Discordance of gravity line and C7PL in patient with adult spinal deformity--factors affecting the occiput-trunk sagittal discordance.
During quiet standing, the gravity line (GL) can be located according to the sum of the ground reaction forces (GRFs) measured with a force platform. C7 plumb line (C7PL) is an easy method to estimate sagittal trunk balance, but discordance between C7PL and the GL is widely recognized. However, the prevalence of occiput-trunk (O-T) discordance (GL-C7PL>3 cm) and the factors affecting this type of discordance have not yet been determined. The purpose of this study was to report the prevalence of O-T discordance in adult spinal deformity (ASD) patients and identify the factors affecting this type of discordance. This was a retrospective consecutive case series of ASD. This retrospective case series included 300 consecutive ASD patients. The inclusion criteria were age more than 50 years, Cobb angle of the main curve more than 20°, and C7PL more than 5 cm. The exclusion criteria consisted of inappropriate radiography; syndromic, neuromuscular, or other pathologic conditions; and previous joint replacement. The outcome measures included self-reported (Scoliosis Research Society 22 and Oswestry disability index [ODI]) and radiographic measures. In a retrospective consecutive case series, demographic and radiographic patient data were reviewed. Demographic data included age, gender, curve type, SRS22, and ODI. Radiographic data included GL, C7PL, C2-C7, T2-T5, T5-T12, T10-L2, T2 tilt, lumbar lordosis (LL), sacrum slope (SS), pelvic tilt (PT), and pelvic incidence (PI). Global sagittal and spinopelvic alignments were also reviewed. Patients were categorized in either a O-T concordance (C group, GL-C7PL<3 cm) or a O-T discordance (D group, GL-C7PL more than +3 cm) group, and the demographic, radiographic, and clinical outcome data were compared between these groups. One-way analysis of variance, correlation coefficient tests, and multiple regression and logistic regression analyses were performed for statistical analysis. p Value less than .01 was considered statistically significant. Force platform analysis was performed to assess the relationship among GRF, GL, and C7PL. Among 300 consecutive ASD patients, 72 (24%) were categorized in the D group. There was no significant difference in terms of demographic data between the C and D groups. The SRS and ODI of patients with GL more than 10 cm were significantly lower than those of patients with GL less than 10 cm. Comparisons of regional sagittal alignment showed significantly higher T5-T12 values in the D group, and multiple regression analysis revealed significant correlations among T2-T5, T5-T12, and GL-C7PL. In contrast, the analysis of global sagittal alignment revealed a significantly large T2 sagittal tilt in the D group compared with the C group. Force platform analysis showed concordance between GRF and GL, whereas discordance was observed between GRF and C7PL. The D group could be classified into 2 groups based on the global sagittal alignment: 10 patients were classified as the hypo-compensation type (small SVA, small CL, small TK, and normal-to-small LL), whereas 62 were classified as the forward-leaning type (large sagittal vertical axis, large cervical lordosis, large thoracic kyphosis, and small LL). The prevalence of discordance between GL and C7PL in ASD patients was 24%, and thoracic kyphosis and global sagittal alignment were significantly correlated with this discordance. The concordance of GRF and GL and the discordance of GRF and C7PL highlight the importance and necessity of accounting for GL when considering surgical treatment.


The chiropractic care of children.
The objective of this study was to characterize the practice of pediatric chiropractic. The study design was a cross-sectional descriptive survey. The settings were private practices throughout the United States, Canada, and Europe. The participants were 548 chiropractors, the majority of whom are practicing in the United States, Canada, and Europe. Practitioner demographics (i.e., gender, years in practice, and chiropractic alma mater), practice characteristics (i.e., patient visits per week, practice income reimbursement), and chiropractic technique were surveyed. The practitioners were also asked to indicate common indicators for pediatric presentation, their practice activities (i.e., use of herbal remedies, exercise and rehabilitation, prayer healing, etc.), and referral patterns. A majority of the responders were female with an average practice experience of 8 years. They attended an average of 133 patient visits per week, with 21% devoted to the care of children (<18 years of age). Practice income was derived primarily from out-of-pocket reimbursement with charges of an average of $127 and $42 for the first and subsequent visits, respectively. These visits were reimbursed to address common conditions of childhood (i.e., asthma, ear infections, etc.). Approach to patient care was spinal manipulative therapy (SMT) augmented with herbal remedies, exercises, rehabilitation, and so on. Wellness care also figured prominently as a motivator for chiropractic care. Fifty-eight percent (58%) indicated an established relationship with an osteopathic or medical physician. Eighty percent (80%) of the responders indicated referring patients to medical practitioners while only 29% indicated receiving a referral from a medical/osteopathic physician. The chiropractic care of children is a significant aspect of the practice of chiropractic. Further research is warranted to examine the safety and effectiveness of this popular nonallopathic approach to children's health.


APC and the three-hit hypothesis.
The seminal 'two-hit hypothesis' implicitly assumes that bi-allelic tumour suppressor gene (TSG) mutations cause loss of protein function. All subsequent events in that tumour therefore take place on an essentially null background for that TSG protein. We have shown that the two-hit model requires modification for the APC TSG, because mutant APC proteins probably retain some function and the two hits are co-selected to produce an optimal level of Wnt activation. We wondered whether the optimal Wnt level might change during tumour progression, leading to selection for more than two hits at the APC locus. Comprehensive screening of a panel of colorectal cancer (CRC) cell lines and primary CRCs showed that some had indeed acquired third hits at APC. These third hits were mostly copy number gains or deletions, but could be protein-truncating mutations. Third hits were significantly less common when the second hit at APC had arisen by copy-neutral loss of heterozygosity. Both polyploid and near-diploid CRCs had third hits, and the third hits did not simply arise as a result of acquiring a polyploid karyotype. The third hits affected mRNA and protein levels, with potential functional consequences for Wnt signalling and tumour growth. Although some third hits were probably secondary to genomic instability, others did appear specifically to target APC. Whilst it is generally believed that tumours develop and progress through stepwise accumulation of mutations in different functional pathways, it also seems that repeated targeting of the same pathway and/or gene is selected in some cancers.


High- versus standard-dose ranitidine for control of heartburn in poorly responsive acid reflux disease: a prospective, controlled trial.
H2-receptor antagonists are commonly used as initial therapy in patients with gastroesophageal reflux disease (GERD) and are frequently continued at the same or higher doses when symptoms persist after 4-6 wk of therapy. Our objective was to compare the efficacy of twice-daily treatment with either ranitidine 150 mg b.i.d. or 300 mg b.i.d. in resolving heartburn in GERD patients who remained symptomatic after 6 wk of therapy with ranitidine 150 mg b.i.d. This was a two-phase, prospective study. In the first phase, GERD patients with heartburn on > or = 4 of the 7 days before entry were treated with open-label ranitidine 150 mg b.i.d. for 6 wk. In the second phase, patients who were still symptomatic were randomized to 8 wk of double-blind ranitidine therapy at either the same (150 mg b.i.d.) or a higher dose (300 mg b.i.d.). The primary efficacy variable was the resolution of heartburn at wk 4 and 8, as monitored through diary cards. Of the 481 patients treated for 6 wk in phase I, 285 (59%) were still symptomatic; 271 patients (95% of those still symptomatic) were randomized to double-blind treatment with ranitidine. In phase II, 45% of the patients in each treatment group experienced no more than mild heartburn; complete heartburn resolution was observed in <20% of patients in either group at wk 4 and 8. There were no significant differences in efficacy between the two treatment groups. These results indicate that the majority of GERD patients still have symptoms of heartburn after 6 wk of ranitidine therapy. Only a minority of these patients experience complete relief of heartburn after an additional 8 wk of treatment, which demonstrates that doubling the dose of ranitidine is not efficacious.


Dual action of vitamin C in iron supplement therapeutics for iron deficiency anemia: prevention of liver damage induced by iron overload.
Vitamin C, an excellent reducing agent, aids in increasing absorbable ferrous iron in iron deficiency anemia. As an efficient antioxidant, it is still unknown whether vitamin C exerts protective effects against liver damage caused by iron excess and whether mitochondria are the target effectors of the above effects. In this study, 48 mice were randomly divided into a control group, iron-overload group, TAU-treated + iron-overload group and vitamin C-treated + iron-overload group with 12 mice per group. The mice were fed 4 months on pellet diets supplemented with iron in the form of ferrocene. The iron ratio in the diet was maintained at 0.2% (w/w) for 90 days and then 0.4% (w/w) for the remaining 30 days. Furthermore, 2 g kg-1 vitamin C and 20 mg kg-1 TAU were administered daily by oral gavage prior to iron-overload administration at 6 weeks and throughout the course of the experiments. We investigated the protective effects of vitamin C against liver damage by assessing the liver weight to body weight ratio (LW/BW), serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, and histological changes. In addition, enzymatic and non-enzymatic antioxidants, reactive oxygen species (ROS) generation, mitochondrial swelling, and mitochondrial membrane potential (MMP) were evaluated to clarify the antioxidant effects of vitamin C. We found that vitamin C significantly attenuated impaired liver function in mice induced by iron overload via antioxidation, whereas no significant effect on iron uptake was observed. Vitamin C targeted the mitochondria, preventing mitochondrial swelling, MMP dissipation, and ROS burst, thus inhibiting hepatic apoptosis. Collectively, our results suggest that vitamin C acts as a "double agent" in iron supplementation therapy for iron deficiency anemia, boosting iron absorption for preventing iron deficiency and preventing liver damage due to excessive iron intake during treatment.


Tryptophan via serotonin/kynurenine pathways abnormalities in a large cohort of aggressive inmates: markers for aggression.
Aggressive behavior is one of the most challenging symptoms in psychiatry, and biological markers for aggression lack of large sample validations. Serotonin (5-HT) and other neuroactive compounds deriving from Tryptophan (Trp), including kynurenine (Kyn), have not yet been investigated in large cohorts of aggressive individuals to validate their potential as biomarkers of aggression. In 361 male inmates we measured serum levels of Trp, 5-hydroxytryptophan, 5-HT, Kyn, the ratios 5-HT/Trp∗1000 and Kyn/Trp∗1000, and performed Structured Clinical Interview for DSM-IV Axis-I and -II Disorders (SCID-I and -II), global assessment of functioning (GAF), and scales for aggressive behavior, impulsivity, adult attention-deficit/hyperactivity disorder (ADHD), and intelligent quotient (IQ). Aggressive compared to non-aggressive inmates exhibited lower Trp and Kyn serum levels but higher levels of 5-HT and 5-HT/Trp∗1000, higher levels of impulsivity and ADHD indices, lower IQ and GAF, higher prevalence of mood disorders, drug abuse/dependence, and borderline, conduct and antisocial behaviors. Interestingly, Kyn/Trp∗1000 was positively correlated to the number of severe aggressive acts (r=0.593, P<0.001). After adjusting for confounding factors, logistic regression analysis indicated that 5-HT/Trp∗1000, antisocial behavior, and GAF were predictors of aggressive behavior. The model combining these three predictors had an area under the ROC curve of 0.851 (95% CI 0.806-0.895). This study indicates that while circulating Trp is reduced in aggressive individuals, the combination of biological (5-HT/Trp ratio) and psychopathological (antisocial behavior and GAF) markers discriminates between aggressive and non-aggressive behavior suggesting the potential of a multi-marker approach in psychiatry given the heterogenic nature of mental diseases.


Bacterial communities of diverse Drosophila species: ecological context of a host-microbe model system.
Drosophila melanogaster is emerging as an important model of non-pathogenic host-microbe interactions. The genetic and experimental tractability of Drosophila has led to significant gains in our understanding of animal-microbial symbiosis. However, the full implications of these results cannot be appreciated without the knowledge of the microbial communities associated with natural Drosophila populations. In particular, it is not clear whether laboratory cultures can serve as an accurate model of host-microbe interactions that occur in the wild, or those that have occurred over evolutionary time. To fill this gap, we characterized natural bacterial communities associated with 14 species of Drosophila and related genera collected from distant geographic locations. To represent the ecological diversity of Drosophilids, examined species included fruit-, flower-, mushroom-, and cactus-feeders. In parallel, wild host populations were compared to laboratory strains, and controlled experiments were performed to assess the importance of host species and diet in shaping bacterial microbiome composition. We find that Drosophilid flies have taxonomically restricted bacterial communities, with 85% of the natural bacterial microbiome composed of only four bacterial families. The dominant bacterial taxa are widespread and found in many different host species despite the taxonomic, ecological, and geographic diversity of their hosts. Both natural surveys and laboratory experiments indicate that host diet plays a major role in shaping the Drosophila bacterial microbiome. Despite this, the internal bacterial microbiome represents only a highly reduced subset of the external bacterial communities, suggesting that the host exercises some level of control over the bacteria that inhabit its digestive tract. Finally, we show that laboratory strains provide only a limited model of natural host-microbe interactions. Bacterial taxa used in experimental studies are rare or absent in wild Drosophila populations, while the most abundant associates of natural Drosophila populations are rare in the lab.


Deaths from pesticide poisoning in South Korea: trends over 10 years.
Pesticide poisoning is a major cause of death in the world. The objective of this study was to examine the trends of pesticide poisoning deaths and their epidemiologic characteristics in South Korea. We evaluated the age-standardized mortality rates from pesticide-related deaths (intentional self-poisoning, accidental poisoning, assault, undetermined intent poisoning) in South Korea from 1996 through 2005, using registered death data obtained from the Korea National Statistical Office. The regional rurality index was calculated and correlation analyses were used to estimate the association with pesticide poisoning mortality. The number of pesticide poisoning deaths from 1996 through 2005 was 25,360, which accounted for 58.3% of the total poisoning fatalities. The age-standardized mortality rates by pesticide poisoning significantly increased from 4.42 to 6.42 per 100,000 population, whereas the total death rate was decreased in the same period. Intentional self-poisoning was the majority cause of death from pesticides (84.8% of total pesticide poisoning deaths). The majority of the pesticide poisoning deaths were men, over 50 years old, with education less than middle school, and residing in rural areas. The rate of pesticide poisoning deaths was the highest in the farming period and was significantly correlated with the rurality index of each region. Pesticide poisoning deaths substantially increased during the 10-year study period, and showed demographic, seasonal and regional variations. More intensive intervention efforts to reduce pesticide mortality should become a public health priority in South Korea.


Cellular and synaptic actions of acetylcholine in the lamprey spinal cord.
This study investigated cellular and synaptic mechanisms of cholinergic neuromodulation in the in vitro lamprey spinal cord. Most spinal neurons tested responded to local application of acetylcholine (ACh) with depolarization and decreased input resistance. The depolarization persisted in the presence of either tetrodotoxin or muscarinic antagonist scopolamine and was abolished with nicotinic antagonist mecamylamine, indicating a direct depolarization through nicotinic ACh receptors. Local application of muscarinic ACh agonists modulated synaptic strength in the spinal cord by decreasing the amplitude of unitary excitatory and inhibitory postsynaptic potentials. The postsynaptic response to direct application of glutamate was unchanged by muscarinic agonists, suggesting a presynaptic mechanism. Cholinergic feedback from motoneurons was assessed using stimulation of a ventral root in the quiescent spinal cord while recording intracellularly from spinal motoneurons or interneurons. Mainly depolarizing potentials were observed, a portion of which was insensitive to removal of extracellular Ca2+, indicating electrotonic coupling. Hyperpolarizing potentials were also observed and were attenuated by the glycinergic antagonist strychnine, whereas depolarizing responses were potentiated by strychnine. Mecamylamine also reduced hyperpolarizing responses. The pharmacology of these responses suggests a Renshaw-like feedback pathway in lamprey. Immunohistochemistry for choline acetyltransferase, performed in combination with retrograde filling of motoneurons, demonstrated a population of nonmotoneuron cholinergic cells in the lamprey spinal cord. Thus endogenous cholinergic modulation of the lamprey spinal locomotor network is likely produced by both motoneurons and cholinergic interneurons acting via combined postsynaptic and presynaptic actions.


The experience of childbirth for survivors of incest.
The purpose of this research was to discover what experiences, including feelings, women who are survivors of incest have during childbirth. The recognition that the childbirth experience may trigger memories of incest that will then alter the course of childbirth was the framework for the study. This exploratory research was undertaken using a phenomenological approach. The six participants were obtained through networking and issues surrounding confidentiality and consent were of a high priority. The data consists of transcripts of taped, one to two hour, in-depth interviews. During interviews each woman recalled her experiences and feelings during and surrounding childbirth. Qualitative analysis was undertaken, the findings indicating that memories of previous abuse may be provoked by childbirth but that it is a very individualised response. The effect of these memories on the childbirth experience is also variable but does occur. Privacy, control and touch are important aspects of these women's childbirth experiences. The results and interpretation of findings presented in this report centre around the labour and delivery experiences of the women. The variety of women's needs demonstrated in this project emphasise the importance of individualised care by midwives and doctors involved in the care of childbearing women.


Alopecia as a rare but distinct manifestation of pemphigus vulgaris.
Pemphigus vulgaris (PV) patients may develop scalp erosions, however, the development of alopecia has been reported to be extremely rare. To delineate the clinicopathological features of alopecia in PV and provide insight into the pathogenesis of this rarely observed manifestation. A retrospective case note review was performed on five PV patients presenting with progressive hair loss and alopecic patches. Data were collected on demographics and clinical findings. Results for hair pull tests, direct immunofluorescence study of plucked hairs, established laboratory tests to detect anti-desmoglein 1 and 3 autoantibodies and scalp swab culture were recorded. A combination of vertical and horizontal sectioning technique enabled detailed histopathological analysis of alopecic patches. Clinical course was monitored. Anagen hair follicles with the outer root sheath structure were easily pulled from perilesional scalp, with intercellular IgG deposition on the outer root sheath keratinocytes. Acantholysis between outer root sheath keratinocytes extending from the infundibulum to suprabalbar level was evident in anagen hair follicles of affected lesions. Perifollicular cell infiltration was observed in the lesions where scalp swabs detected micro-organisms. The bulge stem cell area was mostly intact. Alopecia was non-scarring and following 4 weeks of therapy hair re-growth was seen in all patients. In PV, the combination of anti-desmoglein autoantibody-mediated acantholysis in conjunction with secondary factors, such as inflammatory changes due to infection, may cause weakening of hair follicle anchorage resulting in hair loss and alopecic patches. This unusual clinical phenotype should alert physicians to PV as a potential diagnosis.


Relative expression of a dominant mutated ABCC8 allele determines the clinical manifestation of congenital hyperinsulinism.
Congenital hyperinsulinism (CHI) is most commonly caused by mutations in the β-cell ATP-sensitive K(+) (K(ATP)) channel genes. Severe CHI was diagnosed in a 1-day-old girl; the mother's cousin and sister had a similar phenotype. ABCC8 gene sequencing (leukocyte DNA) revealed a heterozygous, exon 37, six-base pair in-frame insertion mutation in the affected patient and aunt but also in her unaffected mother and grandfather. In expression studies using transfected COSm6 cells, mutant sulfonylurea receptor 1 (SUR1) protein was expressed on the cell surface but failed to respond to MgADP even in the heterozygous state. mRNA expression in lymphocytes determined by sequencing cDNA clones and quantifying 6FAM-labeled PCR products found that although the healthy mother predominantly expressed the normal transcript, her affected daughter, carrying the same mutant allele, primarily transcribed the mutant. The methylation pattern of the imprinting control region of chromosome 11p15.5 and ABCC8 promoter was similar for all family members. In conclusion, differences in transcript expression may determine the clinical phenotype of CHI in this maternally inherited dominant mutation. The use of peripheral lymphocytes as a peripheral window to the β-cell transcription profile can serve in resolving β-cell phenotypes. The severe, dominant-negative nature of the 1508insAS mutation suggests that it affects the functional stoichiometry of SUR1-regulated gating of K(ATP) channels.


Including Bioconcentration Kinetics for the Prioritization and Interpretation of Regulatory Aquatic Toxicity Tests of Highly Hydrophobic Chemicals.
Worldwide, regulations of chemicals require short-term toxicity data for evaluating hazards and risks of the chemicals. Current data requirements on the registration of chemicals are primarily based on tonnage and do not yet consider properties of chemicals. For example, short-term ecotoxicity data are required for chemicals with production volume greater than 1 or 10 ton/y according to REACH, without considering chemical properties. Highly hydrophobic chemicals are characterized by low water solubility and slow bioconcentration kinetics, which may hamper the interpretation of short-term toxicity experiments. In this work, internal concentrations of highly hydrophobic chemicals were predicted for standard acute ecotoxicity tests at three trophic levels, algae, invertebrate, and fish. As demonstrated by comparison with maximum aqueous concentrations at water solubility, chemicals with an octanol-water partition coefficient (Kow) greater than 106 are not expected to reach sufficiently high internal concentrations for exerting effects within the test duration of acute tests with fish and invertebrates, even though they might be intrinsically toxic. This toxicity cutoff was explained by the slow uptake, i.e., by kinetics, not by thermodynamic limitations. Predictions were confirmed by data entries of the OECD's screening information data set (SIDS) (n = 746), apart from a few exceptions concerning mainly organometallic substances and those with inconsistency between water solubility and Kow. Taking error propagation and model assumptions into account, we thus propose a revision of data requirements for highly hydrophobic chemicals with log Kow > 7.4: Short-term toxicity tests can be limited to algae that generally have the highest uptake rate constants, whereas the primary focus of the assessment should be on persistence, bioaccumulation, and long-term effects.


Interaction between DMRT1 function and genetic background modulates signaling and pluripotency to control tumor susceptibility in the fetal germ line.
Dmrt1 (doublesex and mab-3 related transcription factor (1) is a regulator of testis development in vertebrates that has been implicated in testicular germ cell tumors of mouse and human. In the fetal mouse testis Dmrt1 regulates germ cell pluripotency in a strain-dependent manner. Loss of Dmrt1 in 129Sv strain mice results in a >90% incidence of testicular teratomas, tumors consisting cells of multiple germ layers; by contrast, these tumors have never been observed in Dmrt1 mutants of C57BL/6J (B6) or mixed genetic backgrounds. To further investigate the interaction between Dmrt1 and genetic background we compared mRNA expression in wild type and Dmrt1 mutant fetal testes of 129Sv and B6 mice at embryonic day 15.5 (E15.5), prior to overt tumorigenesis. Loss of Dmrt1 caused misexpression of overlapping but distinct sets of mRNAs in the two strains. The mRNAs that were selectively affected included some that changed expression only in one strain or the other and some that changed in both strains but to a greater degree in one versus the other. In particular, loss of Dmrt1 in 129Sv testes caused a more severe failure to silence regulators of pluripotency than in B6 testes. A number of genes misregulated in 129Sv mutant testes also are misregulated in human testicular germ cell tumors (TGCTs), suggesting similar etiology between germ cell tumors in mouse and man. Expression profiling showed that DMRT1 also regulates pluripotency genes in the fetal ovary, although Dmrt1 mutant females do not develop teratomas. Pathway analysis indicated disruption of several signaling pathways in Dmrt1 mutant fetal testes, including Nodal, Notch, and GDNF. We used a Nanos3-cre knock-in allele to perform conditional gene targeting, testing the GDNF coreceptors Gfra1 and Ret for effects on teratoma susceptibility. Conditional deletion of Gfra1 but not Ret in fetal germ cells of animals outcrossed to 129Sv caused a modest but significant elevation in tumor incidence. Despite some variability in genetic background in these crosses, this result is consistent with previous genetic mapping of teratoma susceptibility loci to the region containing Gfra1. Using Nanos3-cre we also uncovered a strong genetic interaction between Dmrt1 and Nanos3, suggesting parallel functions for these two genes in fetal germ cells. Finally, we used chromatin immunoprecipitation (ChIP-seq) analysis to identify a number of potentially direct DMRT1 targets. This analysis suggested that DMRT1 controls pluripotency via transcriptional repression of Esrrb, Nr5a2/Lrh1, and Sox2. Given the strong evidence for involvement of DMRT1 in human TGCT, the downstream genes and pathways identified in this study provide potentially useful candidates for roles in the human disease.


Addressing the determinants of child mental health: intersectionality as a guide to primary health care renewal.
Primary health care (PHC) renewal was designed explicitly to attend to the multidimensional factors impacting on health, including the social determinants of health. These determinants are central considerations in the development of integrated, cross-sectoral, and multi-jurisdictional policies such as those that inform models of shared mental health care for children. However, there are complex theoretical challenges in translating these multidimensional issues into policy. One of these is the rarely discussed interrelationships among the social determinants of health and identities such as race, gender, age, sexuality, and social class within the added confluence of geographic contexts. An intersectionality lens is used to examine the complex interrelationships among the factors affecting child mental health and the associated policy challenges surrounding PHC renewal. The authors argue that an understanding of the intersections of social determinants of health, identity, and geography is pivotal in guiding policy-makers as they address child mental health inequities using a PHC renewal agenda.


Downregulated expression of peroxiredoxin 4 in granulosa cells from polycystic ovary syndrome.
Peroxiredoxin 4 (PRDX4), a member of Peroxiredoxin (PRDX) family, is a typical 2-Cys PRDX. PRDX4 monitors the oxidative burden within cellular compartment and reduces hydrogen peroxide and alkyl hydroperoxide related to oxidative stress and apoptosis. Antioxidant, like PRDX4, may promote follicle development and participate in the pathophysiology of PCOS. In our previous study, we found that PRDX4 was expressed in mice oocyte cumulus oophorus complex, and that PRDX4 could be associated with follicle development. In this study, we explored the expression of PRDX4 in human follicles and possible role of PRDX4 in PCOS pathophysiology. Our data showed that PRDX4 was mainly expressed in granulosa cells in human ovaries. When compared to control group, both PRDX4 mRNA level and protein level decreased in PCOS group. The lowered levels of PRDX4 may relate to oxidative stress in the pathophysiologic progress of PCOS. Furthermore, expression of PRDX4 in the granulosa cells of in vivo or in vitro matured follicles was higher than that in immatured follicles, which suggested that PRDX4 may have a close relationship with follicular development. Altogether, our findings may provide new clues of the pathophysiologic mechanism of PCOS and potential therapeutic strategy using antioxidant, like PRDX4.


Healthy lifestyle and life expectancy at age 30 years in the Chinese population: an observational study.
The improvement of life expectancy is one of the aims of the Healthy China 2030 blueprint. We aimed to investigate the extent to which healthy lifestyles are associated with life expectancy in Chinese adults. We used the prospective China Kadoorie Biobank (CKB) study to examine the relative risk of mortality associated with individual and combined lifestyle factors (never smoking or quitting not for illness, no excessive alcohol use, being physically active, healthy eating habits, and healthy body shape). Participants with coronary heart disease, stroke, cancer, or missing values for body-mass index were excluded. For analysis of chronic respiratory diseases, participants with chronic obstructive pulmonary disease or asthma were excluded. We estimated the national prevalence of lifestyle factors using data from the China Nutrition and Health Surveillance (CNHS; 2015) and derived mortality rates from the Global Burden of Diseases, Injuries, and Risk Factors Study (2015). All three data sources were combined to estimate the life expectancy of individuals at age 30 years following different levels of lifestyle factors by using the life table method. The cause-specific decomposition of the life expectancy differences was analysed using Arriaga's method. After the exclusion of CKB participants with coronary heart disease, stroke, cancer, or missing BMI data at baseline, 487 209 were included in the primary analysis. Participants with COPD or asthma at baseline were additionally excluded for chronic respiratory disease-related analysis, leaving 451 233 participants with data available for analysis. Data from 171 127 adults aged 30-84 years from the CNHS 2015 were used to estimate the sex-specific and age-specific prevalence of lifestyle-related factors. There were 42 496 deaths documented over a median follow-up of 11·1 years (IQR 10·2-12·1) in CKB. The adjusted hazard ratios (aHRs) of participants adopting five versus 0-1 low-risk factors was 0·38 (95% CI 0·34-0·43) for all-cause mortality, aHR 0·37 (0·30-0·46) for cardiovascular disease mortality, aHR 0·47 (0·39-0·56) for cancer mortality, and aHR 0·30 (0·14-0·64) for chronic respiratory disease mortality. The life expectancy at age 30 years for individuals with 0-1 low-risk factors was on average 41·7 years (95% CI 41·5-42·0) for men and 47·3 years (46·6-48·0) for women. For individuals with all five low-risk factors, the life expectancy at age 30 was 50·5 years (95% CI 48·5-52·4) for men and 55·4 years (53·5-57·4) for women; meaning a difference of 8·8 years (95% CI 6·8-10·7) for men and 8·1 years (6·5-9·9) for women. The estimated extended life expectancy for men and women was mainly attributable to reduced death from cardiovascular disease (2·4 years [27% of the total extended life expectancy] for men and 3·7 years [46%] for women), cancer (2·6 years [30%] for men and 0·9 years [11%] for women), and chronic respiratory disease (0·6 years [7%] for men and 1·2 years [15%] for women). Our findings suggest that increasing the adoption of these five healthy lifestyle factors through public health interventions could be associated with substantial gains in life expectancy in the Chinese population. National Natural Science Foundation of China, National Key Research and Development Program of China, Kadoorie Charitable Foundation, UK Wellcome Trust.


Evaluation of Interleukin-10 Levels in Patients Diagnosed with Chronic Hepatitis.
One of the most important factors playing a role in chronic hepatitis B pathogenesis is cytokine release and one of the cytokines with anti-inflammatory characteristic is interleukin-10 (IL-10). The aim of the present study is to examine IL-10 levels in patients with chronic hepatitis B. Sixty-three patients with chronic hepatitis B disease who had not received any antiviral treatment were included in the study. Serum IL-10 level was investigated by enzyme-linked immunosorbent assay (ELISA) method. In the control group, 25 healthy individuals with mean age similar to the patient population were included. Control and patient groups were compared and data were statistically analysed. Interleukin-10 levels of 25 patients with hepatitis B virus (HBV) DNA levels between 2000 and 20 000 IU/mL were compared with those of 25 subjects in the control group, and the level in the chronic hepatitis B group was statistically significantly higher (p < 0.05). Interleukin-10 levels of 38 patients with HBV DNA > 20 000 IU/mL were statistically significantly higher than those in the control group. When chronic hepatitis B patients were compared among themselves, IL-10 levels increased as HBV DNA levels increased. Also, when IL-10 levels of hepatitis B 'e' antigen (HBeAg) positive patients were compared with those of HBeAg negative patients, the difference was not statistically significant. It is believed that decreasing IL-10 levels by various methods would have significant contributions in disease progression and treatment. Moreover, IL-10 level may be an important marker in HBeAg seroconversion and evaluation of treatment response.


Effects of different soil ameliorants on karee trees (Searsia lancea) growing on mine tailings dump soil-part I: pot trials.
Rehabilitation of mine tailings dams is often a challenge due to a lack of nutrients and a poor humus reservoir prevailing in tailings soils. This is especially true for establishing longer lived species such as trees. For these reasons the effects of different soil ameliorants (woodchips compost, vermicompost, mature sewage sludge), added to the root system of Karee (Searsia lancea) saplings were tested in pot trials. Those pots were filled with platinum and gold tailings substrate as well as red clay soil, respectively. For three months plants remained in a greenhouse and were subsequently moved to a test field outside. Throughout the test period regular chl a fluorescence measurements were taken and subjected to JIP-test quantifying changes in photosynthetic vitality status. Additionally, growth measurements and one-off leaf analysis were carried out. Test plants growing on mine tailings experienced an up to 35% higher average photosynthetic vitality (PI(ABS)) and improved nutrient supply, when treated with mature sewage sludge. Consequently, sewage sludge treated plants showed a higher biomass build-up rate and an up to 55% higher diameter growth, compared to control. In summary the experiments present a low cost alternative for reforestation enterprises on platinum and gold tailings dams in South Africa.


Fractional Fourier transform: simulations and experimental results.
Recently two optical interpretations of the fractional Fourier transform operator were introduced. We address implementation issues of the fractional-Fourier-transform operation. We show that the original bulk-optics configuration for performing the fractional-Fourier-transform operation [J. Opt. Soc. Am. A 10, 2181 (1993)] provides a scaled output using a fixed lens. For obtaining a non-scaled output, an asymmetrical setup is suggested and tested. For comparison, computer simulations were performed. A good agreement between computer simulations and experimental results was obtained.


Inhibitory effects of kaempferol on the invasion of human breast carcinoma cells by downregulating the expression and activity of matrix metalloproteinase-9.
Matrix metalloproteinases (MMPs) have been regarded as major critical molecules assisting tumor cells during metastasis, for excessive ECM (ECM) degradation, and cancer cell invasion. In the present study, in vitro and in vivo assays were employed to examine the inhibitory effects of kaempferol, a natural polyphenol of flavonoid family, on tumor metastasis. Data showed that kaempferol could inhibit adhesion, migration, and invasion of MDA-MB-231 human breast carcinoma cells. Moreover, kaempferol led to the reduced activity and expression of MMP-2 and MMP-9, which were detected by gelatin zymography, real-time PCR, and western blot analysis, respectively. Further elucidation of the mechanism revealed that kaempferol treatment inhibited the activation of transcription factor activator protein-1 (AP-1) and MAPK signaling pathway. Moreover, kaempferol repressed phorbol-12-myristate-13-acetate (PMA)-induced MMP-9 expression and activity through suppressing the translocation of protein kinase Cδ (PKCδ) and MAPK signaling pathway. Our results also indicated that kaempferol could block the lung metastasis of B16F10 murine melanoma cells as well as the expression of MMP-9 in vivo. Taken together, these results demonstrated that kaempferol could inhibit cancer cell invasion through blocking the PKCδ/MAPK/AP-1 cascade and subsequent MMP-9 expression and its activity. Therefore, kaempferol might act as a therapeutic potential candidate for cancer metastasis.


Discovery and mode of action of a novel analgesic β-toxin from the African spider Ceratogyrus darlingi.
Spider venoms are rich sources of peptidic ion channel modulators with important therapeutical potential. We screened a panel of 60 spider venoms to find modulators of ion channels involved in pain transmission. We isolated, synthesized and pharmacologically characterized Cd1a, a novel peptide from the venom of the spider Ceratogyrus darlingi. Cd1a reversibly paralysed sheep blowflies (PD50 of 1318 pmol/g) and inhibited human Cav2.2 (IC50 2.6 μM) but not Cav1.3 or Cav3.1 (IC50 > 30 μM) in fluorimetric assays. In patch-clamp electrophysiological assays Cd1a inhibited rat Cav2.2 with similar potency (IC50 3 μM) without influencing the voltage dependence of Cav2.2 activation gating, suggesting that Cd1a doesn't act on Cav2.2 as a classical gating modifier toxin. The Cd1a binding site on Cav2.2 did not overlap with that of the pore blocker ω-conotoxin GVIA, but its activity at Cav2.2-mutant indicated that Cd1a shares some molecular determinants with GVIA and MVIIA, localized near the pore region. Cd1a also inhibited human Nav1.1-1.2 and Nav1.7-1.8 (IC50 0.1-6.9 μM) but not Nav1.3-1.6 (IC50 > 30 μM) in fluorimetric assays. In patch-clamp assays, Cd1a strongly inhibited human Nav1.7 (IC50 16 nM) and produced a 29 mV depolarising shift in Nav1.7 voltage dependence of activation. Cd1a (400 pmol) fully reversed Nav1.7-evoked pain behaviours in mice without producing side effects. In conclusion, Cd1a inhibited two anti-nociceptive targets, appearing to interfere with Cav2.2 inactivation gating, associated with the Cav2.2 α-subunit pore, while altering the activation gating of Nav1.7. Cd1a was inactive at some of the Nav and Cav channels expressed in skeletal and cardiac muscles and nodes of Ranvier, apparently contributing to the lack of side effects at efficacious doses, and suggesting potential as a lead for development of peripheral pain treatments.


Diversity of nicotinic acetylcholine receptors in rat hippocampal neurons. III. Agonist actions of the novel alkaloid epibatidine and analysis of type II current.
The nicotinic-agonist properties of epibatidine, an alkaloid originally isolated from the Ecuadorian frog Epipedobates tricolor, was investigated in rat hippocampal neurons grown in culture. The ability of epibatidine enantiomers to evoke nicotinic whole-cell currents of type IA, sensitive to blockade by alpha-bungaro-toxin and methyllycaconitine and presumably subserved by an alpha-7-based nAChR, and of type II, sensitive to blockade by dihydro-beta-erythroidine and presumably subserved by an alpha 4 beta 2-based nAChR, was studied and compared with that of other nicotinic agonists. Epibatidine elicited type IA currents with EC50 values of 2.9 and 4.3 microM for the (-)- and (+)- enantiomers, respectively. The potency of the agonists in evoking type IA currents was as follows: (-)-epibatidine > (+)- anatoxin-a > (+)-epibatidine > (-)-nicotine > DMPP > cytisine > acetylcholine. The EC50 values of (-)- and (+)- epibatidine in eliciting type II currents were 19 and 15 nM, respectively. The order of potency of the agonists in evoking type II currents was: (+)-epibatidine > (-)-epibatidine > cytisine > (+)-anatoxin-a > (-)-nicotine > acetylcholine > DMPP. Although these agonists produced nearly identical maximal type IA responses, the size of the maximal type II responses varied with the agonist. Cytisine and (+)-anatoxin-a were the least efficacious agonists in inducing type II currents. Enantiomers of epibatidine showed the least stereoselectivity, those of nicotine showed a moderate stereoselectivity, and those of anatoxin-a exhibited the highest stereoselectivity in inducing either type of currents. In summary, these results suggest that epibatidine can be used as a novel nicotinic agonist for the study of type II currents, and that the nicotinic acetylcholine receptor related to type II currents may be one of the key target sites through which agonists such as epibatidine and nicotine elicit their behavioral actions in vivo.


Enhanced viral intergenic region-specific short interfering RNA accumulation and DNA methylation correlates with resistance against a geminivirus.
RNA silencing mediated by short-interfering RNA (siRNA) is used by plants as a defense against viruses. In the case of geminiviruses, viral DNA is targeted at the transcriptional level, while virus-derived transcripts are targeted by posttranscriptional silencing. Mungbean yellow mosaic India virus (MYMIV), a bipartite geminivirus, causes yellow mosaic disease in soybean (Glycine max). A soybean variety resistant to this disease has been identified (line PK416). To understand the molecular mechanism underlying this resistance, distribution of MYMIV-derived siRNAs along the viral genome was compared in resistant and susceptible plants, using samples obtained in the first few days following inoculation. We observed that, in the resistant soybean variety, most of the virus-derived siRNAs were complementary to the intergenic region (IR), while in the susceptible variety (line JS335), a majority of the siRNAs corresponded to coding regions of the viral genome. Most of the IR-specific siRNA molecules produced in the resistant plants were 24 nt in size. Bisulfite sequencing showed that, in the resistant plants, a higher level of methylation occurred in the IR of viral DNA.


Diagnostic yield of renal biopsies: a retrospective single center review.
Previous studies have examined the spectrum of diseases identified with a kidney biopsy and the complications of the procedure. However, few studies have examined the utility of the test to clarify the diagnosis and guide treatment of pediatric patients. This retrospective, single-center chart review was performed to test the hypothesis that at least 80% of native kidney biopsies provide clinically valuable information that rationally guides diagnosis and patient management. 200 biopsies performed between January 1, 2000 and June 30, 2008 were reviewed. A scheme composed of six categories was devised to classify the utility of each kidney biopsy. 196 complete case files were available for review. Twenty-four (12.2%) biopsies did not shed light on the diagnosis and were unhelpful in patient management - 21 biopsies (10.7%) were non-diagnostic and 3 (1.5%) failed to yield enough tissue for examination. The number of unhelpful biopsies did not cluster in any specific disease entity. Our findings provide guidance to nephrologists about the total risk of a kidney biopsy, including uninformative results, when seeking informed consent for the procedure. The results suggest an appropriate balance has been reached which maximizes the use of kidney biopsies while minimizing the risk of this invasive procedure (word count: 202).


Ultrahigh strength and high electrical conductivity in copper.
Methods used to strengthen metals generally also cause a pronounced decrease in electrical conductivity, so that a tradeoff must be made between conductivity and mechanical strength. We synthesized pure copper samples with a high density of nanoscale growth twins. They showed a tensile strength about 10 times higher than that of conventional coarse-grained copper, while retaining an electrical conductivity comparable to that of pure copper. The ultrahigh strength originates from the effective blockage of dislocation motion by numerous coherent twin boundaries that possess an extremely low electrical resistivity, which is not the case for other types of grain boundaries.


Prevalence of Vibrio cholerae O1 El Tor variant in a cholera-endemic zone of Kenya.
Since 2007, Kenya has experienced an increase in cholera outbreaks characterized by a high fatality rate. In this study, we characterized 81 Vibrio cholerae isolates from diarrhoeal stool samples in Nyanza, a cholera-endemic lake region of Kenya, for virulence properties, clonality and antibiotic susceptibility. Eighty of these isolates were V. cholerae O1 El Tor variants carrying the classical ctxB gene sequence, while one isolate was V. cholerae non-O1/O139. All of the El Tor variants were of clonal origin, as revealed by PFGE, and were susceptible to ampicillin, tetracycline, ciprofloxacin, fosfomycin, kanamycin and norfloxacin. However, the isolates showed resistance to sulfamethoxazole/trimethoprim and streptomycin, and intermediate resistance to nalidixic acid, chloramphenicol and imipenem. The non-O1/O139 isolate carried the cholix toxin II gene (chxA II) and was susceptible to all antimicrobials tested except ampicillin. We propose that an El Tor variant clone caused the Nyanza cholera outbreak of 2007-2008.


Comprehensive Protocol to Sample and Process Bone Marrow for Measuring Measurable Residual Disease and Leukemic Stem Cells in Acute Myeloid Leukemia.
Response criteria in acute myeloid leukemia (AML) has recently been re-established, with morphologic examination utilized to determine whether patients have achieved complete remission (CR). Approximately half of the adult patients who entered CR will relapse within 12 months due to the outgrowth of residual AML cells in the bone marrow. The quantitation of these remaining leukemia cells, known as minimal or measurable residual disease (MRD), can be a robust biomarker for the prediction of these relapses. Moreover, retrospective analysis of several studies has shown that the presence of MRD in the bone marrow of AML patients correlates with poor survival. Not only is the total leukemic population, reflected by cells harboring a leukemia associated immune-phenotype (LAIP), associated with clinical outcome, but so is the immature low frequency subpopulation of leukemia stem cells (LSC), both of which can be monitored through flow cytometry MRD or MRD-like approaches. The availability of sensitive assays that enable detection of residual leukemia (stem) cells on the basis of disease-specific or disease-associated features (abnormal molecular markers or aberrant immunophenotypes) have drastically improved MRD assessment in AML. However, given the inherent heterogeneity and complexity of AML as a disease, methods for sampling bone marrow and performing MRD and LSC analysis should be harmonized when possible. In this manuscript we describe a detailed methodology for adequate bone marrow aspirate sampling, transport, sample processing for optimal multi-color flow cytometry assessment, and gating strategies to assess MRD and LSC to aid in therapeutic decision making for AML patients.


Plasmacytoid dendritic cells play a role for effective innate immune responses during Chlamydia pneumoniae infection in mice.
Plasmacytoid dendritic cells (pDCs) are known for their robust antiviral response and their pro-tolerance effects towards allergic diseases and tissue engraftments. However, little is known about the role pDCs may play during a bacterial infection, including pulmonary Chlamydia pneumoniae (CP). In this study, we investigated the role of pDCs during pulmonary CP infection. Our results revealed that depletion of pDCs during acute CP infection in mice results in delayed and reduced lung inflammation, with an early delay in cellular recruitment and significant reduction in early cytokine production in the lungs. This was followed by impaired and delayed bacterial clearance from the lungs which then resulted in a severe and prolonged chronic inflammation and iBALT like structures containing large numbers of B and T cells in these animals. We also observed that increasing the pDC numbers in the lung by FLT3L treatment experimentally results in greater lung inflammation during acute CP infection. In contrast to these results, restimulation of T-cells in the draining lymph nodes of pDC-depleted mice induced greater amounts of proinflammatory cytokines than we observed in control mice. These results suggest that pDCs in the lung may provide critical proinflammatory innate immune responses in response to CP infection, but are suppressive towards adaptive immune responses in the lymph node. Thus pDCs in the lung and the draining lymph node appear to have different roles and phenotypes during acute CP infection and may play a role in host immune responses.


Involving practice nurses in primary care research: the experience of multiple and competing demands.
This paper is a report of a study of the experiences of practice nurses delivering a complex research intervention in an exploratory randomized controlled trial in primary care. As practice nurses increasingly become involved in primary care research, it is important to understand not only what impact this may have on their existing role but also equally on what their potential contribution might be. Fourteen of the 15 practice nurses involved in the delivery of a complex intervention were purposively sampled and interviewed in their workplace between June and October 2007. Interviews were audio-recorded, transcribed and analysed using a Framework Approach and NVivo software. Findings. Time influenced the nurses' engagement with the various aspects of the trial and meant that they constantly had to make judgments and decisions in response to the multiple agendas presented to them: they had to negotiate a range of competing loyalties between their professional clinical role, their role in the research and practice teams and their relationship with patients. The nurses' accounts consequently provide insight into the active role they played both in the trial process and the delivery of the complex intervention. The nurses were key to the delivery of the trial. If practice nurses are to develop a research role in their professional work, it is important to understand their perceptions and the impact such involvement has on them and their practice. Consideration of these factors is consequently valuable when developing research in primary care settings.


Heterodimeric protein complex identification by naïve Bayes classifiers.
Protein complexes are basic cellular entities that carry out the functions of their components. It can be found that in databases of protein complexes of yeast like CYC2008, the major type of known protein complexes is heterodimeric complexes. Although a number of methods for trying to predict sets of proteins that form arbitrary types of protein complexes simultaneously have been proposed, it can be found that they often fail to predict heterodimeric complexes. In this paper, we have designed several features characterizing heterodimeric protein complexes based on genomic data sets, and proposed a supervised-learning method for the prediction of heterodimeric protein complexes. This method learns the parameters of the features, which are embedded in the naïve Bayes classifier. The log-likelihood ratio derived from the naïve Bayes classifier with the parameter values obtained by maximum likelihood estimation gives the score of a given pair of proteins to predict whether the pair is a heterodimeric complex or not. A five-fold cross-validation shows good performance on yeast. The trained classifiers also show higher predictability than various existing algorithms on yeast data sets with approximate and exact matching criteria. Heterodimeric protein complex prediction is a rather harder problem than heteromeric protein complex prediction because heterodimeric protein complex is topologically simpler. However, it turns out that by designing features specialized for heterodimeric protein complexes, predictability of them can be improved. Thus, the design of more sophisticate features for heterodimeric protein complexes as well as the accumulation of more accurate and useful genome-wide data sets will lead to higher predictability of heterodimeric protein complexes. Our tool can be downloaded from http://imi.kyushu-u.ac.jp/~om/.


A typology of time-scale mismatches and behavioral interventions to diagnose and solve conservation problems.
Ecological systems often operate on time scales significantly longer or shorter than the time scales typical of human decision making, which causes substantial difficulty for conservation and management in socioecological systems. For example, invasive species may move faster than humans can diagnose problems and initiate solutions, and climate systems may exhibit long-term inertia and short-term fluctuations that obscure learning about the efficacy of management efforts in many ecological systems. We adopted a management-decision framework that distinguishes decision makers within public institutions from individual actors within the social system, calls attention to the ways socioecological systems respond to decision makers' actions, and notes institutional learning that accrues from observing these responses. We used this framework, along with insights from bedeviling conservation problems, to create a typology that identifies problematic time-scale mismatches occurring between individual decision makers in public institutions and between individual actors in the social or ecological system. We also considered solutions that involve modifying human perception and behavior at the individual level as a means of resolving these problematic mismatches. The potential solutions are derived from the behavioral economics and psychology literature on temporal challenges in decision making, such as the human tendency to discount future outcomes at irrationally high rates. These solutions range from framing environmental decisions to enhance the salience of long-term consequences, to using structured decision processes that make time scales of actions and consequences more explicit, to structural solutions aimed at altering the consequences of short-sighted behavior to make it less appealing. Additional application of these tools and long-term evaluation measures that assess not just behavioral changes but also associated changes in ecological systems are needed.


Functional Movement Screen Items Predict Dynamic Balance Under Military Torso Load.
Musculoskeletal injuries threaten military readiness and impose a significant financial burden. The functional movement screen (FMS), a 7-item, preparticipation screening tool, may aid in identifying compensatory movement patterns that can hinder physical performance and lead to injuries in active-duty military personnel. The primary aim of our study was to determine if items scores from the traditional FMS or a modified FMS (mFMS) obtained under loaded conditions can predict dynamic balance scores while wearing a military load. Thirty physically active adults (19 males and 11 females) who qualified for Army basic training completed unloaded and loaded FMS testing. Loaded balance was assessed using the Y balance test and the Biodex balance system. The mFMS and both loaded balance assessment protocols included a military load consisting of a standard issue rucksack (M.O.L.L.E.), kevlar helmet, and weighted vest (mass = 24.2 kg). mFMS item scores were significantly lower than FMS scores for six of the seven movement items. Both FMS and mFMS composite scores were correlated with Y balance scores (FMS: r = 0.53, P = 0.003; mFMS: r = 0.37, P = 0.043). Participants with higher composite scores (≥15) outperformed those with lower composite scores (≤14) for the FMS (P =0.006, d = 1.16) and mFMS (P = 0.031, d = 0.75). Lasso penalized regression analyses revealed that (1) higher unloaded in-line lunge subscores predicted better Y balance scores, (2) loaded trunk stability push-up scores of three predicted worse balance on the Biodex, and (3) unloaded and loaded shoulder mobility scores of 3 predicted better performance on the Biodex balance system. The in-line lunge subscore from the FMS was the strongest predictor of torso-loaded balance, and the FMS may be more appropriate than a torso-loaded FMS battery when predicting torso-loaded balance among adults with body composition and fitness profiles similar to those of entering military recruits. These findings provide evidence supporting the use of the conventional FMS to identify active-duty personnel who exhibit greater balance deficits and may develop related musculoskeletal injuries while performing operational tasks that require the transport of heavy loads over long distances.


Dietary exposure to soy or whey proteins alters colonic global gene expression profiles during rat colon tumorigenesis.
We previously reported that lifetime consumption of soy proteins or whey proteins reduced the incidence of azoxymethane (AOM)-induced colon tumors in rats. To obtain insights into these effects, global gene expression profiles of colons from rats with lifetime ingestion of casein (CAS, control diet), soy protein isolate (SPI), and whey protein hydrolysate (WPH) diets were determined. Male Sprague Dawley rats, fed one of the three purified diets, were studied at 40 weeks after AOM injection and when tumors had developed in some animals of each group. Total RNA, purified from non-tumor tissue within the proximal half of each colon, was used to prepare biotinylated probes, which were hybridized to Affymetrix RG_U34A rat microarrays containing probes sets for 8799 rat genes. Microarray data were analyzed using DMT (Affymetrix), SAM (Stanford) and pair-wise comparisons. Differentially expressed genes (SPI and/or WPH vs. CAS) were found. We identified 31 induced and 49 repressed genes in the proximal colons of the SPI-fed group and 44 induced and 119 repressed genes in the proximal colons of the WPH-fed group, relative to CAS. Hierarchical clustering identified the co-induction or co-repression of multiple genes by SPI and WPH. The differential expression of I-FABP (2.92-, 3.97-fold down-regulated in SPI and WPH fed rats; P = 0.023, P = 0.01, respectively), cyclin D1 (1.61-, 2.42-fold down-regulated in SPI and WPH fed rats; P = 0.033, P = 0.001, respectively), and the c-neu proto-oncogene (2.46-, 4.10-fold down-regulated in SPI and WPH fed rats; P < 0.001, P < 0.001, respectively) mRNAs were confirmed by real-time quantitative RT-PCR. SPI and WPH affected colonic neuro-endocrine gene expression: peptide YY (PYY) and glucagon mRNAs were down-regulated in WPH fed rats, whereas somatostatin mRNA and corresponding circulating protein levels, were enhanced by SPI and WPH. The identification of transcripts co- or differentially-regulated by SPI and WPH diets suggests common as well as unique anti-tumorigenesis mechanisms of action which may involve growth factor, neuroendocrine and immune system genes. SPI and WPH induction of somatostatin, a known anti-proliferative agent for colon cancer cells, would inhibit tumorigenesis.


The immune response to melanoma is limited by thymic selection of self-antigens.
The expression of melanoma-associated antigens (MAA) being limited to normal melanocytes and melanomas, MAAs are ideal targets for immunotherapy and melanoma vaccines. As MAAs are derived from self, immune responses to these may be limited by thymic tolerance. The extent to which self-tolerance prevents efficient immune responses to MAAs remains unknown. The autoimmune regulator (AIRE) controls the expression of tissue-specific self-antigens in thymic epithelial cells (TECs). The level of antigens expressed in the TECs determines the fate of auto-reactive thymocytes. Deficiency in AIRE leads in both humans (APECED patients) and mice to enlarged autoreactive immune repertoires. Here we show increased IgG levels to melanoma cells in APECED patients correlating with autoimmune skin features. Similarly, the enlarged T cell repertoire in AIRE(-/-) mice enables them to mount anti-MAA and anti-melanoma responses as shown by increased anti-melanoma antibodies, and enhanced CD4(+) and MAA-specific CD8(+) T cell responses after melanoma challenge. We show that thymic expression of gp100 is under the control of AIRE, leading to increased gp100-specific CD8(+) T cell frequencies in AIRE(-/-) mice. TRP-2 (tyrosinase-related protein), on the other hand, is absent from TECs and consequently TRP-2 specific CD8(+) T cells were found in both AIRE(-/-) and AIRE(+/+) mice. This study emphasizes the importance of investigating thymic expression of self-antigens prior to their inclusion in vaccination and immunotherapy strategies.


Pseudomonas aeruginosa invasion of and multiplication within corneal epithelial cells in vitro.
Pseudomonas aeruginosa is usually considered an extracellular pathogen. Using assays to determine intracellular survival in the presence of gentamicin, we have previously demonstrated that P. aeruginosa is able to invade corneal cells during infectious keratitis in mice. In vitro, P. aeruginosa was found to enter the following cells: human corneal cells removed by irrigation; epithelial cells in the cornea of rats, mice, and rabbits; and primary corneal epithelial cells cultured from rat and rabbit eyes. The level of invasion was related to the level of adherent or associated bacteria. In general, invasion was more efficient with cultured epithelial cells than with cells tested in situ. Invasion did not occur when assays were performed at 4 degrees C. Cytochalasin D but not colchicine inhibited bacterial invasion, suggesting that bacterial entry was an endocytic process dependent on actin microfilaments but not microtubules. Bacteria that invaded cultured corneal epithelial cells were found to multiply within cells. The ability of P. aeruginosa to invade and multiply within corneal epithelial cells may contribute to the virulence of this organism during infectious keratitis, since intracellular bacteria can evade host immune effectors and antibiotics commonly used to treat infection.


FGF19/FGFR4 signaling contributes to the resistance of hepatocellular carcinoma to sorafenib.
Sorafenib, a multi-kinase inhibitor, is used as a standard therapy for advanced hepatocellular carcinoma (HCC). However, complete remission has not been achieved and the molecular basis of HCC resistance to sorafenib remains largely unknown. Previous studies have shown that fibroblast growth factor 19 (FGF19) expression correlates with tumor progression and poor prognosis of HCC. Here, we demonstrate the novel role of FGF19 in HCC resistance to sorafenib therapy. FGF19 Knockdown cells were achieved by lentiviral-mediated interference, and FGFR4 knockout cells were achieved by CRISPR-Cas9. Protein levels of FGF19, FGFR4 and c-PARP in various HCC cell lines were measured by Western blotting analysis. Cell viability was determined by MTS assay, apoptosis was determined by DAPI nuclear staining and Western blot of c-PRAP, and ROS generation was determined by DCFH-DA staining and electrochemical biosensor. We showed that FGF19, when overexpressed, inhibited the effect of sorafenib on ROS generation and apoptosis in HCC. In contrast, loss of FGF19 or its receptor FGFR4 led to a remarkable increase in sorafenib-induced ROS generation and apoptosis. In addition, knockdown of FGF19 in sorafenib-resistant HCC cells significantly enhanced the sensitivity to sorafenib. Importantly, targeting FGF19/FGFR4 axis by ponatinib, a third-generation inhibitor of chronic myeloid leukemia, overcomes HCC resistance of sorafenib by enhancing ROS-associated apoptosis in sorafenib-treated HCC. Our results provide the first evidence that inhibition of FGF19/FGFR4 signaling significantly overcomes sorafenib resistance in HCC. Co-treatment of ponatinib and sorafinib may represent an effective therapeutic approach for eradicating HCC.


Robot-assisted therapy for balance function rehabilitation after stroke: A systematic review and meta-analysis.
To identify the rehabilitative effects of robot-assisted therapy on balance function among stroke patients. A systematic review and meta-analysis of randomized controlled trials. Thirteen electronic databases were systematically searched from inception to March 2018: Web of Science, PubMed, EMBase, The Cochrane Library, Science Direct, CINAHL, MEDLINE, AMED, Physiotherapy Evidence Database, SPORTDiscus, WanFang Data, China National Knowledge Infrastructure and Chinese Scientific Journal Database. Randomized controlled trials were retrieved for identifying the effects of robot-assisted therapy on balance function among stroke patients. Two authors independently searched databases, screened studies, extracted data, and evaluated the methodological quality and risk bias of each included study. A standardized protocol and data-collection form were used to extract information. Effect size was evaluated by mean difference with corresponding 95% confidence intervals. Methodological quality and risk bias evaluation for each included study followed the quality appraisal criteria for randomized controlled trials that were recommended by Cochrane Handbook. Meta-analysis was conducted by utilizing Review Manager 5.3, a Cochrane Collaboration tool. Data was synthesized with descriptive analysis instead of meta-analysis where comparisons were not possible to be conducted with a meta-analysis. Thirty-one randomized controlled trials with a total of 1249 participants were included. The majority of the included studies contained some methodological flaws. The results of the meta-analysis indicated that robot-assisted therapy produced positive effects on balance function, as shown by an increase in the Berg balance scale score [random effects model, mean difference = 4.64, 95%CI = 3.22-6.06, P<0.01], as well as Fugl-Meyer balance scale scores [fixed effects model, mean difference = 3.57, 95%CI = 2.81-4.34, P<0.01]. After subgroup and sensitivity analyses, the positive effects were not influenced by different types of robotic devices, by whether robot-assisted therapy was combined with another intervention or not, or by differences in duration and intensity of intervention. Evidence in the present systematic review indicates that robot-assisted therapy may produce significantly positive improvements on balance function among stroke patients compared with those not using this method. More multi-center, high-quality and large-scale randomized controlled trials following the guidelines of CONSORT are necessary to generate high-quality evidence in further research.


Molecular characterization of clinical isolates from vascular access infection: A single-institution study.
Hemodialysis requires repeated, reliable access to the systemic circulation; therefore, a well-functioning vascular access (VA) procedure is crucial for stable hemodialysis. VA infections (VAIs) constitute the most challenging complication and cause considerable morbidity, loss of access, and even death. In this study, we investigated the molecular profiles of different bacterial isolates retrieved from various types of VA grafts. We collected clinical isolates from hemodialysis patients with VAIs in our institution for the period between 2013 and 2018. We identified the bacterial isolates using standard biochemical procedures; we used a polymerase chain reaction for coagulase-negative staphylococci (CoNS) and Burkholderia cepacia complex (BCC) species identification. The antibiotic resistance and molecular profile were analyzed using the disk diffusion method and multilocus sequence typing, respectively. We studied 150 isolates retrieved from patients with VAI and observed that Staphylococcus aureus was the predominant bacterial species, followed by S. argenteus, BCC, and CoNS. According to multilocus sequence typing data, we identified a wide variety of sequence types (STs) in S. aureus isolates, with ST59, ST45, and ST239 being the predominant types. Burkholderia cepacia with two new ST types, namely ST1723 and ST1724, accounted for most of the BCC infections, along with ST102 B. contaminans, which were mainly isolated from infected tunneled-cuffed catheters. In summary, the increased incidence of S. argenteus and BCC infections provides insights into their potential clinical effects in VAIs. The various STs identified in different bacterial species indicate the high genetic diversity of bacterial species isolated from VAIs in our institution.


Outcomes of IED foot and ankle blast injuries.
Improvements in protection and medical treatments have resulted in increasing numbers of modern-warfare casualties surviving with complex lower-extremity injuries. To our knowledge, there has been no prior analysis of foot and ankle blast injuries as a result of improvised explosive devices (IEDs). The aims of this study were to report the pattern of injury and determine which factors are associated with a poor clinical outcome. U.K. service personnel who had sustained lower leg injuries following an under-vehicle explosion from January 2006 to December 2008 were identified with the use of a prospective trauma registry. Patient demographics, injury severity, the nature of the lower leg injury, and the type of clinical management were recorded. Clinical end points were determined by (1) the need for amputation and (2) ongoing clinical symptoms. Sixty-three U.K. service personnel (eighty-nine injured limbs) with lower leg injuries from an explosion were identified. Fifty-one percent of the casualties sustained multisegmental injuries to the foot and ankle. Twenty-six legs (29%) required amputation, with six of them amputated because of chronic pain eighteen months following injury. Regression analysis revealed that hindfoot injuries, open fractures, and vascular injuries were independent predictors of amputation. At the time of final follow-up, sixty-six (74%) of the injured limbs had persisting symptoms related to the injury, and only nine (14%) of the service members were fit to return to their preinjury duties. This study demonstrates that foot and ankle injuries from IEDs are associated with a high amputation rate and frequently with a poor clinical outcome. Although not life-threatening, they remain a source of long-term morbidity in an active population.


Maternal major depression and synchrony of facial affect during mother-child interactions.
Maternal history of Major Depressive Disorder (MDD) dramatically increases children's risk for developing depression, highlighting the critical need for further research on the specific processes involved in the intergenerational transmission of depression. Although previous research suggests that maternal depression may adversely affect the quality of mother-child interactions, less is known about the role of maternal MDD in the moment-to-moment changes in affect that occur during these interactions. The goal of this project, therefore, was to examine synchrony of facial displays of affect during a positive (Vacation Planning) and a negative (Issues Discussion) mother-child interaction, and how this synchrony may be impacted by maternal history of MDD. In doing so, we examined both concurrent and lagged synchrony of facial affect. We recruited 341 mother-child dyads (child average age = 9.30 years; 50.1% girls; 71.6% Caucasian) with and without a maternal history of MDD. Facial electromyography (EMG), continuously recorded during those tasks, was used to index mother and child facial affect. We found that a maternal history of MDD was associated with reduced concurrent synchrony and lagged synchrony (mother facial affect predicting changes in child facial affect) of positive affect during Vacation Planning. Reduced concurrent mother-child synchrony of positive affect during the discussion was also associated with an increase in child self-reported sad affect from before to after the discussion. These findings provide promising initial evidence for how the dynamic exchange of positive affect during mother-child interactions may be disrupted in families with maternal MDD history. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


IL12RB2 gene is associated with the age of type 1 diabetes onset in Croatian family Trios.
Common complex diseases are influenced by both genetic and environmental factors. Many genetic factors overlap between various autoimmune diseases. The aim of the present study is to determine whether four genetic variants known to be risk variants for several autoimmune diseases could be associated with an increased susceptibility to type 1 diabetes mellitus. We genotyped four genetic variants (rs2358817, rs1049550, rs6679356, rs9865818) within VTCN1, ANXA11, IL12RB2 and LPP genes respectively, in 265 T1DM family trios in Croatian population. We did not detect association of these polymorphisms with T1DM. However, quantitative transmission disequilibrium test (QTDT, orthogonal model) revealed a significant association between the age of onset of T1DM and IL12RB2 rs6679356 variant. An earlier onset of T1DM was associated with the rs6679356 minor dominant allele C (p = 0.005). The association remained significant even after the Bonferroni correction for multiple testing and permutation. Variants originally associated with juvenile idiopathic arthritis (VTCN1 gene), sarcoidosis (ANXA11 gene), primary biliary cirrhosis (IL12RB2 gene) and celiac disease (LPP gene) were not associated with type 1 diabetes in our dataset. Nevertheless, association of IL12RB2 rs6679356 polymorphism with the age of T1DM onset suggests that this gene plays a role in defining the time of disease onset.


Modulation of ErbB2 blockade in ErbB2-positive cancers: the role of ErbB2 Mutations and PHLDA1.
We set out to study the key effectors of resistance and sensitivity to ErbB2 tyrosine kinase inhibitors, such as lapatinib in ErbB2-positive breast and lung cancers. A cell-based in vitro site-directed mutagenesis lapatinib resistance model identified several mutations, including the gatekeeper ErbB2 mutation ErbB2-T798I, as mediating resistance. ErbB2-T798I engineered cell models indeed show resistance to lapatinib but remain sensitive to the irreversible EGFR/ErbB2 inhibitor, PD168393, suggestive of potential alternative treatment strategies to overcome resistance. Gene expression profiling studies identified a select group of downstream targets regulated by ErbB2 signaling and define PHLDA1 as an immediately downregulated gene upon oncogenic ErbB2 signaling inhibition. We find significant down-regulation of PHLDA1 in primary breast cancer and PHLDA1 is statistically significantly less expressed in ErbB2 negative compared with ErbB2 positive tumors consistent with its regulation by ErbB2. Lastly, PHLDA1 overexpression blocks AKT signaling, inhibits cell growth and enhances lapatinib sensitivity further supporting an important negative growth regulator function. Our findings suggest that PHLDA1 might have key inhibitory functions in ErbB2 driven lung and breast cancer cells and a better understanding of its functions might point at novel therapeutic options. In summary, our studies define novel ways of modulating sensitivity and resistance to ErbB2 inhibition in ErbB2-dependent cancers.


Comparative performance of AnTicoagulation and Risk factors In Atrial fibrillation and Global Registry of Acute Coronary Events risk scores in predicting long-term adverse events in patients with acute myocardial infarction.
This study is designed to evaluate the recently developed AnTicoagulation and Risk factors In Atrial fibrillation (ATRIA) risk score (RS), which determines the predisposition to thromboembolic and hemorrhagic events in atrial fibrillation, as a predictor of prognosis in patients having acute myocardial infarction (AMI), and to compare the predictive ability of ATRIA RS with GRACE RS. We analyzed 1627 patients having AMI who underwent coronary angiography and/or percutaneous coronary intervention (PCI) between January 2011 and February 2015. The primary endpoints included all-cause mortality, non-fatal MI, and cerebrovascular events during follow-up. Multivariate Cox regression analysis showed that the ATRIA RS>3 was an independent predictor of major adverse cardiac events in patients with AMI [hazard ratio, 2.00, 95% confidence interval, 1.54 to 2.60, p<0,001]. The area under the curve (AUC) for ATRIA RS and GRACE RS was 0.66 and 0.67 (p<0.001, and p<0.001), respectively. We performed a pair-wise comparison of receiver operating characteristic curves,and noted the predictive value of ATRIA RS with regard to primary endpoints was similar to that of GRACE RS (By DeLong method, AUCATRIA vs. AUCGRACE z test=0.64, p=0.52). ATRIA RS may be useful in predicting prognosis in patients having AMI during long-term follow-up.


Predictors of positive outcomes from 'Early Intervention in Psychosis': protocol for a national retrospective cohort study.
Psychotic disorders are severe and prevalent mental health conditions associated with long-term disability, reduced quality of life, and substantial economic costs. Early Intervention in Psychosis (EIP) services aim to provide timely and comprehensive treatment for psychotic disorders, and EIP service input is associated with improved outcomes. However, there is limited understanding of the specific components of EIP care that contribute to these improvements. There is significant nationwide variability in the commissioning and delivery of EIP, with individuals receiving different packages of components from different services. In this study, we seek to explore associations between EIP components and clinically significant outcomes, in order to understand the mechanisms underlying improved psychosis care. This national retrospective cohort study will utilize data from the 2019 National Clinical Audit of Psychosis (NCAP), examining the care received by 10,560 individuals treated by EIP services in England. Exposure data from the NCAP, capturing the components of care delivered by EIP services, will be linked with outcome data from routine NHS Digital datasets over a three-year follow-up period. This will be the first study to use this method to examine this population in England. The primary outcomes will be surrogate measures of relapse of psychosis (hospital admission and referral to community-based crisis intervention services). Secondary outcomes include duration of admissions, emergency hospital attendances, episodes of detention under the Mental Health Act, and all-cause mortality. We will use multilevel regression to examine associations between exposures and outcome events. We will handle missing data using appropriate imputation techniques. This study aims to provide valuable insights into the long-term effects of variations in EIP service delivery. The study involves a large, diverse cohort including individuals treated by every EIP service in England. While there are limitations inherent in the observational nature of the study, any associations identified will be of great relevance to clinicians, researchers, and policymakers seeking to optimize EIP care. The results will enable more targeted treatment planning, resource allocation, and potential innovations in EIP care, ultimately leading to improved prognoses for people experiencing psychosis.


Effect of neovestitol-vestitol containing Brazilian red propolis on accumulation of biofilm in vitro and development of dental caries in vivo.
The present study examined the influences of the neovestitol-vestitol (NV) containing fraction isolated from Brazilian red propolis on the development of biofilm and expression of virulence factors by Streptococcus mutans using saliva-coated surfaces of hydroxyapatite. In addition, NV was tested in a rodent model of dental caries to assess its potential effectiveness in vivo. Topical applications of NV (800 μg ml(-1)) significantly impaired the accumulation of biofilms of S. mutans by largely disrupting the synthesis of glucosyltransferase-derived exopolysaccharides and the expression of genes associated with the adaptive stress response, such as copYAZ and sloA. Of even greater impact, NV was as effective as fluoride (positive control) in reducing the development of carious lesions in vivo. NV is a promising natural anti-biofilm agent that targets essential virulence traits in S. mutans, which are associated with the formation of cariogenic biofilm and the subsequent onset of dental caries disease.


Naked eye visibility of Sirius in broad daylight.
Sirius was spotted with the naked eye at broad daylight by looking along the finder of a 1 m telescope on La Palma Observatory at a 2370 m height. Sun elevation was 73°; Sirius was nearly straight under the Sun at 37° elevation. The sky radiance, although not recorded directly, could be determined from the simultaneously obtained high-precision wavelength-dependent sky polarization data near Sirius. This was done by fitting the polarization data with the doubling-adding KNMI (DAK) radiative transfer model, which provided the values of the surface albedo and of the aerosol optical thickness required for determining the absolute sky radiance. Our analysis implies that Sirius, when positioned overhead, can be a daytime naked eye object from sea level even if its culmination occurs at solar noon. It also suggests that the second-brightest star (Canopus), if positioned overhead, could be perceptible even at solar noon.


Activity of the type I signal peptidase inhibitor MD3 against multidrug-resistant Gram-negative bacteria alone and in combination with colistin.
Effective treatment of Gram-negative bacterial infections is increasingly challenging due to the spread of multidrug-resistant strains and a lack of new antimicrobials in development. Bacterial type I signal peptidases (SPases) represent a highly conserved and essential target for inhibition by novel compounds. SPases are required for the effective processing of membrane translocated proteins involved in core functions related to metabolism, virulence and resistance. In this study we assessed the biochemical and functional activity of a novel synthetic inhibitor (MD3) of SPases against a wide range of Gram-negative pathogens. The activity and specificity of MD3 for recombinant Pseudomonas aeruginosa SPase (LepB) and a genetically engineered LepB-regulatable strain were investigated. Antimicrobial activity of the compound alone and in combination with outer membrane-permeabilizing agents (sodium hexametaphosphate, colistin) was also determined against a collection of P. aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae and Stenotrophomonas maltophilia isolates. MD3 was found to inactivate the P. aeruginosa LepB protein (IC50 10 μM), resulting in antimicrobial effects potentiated in the presence of colistin. MD3 also demonstrated potent activity against wild-type and multidrug-resistant strains of A. baumannii and S. maltophilia with MICs ranging from 0.5 to 14 mg/L in the presence of subinhibitory concentrations of colistin. MD3 is a novel inhibitor of bacterial SPase in a range of non-fermentative Gram-negative bacteria. The antimicrobial activity is potentiated in combination with colistin and suggests that SPase inhibition warrants further exploration as a basis for future mono or combination therapies.


Use of reverse genetics to enhance the oncolytic properties of Newcastle disease virus.
Naturally occurring strains of Newcastle disease virus (NDV) have shown oncolytic therapeutic efficacy in preclinical studies and are currently in clinical trials. Here, we have evaluated the possibility to enhance the cancer therapeutic potential of NDV by means of reverse genetics. Mice bearing s.c. implanted CT26 tumors were treated with intratumoral (i.t.) injections of a recombinant NDV modified to contain a highly fusogenic F protein. These treated mice exhibited significant reduction in tumor development compared with mice treated with the unmodified virus. Furthermore, mice in a CT26 metastatic tumor model treated with an i.v. injection of the genetically engineered NDV exhibited prolonged survival compared with wild-type control virus. In addition, we examined whether the oncolytic properties of NDV could be improved by expression of immunostimulatory molecules. In this regard, we engineered several NDVs to express granulocyte macrophage colony-stimulating factor, IFN-gamma, interleukin 2 (IL-2), or tumor necrosis factor alpha, and evaluated their therapeutic potential in an immunocompetent colon carcinoma tumor model. Mice bearing s.c. CT26 tumors treated with i.t. injections of recombinant NDV expressing IL-2 showed dramatic reductions in tumor growth, with a majority of the mice undergoing complete and long-lasting remission. Our data show the use of reverse genetics to develop enhanced recombinant NDV vectors as effective therapeutic agents for cancer treatment.


Assessment of Application Timing of Bacillus spp. to Suppress Pre- and Postharvest Diseases of Apple.
Four isolates of Bacillus spp. were tested in a 2-year field study for biological control of pre- and postharvest diseases of apple. For the preharvest test, bacteria were applied to 'Golden Delicious' and 'Rome Beauty' trees in May or May and June. Foliar apple scab severity was assessed weekly. After harvest, fruit were wounded and then either left untreated or given a postharvest application of the bacteria. Wounded fruit were then inoculated with the bitter rot pathogen and lesion size was measured over 8 days. Bacillus megaterium isolate A3-6, B. mycoides isolate A1-1, and B. cereus isolate FLS-5 applied in May or May and June significantly reduced fruit and foliar apple scab severity in both years. A postharvest application of the bacteria (alone or in combination with a preharvest application) resulted in the greatest suppression of bitter rot on both cultivars (P < 0.04). The May + June + postharvest application of isolate A3-6 resulted in the greatest suppression of bitter rot, with an average of 45 and 95% reduction in lesion size compared with nontreated controls on 'Golden Delicious' and 'Rome Beauty' fruit, respectively. Results from this study indicate that preharvest applications of the bacteria were able to reduce foliar and fruit scab and an additional postharvest application was effective in reducing bitter rot severity.


Production of water-soluble few-layer graphene mesosheets by dry milling with hydrophobic drug.
A novel, fast, and easy mechano-chemistry-based (dry milling) method has been developed to exfoliate graphene with hydrophobic drugs generating few-layer graphene mesosheets (< 10 nm in thickness and ∼1 μm in width). The electronic properties of the graphitic structure were partially preserved after the milling treatment compared with graphene oxide prepared by Hummers' method. Several characterization techniques such as thermogravimetric analysis, Raman spectroscopy, atomic force microscopy, electron microscopy, and molecular dynamics simulation were used to characterize this material. The drug-exfoliated mesosheets were pharmacologically inactive, offering a new approach for making water-soluble few-layer graphene mesosheets upon dry milling with hydrophobic drugs, mainly used as exfoliating agents.


N-methyl-D-aspartate Receptor Antagonists may Ameliorate Spinal Cord Injury by Inhibiting Oxidative Stress: An Experimental Study in Rats.
To investigate the neuroprotective effect of a N-methyl-D-aspartate (NMDA) receptor antagonist (amantadine) in an experimental spinal cord injury (SCI) model. Thirty male Spragueâ€"Dawley rats were divided into three groups: control (I), SCI (II), and SCI + amantadine (III). SCI was created using clip compression technique. At the end of day 7, blood samples were obtained from the rats and analyzed using various biochemical markers. Histological examination was also performed. MDA, GSH, and MPO assays were done. VEGF, TNF-α, and Baxexpressions were also analyzed. The group III had several inflammatory cells in the gray and white matter, with mildly degenerated multipolar and bipolar cells. Some bipolar and multipolar neurons showed TNF-α expression; however, TNF-α was found to be weak in small groups of inflammatory cells around the blood vessels in the substantia grisea and alba. Positive Bax expression was observed in the substantia grisea layer, particularly in the membrane of some bipolar neurons and glial cells; however, negative Bax expression was observed in neuron and glial cells and showed positive VEGF expression in the vascular endothelium in the group III. NMDA receptor antagonists, especially amantadine, may ameliorate SCI by inducing angiogenesis, affecting inflammation and apoptosis. It inhibits oxidative stress and the signaling pathways following SCI in rats.


Real-time chemical imaging of bacterial activity in biofilms using open-channel microfluidics and synchrotron FTIR spectromicroscopy.
Real-time chemical imaging of bacterial activities can facilitate a comprehensive understanding of the dynamics of biofilm structures and functions. Synchrotron-radiation-based Fourier transform infrared (SR-FTIR) spectromicroscopy can yield high spatial resolution and label-free vibrational signatures of chemical bonds in biomolecules, but the abundance of water in biofilms has hindered SR-FTIR's sensitivity in investigating bacterial activity. We developed a simple open-channel microfluidic system that can circumvent the water-absorption barrier for chemical imaging of the developmental dynamics of bacterial biofilms with a spatial resolution of several micrometers. This system maintains a 10 microm thick laminar-flow-through biofilm system that minimizes both the imaging volume in liquid and the signal interference from geometry-induced fringing. Here we demonstrate the ability of the open-channel microfluidic platform to maintain the functionality of living cells while enabling high-quality SR-FTIR measurements. We include several applications that show how microbes in biofilms adapt to their immediate environments. The ability to directly monitor and map bacterial changes in biofilms can yield significant insight into a wide range of microbial systems, especially when coupled to more sophisticated microfluidic platforms.


Immunosuppressive therapy reduces axonal damage in progressive multiple sclerosis.
In progressive multiple sclerosis (PMS), disease-modifying therapies have not been shown to reduce disability progression. The impact from immunosuppressive therapy in PMS was explored by analyzing cerebrospinal fluid (CSF) biomarkers of axonal damage (neurofilament light protein, NFL), astrogliosis (glial fibrillary acidic protein, GFAP), and B-cell regulation (CXCL13). CSF was obtained from 35 patients with PMS before and after 12-24 months of mitoxantrone (n=30) or rituximab (n=5) treatment, and from 14 age-matched healthy control subjects. The levels of NFL, GFAP, and CXCL13 were determined by immunoassays. The mean NFL level decreased by 51% (1781 ng/l, SD 2018 vs. 874 ng/l, SD 694, p=0.007), the mean CXCL13 reduction was 55% (9.71 pg/ml, SD 16.08, vs. 4.37 pg/ml, SD 1.94, p=0.008), while GFAP levels remained unaffected. Subgroup analysis showed that the NFL reduction was confined to previously untreated patients (n=20) and patients with Gd-enhancing lesions on magnetic resonance imaging (n=12) prior to study baseline. Our data imply that 12-24 months of immunosuppressive therapy reduces axonal damage in PMS, particularly in patients with ongoing disease activity. Determination of NFL levels in CSF is a potential surrogate marker for treatment efficacy and as endpoint in phase II trials of MS.


Effect of additional testosterone on purinergic responses in isolated rabbit corpus cavernosum strips.
Recently, it has been demonstrated that the endothelium of corpus cavernosum (CC) plays an important role on smooth muscle relaxation, which is crucial to initiate and maintain erection. We investigated the effect of long-lasting additional testosterone propionate (TP) therapy on endothelium-dependent and -independent relaxations of isolated rabbit CC. Isolated CC strips were mounted in organ baths and isometric tension was recorded. Addition of a specific inhibitor of nitric oxide synthesis, NG-nitro-L-arginin methyl ester (L-NAME), into the organ bath had no effect on the relaxation responses to adenosine (ADO), adenosine 5'-triphosphate (ATP) and sodium nitroprusside (SNP) in isolated CC strips precontracted with phenylepherine, but completely inhibited relaxation responses produced by ADP. Adenosine and adenine nucleotides relaxed the phenylepherine-induced contractile response in control strips with the potency order: ADO (62.8 +/- 3.2%) > ATP (37.1 +/- 5.2%) > ADP (25.8 +/- 2.5%). The relaxation responses to ADO, ATP and SNP in isolated rabbit CC strips were not significantly altered by additional TP therapy. The relaxation responses produced by ADP were significantly enhanced following 1 and 2 months TP therapy as compared with controls. However, in the group treated with TP for 2 months followed by a 2 months drug-free period, relaxation responses were significantly reduced compared to 1 and 2 months treatment groups, and approached control values. Increased relaxation responses to ADP following 1 and 2 months additional TP therapy may be a result of increased endothelial purinergic receptor density, or it may be due to stimulation and/or release of endothelial nitric oxide (NO) by TP.


The trimer to monomer transition of Tumor Necrosis Factor-Alpha is a dynamic process that is significantly altered by therapeutic antibodies.
The cytokine tumor necrosis factor-alpha (TNF-α) readily forms homotrimers at sub-nM concentrations to promote inflammation. For the treatment of inflammatory diseases with upregulated levels of TNF-α, a number of therapeutic antibodies are currently used as scavengers to reduce the active TNF-α concentration in patients. Despite their clinical success, the mode-of-action of different antibody formats with regard to a stabilization of the trimeric state is not entirely understood. Here, we use a biosensor with dynamic nanolevers to analyze the monomeric and trimeric states of TNF-α together with the binding kinetics of therapeutic biologics. The intrinsic trimer-to-monomer decay rate k = 1.7 × 10-3 s-1 could be measured directly using a microfluidic system, and antibody binding affinities were analyzed in the pM range. Trimer stabilization effects are quantified for Adalimumab, Infliximab, Etanercept, Certolizumab, Golimumab for bivalent and monovalent binding formats. Clear differences in trimer stabilization are observed, which may provide a deeper insight into the mode-of-action of TNF-α scavengers.