plasma cholesterol levels and vascular reactivity in monkeys with hypercholesterolemia. We observed large reductions in both total cholesterol and the very-low-density lipoprotein (VLDL) + low-density lipoprotein (LDL) fraction of cholesterol in these animals during treatment with prazosin. Thus, in addition to its sympatholytic action on the vasculature, alpha 1-adrenergic receptor inhibition appears to exert a beneficial effect in reducing total cholesterol levels.


Characteristics of mental health trials registered in ClinicalTrials.gov.
The ClinicalTrials.gov registry was established in 2000 to address concerns about publication bias and public access to information about clinical trials. We aimed to evaluate differences between for-profit and non-profit sponsors of efficacy mental health trials registered in ClinicalTrials.gov on key trial characteristics that relate to data integrity. We also sought to evaluate whether the registry is fulfilling its purpose as a means of promoting transparency between researchers and the public by providing complete and quality information about the trials it contains. We found that trials tend to be small, use a placebo instead of an active comparator, and employ randomization and blinding. We discuss the implications of these design characteristics and the limitations of the registry.


[Food security in Mexico].
An overview of food security and nutrition in Mexico is presented, based on the analysis of the four pillars of food security: availability, access, utilization of food, and stability of the food supply. In addition, the two faces of malnutrition in Mexico were analyzed: obesity and undernourishment. Data were gathered from the food security indicators of the United Nations's Food and Agriculture Organization, from the Mexican Scale of Food Security, and from the National Health and Nutrition Survey. Mexico presents an index of availability of 3 145 kilocalories per person per day, one of the highest indexes in the world, including both food production and imports. In contrast, Mexico is affected by a double burden of malnutrition: whereas children under five present 14% of stunt, 30% of the adult population is obese. Also, more than 18% of the population cannot afford the basic food basket (food poverty). Using perception surveys, people reports important levels of food insecurity, which concentrates in seven states of the Mexican Federation. The production structure underlying these indicators shows a very heterogeneous landscape, which translates in to a low productivity growth across the last years. Food security being a multidimensional concept, to ensure food security for the Mexican population requires a revision and redesign of public productive and social policies, placing a particular focus on strengthening the mechanisms of institutional governance.


Evaluating the role of spatial landscape literacy in public participation processes and opinions on environmental issues and ecosystem services.
Little is known about how spatial literacy influences mapping and decision-making related to environmental management and ecosystem services. We assessed the role of spatial landscape literacy in participatory environmental management and ecosystem service assessments in peri-urban Bogota, Colombia. Spatial landscape literacy (SL) was evaluated by testing stakeholder's ability to locate specific landscapes and landmarks. We then assessed if opinions on environmental decisions and ecosystem services were significantly related to SL. We used an online instrument to capture 2,397 respondent's socioeconomic characteristics, SL, ecosystem service perceptions, and opinions concerning relevant environmental issues. We evaluated and measured respondents' self-perceived SL and ability to locate four landscapes in an integrated online map. Positional accuracy was calculated using a Spatial Landscape Literacy Index (SLI). We then tested for effects of socio-demographics on SLI, modeled the relationship between socio-demographics and SL, and tested the relationship between respondents' SL and their opinions on relevant environmental issues and ecosystem services. We found that about 75% of the respondents correctly located 2 of the 4 landscapes. The SLI model was also poorly predicted by socio-demographic variables. However, we found significant relationships between SLI and opinions concerning the environment. No relationship was found between respondents' levels of active participation in local governance and SLI. Overall, SL was little affected by education levels. Participatory processes using maps should ideally measure SL and not assume a priori that participants are spatially literate. Further research is needed to evaluate how spatial technologies and understanding stakeholder's values towards the environment can democratize participatory-based decision-making.


Modulation of pulmonary defense mechanisms against viral and bacterial infections by acute exposures to nitrogen dioxide.
The scientific literature suggests that ambient levels of nitrogen dioxide increase susceptibility to respiratory infections. However, this association has not been conclusively demonstrated. The epidemiologic data regarding this relationship are inconclusive because these studies have used parameters of "acute respiratory illness" that are not necessarily related to infectious episodes. Previous animal studies have used either mortality after bacterial infection with virulent bacteria or decreased rate of intrapulmonary killing of bacteria with low virulence. Studies using appropriate bacterial and viral challenge organisms, with morbidity as an endpoint, provide a better basis for extrapolation to humans. The investigations in animals suggest a relationship between nitrogen dioxide and increased susceptibility to respiratory infection, but studies in which functional parameters of host resistance to such infections have been used are few. The aim of this work was to determine the threshold level of acute nitrogen dioxide exposure that would induce increased susceptibility to, and increased severity of, viral and bacterial infections. Physiologic parameters of host resistance to respiratory infections were used as endpoints. A composite picture was developed of dose-response relationships between nitrogen dioxide and the impairment of a spectrum of defense parameters in the murine respiratory tract against viral and bacterial challenges. The salient findings of this study are as follows: (1) the intrapulmonary killing of Staphylococcus aureus was impaired at 5 ppm of nitrogen dioxide; (2) this effect was found at 2.5 ppm or less when nitrogen dioxide exposure was superimposed on lungs predisposed to lowered resistance through immunosuppression with corticosteroids; (3) the adverse effect of nitrogen dioxide occurred at lower concentrations when exposure followed bacterial challenge; and (4) during the course of murine Sendai virus infection, exposure to nitrogen dioxide for four hours per day did not alter the infection in the lungs, but rather it enhanced lung pathology. The implications of these findings are that the antibacterial defenses of the lungs are susceptible to the inhibiting effects of short acute exposures of lower concentrations of nitrogen dioxide when the lungs are predisposed by bacteria present or, even more so, by immunosuppression. The alveolar macrophage phagocytic system is the defense component of the lungs that is most susceptible to the adverse effects of nitrogen dioxide. The finding that nitrogen dioxide increases virus-associated lung damage suggests that the increased severity of the disease process results from the proliferation of the virus to high titers, rather than from alterations of the infective process.


Delayed correlation of mRNA and protein expression in rapamycin-treated cells and a role for Ggc1 in cellular sensitivity to rapamycin.
To identify new molecular targets of rapamycin, an anticancer and immunosuppressive drug, we analyzed temporal changes in yeast over 6 h in response to rapamycin at the transcriptome and proteome levels and integrated the expression patterns with functional profiling. We show that the integration of transcriptomics, proteomics, and functional data sets provides novel insights into the molecular mechanisms of rapamycin action. We first observed a temporal delay in the correlation of mRNA and protein expression where mRNA expression at 1 and 2 h correlated best with protein expression changes after 6 h of rapamycin treatment. This was especially the case for the inhibition of ribosome biogenesis and induction of heat shock and autophagy essential to promote the cellular sensitivity to rapamycin. However, increased levels of vacuolar protease could enhance resistance to rapamycin. Of the 85 proteins identified as statistically significantly changing in abundance, most of the proteins that decreased in abundance were correlated with a decrease in mRNA expression. However, of the 56 proteins increasing in abundance, 26 were not correlated with an increase in mRNA expression. These protein changes were correlated with unchanged or down-regulated mRNA expression. These proteins, involved in mitochondrial genome maintenance, endocytosis, or drug export, represent new candidates effecting rapamycin action whose expression might be post-transcriptionally or post-translationally regulated. We identified GGC1, a mitochondrial GTP/GDP carrier, as a new component of the rapamycin/target of rapamycin (TOR) signaling pathway. We determined that the protein product of GGC1 was stabilized in the presence of rapamycin, and the deletion of the GGC1 enhanced growth fitness in the presence of rapamycin. A dynamic mRNA expression analysis of Deltaggc1 and wild-type cells treated with rapamycin revealed a key role for Ggc1p in the regulation of ribosome biogenesis and cell cycle progression under TOR control.


Celiac disease is diagnosed less frequently in young adult males.
The female predominance in celiac disease is difficult to explain because population-based screening studies reveal similar rates for celiac disease-specific autoantibodies in males and females. The aim of this study was to explore the role of age and gender in the presentation of celiac disease. The frequency of presentation according to age, gender and mode of presentation was determined by analysis of a prospectively maintained database of children and adults seen at a tertiary medical center. Of 1,682 patients (68 % female) aged 3 months to 86 years who were diagnosed with celiac disease, age at diagnosis in females peaked at 40-45 years, whereas the age at diagnosis for males had two peaks: 10-15 and 35-40 years. A significantly lower percentage of males in early adulthood were diagnosed compared with males in all other age groups (P < 0.0001). The young and elderly had a more even gender distribution. Based on our analysis, males are diagnosed with celiac disease less frequently than females, especially in early adulthood. There should be more emphasis on the diagnosis of celiac disease among young adult males.


Advancements in the management of pancreatic cancer: 2013.
Pancreatic cancer still remains a significant, unresolved therapeutic challenge and is the most lethal type of gastrointestinal cancer with a 5-year survival rate of 5%. Adjuvant chemotherapy remains to be gemcitabine alone, though fluorouracil offers the same survival and role of radiation remains controversial. Nevertheless, only a few patients survive for at least 5 years after R0 resection and adjuvant therapy. Borderline resectable pancreatic cancer remains an area that requires multi-disciplinary approach. Neo-adjuvant therapy very likely plays a role to downstage to a resectable state in these subgroup patients. There are different treatment approaches to locally advanced pancreatic cancer management, including single or multi-agent chemotherapy, chemotherapy followed by chemoradiation, or immediate concurrent chemoradiation. Most patients need palliative treatment. Once pancreatic cancer becomes metastatic, it is uniformly fatal with an overall survival of generally 6 months from time of diagnosis. Gemcitabine has been the standard since 1997. FOLFIRINOX (5-fluorouracil, oxaliplatin, irinotecan, leucovorin) has already shown superiority over gemcitabine in both progression-free survival and overall survival, but this regimen is suitable only for selected patients in ECOG performance status 0-1. FOLFIRINOX has already trickled down to the clinic in various modifications and in different patient groups, both locally advanced and metastatic. Many targeted agents, including bevacizumab, cetuximab showed negative results, except mild benefit with addition of erlotinib with gemcitabine, which was not considered clinically significant. There is no consensus regarding treatment in the second-line setting. It will be true to say that there was a real medical breakthrough with regards to improving the prognosis of pancreatic cancer as of 2013 with the results of MPACT study. In this study, patients who received nab-paclitaxel plus gemcitabine lived a median of 8.5 months, compared with 6.7 months for those who received gemcitabine alone. At the end of one year, 35% of those getting nab-paclitaxel were alive, compared with 22% of those getting only gemcitabine. After two years, the figures were 9% for those getting nab-paclitaxel and 4% for those who received gemcitabine.


The C-terminus of substrates is critical but not sufficient for their degradation by the Pseudomonas aeruginosa CtpA protease.
Bacterial carboxyl-terminal processing proteases (CTPs) are widely conserved and have been linked to important processes including signal transduction, cell wall metabolism, and virulence. However, the features that target proteins for CTP-dependent cleavage are unclear. Studies of the Escherichia coli CTP Prc suggested that it cleaves proteins with non-polar and/or structurally unconstrained C-termini, but it is not clear if this applies broadly. Pseudomonas aeruginosa has a divergent CTP, CtpA, which is required for virulence. CtpA works in complex with the outer membrane lipoprotein LbcA to degrade cell wall hydrolases. Here, we investigated if the C-termini of two non-homologous CtpA substrates are important for their degradation. We determined that these substrates have extended C-termini, compared to their closest E. coli homologs. Removing seven amino acids from these extensions was sufficient to reduce their degradation by CtpA both in vivo and in vitro Degradation of one truncated substrate was restored by adding the C-terminus from the other, but not by adding an unrelated sequence. However, modification of the C-terminus of non-substrates, by adding the C-terminal amino acids from a substrate, did not cause their degradation by CtpA. Therefore, the C-termini of CtpA substrates are required but not sufficient for their efficient degradation. Although C-terminal truncated substrates were protected from degradation, they still associated with the LbcA•CtpA complex in vivo Therefore, degradation of a protein by CtpA requires a C-terminal-independent interaction with the LbcA•CtpA complex, followed by C-terminal-dependent degradation, perhaps because CtpA normally initiates cleavage at a C-terminal site.IMPORTANCE Carboxyl-terminal processing proteases (CTPs) are found in all three domains of life, but exactly how they work is poorly understood, including how they recognize substrates. Bacterial CTPs have been associated with virulence, including CtpA of Pseudomonas aeruginosa, which works in complex with the outer membrane lipoprotein LbcA to degrade potentially dangerous peptidoglycan hydrolases. We report an important advance by revealing that efficient degradation by CtpA requires at least two separable phenomena, and that one of them depends on information encoded in the substrate C-terminus. A C-terminal-independent association with the LbcA•CtpA complex is followed by C-terminal-dependent cleavage by CtpA. Increased understanding of how CTPs target proteins is significant, due to their links to virulence, peptidoglycan remodeling, and other important processes.