Epigallocatechin-3-gallate suppresses hemin-aggravated colon carcinogenesis through Nrf2-inhibited mitochondrial reactive oxygen species accumulation.
Previous studies have presented evidence to support the significant association between red meat intake and colon cancer, suggesting that heme iron plays a key role in colon carcinogenesis. Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, exhibits anti-oxidative and anti-cancer effects. However, the effect of EGCG on red meat-associated colon carcinogenesis is not well understood. We aimed to investigate the regulatory effects of hemin and EGCG on colon carcinogenesis and the underlying mechanism of action. Hemin and EGCG were treated in Caco2 cells to perform the water-soluble tetrazolium salt-1 assay, lactate dehydrogenase release assay, reactive oxygen species (ROS) detection assay, real-time quantitative polymerase chain reaction and western blot. We investigated the regulatory effects of hemin and EGCG on an azoxymethane (AOM) and dextran sodium sulfate (DSS)-induced colon carcinogenesis mouse model. In Caco2 cells, hemin increased cell proliferation and the expression of cell cycle regulatory proteins, and ROS levels. EGCG suppressed hemin-induced cell proliferation and cell cycle regulatory protein expression as well as mitochondrial ROS accumulation. Hemin increased nuclear factor erythroid-2-related factor 2 (Nrf2) expression, but decreased Keap1 expression. EGCG enhanced hemin-induced Nrf2 and antioxidant gene expression. Nrf2 inhibitor reversed EGCG reduced cell proliferation and cell cycle regulatory protein expression. In AOM/DSS mice, hemin treatment induced hyperplastic changes in colon tissues, inhibited by EGCG supplementation. EGCG reduced the hemin-induced numbers of total aberrant crypts and malondialdehyde concentration in the AOM/DSS model. We demonstrated that EGCG reduced hemin-induced proliferation and colon carcinogenesis through Nrf2-inhibited mitochondrial ROS accumulation.


Regulation of choline sulphatase synthesis and activity in Aspergillus nidulans.
1. Choline O-sulphate is taken up from the growth medium to the same extent by sulphur-deficient and sulphur-sufficient mycelia of Aspergillus nidulans, but hydrolysis of the transported sulphate ester in vivo only occurs in the sulphur-deficient mycelia. 2. Choline sulphatase activity could not be detected in vitro in sulphur-sufficient mycelia of wild-type and sulphur mutants of A. nidulans, but after sulphur starvation all strains showed appreciable activity of this enzyme. 3. Optimum activity of choline sulphatase in an ultrasonically treated preparation of sulphur-deficient mycelia was at pH7.5. The optimum substrate concentration was in excess of 25mm and K(m) was 0.035m. The enzyme was completely inhibited by 10mm-SO(3) (2-), PO(4) (3-), CN(-) and cysteine. 4. Growth of sulphur-deficient mycelia on various sulphur sources resulted in a decrease of choline sulphatase activity in vitro. The decrease appeared to be due to a repression of choline sulphatase synthesis rather than to inhibition of activity. De-repression by growth on a sulphur-deficient medium was prevented by cycloheximide. Unlike the choline sulphatase of bacteria the fungal enzyme did not need to be substrate-induced. 5. By using sulphur mutants the identity of the co-repressor was limited to S(2)O(3) (2-), cysteine-S-sulphonate, cysteine or compounds derived directly from them. Circumstantial evidence suggests that the co-repressor is cysteine. 6. Inhibition of choline sulphatase activity in vivo was demonstrated with cysteine as the sulphur source for growth.


Is procedural memory relatively spared from age effects?
Numerous types of age-related deficits in the nervous system have been well documented. While a distinction between general types of memories that are susceptible to compromise with advanced age has been fairly well agreed upon, it is often difficult to determine exactly which specific processes are detrimentally influenced. In this study, we used a paradigm that enabled us to distinguish between effects associated with gross motor deficits and those due to learning and memory of a motor skill, per se. In terms of both latency and errors, senescent animals were, on average, impaired in their ability to traverse an elevated obstacle course, compared to younger animals. Yet, if gross motor abilities are accounted for, a fraction of these deficits is readily explained. Moreover, if individual baseline performance differences are normalized, no memory differences are evident across age groups. These observations suggest that memory for a procedural task is not impaired with advanced age.


Socio-demographic factors and health status of adults with disability in Enugu Metropolis, Nigeria.
Disability is a major determinant of impaired health and nutritional status. This study aims to assess the health and nutritional status of adults with disability and their relationship with socio-demographic factors. A descriptive cross-sectional study of 323 adults with disability in support-centers/schools of disability in Enugu Metropolis, Nigeria was conducted. The participants' socio-demographic factors, behavioural characteristics and 24-hour dietary recall were recorded. Blood pressure and anthropometric measurements of height, weight, waist and hip circumference were obtained. The height and weight measurements of non-ambulatory participants were estimated from knee height and mid-arm circumference. Biochemical analyses of blood samples were also performed. Multiple logistic regression was used to assess the effect of socio-demographic factors on health and nutritional status. The participants consisted of females (59.3%) within the age of 20 to 30 years (59.1%). The major area of difficulty was in physical mobility (51.1%) and this occurred mostly in females (26.9%). The participants' mean daily intakes of calorie, protein and fat were below the recommended dietary allowances. The participants were overweight (49.2%), obese (4.6%), hypertensive (29.7%) and diabetic (12.1%). Dyslipidemia (81.8%), anemia (63.6%) and zinc deficiency (51.1%) were highly prevalent among the study group. Gender difference was observed in alcohol consumption (p=0.000), smoking habit (p=0.001), waist circumference (WC)(p=0.000), waist-hip-ratio (WHR) (p=0.000), triglyceride (p=0.026) and haemoglobin concentration (p=0.007). Being boarder was a positive predictor of overweight/obesity (OR= 2.974, 95% CI=1.449-6.104), abnormal WHR (OR=2.893, 95% CI = 1.073-7.801) and hypertension (OR=8.381, 95% CI=1.598-13.959). Female gender was associated with abnormal WC (OR=7.219, 95% CI=3.116-14.228) and WHR (OR=3.590, 95% CI=2.095-6.150) whereas older age-group was associated with overweight/obesity (OR=1.908, 95% CI=1.137- 3.202). Being employed was a negative predictor of hypertension. Overweight/obesity, anemia, zinc deficiency and dyslipidemia were highly prevalent among persons living with disability in Enugu Metropolis.


MiR-140-3p Impedes Gastric Cancer Progression and Metastasis by Regulating BCL2/BECN1-Mediated Autophagy.
MiRNAs have been proven to modulate the progression of gastric cancer (GC). In this field, we evaluated the role and mechanism of miR-140-3p in GC. Western blotting and qRT-PCR were used to detect the levels of miR-140-3p and BCL2. The interaction of miR-140-3p and BCL2 was confirmed by dual-luciferase reporter and miRNA pull-down assays. CCK-8, EdU, wound healing, and Transwell invasion assays were performed to evaluate cell proliferation, migration and invasion. Autophagy was analyzed using Western blot analysis of the LC3-II/I ratio and immunofluorescence staining. A xenograft model was established to reveal the role of miR-140-3p in tumorigenesis. In GC cell lines and tissues, miR-140-3p was highly expressed, and BCL2 was expressed at low levels. MiR-140-3p directly inhibited BCL2 expression and indirectly promoted BECN1 expression, and BCL2 inhibited BECN1 expression. MiR-140-3p overexpression or silencing restrained or facilitated migration, invasion and EMT in GC cells. Moreover, we noticed that overexpression or downregulation of miR-140-3p promoted or suppressed BECN1-dependent autophagy in GC cells. BCL2 introduction or BECN1 silencing in GC cells partially blocked the effects of miR-140-3p. In conclusion, miR-140-3p directly downregulated the expression of BCL2, BCL2 downregulation further activated BECN1-dependent autophagy, and autophagy activation further inhibited EMT. miR-140-3p may act as a tumor suppressor by targeting BCL2 and regulating downstream BECN1-induced autophagy and metastasis in GC progression.


Calling, texting, and searching for information while riding a motorcycle: A study of university students in Vietnam.
The objective of this study was to investigate the prevalence of calling, texting, and searching for information while riding a motorcycle among university students and the influences of sociodemographic characteristics, social norms, and risk perceptions on these behaviors. Students at 2 university campuses in Hanoi and Ho Chi Minh City, the 2 largest cities in Vietnam, were invited to participate in an anonymous online survey. Data collection was conducted during March and May 2016. There were 741 respondents, of whom nearly 90% of students (665) were motorcycle riders. Overall prevalence of mobile phone use while riding is 80.9% (95% confidence interval [CI], 77.9-83.9%) with calling having a higher level of prevalence than texting or searching for information while riding: 74% (95% CI, 70.7-77.3%) vs. 51.7% (95% CI, 47.9-55.5%) and 49.9% (95% CI, 46.1-53.7%), respectively. Random parameter ordered probit modeling results indicate that mobile phone use while riding is associated with gender, motorcycle license duration, perceived crash risk, perceived risk of mobile phone snatching, and perceptions of friends' mobile phone use while riding. Mobile phone use while riding a motorcycle is highly prevalent among university students. Educational programs should focus on the crash and economic risk of all types of mobile phone use while riding, including calling, texting, and searching for information. In addition, they should consider targeting the influence of social norms and peers on mobile phone use while riding.


Contributions of supercritical fluid technology for advancing decellularization and postprocessing of viable biological materials.
The demand for tissue and organ transplantation worldwide has led to an increased interest in the development of new therapies to restore normal tissue function through transplantation of injured tissue with biomedically engineered matrices. Among these developments is decellularization, a process that focuses on the removal of immunogenic cellular material from a tissue or organ. However, decellularization is a complex and often harsh process that frequently employs techniques that can negatively impact the properties of the materials subjected to it. The need for a more benign alternative has driven research on supercritical carbon dioxide (scCO2) assisted decellularization. scCO2 can achieve its critical point at relatively low temperature and pressure conditions, and for its high transfer rate and permeability. These properties make scCO2 an appealing methodology that can replace or diminish the exposure of harsh chemicals to sensitive materials, which in turn could lead to better preservation of their biochemical and mechanical properties. The presented review covers relevant literature over the last years where scCO2-assisted decellularization is employed, as well as discussing major topics such as the mechanism of action behind scCO2-assisted decellularization, CO2 and cosolvents' solvent properties, effect of the operational parameters on decellularization efficacy and on the material's properties.


[Characterization of bacteria of the genera Mycobacterium, Rhodococcus and Nocardia isolated from environmental objects in the Republic of Daghestan].
The bacteriological study of specimens taken from environmental objects, biomaterials from animals and sputa from tuberculosis patients revealed the ubiquity of Mycobacterium, Nocardia and rhodococci obtained from different kinds of environment in the Republic of Daghestan. In the identification 83.6% out of the isolated atypical Mycobacterium cultures were classified with Ranion groups II and IV. Among Nocardia, the most widespread organisms were N. asteroides (62.7%) and among rhodococci, R. erithropolis (61.9%).


E-cigarette use and respiratory disorder in an adult sample.
Little evidence is available on the association of e-cigarettes with health indices. We investigated the association of e-cigarette use with diagnosed respiratory disorder among adults in data from the Behavioral Risk Factor Surveillance Survey (BRFSS). The 2016 Hawaii BRFSS, a cross-sectional random-dial telephone survey, had 8087 participants; mean age was 55 years. Items asked about e-cigarette use, cigarette smoking, and being diagnosed by a health professional with (a) asthma or (b) chronic obstructive pulmonary disease. Multivariable analyses tested associations of e-cigarette use with the respiratory variables controlling for smoking and for demographic, physical, and psychosocial variables. Controlling for the covariates and smoking there was a significant association of e-cigarette use with chronic pulmonary disorder in the total sample (AOR = 2.58, CI 1.36-4.89, p < 0.01) and a significant association with asthma among nonsmokers (AOR = 1.33, CI 1.00-1.77, p < 0.05). The associations were stronger among nonsmokers than among smokers. Results were similar for analyses based on relative risk and absolute risk. There was also a greater likelihood of respiratory disorder for smokers, females, and persons with overweight, financial stress, and secondhand smoke exposure. This is the first study to show a significant independent association of e-cigarette use with chronic respiratory disorder. Several aspects of the data are inconsistent with the possibility that e-cigarettes were being used for smoking cessation by persons with existing respiratory disorder. Theoretical mechanisms that might link e-cigarettes use and respiratory symptoms are discussed.


Symptomatic Outcome after Bone-only Suboccipital Decompression in Adult Patients with Chiari Type I Malformations in the Absence of Hydromyelia or Hydrocephalus.
Background Type I Chiari malformation presents without an associated hydromyelia in 30 to 70% of cases, yet there is no agreement regarding the optimal surgical treatment for these patients. We review our experience for treating symptomatic adult type I Chiari malformation without hydromyelia using a suboccipital bone decompression of the hindbrain and no duraplasty in 12 adult patients. Outcome was measured according to the Chicago Chiari Outcome Scale (CCOS). Results Nine of 12 patients were female; average age at surgery was 34.4 years (range: 17-67 years). Average duration of symptoms prior to surgery was 9.6 years (2 months-29 years). The most common symptom was head and/or neck pain (11/12 patients). All patients additionally presented with at least one non-pain symptom. Mean degree of tonsillar herniation on magnetic resonance imaging was 6.8 mm (range: 5-12 mm) below McRae's line. Operative time was on average 68 minutes (range: 47-120 minutes). No surgical complications were noted in any patient. Length of hospital stay was 2 days (1 overnight) for all patients. Mean follow-up was 167 weeks (range: 13-378 weeks). CCOS for all patients on average was 14.50 (range: 12-16). Pain symptoms underwent improvement (7/11 [63.6%]) or complete resolution (4/11 [36.4%]) in all affected patients. A shorter duration of preoperative symptoms significantly correlated with a better CCOS (p = 0.03). Degree of tonsillar herniation had no significant effect on CCOS (p = 0.67). Of non-pain symptoms, paresthesias/dysesthesias and visual symptoms improved or resolved completely in all affected patients. No patient experienced a worsening of either pain or non-pain symptoms. Conclusion In the subset of adult patients with a type I Chiari malformation and no associated hydromyelia, a craniectomy without an additional opening of the dura may achieve good overall results according to the CCOS.


Exposure to violence and psychological well-being over time in children affected by HIV/AIDS in South Africa and Malawi.
Many of the risk factors for violence against children are particularly prevalent in families and communities affected by HIV/AIDS. Yet, in sub-Saharan Africa, where HIV rates are high, efforts to prevent or address violence against children and its long-lasting effects are hampered by a lack of evidence. We assessed the relationship between violence exposure and mental health among HIV-affected children attending community-based organisations in South Africa (n = 834) and Malawi (n = 155, total sample n = 989) at baseline and 12-15-month follow-up. Exposure to violence in the home and in the community was high. HIV-negative children who lived with an HIV-positive person experienced most violence overall, followed by HIV-positive children. Children unaffected by HIV experienced least violence (all p < .05). Interpersonal violence in the home predicted child depression (β = 0.17, p < .001), trauma symptoms (β = 0.17, p < .001), lower self-esteem (β = -0.17, p < .001), and internalising and externalising behavioural problems (β = 0.07, p < .05), while exposure to community violence predicted trauma symptoms (β = 0.16, p < .001) and behavioural problems (β = 0.07, p < .05). Harsh physical discipline predicted lower self-esteem (β = -0.18, p < .001) and behavioural problems for children (β = 0.24, p < .001). Exposure to home (OR: 1.89, 95% CI: 1.23-2.85) and community violence predicted risk behaviour (OR: 2.39, 95% CI: 1.57-3.62). Over time, there was a decrease in depressed mood and problem behaviours, and an increase in self-esteem for children experiencing different types of violence at baseline. This may have been due to ongoing participation in the community-based programme. These data highlight the burden of violence in these communities and possibilities for programmes to include violence prevention to improve psychosocial well-being in HIV-affected children.


Effects of predation pressure and resource use on morphological divergence in omnivorous prey fish.
Body shape is one of the most variable traits of organisms and responds to a broad array of local selective forces. In freshwater fish, divergent body shapes within single species have been repeatedly observed along the littoral-pelagic axes of lakes, where the structural complexity of near shore habitats provides a more diverse set of resources compared to the open-water zones. It remains poorly understood whether similar resource-driven polymorphism occurs among lakes that vary in structural complexity and predation pressure, and whether this variation is heritable. Here, we analyzed body shape in four populations of omnivorous roach (Rutilus rutilus) inhabiting shallow lakes. We tested the relationship between body shape, gradients of resources, predation pressure, and, in a subset of two lakes, diet composition. We used genome scans of 331 polymorphic AFLP markers to test whether there was a heritable component to the observed morphological diversification. Body shape differed among lakes and was significantly correlated to differences in predation pressure. Roach from the lake with highest predation pressure were most divergent from the average body shape of all populations, characterized by a more streamlined body and caudally inserted dorsal fins; features that facilitate predator escape. Surprisingly, diet composition was not associated with morphology. AFLP analysis revealed weak genetic differentiation among lakes and no isolation by distance (IBD). Outlier analysis detected three loci under positive selection with differing frequencies in the four populations. General linear models did not support an association of lake-specific genotypes with morphological variation. Body shape was divergent among lakes, suggesting that processes previously reported from within single lakes may also be operating at the scale of whole lakes. We found no evidence for body shape being heritable, although sample size was small in these natural populations. Rather than habitat structure and diet, we conclude that predation had a stronger effect on the prevalence of local morphotypes. A variable morphotype facilitating the efficient uptake of a variety of spatially and temporarily scattered resources seems to be favored in these small aquatic systems.


Early predictors of short term neurodevelopmental outcome in asphyxiated cooled infants. A combined brain amplitude integrated electroencephalography and near infrared spectroscopy study.
Brain Cooling (BC) represents the elective treatment in asphyxiated newborns. Amplitude Integrated Electroencephalography (aEEG) and Near Infrared Spectroscopy (NIRS) monitoring may help to evaluate changes in cerebral electrical activity and cerebral hemodynamics during hypothermia. To evaluate the prognostic value of aEEG time course and NIRS data in asphyxiated cooled infants. Twelve term neonates admitted to our NICU with moderate-severe Hypoxic-Ischemic Encephalopathy (HIE) underwent selective BC. aEEG and NIRS monitoring were started as soon as possible and maintained during the whole hypothermic treatment. Follow-up was scheduled at regular intervals; adverse outcome was defined as death, cerebral palsy (CP) or global quotient <88.7 at Griffiths' Scale. 2/12 Infants died, 2 developed CP, 1 was normal at 6 months of age and then lost at follow-up and 7 showed a normal outcome at least at 1 year of age. The aEEG background pattern at 24 h of life was abnormal in 10 newborns; only 4 of them developed an adverse outcome, whereas the 2 infants with a normal aEEG developed normally. In infants with adverse outcome NIRS showed a higher Tissue Oxygenation Index (TOI) than those with normal outcome (80.0±10.5% vs 66.9±7.0%, p=0.057; 79.7±9.4% vs 67.1±7.9%, p=0.034; 80.2±8.8% vs 71.6±5.9%, p=0.069 at 6, 12 and 24 h of life, respectively). The aEEG background pattern at 24h of life loses its positive predictive value after BC implementation; TOI could be useful to predict early on infants that may benefit from other innovative therapies.


Improved sauerkraut production with probiotic strain Lactobacillus plantarum L4 and Leuconostoc mesenteroides LMG 7954.
Probiotic strain Lactobacillus plantarum L4 and strain Leuconostoc mesenteroides LMG 7954 were applied for the controlled fermentation of cabbage heads. The parameters of the controlled and spontaneous fermentations, including antimicrobial effect of cabbage brines obtained at the end of both fermentations, were monitored. To check out the influence of starter culture strains, 10 randomly chosen lactic acid bacteria, isolated at the end of controlled cabbage heads fermentation were identified by API 50 CHL test, and the presence of the probiotic culture was confirmed by Pulsed Field Gel Electrophoresis. The starter cultures applied for cabbage heads fermentation allowed lowering of NaCl concentrations from 4.0% to 2.5% (w/v), considerably accelerated fermentation process by 14 d, and improved the product quality. The produced sauerkraut heads are considered probiotic product as viable probiotic cells count in final product was higher than 10(6) colony-forming units (CFU) per gram of product. The results of this research could be applied in the production of fermented cabbage heads with added functional (probiotic) value and with lower NaCl concentration with expected shortened fermentation time. This could not only be of economic but also of ecological importance.


Using the Health Belief Model to explain patient involvement in patient safety.
With the knowledge that patient safety incidents can significantly impact patients, providers and health-care organizations, greater emphasis on patient involvement as a means to mitigate risks warrants further research. To understand whether patient perceptions of patient safety play a role in patient involvement in factual and challenging patient safety practices and whether the constructs of the Health Belief Model (HBM) help to explain such perceptions. Partial least squares (PLS) analysis of survey data. Four inpatient units located in two tertiary hospitals in Atlantic Canada. Patients discharged from participating units between November 2010 and January 2011. None. A total of 217 of the 587 patient surveys were returned for a final response rate of 37.0%. The PLS analysis revealed relationships between patient perceptions of threat and self-efficacy and the performance of factual and challenging patient safety practices, explaining 46 and 42% of the variance, respectively. The results from this study provide evidence for the constructs and relationships set forth by the HBM. Perceptions of patient safety were shown to influence patient likelihood for engaging in selected patient safety practices. While perceptions of barriers and benefits and threats were found to be a contributing factor to patient involvement in patient safety practices, self-efficacy plays an important role as a mediating factor. Overall, the use of the HBM within patient safety provides for increased understanding of how such perceptions can be influenced to improve patient engagement in promoting safer health care.


Economic analysis of nosocomial infections in a Turkish university hospital.
Nosocomial infections constitute an important health problem with morbidity, and high mortality, prolongation of hospital stay, and increased costs of direct patient care. This study was conducted in Hacettepe University Hospital in order to determine the cost of nosocomial infections and length of hospitalization by matching infected patients with uninfected controls. Data collected from 102 individuals with nosocomial infection (group A) and 102 controls without nosocomial infection (group B) were recorded by using a computer program (dbase IV). Urinary tract infections, surgical wound infections and bacteremias were the most common nosocomial infections. Patients were generally located in Internal Medicine, Neurosurgery, General Surgery Clinics. The mean total length of stay was 35.1 and 14.8 days for group A and group B, respectively (p<0.001). The average hospital cost was US $2280 for group A, and US $698 for group B (p<0.001). Mortality rates were 19.6% for patients with nosocomial infections and 2.9% for uninfected controls (p<0.0001). We conclude that the high economic expense which nosocomial infection represents justifies measures to control this entity.


Linking human behaviours and malaria vector biting risk in south-eastern Tanzania.
To accelerate malaria elimination in areas where core interventions such as insecticide-treated nets (ITNs) are already widely used, it is crucial to consider additional factors associated with persistent transmission. Qualitative data on human behaviours and perceptions regarding malaria risk was triangulated with quantitative data on Anopheles mosquito bites occurring indoors and outdoors in south-eastern Tanzania communities where ITNS are already used but lower level malaria transmission persists. Each night (18:00h-07:00h), trained residents recorded human activities indoors, in peri-domestic outdoor areas, and in communal gatherings. Host-seeking mosquitoes were repeatedly collected indoors and outdoors hourly, using miniaturized exposure-free double net traps (DN-Mini) occupied by volunteers. In-depth interviews were conducted with household representatives to explore perceptions on persistent malaria and its control. Higher proportions of people stayed outdoors than indoors in early-evening and early-morning hours, resulting in higher exposures outdoors than indoors during these times. However, exposure during late-night hours (22:00h-05:00h) occurred mostly indoors. Some of the popular activities that kept people outdoors included cooking, eating, relaxing and playing. All households had at least one bed net, and 83.9% of people had access to ITNs. Average ITN use was 96.3%, preventing most indoor exposure. Participants recorgnized the importance of ITNs but also noted that the nets were not perfect. No complementary interventions were reported being used widely. Most people believed transmission happens after midnight. We conclude that insecticide-treated nets, where properly used, can still prevent most indoor exposures, but significant risk continues unabated before bedtime, outdoors and at communal gatherings. Such exposure is greatest for rural and low-income households. There is therefore an urgent need for complementary interventions, particularly those targeting outdoor-biting and are applicable for all people including the marginalised populations such as migratory farmers and fishermen. Besides, the differences in community understanding of ongoing transmission, and feedback on imperfections of ITNs should be considered when updating malaria-related communication and interventions.


A novel mutation in CD83 results in the development of a unique population of CD4+ T cells.
Using a mouse mutagenesis screen, we have identified CD83 as being critical for the development of CD4(+) T cells and for their function postactivation. CD11c(+) dendritic cells develop and function normally in mice with a mutated CD83 gene but CD4(+) T cell development is substantially reduced. Additionally, we now show that those CD4(+) cells that develop in a CD83 mutant animal fail to respond normally following allogeneic stimulation. This is at least in part due to an altered cytokine expression pattern characterized by an increased production of IL-4 and IL-10 and diminished IL-2 production. Thus, in addition to its role in selection of CD4(+) T cells, absence of CD83 results in the generation of cells with an altered activation and cytokine profile.


Splenic injury: trends in evaluation and management.
Changing methods of evaluating blunt abdominal trauma and expanding selection criteria for nonoperative management (NOM) of splenic injury can increase the number of patients managed nonoperatively without affecting success rates. The charts of 164 patients with blunt splenic injuries from July 1, 1991, to June 30, 1996, were reviewed. Thirty-eight patients were excluded because of immediate laparotomy without adjunctive tests or expiration in the resuscitative period. Injuries were graded according to the Organ Injury Scale. Overall, successful NOM occurred in 84% of patients (73 of 87). NOM was successful in 5 of 7 patients > 55 years old and in 14 of 15 patients with Glasgow Coma Scale scores < 13. Use of computed tomography increased NOM of splenic trauma from 11 to 71% during the 5-year period for injuries of equivalent severity. Age > 55 years or abnormal neurologic status should not preclude NOM, because success was related only to injury grade.


Cartilaginous compression of the liver - clinical and ultrasonographic aspects.
The musculocartilaginous complex is a structure composed of cartilaginous, osseous and muscular elements, which is located at the thoracoabdominal junction, at the level of the right costal arch. To determine the ultrasonographic characteristics of this complex under normal conditions as well as to demonstrate its effects on the liver depending on the constitutional body built, respiratory phase and patient's body position. All abdominal ultrasound scans were performed between 2006 and 2015. A total of 1000 patients (566 females and 434 males aged between 35 and 82 years, mean age 52 years), who had no significant upper abdominal pathologies identified based on clinical and imaging data, were enrolled for the analysis. In addition to standard internal organ assessment, we also attempted to identify the symptoms of hepatic compression by the musculocartilaginous complex. We used 3-6 MHz convex and 7-12 MHz linear transducers. The degree of musculocartilaginous compression of the liver was assessed during breathing in supine and sitting position, as well as with trunk inclined forward. The study showed that musculocartilaginous compression of the liver mostly affects females (96%) with leptosomatic body build. The complex compressing the liver shows a heterogeneous echostructure. Increased hepatic compression was observed during exhalation, in a sitting position and with trunk inclined forward. Cartilaginous compression of the liver depends on body built and patient's body position. The musculocartilaginous complex may cause focal or segmental compression of the hepatic parenchyma, causing pain in the right upper abdomen in some patients.


Novel Genetic Variants in TP37, PIK3R1, CALM1, and PLCG2 of the Neurotrophin Signaling Pathway Are Associated with the Progression from Mild Cognitive Impairment to Alzheimer's Disease.
Alzheimer's disease (AD) is a common neurodegenerative disease and mild cognitive impairment (MCI) is considered as the prodromal stage of AD. Previous studies showed that changes in the neurotrophin signaling pathway could lead to cognitive decline in AD. However, the association of single nucleotide polymorphisms (SNPs) in genes that are involved in this pathway with AD progression from MCI remains unclear. We investigated the associations between SNPs involved in the neurotrophin signaling pathway with AD progression. We performed single-locus analysis to identify neurotrophin-signaling-related SNPs associated with the AD progression using 767 patients from the Alzheimer's Disease Neuroimaging Initiative study and 1,373 patients from the National Alzheimer's Coordinating Center study. We constructed polygenic risk scores (PRSs) using the identified independent non-APOE SNPs and evaluated its prediction performance on AD progression. We identified 25 SNPs significantly associated with AD progression with Bayesian false-discovery probability ≤0.8. Based on the linkage disequilibrium clumping and expression quantitative trait loci analysis, we found 6 potentially functional SNPs that were associated with AD progression independently. The PRS analysis quantified the combined effects of these SNPs on longitudinal cognitive assessments and biomarkers from cerebrospinal fluid and neuroimaging. The addition of PRSs to the prediction model for 3-year progression to AD from MCI significantly increased the predictive accuracy. Genetic variants in the specific genes of the neurotrophin signaling pathway are predictors of AD progression. eQTL analysis supports that these SNPs regulate expression of key genes involved in the neurotrophin signaling pathway.


Osteolytic lumbar discal cyst: case report.
A 25-year-old man presented with left lumboischialgia refractory to medical treatment. Neurological examination revealed L5 and S1 radiculopathy which rapidly worsened over a short period. Magnetic resonance imaging demonstrated disk bulging with a discal cyst at the L4-5 intervertebral space and disk herniation at the L5-S1 intervertebral space. Computed tomography showed osteolytic change of the L5 vertebral body adjacent to the cyst. Resection of the cyst and removal of the herniated disk were performed following fenestration of the L4-5 and L5-S1 interlaminar spaces. Bloody serous fluid followed by clear serous fluid was recognized during the aspiration and partial resection of the cyst at the L4-5 level. Histological examination demonstrated a cyst wall consisting of fibrous connective tissue without a single-layer lining of cells, and fibrin deposits. The patient's symptoms disappeared immediately after the operation. This osteolytic lumbar discal cyst possibly occurred subsequent to hemorrhage from the epidural venous plexus following intervertebral disk injury, hematoma encapsulation by connective fibrous tissue, and cyst wall formation in reaction to the disk injury and hemorrhage. The cyst may have enlarged due to the inflow of the serous fluid from the water-containing degenerated disk.


Evidence for selection on a chordate histocompatibility locus.
Allorecognition is the ability of an organism to differentiate self or close relatives from unrelated individuals. The best known applications of allorecognition are the prevention of inbreeding in hermaphroditic species (e.g., the self-incompatibility [SI] systems in plants), the vertebrate immune response to foreign antigens mediated by MHC loci, and somatic fusion, where two genetically independent individuals physically join to become a chimera. In the few model systems where the loci governing allorecognition outcomes have been identified, the corresponding proteins have exhibited exceptional polymorphism. But information about the evolution of this polymorphism outside MHC is limited. We address this subject in the ascidian Botryllus schlosseri, where allorecognition outcomes are determined by a single locus, called FuHC (Fusion/HistoCompatibility). Molecular variation in FuHC is distributed almost entirely within populations, with very little evidence for differentiation among different populations. Mutation plays a larger role than recombination in the creation of FuHC polymorphism. A selection statistic, neutrality tests, and distribution of variation within and among different populations all provide evidence for selection acting on FuHC, but are not in agreement as to whether the selection is balancing or directional.


The Focal Play Therapy: A Clinical Approach to Promote Child Health and Family Well-being.
Eating and evacuation disorders can cause serious health problems for children. Early recognition and early treatment require a multifactor intervention based on a collaborative relationship between pediatricians, psychotherapists and other health professionals. In this context the Focal Play Therapy (FPT) with children and parents is a psychodynamic model of intervention that improves parental ability to cope with child's difficulties. Parental engagement in child interventions allows to understand child's symptoms within family dynamics and to build an alliance with parents that represents a crucial variable of an effective psychological support for children and families. In the present study data were collected from 17 parental couples and their preschool children at two time points (1st and 6th FPT sessions) marking the first phase of this intervention. This phase was aimed to the assessment of child's symptoms within family relationships and to the promotion of the alliance with parents. Families were in treatment at the Psychological Consultation Center for Children and Parents located at the Department of Psychology of the University of Bologna (Italy). This Center provides health assessment and intervention services to children and their families. We investigated the alliance from both parents and therapist points of view along with other parental and child outcome variables implied in clinical works with children and families. Alliance scores were obtained through the Working Alliance Inventory and the System for Observing Family Therapy Alliances, two measures used in individual and family settings, respectively. Parenting stress and parent-child interactions were investigated using the Parenting Stress Index and the Emotional Availability Scales. Furthermore, paired t-tests were run to detect changes on parental and child variables. Findings advise that special attention should be paid to the building of an early alliance with parents. In this regard the FPT is specifically designed to promote the parent-therapist alliance in the context of child health and family well-being.


Physical therapy in the postoperative of proximal femur fracture in elderly. Literature review.
The proximal femoral fracture in the elderly is a serious public health problem. Surgical treatment of this fracture is used to reduce morbidity, together with postoperative physical therapy. The objective was to conduct a systematic review of physical therapy protocols in postoperative for fractures of the proximal femur in elderly. We selected randomized controlled trials in elderly in the past 10 years, in Portuguese and English. There were 14 articles in the literature. Physical therapy has an important role in functional recovery of the elderly. Level of Evidence I, Systematic Review RCTs (Study results were homogenous).


Failure mode and effect analysis: improving intensive care unit risk management processes.
Purpose Failure modes and effects analysis (FMEA) is a practical tool to evaluate risks, discover failures in a proactive manner and propose corrective actions to reduce or eliminate potential risks. The purpose of this paper is to apply FMEA technique to examine the hazards associated with the process of service delivery in intensive care unit (ICU) of a tertiary hospital in Yazd, Iran. Design/methodology/approach This was a before-after study conducted between March 2013 and December 2014. By forming a FMEA team, all potential hazards associated with ICU services - their frequency and severity - were identified. Then risk priority number was calculated for each activity as an indicator representing high priority areas that need special attention and resource allocation. Findings Eight failure modes with highest priority scores including endotracheal tube defect, wrong placement of endotracheal tube, EVD interface, aspiration failure during suctioning, chest tube failure, tissue injury and deep vein thrombosis were selected for improvement. Findings affirmed that improvement strategies were generally satisfying and significantly decreased total failures. Practical implications Application of FMEA in ICUs proved to be effective in proactively decreasing the risk of failures and corrected the control measures up to acceptable levels in all eight areas of function. Originality/value Using a prospective risk assessment approach, such as FMEA, could be beneficial in dealing with potential failures through proposing preventive actions in a proactive manner. The method could be used as a tool for healthcare continuous quality improvement so that the method identifies both systemic and human errors, and offers practical advice to deal effectively with them.


The Role of Perceived Organizational Support in Mental Health of Healthcare Workers During the COVID-19 Pandemic: A Cross-Sectional Study.
Background: Data support the link between the coronavirus disease 2019 (COVID-19) pandemic and mental distress in healthcare workers (HCWs). Although previous studies have documented the association between organizational policies and employees' psychological and mental status, there is still scant evidence regarding the effect of perceived organizational support (POS) on mental distress in HCWs during the pandemic. Aims: The present study aimed to assess the association between POS and mental distress in HCWs during the COVID-19 pandemic. The role of POS in stress, depressive and trauma symptoms in HCWs was investigated. Methods: This was an online cross-sectional study in 424 HCWs. Data were collected during the first wave of the pandemic, and included demographics, a 7-item questionnaire assessing POS, the "Patient Health Questionnaire" assessing depressive symptoms, the "Impact of Events Scale Revised," measuring post-traumatic stress disorder (PTSD) symptoms and the "Perceived Stress Scale" assessing perceived stress. Results: The mean POS score was 3.33 [standard deviation:1.85; range 0-7]. Younger (p < 0.001), less experienced (p < 0.001), female (p = 0.002), and non-physician HCWs (p = 0.031) were more likely to report lower self-perceived organizational support than older, male, more experienced physicians. Self-perceived organizational support was significantly and negatively associated with and self-assessed intensity of stress, depressive and traumatic symptoms, after adjusting for putative confounders (p < 0.001). Discussion: Self-perceived organizational support was significantly associated with HCWs' self-assessed mental status during the pandemic. Organizational support and mental distress should be addressed simultaneously in HCWs during the COVID-19 pandemic to increase resilience among them.


Cord shape and measurements in cervical spondylotic myelopathy and radiculopathy.
A combined clinical myelographic and computed myelographic study was performed in 30 patients with cervical spondylotic myelopathy; computed myelography was also performed in 16 control patients. Good correlation was found between degree of deformity of cross-sectional shapes of the cord and the outcome of surgery, and measurements of anteroposterior diameter and area of the cord seemed helpful in this preliminary study. There was also reasonably good correlation between the clinical features when grouped according to specific tract involvement, and the pattern of deformity shown by the abnormal cross-sectional cord shapes.


[Simon Bolivar and the cantharides].
Simon Bolivar, The Liberator, did not trust medical care and if he ever requested a doctor, it seems to have been mainly politically motivated. Unfortunately his fears seem to have been correct as his death was accelerated by Dr. Revenand, a man whose medical title is yet under discussion, who applied an extract of cantharides, a beetle known as "the Spanish fly", employed generally as an aphrodisiac, over Bolivar's neck region. According to popular belief of the time, cantharidin, the insect's active principle, reduced "the excess of humours", such as cerebrospinal fluid, plasma, "pituita" and others. Cantharidin has severe adverse effects and was for sure the origin of great suffering for many unfortunate patients in the past.


Gastric emptying as a determinant of the oral glucose tolerance test.
Intraindividual variation in the results of repeated oral glucose tolerance tests in normal subjects is well recognized but incompletely explained. The present studies show that such variation can be produced by ingestion of the glucose solution during different phases of the normal fasting activity cycle of the upper gut. Such variation is not seen when glucose is administered intraduodenally during the same phases of activity. Gastric emptying shows similar variation with the activity cycle; larger volumes of solution were emptied from the stomach during activity than quiescence, thus presenting greater quantities of glucose solution to the small intestine for absorption. Metoclopramide and hyoscine butylbromide, drugs known to influence the rate of gastric emptying, reduced the variation in the glucose tolerance test. The data suggest na possible use of the glucose tolerance test for the assessment of gastric emptying.


Combination surgical excision and fractional carbon dioxide laser for treatment of rhinophyma.
Rhinophyma is a severe late complication of rosacea, which is characterized by progressive hyperplasia of sebaceous glands and connective tissue involving the lower two-thirds of the nose. It can be an emotionally devastating disorder, and serve as a medium for occult cancers and other health problems. Many surgical treatments have been advocated, as well as dermabrasion and laser therapy. In light of the problems faced with these individual therapy modalities, we advocate a combination therapy of surgical debulking and fractionated Carbon dioxide laser therapy. By excising the tissue first, we not only decreased the amount of time needed for the procedure, we were also able to preserve a histopathologic specimen that can be examined for occult cancers. After debulking, the fractionated carbon dioxide laser is then used to blend. By using fractionated carbon dioxide, we avoided the complications associated with non-fractionated carbon dioxide lasers, such as delayed healing times, hypopigmentation, hyperpigmentation, scarring, and persistent redness. We report our experience in two patients with rhinophyma who underwent a combination of surgical excision and fractional Co2 laser therapy for treatment. Pre- and postoperative protocols and treatment parameters are discussed. Both patients had excellent cosmetic and functional results and were followed for at least one year.


Non-labeling multiphoton excitation microscopy as a novel diagnostic tool for discriminating normal tissue and colorectal cancer lesions.
Multiphoton excitation microscopy (MPM) is regarded as an effective tool that enables the visualization of deep regions within living tissues and organs, with little damage. Here, we report novel non-labeling MPM (NL-MPM) imaging of fresh human colorectal mucosa, which is useful for discriminating cancer lesions from normal tissues quantitatively without any need for resection, fixation, or staining. Using NL-MPM, we visualized three components in human colorectal mucosa, epithelial cells, immune cells, and basement membranes, based on their characteristic patterns of fluorescence. These patterns are characterized by the different auto-fluorescence properties of nicotinamide adenine dinucleotide, nicotinamide adenine dinucleotide phosphate, and flavin adenine dinucleotide and from second harmonic generation (SHG). NL-MPM images were at least as informative to pathologists as were 'conventional' images of fixed tissue sections stained with hematoxylin and eosin. Additionally, two quantitative parameters extracted from NL-MPM images - the nucleus diameter (index N) and the intensity of SHG in the basement membrane (index S) - rendered it possible to diagnose cancer regions effectively. In conclusion, NL-MPM is a novel, promising method for real-time clinical diagnosis of colorectal cancers, and is associated with minimal invasiveness.


Coagulopathy and blood component transfusion in trauma.
Trauma is a serious global health problem, accounting for approximately one in 10 deaths worldwide. Uncontrollable bleeding accounts for 39% of trauma-related deaths and is the leading cause of potentially preventable death in patients with major trauma. While bleeding from vascular injury can usually be repaired surgically, coagulopathy-related bleeding is often more difficult to manage and may also mask the site of vascular injury. The causes of coagulopathy in patients with severe trauma are multifactorial, including consumption and dilution of platelets and coagulation factors, as well as dysfunction of platelets and the coagulation system. The interplay between hypothermia, acidosis and progressive coagulopathy, referred to as the 'lethal triad', often results in exsanguination. Current management of coagulopathy-related bleeding is based on blood component replacement therapy. However, there is a limit on the level of haemostasis that can be restored by replacement therapy. In addition, there is evidence that transfusion of red blood cells immediately after injury increases the incidence of post-injury infection and multiple organ failure. Strategies to prevent significant coagulopathy and to control critical bleeding effectively in the presence of coagulopathy may decrease the requirement for blood transfusion, thereby improving clinical outcome of patients with major trauma.


Recent advances in insect thermoregulation.
Ambient temperature (Ta) is a critical abiotic factor for insects that cannot maintain a constant body temperature (Tb). Interestingly, Ta varies during the day, between seasons and habitats; insects must constantly cope with these variations to avoid reaching the deleterious effects of thermal stress. To minimize these risks, insects have evolved a set of physiological and behavioral thermoregulatory processes as well as molecular responses that allow them to survive and perform under various thermal conditions. These strategies range from actively seeking an adequate environment, to cooling down through the evaporation of body fluids and synthesizing heat shock proteins to prevent damage at the cellular level after heat exposure. In contrast, endothermy may allow an insect to fight parasitic infections, fly within a large range of Ta and facilitate nest defense. Since May (1979), Casey (1988) and Heinrich (1993) reviewed the literature on insect thermoregulation, hundreds of scientific articles have been published on the subject and new insights in several insect groups have emerged. In particular, technical advancements have provided a better understanding of the mechanisms underlying thermoregulatory processes. This present Review aims to provide an overview of these findings with a focus on various insect groups, including blood-feeding arthropods, as well as to explore the impact of thermoregulation and heat exposure on insect immunity and pathogen development. Finally, it provides insights into current knowledge gaps in the field and discusses insect thermoregulation in the context of climate change.


Enhanced ethanol production from paper sludge waste under high-solids conditions with industrial and cellulase-producing strains of Saccharomyces cerevisiae.
Reported ethanol titres from hydrolysis-fermentation of the degraded fibres in paper sludge (PS) waste, generally obtained under fed-batch submerged conditions, can be improved through fermentation processes at high solids loadings, as demonstrated in the present study with two industrial PS wastes at enzyme dosages appropriate for solids loadings up to 40% (w/w). The industrial yeast,Saccharomyces cerevisiaestrain Ethanol Red®, was compared to two genetically engineeredS. cerevisiaestrains, namely Cellusec® 1.0 and Cellusec® 2.0, capable of xylose utilisation, and xylose utilisation and cellulase production, respectively. High-solids batch fermentations were conducted in 3 L horizontal rotating reactors and ethanol titres of 100.8 and 73.3 g/L were obtained for virgin pulp and corrugated recycle PS, respectively, at 40% (w/w) solids loading using Ethanol Red®. Xylose utilisation by Cellusec® 1.0 improved ethanol titres by up to 10.3%, while exogenous cellulolytic enzyme requirements were reduced by up to 50% using cellulase-producing Cellusec® 2.0.


Anti-IgLON5 Disease: A Case With 11-Year Clinical Course and Review of the Literature.
Background: Anti-IgLON5 disease is a novel disorder with a complex interplay between inflammation and neurodegeneration. Patients develop antibodies against IgLON5 but also deposition of neuronal tau protein. Symptoms often have an insidious onset, slow progression and mimic other neurological disorders. Here we report a case with severely prolonged 11-year disease course and provide a review of current reported cases with focus on presentation, work-up, treatment, and outcome. Method: All reported cases of anti-IgLON5 disease were evaluated. Cases reported twice (in case series and as single case reports), were carefully excluded. Results: Most patients display a characteristic sleep disorder with severe insomnia, non rapid eye movement (NREM) parasomnia, with finalistic movements and sleep disordered breathing (stridor and obstructive sleep apnea). Other symptoms are bulbar involvement, gait instability, movement disorders, oculomotor abnormalities, dysautonomia, and peripheral symptoms. Antibodies are present in both serum and CSF and there is a strong correlation with human leukocyte antigen (HLA) DRB1*10:01 and HLA-DQB1*05:01. Neuropathological examination reveals neurodegeneration with neuronal tau deposits in regions that correlate with the clinical presentation (e.g., predominantly hypothalamus and tegmentum of the brain stem). Majority of cases respond partially to immunotherapy. Cases, who received no treatment or treatment with IV corticosteroids alone, had a higher mortality than cases treated with more potent immunotherapy. Conclusion: The clinical spectrum of Anti-IgLON5 disease continues to expand. Further studies are needed to elucidate the pathophysiology, therapeutic strategies and outcome in this novel disorder. Aggressive immunotherapy seems to increase survival.


Growth performance and carcass composition of lambs infused for 28 days with a growth hormone-releasing factor analogue.
A human growth hormone-releasing factor analogue, [DesNH2Tyr1,D-Ala2,Ala15]hGRF(1-29)NH2 (GRF-A), was infused s.c. into lambs for 28 d to determine its effects on growth performance and carcass composition. Twenty crossbred wethers weighing 47.0 +/- .5 kg were implanted with 7-d osmotic minipumps at weekly intervals. Minipumps contained either vehicle (dimethyl sulfoxide:H2O, 1:1) or GRF-A, released at a rate of 208 pmol (or .7 micrograms).h-1.kg-1. During the infusion period, plasma GH levels were increased (P less than .01) in GRF-A-treated wethers compared with control wethers (15.0 vs 9.3 ng/ml) and were higher on days that closely followed minipump implantation. Plasma IGF-I and hepatic IGF-I RNA concentrations were similar in lambs of both groups. Analogue treatment improved feed conversion (4.9 vs 5.8 kg dry matter/kg gain, P less than .05), increased average daily gain (.35 vs .30 kg, P = .05) and had no effect on feed intake, wool growth and body, carcass, selected organ and pituitary weights. Carcasses from GRF-A-infused lambs had less adjusted fat depth, a lower percentage of fat and a higher percentage of protein (P less than .05) than carcasses from control lambs. Magnitude of most effects of GRF-A on carcass measurements were correlated with the mean GH level that a lamb had during the infusion period. In conclusion, s.c. infusion of GRF-A improved feed utilization and altered carcass composition of feeder lambs in a relatively short period of time (28 d).


Is Vitamin A an Antioxidant or a Pro-oxidant?
Antioxidant efficiency of all-trans-retinol has been studied on the basis of characteristic thermochemical properties using density functional theory. The influence of the solvent polarity has also been evaluated. It is found that retinol may act in parallel as an effective antioxidant via H atom donating as well as a pro-oxidant in yielding reactive hydroxyl radical. In fact, the lowest values of bond dissociation enthalpy were found at the C18-H and C18-OH positions. Retinol was also determined to be a good electron donor but bad acceptor in the single electron transfer (ET) reaction with hydroperoxyl (HOO•) radical. In addition, potential energy surfaces of H atom transfer (HAT) and radical adduct formation (RAF) reactions between retinol and HOO• radical were also investigated in the gas phase and in the solvent. The results demonstrated that the RAF mechanism was generally more predominant than ET and HAT ones. The most favored radical addition position was found at the C2═C3 double bond in the cyclohexenyl ring. Moreover, the radical scavenging reactivity via RAF reactions was strongly exergonic and thermodynamically feasible while the ET one was endergonic. Natural bond orbital analysis showed that the lone pairs of electrons on the oxygen atom of the HOO• radical were donated to the unoccupied antibonding orbital of transferred H atom in HAT reactions. In contrast, in the case of RAF reactions, strong interactions between 2p orbitals on oxygen atoms of the radical and the π orbital of the double bond on the retinol molecule were recognized. The results obtained in this work were in agreement with previous experimental observations.


CSF levels of oligomeric alpha-synuclein and beta-amyloid as biomarkers for neurodegenerative disease.
Protein misfolding and aggregation is a critically important feature in many devastating neurodegenerative diseases, therefore characterization of the CSF concentration profiles of selected key forms and morphologies of proteins involved in these diseases, including β-amyloid (Aβ) and α-synuclein (a-syn), can be an effective diagnostic assay for these diseases. CSF levels of tau and Aβ have been shown to have great promise as biomarkers for Alzheimer's disease. However since the onset and progression of many neurodegenerative diseases have been strongly correlated with the presence of soluble oligomeric aggregates of proteins including various Aβ and a-syn aggregate species, specific detection and quantification of levels of each of these different toxic protein species in CSF may provide a simple and accurate means to presymptomatically diagnose and distinguish between these diseases. Here we show that the presence of different protein morphologies in human CSF samples can be readily detected using highly selective morphology specific reagents in conjunction with a sensitive electronic biosensor. We further show that these morphology specific reagents can readily distinguish between post-mortem CSF samples from AD, PD and cognitively normal sources. These studies suggest that detection of specific oligomeric aggregate species holds great promise as sensitive biomarkers for neurodegenerative disease.


A grounded theory of reimaging.
Several cross-sectional, qualitative studies suggest that physical alterations in appearance or functioning have the potential to influence self-esteem. There have been no studies describing the process of adapting to body image disruption. A grounded theory of reimaging is proposed, based on the experiences of 28 participants who had experienced significant weight change, loss or paralysis of body parts, ostomies, scarring from burns or trauma, or surgical reconstruction. Participants were interviewed at 3, 6, 12, and 18 months following the physical alteration. Three phases, action processes, influencing factors, and outcomes, of reimaging are described.


Inter-rater reliability and comfort in the application of a basic life support termination of resuscitation clinical prediction rule for out of hospital cardiac arrest.
This study evaluates inter-rater reliability and comfort of BLS providers with the application of an out-of-hospital Basic Life Support Termination of Resuscitation (BLS TOR) clinical prediction rule. This rule suggests that continued BLS cardiac resuscitation is futile and can be terminated in the field if the following three conditions are met: (1) no return of spontaneous circulation; (2) no shock given prior to transport; (3) cardiac arrest not witnessed by EMS personnel. Providers hypothetically applied the rule and rated their comfort level on a five-point Likert-type scale, from "very comfortable" to "very uncomfortable" during the prospective validation of a BLS TOR clinical prediction rule in out-of-hospital cardiac arrest conducted in 12 rural and urban communities [Morrison LJ, Visentin LM, Kiss A, et al. Validation of a rule for termination of resuscitation in out-of-hospital cardiac arrest. New Engl J Med 2006;355(5):478-87]. A Kappa score measured agreement between providers and compared to the correct interpretation of the rule. We compared mean comfort levels of providers who interpreted the rule correctly versus incorrectly. Of 1240 enrolled cases, 1184 (95.5%) had paramedic attendant forms and 1211 (97.7%) had driver forms and 1175 (94.7%) had both. Kappa for interpretation agreement between driver and attendant was 0.90 (95% CI, 0.87-0.92); between attendant and correct interpretation of the BLS TOR clinical prediction rule, 0.88 (95% CI, 0.85-0.91); between driver and correct interpretation of the BLS TOR clinical prediction rule, 0.88 (95% CI, 0.85-0.91). For instances in which both providers applied the rule correctly (607/635 [95.6%]), the providers were significantly more comfortable (chi(2)(4)=30.5, p<0.0001) than those instances in which they did not (28/635 [4.4%]. The vast majority of providers were able to apply the BLS TOR clinical prediction rule correctly and were comfortable doing so. This suggests that both reliability and comfort will remain high during routine application of the rule when paramedics are well trained as users of the rule.


SARS-CoV-2 Transmission Dynamics in a Sleep-Away Camp.
In late June 2020, a large outbreak of coronavirus disease 2019 (COVID-19) occurred at a sleep-away youth camp in Georgia, affecting primarily persons ≤21 years. We conducted a retrospective cohort study among campers and staff (attendees) to determine the extent of the outbreak and assess factors contributing to transmission. Attendees were interviewed to ascertain demographic characteristics, known exposures to COVID-19 and community exposures, and mitigation measures before, during, and after attending camp. COVID-19 case status was determined for all camp attendees on the basis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) test results and reported symptoms. We calculated attack rates and instantaneous reproduction numbers and sequenced SARS-CoV-2 viral genomes from the outbreak. Among 627 attendees, the median age was 15 years (interquartile range: 12-16 years); 56% (351 of 627) of attendees were female. The attack rate was 56% (351 of 627) among all attendees. On the basis of date of illness onset or first positive test result on a specimen collected, 12 case patients were infected before arriving at camp and 339 case patients were camp associated. Among 288 case patients with available symptom information, 45 (16%) were asymptomatic. Despite cohorting, 50% of attendees reported direct contact with people outside their cabin cohort. On the first day of camp session, the instantaneous reproduction number was 10. Viral genomic diversity was low. Few introductions of SARS-CoV-2 into a youth congregate setting resulted in a large outbreak. Testing strategies should be combined with prearrival quarantine, routine symptom monitoring with appropriate isolation and quarantine, cohorting, social distancing, mask wearing, and enhanced disinfection and hand hygiene. Promotion of mitigation measures among younger populations is needed.


Development and validation of the Health Literacy Scale for community-dwelling older adults in Taiwan.
Health literacy is a crucial factor in determining one's health outcomes. Individuals with high health literacy possess the ability to access, comprehend, evaluate, and utilize health information to maintain, enhance, and advocate for good health. This study aimed to develop and validate a tool called the Health Literacy Questionnaire for Older Adults (HLQ-OA) specifically designed to assess the health literacy of community-dwelling older adults in Taiwan. The development of the HLQ-OA occurred in two phases. Firstly, a systematic literature review was conducted, followed by the evaluation of content validity and face validity through expert and older adult assessments. Subsequently, a cross-sectional study was carried out, involving a convenient sample of 481 older adults from community-dwelling long-term care stations. The final version of the HLQ-OA comprised 16 items that assessed four competencies associated with accessing, understanding, appraising, and applying health-related information. The HLQ-OA demonstrated excellent internal consistency, with an overall Cronbach's α coefficient of 0.94. Additionally, the model fit indices indicated an acceptable fit: RMSEA = 0.046, GFI = 0.909, CFI = 0.945, TLI = 0.939. Furthermore, a high correlation was observed between the HLQ-OA and the HL-SF12 (r = 0.832). These findings support the reliability and validity of the HLQ-OA as a tool for assessing the health literacy of older adults. In conclusion, the HLQ-OA is a reliable and valid instrument that can be effectively utilized to measure the health literacy levels of older adults.


Limited development and progression of resistance of HIV-1 to the nucleoside analogue reverse transcriptase inhibitor lamivudine in human primary macrophages.
The aim of this study was to investigate the development and progression of phenotypic resistance to the HIV-1-reverse transcriptase (RT) inhibitor lamivudine, and genotypic variations of HIV-1-RT occurring under lamivudine treatment in HIV-1-infected human primary monocytes-macrophages (M/M). Cellular passages in the presence of lamivudine were performed every 2 weeks by transferring supernatants of infected M/M to fresh M/M. A fitness assay using wild-type virus and a lamivudine-resistant HIV-1 virus (harbouring the M184V RT mutation) was performed in peripheral blood mononuclear cells. Culture supernatants were tested for p24 antigen production and RT activity. The M184V RT mutant virus was obtained by site-directed mutagenesis on a CCR5-using HIV-1 backbone. The mutagenized M184V RT virus showed full resistance to lamivudine in M/M. However, no detectable phenotypic and genotypic resistance (neither virus breakthrough, nor RT resistance-related mutations) developed in M/M infected by HIV-1 and cultured for up to seven passages in vitro (i.e. 105 days). This inefficiency of M/M to develop M184V RT mutated virus is tightly related to the low 2'-deoxynucleotide (dNTP) pool in such cells, which in turn decreases the kinetics of HIV-1-RT. Despite this, the M184V RT mutant virus replicates in M/M, although with a 30% decreased efficiency compared with the wild-type. Our results show that the chances of development of resistance are far lower in M/M than in lymphocytes. This underlines the importance and the peculiar role of M/M as reservoirs of either wild-type or resistant strains in human organs.


Genomic approaches to coronary artery disease.
Coronary artery disease (CAD) is a leading cause of death and disability worldwide. In addition to lifestyle and environmental factors which are major aetiologic determinants, there is considerable familial clustering of the disease indicating a genetic component in its causation. Although the total genetic contribution to CAD risk can be quantified, the determination of the size and number of contributing effects is impossible without identifying all CAD susceptibility genes. However, despite extensive studies, strong evidence of a molecular genetic association with coronary artery disease or myocardial infarction remains elusive. Genome wide association studies have been successful in identifying robust associations of single nucleotide polymorphisms (SNP) with CAD. Identifying the causal variant and dissecting pathways linking these variants to disease process is a major challenge. Technologies from whole genome sequencing, proteomics, transcriptomics and metabolomics are now available to extend analysis to a more complete range of potential susceptibility variants, and to support more explicit modelling of the joint effects of genes and environment. The availability of these high throughput technologies does not diminish the importance of rigorous phenotyping and appropriate study designs in all the endeavours to understand the aetiopathogenesis of CAD. Combining classical epidemiology with modern genomics will require collaborative efforts within the cardiovascular disease community at both bench and bedside and this will have the potential to expand our understanding of CAD and translate discoveries into clinically useful applications that will have a major impact on public health.


Renin angiotensin aldosterone system and glycemia in pregnancy.
The renin-angiotensin-aldosterone system (RAAS) is involved in the pathogenesis of insulin resistance and type 2 diabetes in the general population. The RAAS is activated during pregnancy. However, it is unknown whether the RAAS contributes to glycemia in pregnant women. Plasma renin activity (PRA) and plasma aldosterone levels were quantified at delivery in 689 Chinese mothers. An oral glucose tolerance test in fasted women was performed in the second trimester of pregnancy. The diagnosis of gestational diabetes mellitus (GDM) and impaired glucose tolerance during pregnancy were made according to the guidelines of the Chinese Society of Obstetrics. Plasma aldosterone was significantly higher in pregnant women with GDM as compared to those without impairment of glycemic control (normal pregnancies: 0.27 +/- 0.21 ng/mL, GDM: 0.36 +/- 0.30 ng/mL; p < 0.05). Regression analyses revealed that PRA was negatively correlated with fasting blood glucose (FBG) (R2 = 0.03, p = 0.007), whereas plasma aldosterone and aldosterone/PRA ratio were positively correlated with FBG (R2 = 0.05, p < 0.001 and R = 0.03, p = 0.007, respectively). Multivariable regression analysis models considering relevant confounding factors confirmed these findings. This study demonstrated that fasting blood glucose in pregnant women is inversely correlated with the PRA, whereas plasma aldosterone showed a highly significant positive correlation with fasting blood glucose during pregnancy. Moreover, plasma aldosterone is significantly higher in pregnant women with GDM as compared to those women with normal glucose tolerance during pregnancy. Although causality cannot be proven in association studies, these data may indicate that the RAAS during pregnancy contributes to the pathogenesis of insulin resistance/new onset of diabetes during pregnancy.


Validation of an objective scar pigmentation measurement by using a spectrocolorimeter.
Scar pigmentation changes throughout its maturation process and it is often used as one of the indicators for scar maturation, yet it is often rated subjectively. The purpose of this study was to investigate the application of a commercial spectrocolorimeter to produce a reliable measurement on scar pigmentation. Commission Internationale de l'Eclairage (CIE) model of color has been adopted in this study for measurement of scar pigmentation 24 patients with hypertrophic scars at different stages of maturation were selected for the study, were inspected by two therapists using the Vancouver scar scale (VSS) and then using spectrocolorimeter for inter-rater reliability. The measurements were taken after 30min by the same group of therapists (test-retest reliability). Results indicated that the inter-rater reliability among the three therapists was satisfactory, with intra-class correlation coefficient (ICC) (2, 2) from 0.50 to 0.99 in all the three color parameters. The test-retest reliability of the spectrocolorimeter was satisfactory with ICC (3, 6) ranged from 0.95 to 0.99. A significant difference was also noted between the measurements of normal skin and hypertrophic scar (P<0.00, t-values: from 2.78 to 0.05, d.f.: from 29.7 to 46.00) in all color parameters, except the chroma C(*). We also found a positive relationship between VSS scores and the spectrocolorimeter readings. The spectrocolorimeter is found to be a reliable instrument to quantify scar pigmentation and to differentiate normal skin and scar tissue. With further studies, the constructs of scar properties could further be explored using this spectrocolorimeter.


PreDTIs: prediction of drug-target interactions based on multiple feature information using gradient boosting framework with data balancing and feature selection techniques.
Discovering drug-target (protein) interactions (DTIs) is of great significance for researching and developing novel drugs, having a tremendous advantage to pharmaceutical industries and patients. However, the prediction of DTIs using wet-lab experimental methods is generally expensive and time-consuming. Therefore, different machine learning-based methods have been developed for this purpose, but there are still substantial unknown interactions needed to discover. Furthermore, data imbalance and feature dimensionality problems are a critical challenge in drug-target datasets, which can decrease the classifier performances that have not been significantly addressed yet. This paper proposed a novel drug-target interaction prediction method called PreDTIs. First, the feature vectors of the protein sequence are extracted by the pseudo-position-specific scoring matrix (PsePSSM), dipeptide composition (DC) and pseudo amino acid composition (PseAAC); and the drug is encoded with MACCS substructure fingerings. Besides, we propose a FastUS algorithm to handle the class imbalance problem and also develop a MoIFS algorithm to remove the irrelevant and redundant features for getting the best optimal features. Finally, balanced and optimal features are provided to the LightGBM Classifier to identify DTIs, and the 5-fold CV validation test method was applied to evaluate the prediction ability of the proposed method. Prediction results indicate that the proposed model PreDTIs is significantly superior to other existing methods in predicting DTIs, and our model could be used to discover new drugs for unknown disorders or infections, such as for the coronavirus disease 2019 using existing drugs compounds and severe acute respiratory syndrome coronavirus 2 protein sequences.


Patient Feedback on a Warfarin Action Plan Used in a Local Australian Physician Practice Setting.
Warfarin is a high-risk medicine, and older persons (those aged 65 years and older)1,2 who take this therapy need medicines information about it that is at a level which is both understandable and comprehensive to improve their knowledge about the risks and benefits of warfarin therapy.3,4 Therefore, the primary objective of this study was to report patient feedback on a Warfarin Action Plan (WAP) (leaflet) and identify patients' preferences regarding its content and format. The secondary objective was to canvass in-depth feedback regarding the participants' information needs and current information-seeking practices with respect to warfarin therapy. In an Australian General Practice medical centre setting, a qualitative study comprising 34 individual interviews was conducted. Emergent themes were elicited via a qualitative analysis using manual inductive coding. The majority of participants gave very positive feedback on the WAP leaflet, stating that it was a useful and concise resource. In canvasing this feedback, 4 themes emerged: (1) the need for information about warfarin therapy, (2) reliance on doctors and/or pharmacists for information, (3) the need for information to normalize their daily life, and (4) patients and carers acting on the new information. The WAP is a simple and well-received tool that meets the knowledge and education needs about warfarin therapy for older people and their carers.


In vivo magnetic resonance evaluation of blood oxygen saturation in the superior mesenteric vein as a measure of the degree of acute flow reduction in the superior mesenteric artery: findings in a canine model.
The authors tested the hypothesis that changes in oxygen saturation (%HbO2) in the superior mesenteric vein (SMV), as measured with in vivo magnetic resonance (MR) oximetry, correlate with the degree of acute superior mesenteric artery (SMA) flow reduction. Ten mongrel dogs were studied. A catheter was inserted into the SMV, and a perivascular ultrasonic flow probe and an adjustable mechanical occluder were placed around the SMA. MR oximetry was carried out at the resting state and after the SMA was constricted to predetermined levels (0%-75% of initial flow). In seven dogs, SMV blood samples were obtained immediately before and after each MR measurement; %HbO2 was measured simultaneously by using an oximeter. With linear regression analysis, the SMV %HbO2 measurements obtained at MR imaging were compared with those obtained at oximetry. With a logistic model, MR imaging changes in SMV %HbO2 were compared with the degree of SMA flow reduction. SMV %HbO2 measurements obtained with MR imaging correlated well with those obtained with oximetry (r = .97). Changes in SMV %HbO2 measured at MR imaging also correlated well with the degree of SMA flow reduction, as determined with a logistic model (P = .01). Noninvasive in vivo MR measurements of SMV %HbO2 can be used to determine the degree of acute SMA flow reduction with a high degree of accuracy in a canine model.


Assembly of TolC, a structurally unique and multifunctional outer membrane protein of Escherichia coli K-12.
TolC is a multifunctional outer membrane protein of Escherichia coli that folds into a novel alpha-beta-barrel conformation absent in the other model outer membrane proteins used in assembly studies. The data presented in this work show that the unique folded structure of TolC reflects a unique assembly pathway. During its assembly, the newly translocated nascent TolC monomers are released in the periplasm. Maturation of these nascent monomers, and possibly their oligomerization, in the periplasm precedes their insertion in the outer membrane. The completion of the assembly process is signaled by the development of a characteristic proteinase K-resistant fragment generated by cleavage at a single, periplasmically exposed, protease-sensitive site of the membrane-anchored trimer. None of the assembly steps of TolC is affected by known folding factors, such as SurA, Skp, and lipopolysaccharide, which have profound effects on the assembly of other model trimeric outer membrane proteins. Two assembly-defective TolC mutants were isolated and characterized. One of the mutants (TolC(I106N)) was defective in the folding of nascent monomers, while the other (TolC(S350F)) was impaired in steps involving trimerization and membrane insertion of folded monomers.


The Sinocyclocheilus cavefish genome provides insights into cave adaptation.
An emerging cavefish model, the cyprinid genus Sinocyclocheilus, is endemic to the massive southwestern karst area adjacent to the Qinghai-Tibetan Plateau of China. In order to understand whether orogeny influenced the evolution of these species, and how genomes change under isolation, especially in subterranean habitats, we performed whole-genome sequencing and comparative analyses of three species in this genus, S. grahami, S. rhinocerous and S. anshuiensis. These species are surface-dwelling, semi-cave-dwelling and cave-restricted, respectively. The assembled genome sizes of S. grahami, S. rhinocerous and S. anshuiensis are 1.75 Gb, 1.73 Gb and 1.68 Gb, respectively. Divergence time and population history analyses of these species reveal that their speciation and population dynamics are correlated with the different stages of uplifting of the Qinghai-Tibetan Plateau. We carried out comparative analyses of these genomes and found that many genetic changes, such as gene loss (e.g. opsin genes), pseudogenes (e.g. crystallin genes), mutations (e.g. melanogenesis-related genes), deletions (e.g. scale-related genes) and down-regulation (e.g. circadian rhythm pathway genes), are possibly associated with the regressive features (such as eye degeneration, albinism, rudimentary scales and lack of circadian rhythms), and that some gene expansion (e.g. taste-related transcription factor gene) may point to the constructive features (such as enhanced taste buds) which evolved in these cave fishes. As the first report on cavefish genomes among distinct species in Sinocyclocheilus, our work provides not only insights into genetic mechanisms of cave adaptation, but also represents a fundamental resource for a better understanding of cavefish biology.


Genetic and molecular requirements for function of the Pto/Prf effector recognition complex in tomato and Nicotiana benthamiana.
The Pto gene of tomato (Solanum lycopersicum) confers specific recognition of the unrelated bacterial effector proteins AvrPto and AvrPtoB. Pto resides in a constitutive molecular complex with the nucleotide binding site-leucine rich repeats protein Prf. Prf is absolutely required for specific recognition of both effectors. Here, using stable transgenic lines, we show that expression of Pto from its genomic promoter in susceptible tomatoes was sufficient to complement recognition of Pseudomonas syringae pv. tomato (Pst) bacteria expressing either avrPto or avrPtoB. Pto kinase activity was absolutely required for specific immunity. Expression of the Pto N-myristoylation mutant, pto(G2A), conferred recognition of Pst (avrPtoB), but not Pst (avrPto), although bacterial growth in these lines was intermediate between resistant and susceptible lines. Overexpression of pto(G2A) complemented recognition of avrPto. Transgenic tomato plants overexpressing wild-type Pto exhibited constitutive growth phenotypes, but these were absent in lines overexpressing pto(G2A). Therefore, Pto myristoylation is a quantitative factor for effector recognition in tomato, but is absolutely required for overexpression phenotypes. Native expression of Pto in the heterologous species Nicotiana benthamiana did not confer resistance to P. syringae pv. tabaci (Pta) expressing avrPto or avrPtoB, but recognition of both effectors was complemented by Prf co-expression. Thus, specific resistance conferred solely by Pto in N. benthamiana is an artefact of overexpression. Finally, pto(G2A) did not confer recognition of either avrPto or avrPtoB in N. benthamiana, regardless of the presence of Prf. Thus, co-expression of Prf in N. benthamiana complements many but not all aspects of normal Pto function.


The differential ability of splenic adherent cells from B6.lpr mice to induce suppressor T cells.
Hapten-coupled splenic adherent cells or resident peritoneal cells from autoimmune B6.lpr mice that are over 5 mo of age fail to induce first-order inducer suppressor T cells (Ts1). However, the same population of hapten-coupled cells can induce both delayed-type hypersensitivity responses and third-order effector suppressor T cells (Ts3). Thus, splenic and peritoneal antigen-presenting cells from B6.lpr mice display a defined defect in the ability to induce certain suppressor T cell responses. The cellular defect in Ts1 induction is controlled by the lpr gene, since age-matched congenic B6 mice do not display this defect. The splenic adherent cell defect is temporarily correlated with the autoimmunity that develops in B6.lpr animals. The antigen-presenting defect in the B6.lpr splenic adherent population for Ts1 induction is reversible by culturing the cells in interferon-gamma. The results are discussed as an illustration of the relationship between experimental models of autoimmunity and defects in a suppressor T cell cascade.


Perceptions of ceremonial and nonceremonial uses of tobacco by American-Indian adolescents in California.
American-Indian adolescents have the highest tobacco use prevalence of all ethnic groups in the United States. Although much has been written about the role of tobacco in traditional Native-American cultures, little is known about modern-day perceptions of tobacco among American-Indian adolescents. This study conducted focus groups of 40 American-Indian adolescents in urban and rural areas of Southern California. Participants discussed the role of traditional ceremonial tobacco use in their lives, the use of commercial tobacco as a substitute for sacred tobacco, the perceived safety of traditional versus commercial tobacco, and the perceptions of American-Indian imagery in tobacco advertising. Many American-Indian adolescents may be introduced to traditional tobacco use at early ages. Smoking is viewed as a sign of respect for the elders, but there are acceptable ways for adolescents to participate in ceremonies without inhaling smoke. Commercial cigarettes often are substituted for homegrown tobacco at ceremonies and events. Traditional tobacco was perceived as less dangerous than commercial tobacco because it does not contain chemical additives. However, respondents still perceived that smoking traditional tobacco and breathing tobacco smoke conferred health hazards. Participants found the use of American-Indian imagery in tobacco advertising offensive and stereotypical. Indian casinos were mentioned frequently as places where smoking occurred. Continued health education efforts are needed to decrease habitual use of commercial tobacco products and secondhand smoke exposure among American-Indian youth. Further research is needed to identify ways for American-Indian youth to participate in their cultural traditions while minimizing their risk for tobacco-related diseases.


Prospective multicenter study of toric IOL outcomes when dual zone automated keratometry is used for astigmatism planning.
To evaluate clinical outcomes when toric intraocular lens (IOL) calculations are based on the keratometric output from the Lenstar LS-900 dual zone automated keratometer (Haag-Streit AG, Koeniz, Switzerland). Eligible subjects presenting for toric IOL implantation at five sites were measured with a dual-zone automated keratometer. The data were used to plan the power and angle of the toric IOL to be implanted. Refractive and visual acuity status were checked at 1 and 3 months postoperatively. A total of 102 eyes had relevant data for analysis. More than 76% of eyes had 0.50 diopter or less of refractive astigmatism at 1 and 3 months, with no difference by level of astigmatism corrected. More than half of the eyes had uncorrected distance visual acuity of 20/20 or better and 78% were 20/25 or better. A new measure of effectiveness of toric correction power is described that suggested lens selection was appropriate. Results appeared better than those obtained in previous studies when the IOL cylinder power and alignment were calculated using manual keratometry. In this series of eyes from multiple centers, the calculation of toric IOL power using dual-zone automated keratometry measurements produced clinical results that were better than results in the literature where manual keratometry was used.


Induction of protective immunity with antibody to herpes simplex virus type 1 glycoprotein H (gH) and analysis of the immune response to gH expressed in recombinant vaccinia virus.
Passive administration of neutralizing monoclonal antibody (MAb) to glycoprotein H (gH) of herpes simplex virus type 1 (HSV-1) was found to protect mice from an HSV-1 strain SC16 challenge infection. To investigate further the protective potential of gH, recombinant vaccinia viruses were constructed which expressed the HSV-1 gH open reading frame under the control of the vaccinia virus 7.5K early/late promoter or the 4b late promoter. Immunization with recombinant viruses, however, did not induce the production of neutralizing antisera and the mice were not protected from zosteriform spread or the establishment of latent infection following viral challenge. The gH produced by the recombinant vaccinia viruses differed in electrophoretic mobility and antigenicity from authentic HSV-1 gH. Only one of three neutralizing MAbs specific for conformational epitopes on gH was able to immunoprecipitate gH synthesized in recombinant vaccinia virus-infected cells. In addition cell surface expression of gH was not detected in cells infected with the recombinant vaccinia viruses.


Chlorpromazine and Amitriptyline Are Substrates and Inhibitors of the AcrB Multidrug Efflux Pump.
Efflux is an important mechanism in Gram-negative bacteria conferring multidrug resistance. Inhibition of efflux is an encouraging strategy to restore the antibacterial activity of antibiotics. Chlorpromazine and amitriptyline have been shown to behave as efflux inhibitors. However, their mode of action is poorly understood. Exposure of Salmonella enterica serovar Typhimurium and Escherichia coli to chlorpromazine selected for mutations within genes encoding RamR and MarR, regulators of the multidrug tripartite efflux pump AcrAB-TolC. Further experiments with S. Typhimurium containing AcrB D408A (a nonfunctional efflux pump) and chlorpromazine or amitriptyline resulted in the reversion of the mutant acrB allele to the wild type. Together, this suggests these drugs are AcrB efflux substrates. Subsequent docking studies with AcrB from S. Typhimurium and E. coli, followed by molecular dynamics simulations and free energy calculations showed that chlorpromazine and amitriptyline bind at the hydrophobic trap, a preferred binding site for substrates and inhibitors within the distal binding pocket of AcrB. Based on these simulations, we suggest that chlorpromazine and amitriptyline inhibit AcrB-mediated efflux by interfering with substrate binding. Our findings provide evidence that these drugs are substrates and inhibitors of AcrB, yielding molecular details of their mechanism of action and informing drug discovery of new efflux inhibitors.IMPORTANCE Efflux pumps of the resistance nodulation-cell division (RND) superfamily are major contributors to multidrug resistance for most of the Gram-negative ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogens. The development of inhibitors of these pumps would be highly desirable; however, several issues have thus far hindered all efforts at designing new efflux inhibitory compounds devoid of adverse effects. An alternative route to de novo design relies on the use of marketed drugs, for which side effects on human health have been already assessed. In this work, we provide experimental evidence that the antipsychotic drugs chlorpromazine and amitriptyline are inhibitors of the AcrB transporter, the engine of the major RND efflux pumps in Escherichia coli and Salmonella enterica serovar Typhimurium. Furthermore, in silico calculations have provided a molecular-level picture of the inhibition mechanism, allowing rationalization of experimental data and paving the way for similar studies with other classes of marketed compounds.


Constructive neural-network learning algorithms for pattern classification.
Constructive learning algorithms offer an attractive approach for the incremental construction of near-minimal neural-network architectures for pattern classification. They help overcome the need for ad hoc and often inappropriate choices of network topology in algorithms that search for suitable weights in a priori fixed network architectures. Several such algorithms are proposed in the literature and shown to converge to zero classification errors (under certain assumptions) on tasks that involve learning a binary to binary mapping (i.e., classification problems involving binary-valued input attributes and two output categories). We present two constructive learning algorithms MPyramid-real and MTiling-real that extend the pyramid and tiling algorithms, respectively, for learning real to M-ary mappings (i.e., classification problems involving real-valued input attributes and multiple output classes). We prove the convergence of these algorithms and empirically demonstrate their applicability to practical pattern classification problems. Additionally, we show how the incorporation of a local pruning step can eliminate several redundant neurons from MTiling-real networks.


Right ventricular and pulmonary function in sickle cell disease patients with pulmonary hypertension.
The effects of sickle cell disease (SCD) on right ventricular (RV) and pulmonary function in SCD patients with pulmonary hypertension is not well-known. The aim of this study was to investigate RV and pulmonary functions in patients suffering from SCD with or without pulmonary hypertension using color tissue Doppler imaging and spirometry. We evaluated 48 asymptomatic patients with SCD. All patients underwent echocardiography with tissue Doppler imaging and pulmonary function test. Patients were divided into two groups: Group 1 consisted of 27 patients (age, 18.1 +/- 7.1 years) with normal pulmonary artery pressure, and group 2 consisted of 21 patients (age, 21.4 +/- 7.4 years) with pulmonary hypertension. Both groups were compared with a sex- and age-matched control group including 24 normal healthy subjects (age, 19.8 +/- 9.2 years). Tricuspid lateral annular systolic (S (m)) and early diastolic velocity (E (m)) were higher in group 1 than group 2 and the control group (p < 0.05). Tricuspid lateral annular late diastolic velocities (A (m)), isovolumetric contraction time, and myocardial performance index (MPI) were higher and the E (m)/A (m) ratio was lower in group 2 than group 1 and the control group (p < 0.05). However, no differences were found in the tricuspid lateral annular E (m) deceleration time, ejection time, and isovolumetric relaxation time between group 1, group 2, and the control group. Tricuspid lateral annular S (m) and E (m) were similar in group 2 and the control group. Forced expiratory volume in 1 second (FEV(1)), forced vital capacity (FVC), and the diffusion capacity of the lung for carbon monoxide were decreased in both groups of patients compared to the control group (p < 0.05). However, there was no difference in respiratory rate, FEV(1)/FVC ratio, peak expiratory flow, and total lung capacity between group 1, group 2, and the control group. There were no differences in any indices of lung function between the two groups of patients. MPI is useful index to evaluate RV function in patients with SCD. RV diastolic function was disturbed in only SCD patients with pulmonary hypertension. On the other hand, the restrictive pattern of pulmonary function abnormalities had developed in both groups of patients.


Progress in contraception: new technology.
Unintended pregnancy continues to be a major public health issue in this country. Approximately 50% of pregnancies in the United States are unintended, and, of these, half end in abortion. Although tubal sterilization is highly effective, many women subsequently express regret and remorse. Oral contraceptives represent an effective, reversible method. However, many women have difficulty using them consistently and correctly. Recently, four new delivery systems for hormonal contraception have become available in the United States: the monthly injection, the levonorgestrel intrauterine system, the combination hormonal contraceptive vaginal ring, and the transdermal contraceptive patch. All four new methods are effective, readily reversible, generally discreet, and reduce daily compliance challenges. The monthly injection, vaginal ring, and transdermal patch can be discontinued without the need for an office visit. This expanded menu of effective contraceptive options should help women find a method that suits their particular life style.