Hypo-phosphataemia in children under five years with kwashiorkor and marasmic kwashiorkor.
Severe malnutrition contributes up to 50% of childhood mortality in developing countries is frequently characterised by electrolyte depletion, including low total body phosphate. During therapeutic re-feeding, electrolyte shift from extracellular to intra-cellular compartments may induce hypo-phosphataemia (hypo-P) with resultant increased morbidity and mortality. This biochemical imbalance is under-recognised, and the frequency of this problem among African malnourished children is unclear. To determine the magnitude of hypo-phosphataemia in children under five years of age presenting to Kenyatta National Hospital with kwashiorkor and marasmic kwashiorkor and to evaluate the relationship between hypo-phosphataemia and nutritional intervention during the first five days of treatment. Short longitudinal survey. The General Paediatric wards of the Kenyatta National Hospital (KNH), Nairobi. Children under five years of age presenting with kwashiorkor or marasmic kwashiorkor at KNH were recruited into the study. Low serum phosphate level (< 1.20 mmol/l) and patient outcome (survival or death) during the first five days of treatment. One hundred and sixty five children were enrolled between June 2005 and February 2006 of which 107 (64%) had kwashiorkor and 58 (36%) had marasmic kwashiorkor. They were of mean age 20 months (range 3-60), and 95 (58%) were male. The prevalence of hypo-phosphataemia was 86% on admission, increased to 90% and 93% on day one and two respectively, and then declined to 90% by the fourth day. At admission 6% were hypo-phosphataemic, increasing to 18% and 22% on day one and two respectively, and declining to 11% by day four. On admission mean serum phosphate was below normal at 0.91 mmol/l, declined significantly to 0.67 mmol/l and to a nadir of 0.63 mmol/l after the first and second day of treatment respectively, then rose slightly to 0.75 mmol/l on the fourth day (p < 0.001 comparing each follow-up mean level with the admission level). There was a positive association between severity of nadir serum phosphate level and mortality (p = 0.028). There were no deaths among children with normal nadir serum phosphate levels. However, among children with mild, moderate and severe nadir hypo-phosphataemia, 8,14 and 21% died respectively. Children with dermatosis and hypomagnesaemia showed a trend for association with mortality (p = 0.082 and 0.099 respectively). Hypo-phosphataemia is frequent among children with kwashiorkor and marasmic kwashiorkor presenting at KNH. Serum phosphate levels decline significantly during the first two days of nutritional intervention, and severity of


Ophthalmic abnormalities secondary to periocular or ocular snakebite (pit vipers) in dogs--11 cases (2012-2014).
To describe ophthalmic abnormalities secondary to periocular and ocular snakebite in dogs. Retrospective review of medical records from dogs presenting to the Small Animal Hospital at University of Florida following snakebites to the face (2012-2014). Two groups were identified: periocular bites (PB) and ocular bites (OB). Records from eleven dogs matched the search criteria and were included in the study (PB=9, 81.8%; OB=2, 18.2%). Both OB cases involved the cornea. Facial edema, blepharospasm, chemosis, and conjunctival hyperemia occurred in all cases (100%). Hemorrhage from the eyelids occurred in eight cases (72.7%; PB=7, OB=1). Subconjunctival hemorrhage occurred in seven cases (63.6%; PB=6, OB=1). Third eyelid laceration and nictitans gland prolapse occurred in 1 case each (9%; PB=1). Lagophthalmia was present in three cases (27.3%; PB=3), with secondary corneal ulcer in two cases (18.2%; PB=2). Corneal ulcer due to direct corneal bite occurred in two cases (18.2%-partial thickness with melting (1) and full thickness (1) ). Uveitis was present in 6 cases (54.5%; PB=4, OB=2), with flare and miosis in 4 cases (36.4%; PB=2, OB=2). Hyphema, fibrin in anterior chamber, and cataract occurred in one case (9%; OB=1). Vision loss occurred in two cases (18.2%; PB=2), secondary to retinal degeneration (PB=1) and amaurosis (PB=1). Mean follow-up time was 7 weeks (range: 3 days-11 months). Most clinical signs had resolved by last examination. Periocular symptoms were more commonly observed than ocular alterations, regardless of bite location. Appropriate supportive therapy should be instituted according to clinical signs.


Effects of buttermilk powders on emulsification properties and acid tolerance of cream.
Emulsifying properties and acid tolerance are 2 of the most important characteristics of cream. The effects of the buttermilk component, especially its phospholipids, on the emulsifying properties and acid tolerance of cream were investigated in this study. Two buttermilks with differing phospholipid contents and skimmed milk were used to evaluate the effects of phospholipids on the aforementioned parameters. The mean diameter of fat globules and the cream viscosity were used as indicators of emulsifying properties. Acid tolerance was evaluated by studying the effect of citric acid on the maximum viscosity of cream. This was tested by adding 400 μL of 10% (w/w) citric acid solution to cream every minute and simultaneously measuring pH and viscosity. In 45% and 40% fat cream systems, buttermilk, and especially that with higher phospholipid content, improved the emulsifying properties and acid tolerance of the cream. The components of buttermilk could alter the properties of the surface of fat globules, thereby altering the emulsification properties of the cream. However, neither of the tested buttermilks affected the emulsifying properties and acid tolerance of lower-fat (35% and 30%) cream systems. Emulsifying components exist in proportionately larger amounts in lower-fat creams, which could render the emulsifying properties resistant to change. The number of fat globules may also influence acid-induced changes in viscosity. The addition of phospholipids or lysophospholipids did not improve the acid tolerance of creams, a finding that may be attributable to the formation of complexes of phospholipids and protein. The findings presented herein demonstrate the ability to improve the acid tolerance of cream using materials derived from milk. Implementing these findings appropriately may result in a high-quality cooking cream.


AKAP12 mediates barrier functions of fibrotic scars during CNS repair.
The repair process after CNS injury shows a well-organized cascade of three distinct stages: inflammation, new tissue formation, and remodeling. In the new tissue formation stage, various cells migrate and form the fibrotic scar surrounding the lesion site. The fibrotic scar is known as an obstacle for axonal regeneration in the remodeling stage. However, the role of the fibrotic scar in the new tissue formation stage remains largely unknown. We found that the number of A-kinase anchoring protein 12 (AKAP12)-positive cells in the fibrotic scar was increased over time, and the cells formed a structure which traps various immune cells. Furthermore, the AKAP12-positive cells strongly express junction proteins which enable the structure to function as a physical barrier. In in vivo validation, AKAP12 knock-out (KO) mice showed leakage from a lesion, resulting from an impaired structure with the loss of the junction complex. Consistently, focal brain injury in the AKAP12 KO mice led to extended inflammation and more severe tissue damage compared to the wild type (WT) mice. Accordingly, our results suggest that AKAP12-positive cells in the fibrotic scar may restrict excessive inflammation, demonstrating certain mechanisms that could underlie the beneficial actions of the fibrotic scar in the new tissue formation stage during the CNS repair process.


Exercise issues in older cancer survivors.
Older cancer survivors experience the combined deleterious effects associated with aging and a cancer diagnosis. The purpose of the present paper is to review the potential role of physical exercise in attenuating the effects of cancer and its treatments in older cancer survivors. Noting the limited direct research on exercise in older cancer survivors, we review the literature on: (a) older adults in general; and (b) cancer survivors in general. We conclude that although there is limited direct evidence on the benefits of exercise in older cancer survivors, there is compelling evidence of the benefits of exercise in cancer survivors in general and other older populations. We also conclude that exercise adherence will be a significant challenge in this population. Based on this evidence, we tentatively recommend exercise to older cancer survivors using the American College of Sports Medicine's [Med Sci Sports Exerc 30 (6) (1998) 992] guidelines for older adults in general. Finally, we offer future research directions that will provide important evidence needed to guide clinical decisions about exercise in older cancer survivors.


[Musculoskeletal pain in pediatric patients. Prevalence and etiology in Primary Care].
1) To determine the number of clinic visits in primary care due to musculoskeletal pain in children > or = 3 years. 2) To describe the demographic characteristics of the population assessed for limb/back pain. 3) To characterize the etiology of musculoskeletal pain in a general pediatric clinic. A prospective lineal study was performed between October 30, 1996 and January 14, 1997 in two urban general pediatric clinics located in the city of Madrid (Spain). During the period of the study, all consultations related to musculoskeletal pain were registered in a protocol form. In addition, a registry of all clinic visits was maintained during the study. During the study period, 43 clinic visits were due to musculoskeletal pain, representing 6.2% (43/692) of the clinic visits in children > or = 3 years of age (95% confidence interval 4.4-7.9%). The mean age was 9.8 years (+/- 3.2 years). These complaints, although statistically non-significant, were more frequent in children > or = 10 years old (53.4%) and in males (65.1%). The most frequent complaints were knee athralgias (30%), arthralgias of other joints (23%) and soft tissue pain (18%). These symptoms were usually due to overuse syndromes and normal skeletal growth variants. The increase of primary pediatric care age from 7 to 14 years of age is associated with an increase in the number of consultations related to musculoskeletal pain. This information should be kept in mind at the time of designing continuing medical education and training programs in pediatrics.


Cognitive status and quality of life in patients with suspected versus proven low-grade gliomas.
The preferred management of patients with suspected low-grade gliomas (S-LGG) remains controversial. The benefits of resection or radiotherapy early in the course of the disease have not been proven in terms of survival. Little is known about the effects of early therapy on quality of life (QOL) and cognitive status. The authors compared functional status, QOL, and cognitive status of patients suspected of having a LGG, in whom treatment was deferred, and patients with proven LGG (P-LGG), who underwent early surgery. The authors recruited 24 patients suspected of having an LGG. These patients were matched with 24 patients with a histologically proven LGG and healthy control subjects for educational level, handedness, age, and gender. The two patient groups were also matched for tumor laterality, use of anticonvulsants, and interval between diagnosis and testing. Functional status was determined in both patient groups. QOL and cognitive status were compared between the three groups. Matching criteria and functional status did not differ significantly between groups. Both patient groups scored worse on QOL scales than healthy control subjects. Unoperated patients with S-LGG scored better on most items than patients with P-LGG. Cognitive status was worse in both groups than in healthy control subjects, but, again, patients with S-LGG performed better than patients with P-LGG. These data suggest that a wait-and-see policy in patients with S-LGG has no negative effect on cognitive performance and QOL.


Effect of dentin age and acid etching time on dentin bonding.
To compare the bond strengths of young and old dentin using a single-bottle etch-and-rinse adhesive system with two acid-etching times (15 s and 30 s), and to examine the correlation between dentin hardness and bond strength. Twenty-four molars from 18- to 22- or 55- to 60-year-old patients were prepared to expose the occlusal dentin, cut into two equal parts, and assigned to four groups (G), varying the etching time (15 s and 30 s) and dentin age (young [Y]or old [O]): G15Y, G30Y, G15O and G30O. After etching, AdperSingleBond (3M ESPE) adhesive was applied and the tooth was built up with a composite (Filtek Z250). The specimens were prepared for the microtensile bond strength test (μTBS) at a crosshead speed of 0.5 mm/min and the interfaces were observed under SEM. Also, the Knoop hardness (KHN) of young vs old dentin was measured. The mean (± SD) μTBS (MPa) were: G15Y = 45.9 (± 10.7)a, G15O = 34.1 (± 9.4)b, G30Y = 48.6 (± 14.3)a and G30O = 47.7 (± 11.0)a. Two-way ANOVA showed no difference in μTBS between young and old dentin. Old dentin acid etched for 15 s (G15O) had a lower μTBS than when acid etched for 30 s (G30O). Dentin hardness was higher for old dentin (67.0 ± 4.8KHN) than young dentin (63.7 ± 2.9KHN) (p < 0.004). No correlation between μTBS and dentin hardness was observed. Resin tags were larger and more numerous for young dentin. The hybrid layer formed in intertubular old dentin (G15O) was very thin. Bonding to old dentin with 30 s of etching time resulted in higher bond strength and more homogeneous hybrid layer formation than dentin acid etched for 15 s.


Managements of placenta accreta.
From January 1986 to June 1990 at Veterans General Hospital-Taipei, 12 cases of placenta accreta were reviewed. The incidence were 1 per 1,101 deliveries. Eight cases were confirmed by pathological examination, and the incidence was 0.06% (1/1,652). Placenta previa was found in 7 cases (58.3%); 6 of them had history of cesarean section. All the cases had history of uterine surgery. Seven cases underwent cesarean section, and the other one received myomectomy. Hysterectomy was performed on 6 patients, and conservative treatments were employed on the other 6 cases. No maternal mortality occurred. Besides one case of missed abortion, there was only one perinatal loss due to immaturity. Accurate prenatal diagnosis of placenta accreta by ultrasound was obtained in 4 cases. Although hysterectomy is the definite treatment of placenta accreta, conservative managements should be tried before this intervention.


Evaluation of the Adult Strabismus-20 (AS-20) questionnaire using Rasch analysis.
To further refine the Adult Strabismus 20 (AS-20) health-related quality of life (HRQOL) questionnaire using Rasch analysis. Rasch analysis was performed independently on the original AS-20 using the following steps: dimensionality, response ordering, local dependence, infit and outfit analyses, differential item functioning, subject targeting, and confirmatory dimensionality. Two subscales were present in each of the original AS-20 subscales, for a total of 4 subscales, which were labeled "self-perception," "interaction," "reading function," and "general function." Response ordering was appropriate for 3 of the subscales but required reduction to 4 response options for the fourth subscale. No notable local dependence was found for any subscale. As a result of fit analysis, 2 items were removed, 1 each from 2 subscales. No significant differential item functioning was seen for sex or age. The resulting 5-item self-perception subscale and 4-item reading function subscale are reliable and target the adult strabismus patient cohort appropriately. The resulting 5-item interaction subscale and 4-item general function subscale have less than optimal reliability. The AS-20 benefits from reduction to 4 subscales (self-perception, interaction, reading function, and general function) and reducing the response options in the general function subscale from 5 to 4 options. The refined AS-20 may prove to be even more responsive to HRQOL changes in adult strabismus following treatment or changes over time.


Arabidopsis DNA ligase IV is induced by gamma-irradiation and interacts with an Arabidopsis homologue of the double strand break repair protein XRCC4.
Rejoining of single- and double-strand breaks (DSBs) introduced in DNA during replication, recombination, and DNA damage is catalysed by DNA ligase enzymes. Eukaryotes possess multiple DNA ligase enzymes, each having distinct roles in cellular metabolism. Double-strand breaks in DNA, which can occur spontaneously in the cell or be induced experimentally by gamma-irradiation, represent one of the most serious threats to genomic integrity. Non-homologous end joining (NHEJ) rather than homologous recombination is the major pathway for repair of DSBs in organisms with complex genomes, including humans and plants. DNA ligase IV in Saccharomyces cerevisiae and humans catalyses the final step in the NHEJ pathway of DSB repair. In this study we identify an Arabidopsis thaliana homologue (AtLIG4) of human and S. cerevisiae DNA ligase IV which is shown to encode an ATP-dependent DNA ligase with a theoretical molecular mass of 138 kDa and 48% similarity in amino-acid sequence to the human DNA ligase IV. Yeast two-hybrid analysis demonstrated a strong interaction between A. thaliana DNA ligase IV and the A. thaliana homologue of the human DNA ligase IV-binding protein XRCC4. This interaction is shown to be mediated via the tandem BRCA C-terminal domains of A. thaliana DNA ligase IV protein. Expression of AtLIG4 is induced by gamma-irradiation but not by UVB irradiation, consistent with an in vivo role for the A. thaliana DNA ligase IV in DSB repair.


Unlinked vital events in census-based longitudinal studies can bias subsequent analysis.
To examine the potential biases arising from the nonlinkage of census records and vital events in longitudinal studies. A total of 56,396 deaths of residents of Northern Ireland in the 4 years after the 2001 Census were linked to the 2001 Census records. The characteristics of matched and nonmatched death records were compared using multivariate logistic regression. Subject attributes were as recorded on the death certificate. In total, 3,392 (6.0%) deaths could not be linked to a census record. Linkage rates were lowest in young adults, males, the unmarried, people living in communal establishments, or living in areas that were more deprived or had recorded low census enumeration. For those aged less than 65 years at census, this linkage would exclude from analysis 20.2% of suicides and 19.7% of deaths by external causes. The nonlinkage of census and death records is a combination of nonenumeration at census and deficient information about the deceased recorded at the time of death. Unmatched individuals may have been more disadvantaged or socially isolated, and analysis based on the linked data set may therefore show some bias and perhaps understate true social gradients.


Aorto-duodenal fistula: a rare but serious complication of gastrointestinal hemorrhage. A case report.
A primary aortoduodenal fistula (PADF) is a rare cause of gastrointestinal bleeding that is difficult to diagnose (and sometimes not diagnosed until a laparotomy.) A PADF is associated with high mortality if undiagnosed and untreated (the mortality rate of nearly 100% in the absence of a surgical intervention). While this condition is extremely rare with an incidence rate at autopsy of 0.04% to 0.07%, a secondary ADF occurs much more commonly (the post-operative incidence of 0.5% to 2.3%) and is due to prior aortic surgery and/or the placement of a synthetic aortic graft. It should be considered in any elderly patient who presents with upper gastrointestinal bleeding in the context of a known abdominal aortic aneurysm or without it when no identifiable source of bleeding is found. We present an autopsy case of a 59-year-old man with no history of an abdominal aortic aneurysm who presented with intermittent massive gastrointestinal bleeding. The autopsy revealed a pinhole fistula. It was identified between an atherosclerotic abdominal aortic aneurysm and the lower horizontal part of the duodenum. Our case indicates that the aortoenteric fistula can result in fatal gastrointestinal bleeding. This case is unique in that the fistula formed as a result of a complex atherosclerotic abdominal aorta and a localized necrotizing granulomatous aortitis the etiology of which was not clear.


Nonverbal teaching behavior and the health educator.
The role of the health educator is rapidly changing. With this change comes the need to analyze the teaching techniques used by the health educator to disseminate health information. There is very little research to date in health education dealing with the issue of teacher-learner interaction. Because of the unique issues raised in this domain, research designs used in other areas of education do not meet the specific needs of health education. The study was designed initially to determine if nonverbal teaching behavior of health educators would be different if they were exposed to a systematic self-analysis process. Utilizing health educators in 28 secondary schools in central New York State, the subjects were videotaped and their behavior analyzed with a self-analysis instrument. The results of the study showed a need for written and oral feedback if health educators are to become aware of their teaching behavior. On-going self-analysis, too, appeared essential in changing teaching behavior as did the use of videotaping. Duplicate studies in this area are essential in determining whether teacher behavior can be altered in a positive direction.


When nature turns up the heat.
Heat-related illness can present as prickly heat, heat cramps, heat exhaustion, or heat stroke. Can you spot the different types and provide the appropriate care? Can you offer advice on how to prevent these disorders in the first place? This article will allow you to answer Yes.


Brain Network Disruption in Whiplash.
Whiplash-associated disorders frequently develop following motor vehicle collisions and often involve a range of cognitive and affective symptoms, though the neural correlates of the disorder are largely unknown. In this study, a sample of participants with chronic whiplash injuries were scanned by using resting-state fMRI to assess brain network changes associated with long-term outcome metrics. Resting-state fMRI was collected for 23 participants and used to calculate network modularity, a quantitative measure of the functional segregation of brain region communities. This was analyzed for associations with whiplash-associated disorder outcome metrics, including scales of neck disability, traumatic distress, depression, and pain. In addition to these clinical scales, cervical muscle fat infiltration was quantified by using Dixon fat-water imaging, which has shown promise as a biomarker for assessing disorder severity and predicting recovery in chronic whiplash. An association was found between brain network structure and muscle fat infiltration, wherein lower network modularity was associated with larger amounts of cervical muscle fat infiltration after controlling for age, sex, body mass index, and scan motion (t = -4.02, partial R 2 = 0.49, P < .001). This work contributes to the existing whiplash literature by examining a sample of participants with whiplash-associated disorder by using resting-state fMRI. Less modular brain networks were found to be associated with greater amounts of cervical muscle fat infiltration suggesting a connection between disorder severity and neurologic changes, and a potential role for neuroimaging in understanding the pathophysiology of chronic whiplash-associated disorders.


SIGLEC-G deficiency increases susceptibility to develop B-cell lymphoproliferative disorders.
The sialic-acid-binding immunoglobulin-like lectin SIGLEC-G is a negative regulator of B-cell receptor-mediated calcium signaling. Its deficiency leads to reduced turnover and increased proliferation and survival of murine B-1a cells. Siglecg(-/-) mice show a premature expansion of polyclonal CD5(+) B cells in the spleen and the peritoneal cavity. Here we studied the fate of B lymphocytes in Siglecg(-/-) mice over time. We demonstrate that in aging animals SIGLEC-G deficiency promotes progressive accumulation of monoclonal B lymphocytes and increases the susceptibility to develop B-cell lymphoproliferative disorders. Lymphoid tumors arising in aged Siglecg(-/-) mice are monoclonal and histologically heterogeneous as they include diffuse large B-cell lymphoma, follicular lymphoma, and medium-to-large B-cell monomorphic lymphoma but surprisingly not chronic lymphocytic leukemia. The tumors express high levels of BCL-2 and are transplantable. In keeping with these findings we have also observed a remarkable down-regulation of the human ortholog SIGLEC10 in human B-cell lymphoma and leukemia cell lines. Taken together, these observations indicate that the down-regulation of negative B-cell receptor regulators such as SIGLEC-G/SIGLEC10 may represent another mechanism relevant to the pathogenesis of B-cell lymphomas.


Altered thymidine-thymidylate kinases from strains of herpes simplex virus with modified drug sensitivities to acyclovir and (E)-5-(2-bromovinyl)-2'-deoxyuridine.
Virus-coded thymidine (dThd) kinases were purified by affinity chromatography from a parental strain (SC16) and two strains (SC16 B3 and SC16 S1) of herpes simplex virus, Type 1, with altered drug sensitivities. These latter two strains were less sensitive, respectively, to E-5-(2-bromovinyl)-2'-deoxyuridine (BrVdUrd) and to both BrVdUrd and 9-(2-hydroxyethoxymethyl)guanine (acyclovir). The enzymes were characterized with respect to physical and catalytic properties. The enzyme from SC16 B3 was very similar to the parental enzyme except in its substrate specificity and kinetic constants. It catalyzed the phosphorylation of BrVdUrd at a relative rate that was 110% of the rate with dThd versus a relative rate of 140% with the parental enzyme. The apparent Km value for BrVdUrd was 6 microM versus 0.1 microM for the parental enzyme. The reaction kinetics with acyclovir were similar for the two enzymes. The SC16 B3 enzyme catalyzed the phosphorylation of dTMP, but at only 2% the efficiency of the parental enzyme; phosphorylation of the monophosphate of BrVdUrd (BrVdUMP) was not detected with the SC16 B3 enzyme. The enzyme from the SC16 S1 variant had a much narrower phosphate acceptor specificity than the enzyme from the parental virus. BrVdUrd was a substrate but with a relative rate of 30% and an apparent Km value of 4 microM; acyclovir was neither detectably phosphorylated nor a good inhibitor. BrVdUMP was not detectably phosphorylated. The relative efficiencies of the two variant enzymes for acyclovir phosphorylation correlated well with the sensitivities of the viruses to this compound. In contrast, the relative efficiencies of the second phosphorylation step (BrVdUMP to BrVdUDP) were most consistent with the sensitivities of the viruses to BrVdUrd.


Neonatal skinfold thickness. Measurement and interpretation at or near term.
Skinfold thickness was measured at five sites in 750 infants. The study population was unselected except that twin pregnancies and the infants of diabetic mothers were excluded, and very preterm infants were under represented. A pilot study had indicated that skinfold measurement was most reproducible at the thigh site. Thigh skinfold correlated better with the sum of other skinfolds than did skinfold measurement at any other site and closely resembled the summed skinfold in correlations with a number of maternal and fetal variables. Median skinfold increased with birthweight and was greater in girls than in boys. 'Corrected skinfold', a mathematical approach to comparing skinfolds in infants of differing sex and birthweight, is suggested as an alternative to absolute skinfold measurement.


Oxygen triggers signal transduction in the DevS (DosS) sensor of Mycobacterium tuberculosis by modulating the quaternary structure.
A major challenge to the control and eventual eradication of Mycobacterium tuberculosis infection is this pathogen's prolonged dormancy. The heme-based oxygen sensor protein DevS (DosS) plays a key role in this phenomenon, because it is a major activator of the transcription factor DevR. When DevS is active, its histidine protein kinase region is ON and it phosphorylates and activates DevR, which can induce the transcription of the dormancy regulon genes. Here, we have investigated the mechanism by which the ligation of molecular oxygen to a heme-binding domain in DevS switches OFF its histidine protein kinase region. To shed light on the oligomerization states of this protein and possible protein-surfaces of interaction, we used analytical gel filtration, together with dynamic light scattering, fluorescence spectroscopy and chemical crosslinking. We found that DevS exists as three major species: an octamer, a tetramer and a dimer. These three states were observed for the concentration range between 0.5 and 20 μm DevS, but not below 0.1 μm. Levels of DevS in M. tuberculosis are expected to range from 5 to 26 μm. When this histidine protein kinase was OFF, the DevS was mainly tetrameric and dimeric; by contrast, when the kinase was ON, the protein was predominantly octameric. The changes in quaternary structure were rapid upon binding to the physiological signal. This finding represents a novel strategy for switching the activity of a two-component heme-based sensor. An enhanced understanding of this process might potentially lead to the design of novel regulatory agents that target the multimer interfaces for treatment of latent tuberculosis.


Effects of Green Tea Gargling on the Prevention of Influenza Infection: An Analysis Using Bayesian Approaches.
The aim of this study is to analyze the data obtained from a randomized trial on the prevention of influenza by gargling with green tea, which gave nonsignificant results based on frequentist approaches, by using Bayesian approaches. The posterior proportion, with 95% credible interval (CrI), of influenza in each group was calculated. The Bayesian index θ is the probability that a hypothesis is true. In this case, θ is the probability that the hypothesis that green tea gargling reduced influenza compared with water gargling is true. Univariate and multivariate logistic regression analyses were also performed by using the Markov chain Monte Carlo method. The full analysis set included 747 participants. During the study period, influenza occurred in 44 participants (5.9%). The difference between the two independent binominal proportions was -0.019 (95% CrI, -0.054 to 0.015; θ = 0.87). The partial regression coefficients in the univariate analysis were -0.35 (95% CrI, -1.00 to 0.24) with use of a uniform prior and -0.34 (95% CrI, -0.96 to 0.27) with use of a Jeffreys prior. In the multivariate analysis, the values were -0.37 (95% CrI, -0.96 to 0.30) and -0.36 (95% CrI, -1.03 to 0.21), respectively. The difference between the two independent binominal proportions was less than 0, and θ was greater than 0.85. Therefore, green tea gargling may slightly reduce influenza compared with water gargling. This analysis suggests that green tea gargling can be an additional preventive measure for use with other pharmaceutical and nonpharmaceutical measures and indicates the need for additional studies to confirm the effect of green tea gargling.


Differential effects of conventional and high-definition transcranial direct-current stimulation of the motor cortex on implicit motor sequence learning.
Conventional transcranial direct-current stimulation (tDCS) delivered to the primary motor cortex (M1) has been shown to enhance implicit motor sequence learning (IMSL). Conventional tDCS targets M1 but also the motor association cortices (MAC), making the precise contribution of these areas to IMSL presently unclear. We aimed to address this issue by comparing conventional tDCS of M1 and MAC to 4 * 1 high-definition (HD) tDCS, which more focally targets M1. In this mixed-factorial, sham-controlled, crossover study in 89 healthy young adults, we used mixed-effects models to analyse sequence-specific and general learning effects in the acquisition and short- and long-term consolidation phases of IMSL, as measured by the serial reaction time task. Conventional tDCS did not influence general learning, improved sequence-specific learning during acquisition (anodal: M = 42.64 ms, sham: M = 32.87 ms, p = .041), and seemingly deteriorated it at long-term consolidation (anodal: M = 75.37 ms, sham: M = 86.63 ms, p = .019). HD tDCS did not influence general learning, slowed performance specifically in sequential blocks across all learning phases (all p's < .050), and consequently deteriorated sequence-specific learning during acquisition (anodal: M = 24.13 ms, sham: M = 35.67 ms, p = .014) and long-term consolidation (anodal: M = 60.03 ms, sham: M = 75.01 ms, p = .002). Our findings indicate that the observed superior conventional tDCS effects on IMSL are possibly attributable to a generalized stimulation of M1 and/or adjacent MAC, rather than M1 alone. Alternatively, the differential effects can be attributed to cathodal inhibition of other cortical areas involved in IMSL by the 4 * 1 HD tDCS return electrodes, and/or more variable electric field strengths induced by HD tDCS, compared with conventional tDCS.


Deletion mapping of functional domains in the rotavirus capsid protein VP6.
VP6, the major capsid protein of rotavirus, oligomerizes into trimers that constitute the intermediate shell of the virions. In order to map functional domains in this protein, we introduced seven internal in-frame deletions within the coding region of gene 6 of human rotavirus strain Wa. Regions of homology among the VP6 proteins of group A and group C rotaviruses were targeted for deletion mutagenesis. The mutant VP6 proteins were expressed in mammalian cells using the recombinant vaccinia virus system and were examined for their ability to oligomerize into trimers as well as to assemble into double-layered virus-like particles upon coexpression with the rotavirus core protein VP2. Deletions that abolished trimerization defined a domain (residues 246 to 314) that maps within a larger region previously found to be critical for oligomerization (amino acids 105 to 328). When the capacity of each mutant to assemble into double-layered virus-like particles was analysed, three different assembly phenotypes were observed. Phenotype I was represented by two deletion mutants lacking residues 246 to 250 and 308 to 314 that produced particles with efficiencies similar to that of wild-type VP6. Phenotype II, characterized by a moderate decrease in the efficiency of particle assembly with respect to that of wild-type VP6, included two mutants with deletions at the C terminus of the protein. Phenotype III was exhibited by three mutants whose abilities to assemble into double-layered virus-like particles were drastically impaired. Two of these mutants define a previously unidentified assembly domain (amino acids 122 to 147) at the N terminus of rotavirus VP6.


Intestinal specific LXR activation stimulates reverse cholesterol transport and protects from atherosclerosis.
Several steps of the HDL-mediated reverse cholesterol transport (RCT) are transcriptionally regulated by the nuclear receptors LXRs in the macrophages, liver, and intestine. Systemic LXR activation via synthetic ligands induces RCT but also causes increased hepatic fatty acid synthesis and steatosis, limiting the potential therapeutic use of LXR agonists. During the last few years, the participation of the intestine in the control of RCT has appeared more evident. Here we show that while hepatic-specific LXR activation does not contribute to RCT, intestinal-specific LXR activation leads to decreased intestinal cholesterol absorption, improved lipoprotein profile, and increased RCT in vivo in the absence of hepatic steatosis. These events protect against atherosclerosis in the background of the LDLR-deficient mice. Our study fully characterizes the molecular and metabolic scenario that elects the intestine as a key player in the LXR-driven protective environment against cardiovascular disease.


Connexin45 interacts with zonula occludens-1 and connexin43 in osteoblastic cells.
The relative expression of connexin43 and connexin45 modulates gap junctional communication and production of bone matrix proteins in osteoblastic cells. It is likely that changes in gap junction permeability are determined by the interaction between these two proteins. Cx43 interacts with ZO-1, which may be involved in trafficking of Cx43 or facilitating interactions between Cx43 and other proteins. In this study we sought to identify proteins that associate with Cx45 by coprecipitation in non-denaturing conditions. Cx45 was isolated with a 220-kDa protein that we identified as ZO-1. Under the same conditions, Cx43 also was isolated with anti-Cx45 antiserum from Cx45-transfected ROS cells (ROS/Cx45 cells). Cx43 antiserum could also coprecipitate ZO-1 in the transfected and untransfected ROS cells. Double label immunofluorescence studies showed that ZO-1, Cx43, and Cx45 colocalized at appositional membranes in ROS/Cx45 cells suggesting that all three proteins are normally associated in the cells. Additionally, we found that in vitro translated ZO-1 binds to the carboxyl-terminal of Cx45 indicating that there is a direct interaction between the carboxyl-terminal of Cx45 and ZO-1. These studies demonstrate that ZO-1 interacts with Cx45 as well as with Cx43, and suggest that the interaction of connexins with ZO-1 may play a role in regulating the composition of the gap junction and may modulate connexin-connexin interactions.


[Structural-functional analysis of diffusion in glucose absorption by rat small intestine enterocytes].
To elucidate mechanisms providing transport of sugars across intestinal epithelium, on taking into account the current hypotheses (active transport, participation of paracellular transport and passive component of transcellular transport), it was important to reveal structural changes of tight junctions and distribution of the carriers of facilitated diffusion of GLUT2 and protein kinase C during absorption of glucose. On using confocal and electron microscopy, ultrastructural and immunocytochemical studies of enterocytes after perfusion of isolated rat small intestine fragment with 75 mM glucose (chronic experiment) have shown: 1) fluorescent labels of transporter GLUT2 and PKCbetaII are located in the apical area of enterocytes situated at the upper half of the villus. Antibodies against GLUT2, conjugated with gold, are revealed at the microvilli or apical membrane and in the area of terminal network; 2) no ultrastructural changes of the tight junction are detected on ultrathin sections and freeze--fracture replics. At the same time, fluorescent and gold labels against actin are concentrated in the vicinity of the lateral membrane in the tight junction area. The results obtained can serve a confirmation of a hypothesis that at high glucose concentrations GLUT2 participates in its transfer across the apical membrane.


Endoscopic hydroxyapatite augmentation for patulous Eustachian tube.
To evaluate the safety and efficacy of endoscopic calcium hydroxyapatite injection in patients with patulous Eustachian tube. Retrospective case series. Fourteen patients diagnosed with patulous Eustachian tube (PET) underwent endoscopic hydroxyapatite injection under general anesthesia. All patients had at least two of three major PET symptoms including voice autophony, breathing autophony, or aural fullness/pressure. Patients were evaluated postoperatively with nasal endoscopy and a symptom questionnaire. Endoscopic Eustachian tube injection was performed in a total of 23 sides in 14 patients with an average volume of 2.1 cc injected per side. Mean follow-up was 17.5 months. The most common symptoms reported preoperatively were voice autophony (96%), breathing autophony (91%), and ear fullness (83%). A complete or significant response to treatment was noted in 13/22 sides with voice autophony (59%), 12/21 sides with breathing autophony (57%), and 12/19 sides with ear fullness (63%). All complete or significant symptom improvements remained durable through the entirety of the follow-up period. Four sides that had temporary or no improvement with treatment underwent repeat injection but did not achieve additional improvement. No intraoperative or postoperative complications were observed. Endoscopic hydroxyapatite injection of the Eustachian tube is a minimally invasive procedure that provided significant or complete relief of autophony and ear fullness in 57% to 63% of sides treated. The procedure is well tolerated and can be performed safely under endoscopic visualization. Hydroxyapatite injection may be a satisfactory alternative to more invasive treatments for PET.


Pharmacological and molecular characterization of 5-hydroxytryptamine(7) receptors in the rat adrenal gland.
Serotonin (5-hydroxytryptamine; 5-HT) is a potent stimulator of aldosterone secretion in the rat adrenal gland but the type of receptor involved in the mechanism of action of 5-HT remains unknown. The aim of the present study was to determine the pharmacological profile and to clone the receptor responsible for the corticotropic effect of 5-HT in rat glomerulosa cells. A series of 10 serotonergic receptor agonists and 12 receptor antagonists was used to characterize the receptor mediating the effect of 5-HT on aldosterone secretion from perifused rat adrenocortical slices. Correlation analysis between the potencies of the different compounds in our model and those previously reported for various recombinant 5-HT receptors showed that the rat adrenal 5-HT receptor exhibits the same pharmacological profile as the 5-HT(7) receptor transiently expressed in COS-7 cells (r = 0.82 for agonists, p <.05; r = 0.83 for antagonists, p <.01). Polymerase chain reaction with specific primers revealed the expression of 5-HT(7) receptor mRNA in the rat adrenal gland. Cloning of the polymerase chain reaction product confirmed that the amplified DNA corresponded to the 5-HT(7) receptor cDNA sequence. Western blot analysis showed the presence of a protein with an apparent molecular mass of 66 kDa in the adrenal cortex but not in the medulla. Taken together, these data demonstrate that the rat adrenal glomerulosa expresses functional 5-HT(7) receptors. Rat glomerulosa cells will thus provide a robust and sensitive bioassay for future studies on native 5-HT(7) receptors.


Monitoring Protein-Unbound Valproic Acid Serum Concentrations in Clinical Practice.
Valproic acid (VPA) is an effective antiepileptic drug and mood stabilizer. A key characteristic of VPA is its high and saturable protein binding at higher concentrations. Although the unbound concentration of VPA is responsible for its pharmacological activity, total drug concentrations are monitored in routine clinical practice. Therapeutic drug monitoring (TDM) of unbound VPA is recommended for specific clinical situations. The goal of this study was to evaluate TDM requests for unbound VPA in clinical practice. All TDM requests at our laboratory for unbound VPA in 2014 and 2015 were evaluated retrospectively. In patients with potentially toxic unbound VPA concentrations (ie, >12 mg/L), we evaluated whether toxicity was noted and whether the dose adjustment advice was followed. Total and unbound VPA concentrations were measured by means of a validated immunoassay. A total of 273 unbound VPA serum concentrations in 132 different patients were analyzed. The main reasons for unbound VPA TDM were decreased renal function (34%) and a low serum albumin (27%). The median (range) unbound VPA concentration was 9.8 (2.5-47.6) mg/L. In 49 patients (37%), the initial unbound VPA concentration was above the threshold of 12 mg/L, potentially resulting in toxicity. Only 6 of these 49 patients had elevated total VPA concentrations. Clinical toxicity was noted in 38 of the 49 patients (77.6%) with elevated unbound VPA concentrations. Toxicities included drowsiness (n = 26), decreased consciousness (n = 4), rigidity (n = 2), and confusion (n = 2). In 36 of the 38 patients with elevated unbound VPA concentrations and clinical toxicity, a dose reduction was applied. In 27 of 36 patients who had their dose reduced, dose reduction was associated with improvement or resolution of VPA toxicity. TDM of unbound VPA is an important tool to manage VPA therapy in selected, vulnerable patients.


Effects of beta-mannanase addition to corn-soybean meal diets on growth performance, carcass traits, and nutrient digestibility of weanling and growing-finishing pigs.
Four experiments were conducted to determine the effects of adding a beta-mannanase preparation (Hemicell, ChemGen, Gaithersburg, MD) to corn-soybean meal-based diets on growth performance and nutrient digestibility of weanling and growing-finishing pigs. In Exp. 1, 156 weanling pigs (20 d, 6.27 kg BW) were allotted to four dietary treatments in a randomized complete block design. Treatments were a factorial arrangement of diet complexity (complex vs simple) and addition of 3-mannanase preparation (0 vs 0.05%). Pigs were fed in three dietary phases (Phase 1, d 0 to 14; Phase 2, d 14 to 28; and Phase 3, d 28 to 42). Pigs fed complex diets gained faster and were more efficient (P < 0.05) during Phase 1 compared with pigs fed simple diets. Overall, gain:feed ratio (G:F) tended to be improved (P < 0.10) for pigs fed complex diets and it was improved (P < 0.01) for those fed diets with beta-mannanase. In Exp. 2, 117 pigs (44 d, 13.62 kg BW) were allotted randomly to three dietary treatments. Dietary treatments were 1) a corn-soybean meal-based control, 2) the control diet with soybean oil added to increase metabolizable energy (ME) by 100 kcal/kg, and 3) the control diet with 0.05% beta-mannanase preparation. Beta-mannanase or soybean oil improved (P < 0.05) G:F compared with pigs fed the control diet. In Exp. 3, 60 pigs (22.5 kg BW) were allotted randomly to the three dietary treatments used in Exp. 2. Dietary treatments were fed in three phases (23 to 53 kg, 53 to 82 kg, and 82 to 109 kg with 0.95, 0.80, and 0.65% lysine, respectively). Overall, the addition of soybean oil tended to improve G:F (P < 0.10) compared with that of pigs fed the control diet, and G:F was similar (P > 0.54) for pigs fed diets with soybean oil or beta-mannanase. Also, addition of beta-mannanase increased ADG (P < 0.05) compared with that of pigs fed the control or soybean oil diets. There were no differences (P > or = 0.10) in longissimus muscle area or backfat; however, on a fat-free basis, pigs fed the diet with beta-mannanase had greater (P < 0.05) lean gain than pigs fed the control or soybean oil diets. In Exp. 4, 12 barrows (93 kg BW) were allotted randomly to one of the three dietary treatments used in Exp. 3. Addition of 3-mannanase had no effect (P > 0.10) on energy, nitrogen, phosphorus, or dry matter digestibility. These results suggest that beta-mannanase may improve growth performance in weanling and growing-finishing pigs but has minimal effects on nutrient digestibility.


Heart Rate Variability and Training Load Among National Collegiate Athletic Association Division 1 College Football Players Throughout Spring Camp.
Flatt, AA, Esco, MR, Allen, JR, Robinson, JB, Earley, RL, Fedewa, MV, Bragg, A, Keith, CM, and Wingo, JE. Heart rate variability and training load among National Collegiate Athletic Association Division 1 college football players throughout spring camp. J Strength Cond Res 32(11): 3127-3134, 2018-The purpose of this study was to determine whether recovery of cardiac-autonomic activity to baseline occurs between consecutive-day training sessions among positional groups of a collegiate football team during Spring camp. A secondary aim was to evaluate relationships between chronic (i.e., 4-week) heart rate variability (HRV) and training load parameters. Baseline HRV (lnRMSSD_BL) was compared with HRV after ∼20 hours of recovery before next-day training (lnRMSSDpost20) among positional groups composed of SKILL (n = 11), MID-SKILL (n = 9), and LINEMEN (n = 5) with a linear mixed model and effect sizes (ES). Pearson and partial correlations were used to quantify relationships between chronic mean and coefficient of variation (CV) of lnRMSSD (lnRMSSD_chronic and lnRMSSDcv, respectively) with the mean and CV of PlayerLoad (PL_chronic and PL_cv, respectively). A position × time interaction was observed for lnRMSSD (p = 0.01). lnRMSSD_BL was higher than lnRMSSDpost20 for LINEMEN (p < 0.01; ES = large), whereas differences for SKILL and MID-SKILL were not statistically different (p > 0.05). Players with greater body mass experienced larger reductions in lnRMSSD (r = -0.62, p < 0.01). Longitudinally, lnRMSSDcv was significantly related to body mass (r = 0.48) and PL_chronic (r = -0.60). After adjusting for body mass, lnRMSSDcv and PL_chronic remained significantly related (r = -0.43). The ∼20-hour recovery time between training sessions on consecutive days may not be adequate for restoration of cardiac-parasympathetic activity to baseline among LINEMEN. Players with a lower chronic training load throughout camp experienced greater fluctuation in lnRMSSD (i.e., lnRMSSDcv) and vice versa. Thus, a capacity for greater chronic workloads may be protective against perturbations in cardiac-autonomic homeostasis among American college football players.


Follicular dendritic cell-derived antigen and accessory activity in initiation of memory IgG responses in vitro.
Follicular dendritic cells (FDC) release Ag in developing germinal centers in the form of immune complex-coated bodies (iccosomes). Iccosomes are endocytosed by B cells, and the B cells process and present the FDC-derived Ag to T cells. By 3 days after Ag challenge, Ab-forming cells producing IgG specific for the iccosomal Ag emerge from the developing germinal centers and home to the bone marrow, where most Ab in a secondary response are produced. In addition to providing Ag, FDC exhibit potent accessory activities that promote B cell proliferation. These observations prompted the hypothesis that both FDC-derived Ag and FDC-derived Ag-independent secondary signal(s) are essential for optimal secondary Ab responses. To test this hypothesis, methods were developed to separate Ag-bearing iccosomes from intact FDC, and we then examined the ability of isolated iccosomes to elicit secondary Ab responses in vitro in the presence and absence of intact FDC. In the absence of FDC, iccosomes bearing the appropriate Ag elicited only minimal levels of specific IgG. Proliferation studies revealed that iccosomes lacked the FDC accessory activity necessary to augment B cell proliferation. When a source of FDC lacking the relevant Ag but exhibiting accessory activity was added to the iccosomal/Ag/lymphocyte mixture, dramatic increases in IgG specific for the iccosomal Ag were obtained (increases were from low ng/ml to microg/ml levels of specific IgG). The results suggest the concept that Ag on FDC or on iccosomes signals B cells through B cell Ag receptor, the iccosome provides these B cells with Ag necessary to process and elicit T cell help, and a secondary signal(s) necessary to optimize the memory response is delivered to B cells by FDC in an Ag-nonspecific fashion.


One-Class Fingerprint Presentation Attack Detection Using Auto-Encoder Network.
Automated Fingerprint Recognition Systems (AFRSs) have been threatened by Presentation Attack (PA) since its existence. It is thus desirable to develop effective presentation attack detection (PAD) methods. However, the unpredictable PAs make PAD be a challenging problem. This paper proposes a novel One-Class PAD (OCPAD) method for Optical Coherence Technology (OCT) images based fingerprint PA detection. The proposed OCPAD model is learned from a training set only consists of Bonafides (i.e. real fingerprints). The reconstruction error and latent code obtained from the trained auto-encoder network in the proposed model is taken as the basis for the following spoofness score calculation. To get more accurate reconstruction error, we propose an activation map based weighting model to further refine the accuracy of reconstruction error. We test different statistics and distance measures and finally use a decision level fusion to make the final prediction. Our experiments are performed using a dataset with 93200 bonafide scans and 48400 PA scans. The results show that the proposed OCPAD can achieve a True Positive Rate (TPR) of 99.43% when the False Positive Rate (FPR) equals to 10% and a TPR of 96.59% when FPR=5%, which significantly outperformed a feature based approach and a supervised learning based model requiring PAs for training.


[The femoral neck fracture in the elderly patient - cemented or cementless hip arthroplasty?].
This is a report on the treatment results of femoral neck fractures with a cemented (group 1) and cementless (group 2) type of hip prosthesis, resp. 72 patients were enrolled. 35 patients were treated with a cemented hip prosthesis (mean age: 78 years), and 37 patients with a cementless modular hip prosthesis (mean age: 77 years). In the cemented group we observed 5 cases of hypotension during insertion of the prosthesis in the femoral shaft. One of these patients required mechanical resuscitation during surgery. In the second group 3 cases of proximal femur fissure and one case of distal femoral fracture were recorded. One year after surgery 43 patients presented for follow up evaluation (cemented group: n = 24; cementless group: n = 19). Both groups revealed comparable results according to the Harris Hip Score (75 versus 78,3 points). No prosthesis loosening was observed in either group. In our view the cemented hip prosthesis is the treatment of choice for femoral neck fractures among the old and very old, if no stabile osteosynthesis can be performed. Patients with cardiopulmonary risk factors, however, may profit from cementless hip arthroplasty to avoid the well known cardiodepression during surgery.


Selective Protein Expression Changes of Leukocyte-Migration-Associated Cluster of Differentiation Antigens at the Blood-Brain Barrier in a Lipopolysaccharide-Induced Systemic Inflammation Mouse Model without Alteration of Transporters, Receptors or Tight Junction-Related Protein.
Leukocyte migration across the blood-brain barrier (BBB) is an important step in the progression of brain dysfunction in systemic inflammation. The purpose of this study was to identify the key regulatory molecule(s) at the BBB among the cluster of differentiation (CD) antigens involved in leucocyte migration in lipopolysaccharide (LPS)-induced systemic inflammation based on their absolute protein expressions. Here, we identified the absolute expression levels of 17 CD antigens in isolated brain capillaries (Bcap) of LPS-administered mice. Among them, the expression levels of CD54 and CD106 were dramatically increased in LPS-administered mice compared to the control by 6.21- and 3.67-fold, respectively. In peripheral blood mononuclear cells, the expression levels of CD11a/CD18, the counter-receptor for CD54, were similar to those of CD54 in Bcap of LPS-administered mice. On the other hand, the expression level of CD49d, part of CD29/CD49d complex, which is the counter-receptor for CD106, was under the limit of quantification. It is thus likely that CD54 at the BBB is predominantly involved in promoting leukocyte migration across the BBB in systemic inflammation. The expression levels of CD9, CD49c and CD97, which are thought to be involved in cell-to-cell interaction, were decreased by 40-60% in Bcap of LPS-administered mice. In contrast, the expression levels of 9 transporters, 2 receptors, and 1 tight junction-related protein in Bcap of LPS-administered mice were essentially unchanged compared to the control. These results suggest that enhancement of leucocyte migration in systemic inflammation involves dynamic changes of CD antigens without alterations of other major functional molecules.


An academic practice's transition to the business of medicine in the community. A case study.
This case study highlights the problems confronting a clinical practice corporation affiliated with a major medical school, and the business realizations it made in the acquisition of a community-based clinic. Launching a financially viable enterprise requires careful planning, determination of formal goals and expectations, an appropriate mix of physicians and services, a specific marketing campaign and community support.


Hybrid fullerene conjugates as vectors for DNA cell-delivery.
The present study reports fullerene conjugates that act as efficient binders of double stranded DNA (dsDNA) into cytofriendly polyplexes. The conjugates are designed to generate dendrimeric structures, having C60 as the core and bearing linear or branched PEI and polyethyleneglycol (PEG) arms (∼2 kDa). Simple and reproducible synthesis pathways provided C60-PEI and C60-PEG-PEI conjugates. They were able to bind linear and plasmidic dsDNA and they form particulate polyplexes of 50 to 200 nm in diameter. The resulted polyplexes toggle between the anionic and cationic state at nitrogen to phosphorous ratios (N/P) of about 5, as revealed by their zeta potential and became colloidally stable at N/P ratios above 10, as determined by atomic force microscopy (AFM). They are electrophoretically unbreakable starting with N/P ratios of 3 and of 5 when salmon sperm DNA and pEYFP-C1 plasmid, respectively are loaded. Both C60-PEI·pEYFP and C60-PEG-PEI·pEYFP polyplexes are non-cytotoxic against HEK 293T cells in culture and exhibit transfection efficiency better than 25% (N/P ratios above 20) and 6% (N/P ratios above 60) respectively, measured by flow cytometry. For comparison, the commercial SuperFect® from Qiagen (positive control) was able to provide an efficiency of 15-20%, under similar conditions. Moreover, the C60-PEG-PEI conjugate is as performant as the positive control in terms of expression of EYFP reporter gene in cultured cells and exhibited high cytocompatibility, determining cell proliferation up to 200%. Our study proved that C60-PEG-PEI is effective vector for DNA delivery being, in addition, easily synthesizable, practically non-cytotoxic and as efficient the commercially available transfection tools.


[Induction of hydrogen ion transport in mitochondrial membranes].
The system of ion transport in mitochondria induced with Brensted acids (H+-acids) was studied. It has been shown that an increase of the activity of hydrogen ions in the incubation medium (pH decrease) along with the earlier obtained [3--5] K+/H+ exchange brings about the appearance of the system of electrogenic transport of potassium ions in mitochondria. A synchronous functioning of the two systems indicated provides the electrogenic transmembrane transport of hydrogen ions through the mitochondrial membrane (Fig. 3, diagram 6). When studying the induction of ionic transport the data were obtained in favour of the concept that the induction resulted from conjugated oxidation and hydrolysis of lipids.


Transforming growth factors-beta as regulators of cellular growth and phenotype.
An increasing number of cDNAs and polypeptides have been ascribed to the transforming growth factor-beta (TGF-beta) superfamily. It has become evident that this evolutionally well-conserved gene family has representatives in many organisms, from invertebrates to mammalians. As new members are being identified, the knowledge of their biological significance is derived not only from studies of their effects in various in vitro models, but also of their organismal expression during different stages of embryogenesis and morphogenesis. The major described functions of the TGF-beta and like proteins seem to be the regulation of cell proliferation, phenotype, and differentiation. Part of these effects may be attributed to changes in the adhesive properties of the cells and integrity of their extracellular matrices modulated by TGF-beta. Identification of the new members of the TGF-beta family, their biological effects, cell-surface receptors, and cell or tissue-specific expression will be invaluable in trying to understand the purpose of many seemingly alike multiregulators.


Prenatal metal(loid) mixtures and birth weight for gestational age: A pooled analysis of three cohorts participating in the ECHO program.
A growing number of studies have identified both toxic and essential metals which influence fetal growth. However, most studies have conducted single-cohort analyses, which are often limited by narrow exposure ranges, and evaluated metals individually. The objective of the current study was to conduct an environmental mixture analysis of metal impacts on fetal growth, pooling data from three geographically and demographically diverse cohorts in the United States participating in the Environmental Influences on Child Health Outcomes program. The pooled sample (N = 1,002) included participants from the MADRES, NHBCS, and PROTECT cohorts. Associations between seven metals (antimony, cadmium, cobalt, mercury, molybdenum, nickel, tin) measured in maternal urine samples collected during pregnancy (median: 16.0 weeks gestation) and birth weight for gestational age z-scores (BW for GA) were investigated using Bayesian Kernel Machine Regression (BKMR). Models were also stratified by cohort and infant sex to investigate possible heterogeneity. Chromium and uranium concentrations fell below the limits of detection for most participants and were evaluated separately as binary variables using pooled linear regression models. In the pooled BKMR analysis, antimony, mercury, and tin were inversely and linearly associated with BW for GA, while a positive linear association was identified for nickel. The inverse association between antimony and BW for GA was observed in both males and females and for all three cohorts but was strongest for MADRES, a predominantly low-income Hispanic cohort in Los Angeles. A reverse j-shaped association was identified between cobalt and BW for GA, which was driven by female infants. Pooled associations were null for cadmium, chromium, molybdenum, and uranium, and BKMR did not identify potential interactions between metal pairs. Findings suggest that antimony, an understudied metalloid, may adversely impact fetal growth. Cohort- and/or sex-dependent associations were identified for many of the metals, which merit additional investigation.


Finding multiple possible critical paths using fuzzy PERT.
Program evaluation and review techniques (PERT) is an efficient tool for large project management. In actual project control decisions, PERT has successfully been applied to business management, industry production, project scheduling control, logistics support, etc. However, classical PERT requires a crisp duration time representation for each activity. This requirement is often difficult for the decision-makers due to the fact that they usually can not estimate these values precisely. In recent years, some fuzzy PERT methods have been proposed based on fuzzy set theory for project management. However, there is a drawback in the existing fuzzy PERT methods, i.e., sometimes they maybe cannot find a critical path in a fuzzy project network. In this paper, we propose a fuzzy PERT algorithm to find multiple possible critical paths in a fuzzy project network, where the duration time of each activity in a fuzzy project network is represented by a fuzzy number. The proposed algorithm can overcome the drawback of the existing fuzzy PERT methods.


Dioctophyme renale (Nematoda, Dioctophymatidae) infection in the crab-eating fox (Cerdocyon thous) from Brazil.
The purpose of this study is to report the first case of parasitism by the giant kidney worm, Dioctophyme renale (Goeze, 1782), in the crab-eating fox, Cerdocyon thous (Linnaeus, 1766), in Brazil. A debilitated adult male fox was taken to the Screening Center for Wild Animals in Rio de Janero (CETAS/RJ) and eventually died of an unknown cause. During necropsy, a brownish-red nematode, 28.9 cm long, was found in the peritoneal cavity. The worm was identified as a male D. renale. This study reports the first case of dioctophymatosis in the crab-eating fox, enlarging the range of wild definitive host species infected by the giant kidney worm in the Neotropical region.


[Pathogenesis of intestinal dysbacteriosis].
Study mechanism of the development of intestine dysbiosis on the experimental model of mice from the position of reinforcement of translocation of opportunistic Gram negative enterobacteria (OGNE) from the intestine lumen into the system blood flow and the possibility of its correction by lactoglobulin preparations. Experimental medicinal dysbacteriosis of the intestine was reproduced in mice by using gentamycin. Changes of immunologic parameters in the animals during translocation of Gram negative enterobacteria and their LPS from the intestine lumen into system blood flow were studied. During intestine dysbioses a pronounced inflammatory reaction was shown to develop at a local level with the increase of the amount of proinflammatory cytokines with the sequence IL-1beta - IFNgamma- IL-6 and reinforcement of formation of anti-endotoxin antibodies. Changes in the parameters of allergization of macroorganism under the influence of LPS occurred synchronously with the increase of amount of (OGNE) including escherichia with altered lactase activity in the background of its translocation into extrinsic biotopes. In decompensated variant of dysbiosis a pronounced activation of immune reactive spleen cells was noted. Immunobiologic preparations (lactoglobulin and low molecular peptides of immune colostrum) administered to the animals in parallel with the antibiotic caused deintoxication and desensitizing effect and reduced the side effect of its action. A scheme of development of dysbiotic disorders in the intestine from the position of reinforcement of translocation of (OGNE) from the intestine lumen into the system blood flow is proposed and mechanisms of positive effect of lactoglobulin preparations and low molecular peptides of immune colostrum that are perspective for the correction of this state are explained.


Effectiveness of an organized cervical cancer screening program in Korea: results from a cohort study.
Although the value of cervical cancer screening is widely acknowledged, the effectiveness of an organized cervical cancer screening program in Korea has never been evaluated. We investigated the associations of the frequency of cervical cancer screening with cervical cancer incidence using data from a large prospective cohort study. In this analysis, 253,472 women without a hysterectomy or previous cancer diagnosis were included. Follow-up was between 1995 and 2002. Frequency of Pap smear testing was determined by searching the National Health Examination Database. Using the Korean Central Cancer Registry, 248 cases of invasive cervical cancer and 346 cases of carcinoma in situ (CIS) of the cervix were identified. Subjects screened 2 or more times showed a 71% (corrected reduction 60%) and a 66% (corrected reduction 53%) reduced risk of invasive cervical cancer and CIS of the cervix, respectively, as compared with unscreened subjects [relative risk (RR) = 0.29; 95% confidence interval (CI) = 0.20-0.45; RR = 0.34; 95% CI = 0.25-0.46, respectively]. Women with a normal or benign pap smear had a statistically significantly lower risk of invasive cervical cancer and CIS of cervix compared with those never screened. In age-stratified analyses, there was a significant reduction in cervical cancer incidence among women aged 30 and over who were screened 2 or more times compared with women never screened. The results of this prospective cohort study show that regular screening of cervical cancer reduces invasive cervical cancer incidence and CIS of the cervix among Korean women.


Risk assessment of vapor cloud explosions in a hydrogen production facility with consequence modeling.
New technologies using hazardous materials usually have certain risks. It is more serious when the technology is supposed to be applied in a large scale and become widely used by many people. The objective of this paper was to evaluate the risk of vapor cloud explosion in a hydrogen production process. Potential hazards were identified using the conventional hazard identification method (HAZID). The frequency of the proposed scenarios was estimated from statistical data and existing records. The PHAST professional software was applied for consequence modeling. Both individual and societal risks were evaluated. This cross-sectional study was conducted from June 2010 to December 2011 in a Hydrogen Production Plant in Tehran. The full bore rupture in heat exchanger had the highest harm effect distance. The full bore rupture in desulphurization reactor had the highest (57% of total) individual risk. Full bore rupture in heat exchanger was the highest contributor to social risk. It carried 64% & 66.7% of total risk in day and night respectively. For the sake of safety, mitigation measures should be implemented on heat exchanger, reformer and hydrogen purification absorbers. The main proposed risk reductive measures included; the increasing of installed equipment elevation, the application of smaller vessels and pipes.


A multifunctional catalyst that stereoselectively assembles prodrugs.
The catalytic stereoselective synthesis of compounds with chiral phosphorus centers remains an unsolved problem. State-of-the-art methods rely on resolution or stoichiometric chiral auxiliaries. Phosphoramidate prodrugs are a critical component of pronucleotide (ProTide) therapies used in the treatment of viral disease and cancer. Here we describe the development of a catalytic stereoselective method for the installation of phosphorus-stereogenic phosphoramidates to nucleosides through a dynamic stereoselective process. Detailed mechanistic studies and computational modeling led to the rational design of a multifunctional catalyst that enables stereoselectivity as high as 99:1.


Mucinous adenocarcinoma of the prostate: histochemical and immunohistochemical studies.
Twelve patients with primary mucinous adenocarcinoma of the prostate were included in a clinicopathologic study; criteria included a total tumor volume more than 25% mucinous and single or clustered tumor cells floating in mucin lakes. Patient ages were 57 to 81 years; tumor stages were C (three), D (five), and unknown (four). Bone was the most frequent metastatic site (usually osteoblastic), followed by lymph nodes and lungs. Serum levels of prostatic acid phosphatase and prostate-specific antigen were frequently elevated (five of 10 and three of three measured, respectively). All mucinous adenocarcinomas also contained other histologic patterns: microglandular (four), cribriform (three), comedo (two), solid (two), and hypernephroid (one). Mucinous components composed less than 50% of three tumors, 50% and 75% of six, and more than 75% of three. No tumor contained signet-ring cells. Immunoperoxidase staining was positive for prostatic acid phosphatase and prostate-specific antigen and negative for carcinoembryonic antigen. Treatment was radiation, estrogen, orchiectomy, or a combination. In two of four patients, serum prostatic acid phosphatase levels normalized after therapy. Seven patients died of disease (mean follow-up, 56 months), and five patients are alive with disease (mean, 32.2 months). The proportion of mucinous component did not affect prognosis.


[Structural-functional motifs of sterol 14-alpha demethylases (CYP51)].
CYP51 family of cytochromes P450 (sterol 14-alpha-demethylases) comprises the representatives from different kingdoms of living world, thus positioning itself as the most ancient member of the superfamily. In the course of the present research the collection of 36 full-length CYP51 amino acid sequences was submitted to cluster analysis. Each node of the clustering dendrogram corresponds to the groups of proteins, located on the branches descending from the node. By making the multiple alignment of each group of protein sequences we obtained the node-specific consensus sequences. The informational content of the consensus was defined as the presence of the compact conserved sites, the motifs. The assessment of informational content was computed using Sherman's non-parametric statistical criterion. The high informational content was observed for the 100% conserved consensus sequences of the following CYP51,s groups: fungi, animal+plant, plant+protista and bacteria. These selected consensus sequences were next aligned all together to get the final consensus for the whole family. To enrich the informational content of the CYP51 consensus the level of its conservation was dropped to 75%. Regions of statistically significant conservation were unraveled in the CYP51 consensus sequence. These regions (motifs) were then correlated with the information on secondary structure elements and substrate recognition sites reported for CYP51 from Mycobacterium tuberculosis. Seven motifs appeared to be obligatory for every CYP51 protein. The motifs thus obtained were searched for among all the known cytochrome P450 proteins. Some motifs were found to be absolutely specific for 14-alpha-demethylases, whereas others were common to different species of cytochromes P450.


Cognitive Behavioral and Cognitive Remediation Strategies for Managing Co-Occurring Anorexia Nervosa and Elevated Autism Spectrum Traits.
Anorexia nervosa (AN) is a potentially severe eating disorder whose core characteristics include energy intake restriction leading to low body weight. Autism spectrum disorder (ASD) is a developmental disorder characterized by deficits in social interaction and communication as well as repetitive, stereotyped behavior and interests. Both high ASD traits and diagnosed ASD are overrepresented among individuals with AN, and AN and ASD appear to share certain neurocognitive features. These features are associated with the severity of eating disorder symptoms and prolongation of AN. Thus, individuals with AN and high ASD traits or ASD may benefit less from traditional treatment when compared to those with low ASD traits. No previous reviews have summarized what is known about treatment adaptations for individuals with AN and high ASD traits or ASD. The purpose of this narrative review was to investigate the feasibility of cognitive remediation therapy (CRT), cognitive remediation and emotional skill training (CREST), and cognitive behavioral therapy (CBT), and give an overview of treatment modifications for individuals with AN and co-occurring ASD or high ASD traits. We found nine studies that fulfilled our inclusion criteria. The combined results suggest that individuals with AN and high ASD traits or ASD benefit less from CRT, CREST, and CBT than those with AN and low ASD traits. However, CRT and CREST administered in individual format may be associated with improved cognitive flexibility, motivation for change, and decreased alexithymia among adults with AN and high ASD traits or ASD. Individuals with comorbid AN and ASD themselves highlight the importance of treatment adaptations that take the characteristics of ASD into account. In the future, controlled studies of the treatment strategies for individuals with AN and ASD/high ASD traits are needed in order to improve the outcome of individuals with this challenging comorbidity.


Protein sialylation by sialyltransferase involves radiation resistance.
Previously, we identified beta-galactoside alpha(2,6)-sialyltransferase (ST6Gal I) as a candidate biomarker for ionizing radiation. The expression of ST6Gal I and the level of protein sialylation increased following radiation exposure in a dose-dependent manner. Radiation induced ST6Gal I cleavage and the cleaved form of ST6Gal I was soluble and secreted. Sialylation of integrin beta1, a glycosylated cell surface protein, was stimulated by radiation exposure and this increased its stability. Overexpression of ST6Gal I in SW480 colon cancer cells that initially showed a low level of ST6Gal I expression increased the sialylation of integrin beta1 and also increased the stability of the protein. Inhibition of sialylation by transfection with neuraminidase 2 or neuraminidase 3 or by treatment with short interfering RNA targeting ST6Gal I reversed the effects of ST6Gal I overexpression. In addition, ST6Gal I overexpression increased clonogenic survival following radiation exposure and reduced radiation-induced cell death and caspase 3 activation. However, removal of sialic acids by neuraminidase 2 or knockdown of expression by short interfering RNA targeting ST6Gal I restored radiation-induced cell death phenotypes. In conclusion, radiation exposure was found to increase the sialylation of glycoproteins such as integrin beta1 by inducing the expression of ST6Gal I, and increased protein sialylation contributed to cellular radiation resistance.


Sp3 is a transcriptional activator of the human alpha2(I) collagen gene.
Sp3 is an ubiquitously expressed transcription factor, closely related to Sp1 but, unlike Sp1, it often functions as a transcriptional repressor. In this study we investigated the role of Sp3 in regulating the transcription of the human alpha2(I) collagen gene. We show that Sp1 and Sp3 specifically bind to three of the previously characterized cis -elements in this promoter, including two positive cis-elements between -303 and -271 and -128 and -123, and a repressor site between -164 and -159, but do not bind to the fourth cis-element bound by CBF. Functional analyses of Sp3 and Sp1 in Drosophila cells indicate that each protein transactivates the human alpha2(I) collagen promoter with equal potency and, when tested together, have an additive effect on the promoter activity. Furthermore, in vitro transcription assays demonstrate that both Sp1 and Sp3 are capable of supporting transcription from the collagen promoter independently of each other. However, when activities of both Sp1 and Sp3 are blocked with specific antibodies, in vitro transcription from this promoter is almost completely abolished. The results of this study demonstrate that Sp3 is as potent an activator of the human alpha2(I) collagen promoter as is Sp1 and that a transcriptional activity of the human alpha2(I) promoter is dependent on both proteins.


Modular assembly using sequential palladium coupling gives easy access to the SMoC class of cellular transporters.
The transducing ability of the third helix of transcription factor homeodomains is effectively mimicked by a biphenyl system displaying guanidine groups. The biphenyl class of small molecule carriers (SMoCs) can carry biomolecules into a wide variety of cell types. A "combinatorial" approach to the synthesis of SMoCs is described using sequential Pd(0) coupling chemistry to assemble the molecules from highly functionalized building blocks. SMoCs coupled to the DNA licensing repressor protein geminin can inhibit DNA replication in vitro. We conducted a structure-activity investigation utilizing a range of SMoC-geminin conjugates and demonstrate that both electrostatic and structural features are important for efficient uptake and functional activity. The best analogue was more efficient than either (Arg)(4) or (Arg)(8) linked to geminin. Effective inhibition of DNA synthesis was achieved in fibroblasts and osteosarcoma cell lines.


Macaques infected with attenuated simian immunodeficiency virus resist superinfection with virulence-revertant virus.
Macaques infected with attenuated simian immunodeficiency virus (SIVmac) can resist superinfection challenge with virulent virus, showing the potential of live attenuated virus as an AIDS vaccine. Superinfection resistance does not, however, prevent the generation of virulent virus in vivo, suggesting that such virus may circumvent the resistance effect. Here, we show that three macaques already infected with the attenuated molecular clone SIVmacC8 were resistant to superinfection with virulent virus that arose in vivo following repair of a 12 bp attenuating lesion in the nef/3' LTR. In contrast, four naive animals became infected following inoculation with blood taken from the macaque in which virulent virus arose. Loss of nef-specific cytotoxic T lymphocyte (CTL) responses followed repair of the attenuating lesion within nef in the donor animal, suggesting the possibility of escape from CTL-driven selection pressure.


Neuropsychiatric diagnosis and management of chronic sequelae of war-related mild to moderate traumatic brain injury.
Soldiers with a traumatic brain injury (TBI) present with an array of neuropsychiatric symptoms that can be grouped into nosological clusters: (1) cognitive dysfunctions: difficulties in memory, attention, language, visuospatial cognition, sensory-motor integration, affect recognition, and/or executive function typically associated with neocortical damage; (2) neurobehavioral disorders: mood, affect, anxiety, posttraumatic stress, and psychosis, as well as agitation, sleep problems, and libido loss, that may have been caused by damage to the cortex, limbic system, and/or brain stem monoaminergic projection systems; (3) somatosensory disruptions: impaired smell, vision, hearing, equilibrium, taste, and somatosensory perception frequently caused by trauma to the sensory organs or their projections through the brain stem to central processing systems; (4) somatic symptoms: headache and chronic pain; and (5) substance dependence. TBI-related cognitive impairment is common in veterans who have served in recent conflicts in the Middle East and is often related to blasts from improvised explosive devices. Although neurobehavioral disorders such as depression and posttraumatic stress disorder commonly occur after combat, the presentation of such disorders in those with head injury may pass undetected with use of current diagnostic criteria and neuropsychological instruments. With a multidimensional approach (such as the biopsychosocial model) applied to each symptom cluster, psychological, occupational, and social dysfunction can be delineated and managed.


Optogenetic Manipulation of Postsynaptic cAMP Using a Novel Transgenic Mouse Line Enables Synaptic Plasticity and Enhances Depolarization Following Tetanic Stimulation in the Hippocampal Dentate Gyrus.
cAMP is a positive regulator tightly involved in certain types of synaptic plasticity and related memory functions. However, its spatiotemporal roles at the synaptic and neural circuit levels remain elusive. Using a combination of a cAMP optogenetics approach and voltage-sensitive dye (VSD) imaging with electrophysiological recording, we define a novel capacity of postsynaptic cAMP in enabling dentate gyrus long-term potentiation (LTP) and depolarization in acutely prepared murine hippocampal slices. To manipulate cAMP levels at medial perforant path to granule neuron (MPP-DG) synapses by light, we generated transgenic (Tg) mice expressing photoactivatable adenylyl cyclase (PAC) in DG granule neurons. Using these Tg(CMV-Camk2a-RFP/bPAC)3Koka mice, we recorded field excitatory postsynaptic potentials (fEPSPs) from MPP-DG synapses and found that photoactivation of PAC during tetanic stimulation enabled synaptic potentiation that persisted for at least 30 min. This form of LTP was induced without the need for GABA receptor blockade that is typically required for inducing DG plasticity. The paired-pulse ratio (PPR) remained unchanged, indicating the cAMP-dependent LTP was likely postsynaptic. By employing fast fluorescent voltage-sensitive dye (VSD: di-4-ANEPPS) and fluorescence imaging, we found that photoactivation of the PAC actuator enhanced the intensity and extent of dentate gyrus depolarization triggered following tetanic stimulation. These results demonstrate that the elevation of cAMP in granule neurons is capable of rapidly enhancing synaptic strength and neuronal depolarization. The powerful actions of cAMP are consistent with this second messenger having a critical role in the regulation of synaptic function.


Ex Vivo Analysis of Tryptophan Metabolism Using 19F NMR.
Tryptophan, an essential amino acid, is metabolized into a variety of small molecules capable of impacting human physiology, and aberrant tryptophan metabolism has been linked to a number of diseases. There are three principal routes by which tryptophan is degraded, and thus methods for measuring metabolic flux through these pathways can be used to understand the factors that perturb tryptophan metabolism and potentially to measure disease biomarkers. Here, we describe a method utilizing 6-fluorotryptophan as a probe for detecting tryptophan metabolites in ex vivo tissue samples via 19F nuclear magnetic resonance. As a proof of concept, we demonstrate that 6-fluorotryptophan can be used to measure changes in tryptophan metabolism resulting from antibiotic-induced changes in gut microbiota composition. Taken together, we describe a general strategy for monitoring amino acid metabolism using 19F nuclear magnetic resonance that is operationally simple and does not require chromatographic separation of metabolites.


A study on rational drug prescribing and dispensing in outpatients in a tertiary care teaching hospital of Western Nepal.
To analyze the prescriptions of out-patients for rational prescribing and dispensing and to evaluate the patient's knowledge regarding use of drugs, using INRUD indicators. A cross-sectional, descriptive study was conducted at the Manipal Teaching Hospital, Pokhara, Nepal during the time period from June 10th to August 19th 2004. Totally 247 prescriptions were randomly selected for analysis, wherein 720 drugs were prescribed. Only 15% of drugs were prescribed by generic name, 21.67% of the total drugs consisted of fixed-dose combinations, only 40% of drugs were from the Essential drug list of Nepal and 29.44% (n=212) were from the WHO Essential drug list. It was found that more than half (54.17%) of the drugs were from Nepalese National Formulary and 35.69% were from WHO model formulary. Dermatological products were most commonly prescribed followed by drugs acting on central nervous system, antimicrobials and drugs acting on cardiovascular system. Among the drugs dispensed, 79.16% were oral followed by topical (18.19%) and parenteral forms (2.98%). Diagnosis was mentioned only in 3.23% (n=8) of the prescriptions and the average cost per prescription was found to be 241.11 Nepalese rupees (US$ 3.26). It was found out that pharmacist labelled only 0.4% of the medication envelopes with the name of the patient. However, 82.6% of the medication envelopes were labelled with name of the drug and 87.0% with drug strength. Only 53.8% (n=133) of the patient knew both the duration of the therapy and administration time of drugs. There is a need for educational intervention for prescribers and both managerial and educational intervention for the hospital pharmacists to improve prescribing and dispensing.


Kinetic Modeling Assisted Analysis of Vitamin C-Mediated Copper Redox Transformations in Aqueous Solutions.
The kinetics of oxidation of micromolar concentrations of ascorbic acid (AA) catalyzed by Cu(II) in solutions representative of biological and environmental aqueous systems has been investigated in both the presence and absence of oxygen. The results reveal that the reaction between AA and Cu(II) is a relatively complex set of redox processes whereby Cu(II) initially oxidizes AA yielding the intermediate ascorbate radical (A•-) and Cu(I). The rate constant for this reaction was determined to have a lower limit of 2.2 × 104 M-1 s-1. Oxygen was found to play a critical role in mediating the Cu(II)/Cu(I) redox cycle and the oxidation reactions of AA and its oxidized forms. Among these processes, the oxidation of the ascorbate radical by molecular oxygen was identified to play a key role in the consumption of ascorbic acid, despite being a slow reaction. The rate constant for this reaction (A•-+O2→DHA+O2•-) was determined for the first time with a calculated value of 54 ± 8 M-1 s-1. The kinetic model developed satisfactorily describes the Cu/AA/O2 system over a range of conditions including different concentrations of NaCl (0.2 and 0.7 M) and pH (7.4 and 8.1). Appropriate adjustments to the rate constant for the reaction between Cu(I) and O2 were found to account for the influence of the chloride ions and pH on the kinetics of the process. Additionally, the presence of Cu(III) as the primary oxidant resulting from the interaction between Cu(I) and H2O2 in the Cu(II)/AA system was confirmed, along with the coexistence of HO•, possibly due to an equilibrium established between Cu(III) and HO•.


Cell signaling and transcriptional regulation via histone phosphorylation.
Regulation of transcription involves a large number of histone lysine and arginine posttranslational modifications found marking gene promoters and gene bodies. Within histones there are abundant accessible serine/threonine/tyrosine residues for potential phosphorylation; however, few sites have been clearly documented with regard to actual modification, relevant physiological pathway, and cognate enzyme. In addition, kinases within signaling pathways are thought to be localized to the cytoplasm and thus not able to directly modify histones within chromatin in the nucleus. However, direct assays in the model eukaryote Saccharomyces cerevisiae via chromatin immunoprecipitation have placed numerous signaling kinases at promoters and within gene bodies. In addition, recent studies in mammalian cells of two signaling pathways place the terminal kinase within the nucleus or directly at genes, have identified histone phosphorylation sites, and furthermore, have uncovered potential mechanisms by which these histone phosphorylation sites activate transcription. These results lead to a gathering appreciation of the potential role of signal transduction kinase-mediated direct histone phosphorylation in regulating transcription.


Fibrinopeptide A, platelet factor 4, and beta-thromboglobulin levels in coronary heart disease.
In vivo platelet alpha-granule release and fibrin I formation were measured in 82 patients with ischemic heart disease by radioimmunoassay of platelet factor 4, beta-thromboglobulin, and fibrinopeptide A. The presence and extent of coronary artery disease were determined by coronary arteriography, and the extent of left ventricular regional dysfunction was assessed by contrast left ventriculography. In patients with abnormal coronary arteriograms without previous myocardial infarction, mean levels of platelet factor 4, beta-thromboglobulin, and fibrinopeptide A were not elevated. In patients in whom myocardial infarction had occurred more than 6 mo previously, platelet factor 4 (8.3 ng/ml; p less than 0.01) and beta-thromboglobulin (33.2 ng/ml; p less than 0.001) levels were significantly elevated, but fibrinopeptide A levels were normal. Levels of platelet factor 4 and beta-thromboglobulin were unrelated to the extent of coronary artery disease. In the patients with prior infarction, beta-thromboglobulin correlated directly with extent of left ventricular regional dysfunction (r = 0.53; p less than 0.01) and inversely with ejection fraction (r = -056; p less than 0.05). In a small group of patients with left ventricular aneurysm, mean fibrinopeptide A levels were also elevated. We interpret these findings as indicating that platelet release in patients with ischemic heart disease results from platelet reaction with previously infarcted myocardium rather than with the atherosclerotic coronary arteries.


Soy allergy in perspective.
The purpose of this paper is to review and discuss studies on soy allergy. In Central Europe soy is a clinically relevant birch pollen-related allergenic food. Crossreaction is mediated by a Bet v 1 homologous protein, Gly m 4. Additionally, birch pollen allergic patients might acquire through Bet v 1 sensitization allergies to mungbean or peanut, in which Vig r 1 and Ara h 8 are the main cross-reactive allergens. Threshold doses in soy allergic individuals range from 10 mg to 50 g of soy and are more than one order of magnitude higher than in peanut allergy. No evidence was found for increased allergenicity of genetically modified soybeans. In Europe, both primary and pollen-related food allergy exist. The diagnosis of legume allergy in birch pollen-sensitized patients should not be excluded on a negative IgE testing to legume extracts. Bet v 1 related allergens are often underrepresented in extracts. Gly m 4 from soy and Ara h 8 from peanut are nowadays commercially available and are recommended in birch pollen allergic patients with suspicion of soy or peanut allergy, but negative extract-based diagnostic tests to screen for IgE specific to these recombinant allergens.


Evolution of eye morphology and rhodopsin expression in the Drosophila melanogaster species subgroup.
A striking diversity of compound eye size and shape has evolved among insects. The number of ommatidia and their size are major determinants of the visual sensitivity and acuity of the compound eye. Each ommatidium is composed of eight photoreceptor cells that facilitate the discrimination of different colours via the expression of various light sensitive Rhodopsin proteins. It follows that variation in eye size, shape, and opsin composition is likely to directly influence vision. We analyzed variation in these three traits in D. melanogaster, D. simulans and D. mauritiana. We show that D. mauritiana generally has larger eyes than its sibling species, which is due to a combination of larger ommatidia and more ommatidia. In addition, intra- and inter-specific differences in eye size among D. simulans and D. melanogaster strains are mainly caused by variation in ommatidia number. By applying a geometric morphometrics approach to assess whether the formation of larger eyes influences other parts of the head capsule, we found that an increase in eye size is associated with a reduction in the adjacent face cuticle. Our shape analysis also demonstrates that D. mauritiana eyes are specifically enlarged in the dorsal region. Intriguingly, this dorsal enlargement is associated with enhanced expression of rhodopsin 3 in D. mauritiana. In summary, our data suggests that the morphology and functional properties of the compound eyes vary considerably within and among these closely related Drosophila species and may be part of coordinated morphological changes affecting the head capsule.


Cognitive processing speed in Lyme disease.
The goal of this study was to more precisely define the nature of the cognitive processing deficits in the patients with Lyme disease. Lyme disease has been associated with cognitive disturbances. Sixteen patients who met the Centers for Disease Control's case definition for Lyme disease and 15 age- and education-matched control subjects completed two computerized assessments. The first was a matching procedure that assessed perceptual/motor speed. The second task was an alphabet-arithmetic (AA) test that measured the speed of mental arithmetic. On the matching task, subjects judged as true or false simple identity equations (e.g., B + 0 = B). On the AA task, subjects indicated the veracity of equations of the same form as those of the matching task but which required mental arithmetic (e.g., A + 3 = D). The use of this paradigm permits sensory or motor slowing to be distinguished from slowed cognitive processing speed. Also, the tests do not involve automated or overlearned responses. Lyme disease patients and healthy controls did not differ in perceptual/motor speed. However, Lyme disease patients' response times were significantly longer than those of healthy controls during the AA task, demonstrating specific impairments in mental activation speed. These results suggest that Lyme disease patients show specific deficits when initiating a cognitive process. These impairments are independent of sensory, perceptual, or motor deficits.


Career choices for geriatric medicine: national surveys of graduates of 1974-2009 from all UK medical schools.
numbers of elderly people are increasing worldwide. This increases the importance of the specialty of geriatric medicine. Recruitment to the specialty may not be keeping pace with need. to report trends in junior doctors' career choices for geriatric medicine, factors that influence career choice, and associations between early career choices and later specialty destinations. questionnaire surveys of all medical qualifiers from all UK medical schools in selected year-of-qualification cohorts (1974-2009). Survey response rates 1, 3 and 5 years after graduation were, respectively, 65.9% (33,972/51,535), 65.5% (29,400/44,879) and 66.1% (22,600/34,197). geriatric medicine was the career choice of 0.9% of medical graduates (0.4% of men, 1.3% of women) 1 year after qualification; and of 1.5% (1.2% of men, 1.9% of women) after 5 years. There was a modest increase in recent cohorts. Important influences on career choice included enthusiasm for and commitment to the specialty, experience of working in geriatric medicine and self-appraisal of own skills. Early career choices were not highly predictive of later destinations. Of practising geriatricians in our surveys, 9% (20/212) had told us that they wanted to be geriatricians in their first year after graduation, as had 36% when in their third year and 74% in their fifth year. a higher percentage of women than men choose geriatric medicine; in recent years its popularity has increased slightly. Early career choice is not highly predictive of an eventual career in the specialty. Flexibility is needed about when doctors can enter training in geriatric medicine.