Leadership for medicine's promising future.
Leadership in improving the education of doctors, while impressive, is not happening fast enough. While there are many obstacles, there is no time to waste in restructuring medical education to repair its present deficiencies, for otherwise outside forces could overwhelm today's education leaders with imperatives to make improvements on their own terms. The first step in addressing present shortcomings is to establish measurable objectives for the education of doctors that are aligned with the legitimate expectations of society and the enduring precepts of the medical profession. To provide guidance in establishing these objectives, the AAMC launched the Medical School Objectives Project (MSOP) two years ago. This project is now close to completing its initial phase, which is to define the knowledge, skills, attitudes, and values every medical student must demonstrate before graduating. Phase Two will be concerned with implementation (e.g., establishing assessment methods; improving faculty development; etc.). As for aligning the outcomes of medical education with the precepts of the profession, nothing is more important: if doctors do not have high standards of professionalism--altruism, respect, compassion, honesty, integrity, and others--medicine's very survival is threatened. Medical educators must insist that their graduates demonstrate these attributes, through more careful admission criteria, more attention to medical professionalism in the curriculum and in the evaluation of students, more community service for students, and improved role modeling by faculty. Leadership for the changes that are needed will not come from a once-in-a-lifetime leader of heroic proportions but from everyone within academic medicine leading the profession to its promising future through quality education.


Magnitude and time course of insulin resistance accumulation with the risk of cardiovascular disease: an 11-years cohort study.
The risk of cardiovascular disease (CVD) depended on the magnitude and exposure duration of insulin resistance (IR). This study aimed to investigate the associations of cumulative metabolic score for IR (cumMETS-IR) with incident CVD, and to further explore the modulated effects of time course of METS-IR accumulation. We enrolled 47,270 participants without CVD and underwent three examinations during 2006-2010 from the Kailuan study. CumMETS-IR from 2006 to 2010 were calculated as the mean values of METS-IR between consecutive examinations multiplying by time intervals between visits. Time course of METS-IR accumulation was calculated as the slope of METS-IR versus time. Hazard ratios (HRs) and 95% confidence intervals (CIs) for CVD risk were calculated with multivariable-adjusted Cox regressions. During a median follow-up of 10.99 years, we identified 3184 cases of incident CVD. The risk of incident CVD increased with increasing cumMETS-IR (HR, 1.77; 95% CI 1.58-1.98 for the Q4 versus Q1 group), exposure duration (HR, 1.60; 95% CI 1.45-1.77 for 6 years versus 0 years), and cumulative burden (HR, 1.49; 95% CI 1.37-1.61 for burden ≥ 0 versus < 0). A positive slope was associated with 14% higher risk of CVD (HR, 1.14; 95% CI 1.07-1.22). When combining cumMETS-IR and slope, those with cumMETS-IR ≥ median (142.78) and slope ≥ 0 had the highest risk of CVD (HR,1.38; 95% CI 1.25-1.53). The risk of CVD increased with elevated cumMETS-IR and an increasing trend over time, emphasizing the importance of maintaining optimal METS-IR levels across life span.


Acute pancreatitis caused by Legionella pneumophila.
A 57-year-old man had Legionella pneumonia and acute pancreatitis. The diagnosis of legionellosis was established by both culture and significant rise in antibody titer to Legionella pneumophila. The pancreatitis was diagnosed by elevated amylase and lipase values, peaking on the fourth hospital day, roentgenologic findings, and a clinical picture compatible with the disease. The patient was not an alcohol consumer and had had no previous pancreatic disease. We conclude that acute pancreatitis can be a manifestation of legionellosis.


Microsystems for biomimetic stimulation of cardiac cells.
The heart is a complex integrated system that leverages mechanoelectrical signals to synchronize cardiomyocyte contraction and push blood throughout the body. The correct magnitude, timing, and distribution of these signals is critical for proper functioning of the heart; aberrant signals can lead to acute incidents, long-term pathologies, and even death. Due to the heart's limited regenerative capacity and the wide variety of pathologies, heart disease is often studied in vitro. However, it is difficult to accurately replicate the cardiac environment outside of the body. Studying the biophysiology of the heart in vitro typically consists of studying single cells in a tightly controlled static environment or whole tissues in a complex dynamic environment. Micro-electromechanical systems (MEMS) allow us to bridge these two extremes by providing increasing complexity for cell culture without having to use a whole tissue. Here, we carefully describe the electromechanical environment of the heart and discuss MEMS specifically designed to replicate these stimulation modes. Strengths, limitations and future directions of various designs are discussed for a variety of applications.


How do Lebanese patients perceive the ideal doctor based on the CanMEDS competency framework?
During their training, Lebanese medical students develop a high medical expertise but are not focusing on other competencies such as communication, collaboration, erudition, professionalism, leadership and health promotion. There is also insufficient data about patients' preference for these skills. This study describes the different weights patients attribute to these physician's competencies. This is a cross-sectional study based on a questionnaire distributed to 133 Lebanese patients. It included 15 questions assessing how patients prioritize the physician's competencies, with open-ended questions asking them to define "the good doctor". Krippendorff's alpha coefficient was used to analyze the reliability of the competencies' classification. One hundred twenty five patients completed the questionnaire in this cross-sectional study. Their mean age was 48 ± 16.76 years. When classifying competencies, 73.6% opted for medical expertise as first choice and 48% put communication as second. Based on the Krippendorff's coefficient, we identified a moderate agreement for the seven choices (alpha = 0.44). In open-ended questions, patients defined the good doctor in 325 answers: 64.3% mentioned medical expertise, 34.1% high ethics and 26.2% communication. This patient-centered study concurs well with the worldwide practice that puts medical expertise at the center of medical education. However Lebanese patients don't perceive equally other competencies and favor professionalism and communication that should be integrated in priority in students' curricula.


Resolving latent conflict: What happens when latent print examiners enter the cage?
Latent print examination traditionally follows the ACE-V process, in which latent prints are first analyzed to determine whether they are suitable for comparison, and then compared to an exemplar and evaluated for similarities and differences. Despite standard operating procedures and quality controls designed, in part, to mitigate differences between examiners, latent print processing and review are inherently subjective. The ACE-V process addresses subjectivity, and the possibility of error, in the verification stage in which a second examiner repeats the analysis, comparison, and evaluation steps in a given case. Other procedures outside the ACE-V framework, such as consultation and conflict resolution, provide further opportunity to understand how differences between latent print examiners emerge. Despite the growing body of research on latent print examination, questions have emerged about how these procedures work in practice. This study reviews case processing data for two years of casework at the Houston Forensic Science Center (HFSC). We describe these data as cases proceed through each step of the ACE-V process, with a particular focus on verification, consultation, and conflict resolution. We discuss trends in these processes regarding modal types of disagreements, modal outcomes, and roles of the examiners involved. Results reveal implications for improving the practice of latent print examination.


Safety and tolerability of denufosol tetrasodium inhalation solution, a novel P2Y2 receptor agonist: results of a phase 1/phase 2 multicenter study in mild to moderate cystic fibrosis.
Denufosol tetrasodium (INS37217) is a selective P2Y(2) agonist that stimulates ciliary beat frequency and Cl(-) secretion in normal and cystic fibrosis (CF) airway epithelia, and is being investigated as an inhaled treatment for CF. The Cl(-) secretory response is mediated via a non-CFTR pathway, and the driving force for Cl(-) secretion is enhanced by the effect of P2Y(2) activation to also inhibit epithelial Na(+) transport. Denufosol is metabolically more stable and better tolerated, and may enhance mucociliary clearance for a longer period of time than previously investigated P2Y(2) agonists. The goal of this phase 1/phase 2 study was to assess the safety and tolerability of single and repeated doses of aerosolized denufosol in subjects with CF. The study was a double-blind, placebo-controlled, multicenter comparison of ascending single doses of denufosol (10, 20, 40, and 60 mg, administered by inhalation via the Pari LC Star nebulizer) vs. placebo (normal saline), followed by a comparison of twice-daily administration of the maximum tolerated dose (MTD) of denufosol or placebo for 5 days. Thirty-seven adult (18 years of age or older) and 24 pediatric (5-17 years of age) subjects with CF were evaluated in five cohorts. Subjects were randomized in a 3:1 ratio to receive either denufosol or placebo within each cohort. The percent of subjects experiencing adverse events was similar between the denufosol and placebo groups. The most common adverse event in subjects receiving denufosol was chest tightness in adult subjects (39%) and cough in pediatric subjects (56%). Three (7%) subjects receiving denufosol and one (7%) subject receiving placebo experienced a serious adverse event. Forced expiratory volume in 1 sec (FEV(1)) profiles following dosing were similar across treatment groups, with some acute, reversible decline seen in both groups, most notably in subjects with lower lung function at baseline. In conclusion, doses up to 60 mg of denufosol inhalation solution were well-tolerated in most subjects. Some intolerability was noted among subjects with lower baseline lung function. Based on the results of this phase 1/phase 2 study, the Therapeutics Development Network (TDN) of the Cystic Fibrosis Foundation (CFF) and Inspire Pharmaceuticals, Inc., recently completed a multicenter, 28-day, phase 2 safety and efficacy clinical trial of denufosol inhalation solution in CF subjects with mild lung disease.


Phosphate kinetic modeling as an estimate of daily ingested phosphate in hemodialysis patients with or without residual kidney function.
Daugirdas suggested a 2-pool phosphate kinetic model based on his previously established urea kinetic model. The current study aims to assess the level of agreement between the modeled daily ingested phosphorus (DIP) values and the routine method of dietary recall calculations in hemodialysis patients. The study was conducted on 100 hemodialysis patients; 50 were anuric, and the others had residual kidney function (RKF). The level of correlation and agreement between the dietary calculated and modeled DIP were assessed in both study groups. A statistically significant positive correlation existed between the calculated and modeled DIP (r = 0.79 for the anuric group, r = 0.84 for the RKF group, p < 0.001). There was a significant level of agreement between calculated and modeled DIP in RKF patients only. These findings suggest that phosphate modeling can estimate phosphate intake in RKF patients and be cost-effective in their management.


Thyroid nodules and cancers in children.
The incidence of thyroid nodules in children is estimated to be 1 to 1.5% based on clinical examination. Children with thyroid nodules, compared to adults with thyroid nodules, have a fourfold greater risk of developing malignant thyroid disease. Differentiated thyroid carcinoma is the most common pediatric endocrine tumor, constituting 0.5-3% of all childhood malignancies. The thyroid is one of the most frequent sites of secondary neoplasm in children who receive radiation therapy for other malignancies. Thyroid carcinoma has been studied extensively in adults. However, the pediatric literature on this subject is much less complete, owing to the rarity of its diagnosis. This article reviews the predisposing factors, genetics, pathology, pathogenesis , clinical presentation, detailed treatment and follow-up management of children with thyroid carcinoma. Additionally, a discussion regarding the controversial aspects of radioiodine therapy in children is included.


Complete Genome Sequence of Lactobacillus oris J-1, a Potential Probiotic Isolated from the Human Oral Microbiome.
Lactobacilli can exert health-promoting effects in the human oral microbiome through many mechanisms, including pathogen inhibition, maintenance of microbial balance, immunomodulation, and enhancement of the epithelial barrier function. Here, we present the complete genome sequence of a potential probiotic, Lactobacillus oris J-1, that was isolated from the oral cavity of a health child.


Fatalistic responses to different types of genetic risk information: exploring the role of self-malleability.
Providing people with genetic risk information may induce a sense of fatalism, the belief that little can be done to reduce the risk. We postulated that fatalism is a function of health risk information and individual differences in self-perception. DNA-based risk information was hypothesised to generate more fatalism than risk information based on family history or non-genetic risk information. Moreover, people who view themselves as more rather than less able to change self-attributes were hypothesised to respond least fatalistically. Factor analyses in separate samples were used to construct a five-item 'Malleability of self' measure. Predictive validity of the measure was tested using a within-subjects analogue design. Participants responded to three scenario vignettes in which they were informed of an increased risk of cardiovascular disease (CVD). In Scenario 1, risk was ascertained by DNA testing, family history and cholesterol testing; in Scenario 2, it was ascertained by family history and cholesterol testing; in Scenario 3, risk was ascertained by cholesterol testing alone. Scenario 1 was associated with least perceived control over cholesterol level and CVD risk. People who viewed themselves as more able to change self-attributes experienced more control in all three scenarios.


The cholinergic antagonist alpha-bungarotoxin also binds and blocks a subset of GABA receptors.
The polypeptide snake toxin alpha-bungarotoxin (BTX) has been used in hundreds of studies on the structure, function, and development of the neuromuscular junction because it binds tightly and specifically to the nicotinic acetylcholine receptors (nAChRs) at this synapse. We show here that BTX also binds to and blocks a subset of GABA(A) receptors (GABA(A)Rs) that contain the GABA(A)R beta3 subunit. These results introduce a previously unrecognized tool for analysis of GABA(A)Rs but may complicate interpretation of some studies on neuronal nAChRs.


An in vitro investigation of the astringency property of certain anhidrotic solutions.
Compounds effective in vivo in the rat as anhidrotic wee found to precipitate egg white protein. There was little correlation between the effectiveness of a compound as an antiperspirant and its ability to precipitate protein. Lanthanum chloride was an exception since it was found to be the most effective anhidrotic in the rat and also produced the highest levels of precipitate. The addition of calcium or magnesium chloride increased the quantity of protein precipitated by the anhidrotic solutions, in contrast to their effect in vivo.


Troglitazone: antihyperglycemic activity and potential role in the treatment of type 2 diabetes.
Insulin resistance is a major component of type 2 diabetes; therefore, an insulin sensitizer agent like the thiazolidinedione compound troglitazone is considered a very promising drug. Troglitazone exerts an antihyperglycemic activity in a dose-dependent manner between 200 and 600 mg/day in type 2 diabetic patients treated with diet alone, sulfonylureas, or insulin. Additive antihyperglycemic effect may also be obtained by combining troglitazone and metformin. The antihyperglycemic effect of troglitazone as monotherapy is rather modest (reduction of HbA1c by 0.5-1.0%), but it appears to be somewhat greater when it is combined with other antidiabetic drugs. No double-blind studies have directly compared the activity of troglitazone with that of sulfonylureas or metformin. Troglitazone has been shown to exert additional beneficial effects on serum lipid profile and arterial blood pressure. It may be considered as a valuable alternative in insulin-resistant (obese and hyperinsulinemic) diabetic patients who appear to be the best responders to the drug. However, the efficacy of troglitazone is challenged by its safety profile, and the risk of hepatotoxicity still remains a major concern in clinical practice.


Emended description of Actinobacillus capsulatus Arseculeratne 1962, 38AL.
The taxonomic position of Actinobacillus capsulatus, a member of the family Pasteurellaceae found in rabbits, hares and hamsters, has been challenged. 16S rRNA gene (rrs) sequence data show the species to be heterogeneous. Using a polyphasic approach, 23 strains that were identified previously as belonging, or closely related, to A. capsulatus were analysed. Eighty characters were included in the phenotypic analysis. Phylogenetic analysis was done based on rrs, rpoB, infB and recN sequences. In addition, the recN sequence similarities were used to calculate the whole-genome sequence relatedness of all strains investigated as well as that with other members of the family Pasteurellaceae. The phenotypic analysis allowed identification of five groups. The major group of 17 strains could be classified as A. capsulatus. Two hamster isolates were closely related to A. capsulatus but differed in a few characters. Single isolates from a rabbit and snowshoe-hare were phenotypically related to Actinobacillus suis. One rabbit isolate was related to the genus Mannheimia, while another isolate could not be classified phenotypically with known taxa. The phylogenetic analysis confirmed the phenotypic grouping. In contrast to the rrs-based tree, the A. capsulatus strains clustered unambiguously with the type species and related species of the genus Actinobacillus in the rpoB-, infB- and recN-based trees. Genome similarity comparison using recN finally confirmed the high genomic relationship of the A. capsulatus strains with the type species and related species of the genus Actinobacillus and allowed a clear assignment of the other unrelated strains to the phenotypic and phylogenetic clusters outlined. The present findings allow the description of A. capsulatus to be emended and separate it more clearly from other species, both phenotypically and genotypically. The type strain of A. capsulatus is CCUG 12396(T) (=Frederiksen 243(T)=ATCC 51571(T)=NCTC 11408(T)=CIP 103283(T)).


Development of a Stable Respiratory Syncytial Virus Pre-Fusion Protein Powder Suitable for a Core-Shell Implant with a Delayed Release in Mice: A Proof of Concept Study.
Currently, there is an increasing interest to apply pre-fusion (pre-F) protein of respiratory syncytial virus (RSV) as antigen for the development of a subunit vaccine. A pre-F-containing powder would increase the flexibility regarding the route of administration. For instance, a pre-F-containing powder could be incorporated into a single-injection system releasing a primer, and after a lag time, a booster. The most challenging aspect, obtaining the booster after a lag time, may be achieved by incorporating the powder into a core encapsulated by a nonporous poly(dl-lactic-co-glycolic acid) (PLGA) shell. We intended to develop a stable freeze-dried pre-F-containing powder. Furthermore, we investigated whether incorporation of this powder into the core-shell implant was feasible and whether this system would induce a delayed RSV virus-neutralizing antibody (VNA) response in mice. The developed pre-F-containing powder, consisting of pre-F in a matrix of inulin, HEPES, sodium chloride, and Tween 80, was stable during freeze-drying and storage for at least 28 days at 60 °C. Incorporation of this powder into the core-shell implant was feasible and the core-shell production process did not affect the stability of pre-F. An in vitro release study showed that pre-F was incompletely released from the core-shell implant after a lag time of 4 weeks. The incomplete release may be the result of pre-F instability within the core-shell implant during the lag time and requires further research. Mice subcutaneously immunized with a pre-F-containing core-shell implant showed a delayed RSV VNA response that corresponded with pre-F release from the core-shell implant after a lag time of approximately 4 weeks. Moreover, pre-F-containing core-shell implants were able to boost RSV VNA titers of primed mice after a lag time of 4 weeks. These findings could contribute to the development of a single-injection pre-F-based vaccine containing a primer and a booster.


An ad libitum, very low-fat diet results in weight loss and changes in nutrient intakes in postmenopausal women.
To determine whether a very low-fat diet (<15% of energy intake) consumed ad libitum during an 8-month period can achieve weight loss of 5% to 10% of initial body weight while still providing adequate intakes of other essential nutrients. Longitudinal, 8-month, ad libitum, free living, very low-fat diet trial. Fifty-four of the sixty-four healthy postmenopausal women recruited completed the entire study (age 59+/-8 years, BMI=29.6+/-6.3). Twenty-four of these women used hormone replacement therapy, thirty women did not. Weekly sessions aimed at teaching and reinforcing a very low-fat intake diet for eight months. MAIN: outcome measures Body weight, percent body fat, waist-to-hip ratio, resting energy expenditure, respiratory quotient, and nutrient intakes derived from 7-day food records at the beginning and at 2, 4, 6, and 8 months of the study. Statistical analysis performed Repeated measures analysis of variance and Tukey post hoc analysis were used to analyze significant differences in mean data (P<.05). Fat intake decreased from 33.2+/-7.5% to 11+/-4% over the 8-month intervention period (P<.00001). Weight loss was 6.0 kg+/- 4.2 kg (P<.000038), an 8% weight change, and decrease in percent body fat of 2.7%+/-0.2% (P< or =.000046). Weight correlated better with the self-reported fat intake (r=0.321, P<.01) than the energy intake (r=0.263, P<.05) at baseline. Fiber intake increased from 16 g+/-0.6 g to 23 g+/-0.2 g (P<.0005). All micronutrient intakes remained at or above preintervention ranges, except for a decrease in vitamin E intake from 8.1 mg+/-4.0 mg to 3.7 mg+/-1.1 mg (P<.0005) on the very low-fat diet and linoleic acid from 6.3%+/-1.5% to 2.5%+/-0.7% (P<.000001) with no significant reduction in linolenic acid. Hormone replacement was not associated with the amount of weight loss. This study demonstrates that adherence to a very low-fat diet consumed ad libitum causes weight loss in the 5% to 10% range and a reduction of body fat. These reductions, along with the observed decreases in fat intake, are associated with improved health outcomes. Because of the decreased vitamin E and n-3 fatty acid intake, emphasis on foods high in these nutrients may need to be encouraged for those consuming a very low-fat diet.


[Combined effect of metronidazole, interferon synthesis inducers and DNA repair inhibitors].
In experiments on mice with solid sarcoma 37, a study was made of modification of the radiosensitizing effect of metronidazole. The increase in the leucotic interferon titre, induced by the interferon synthesis inductors, did not influence the radiosensitizing efficiency of metronidazole: after the administration of the repair inhibitor, 8-bromocaffeine, the coefficient of suppression of tumor growth was higher and the number of mice with the regressed tumors greater as compared to animals affected by metronidazole and radiation.


Different Structures-Similar Effect: Do Substituted 5-(4-Methoxyphenyl)-1H-indoles and 5-(4-Methoxyphenyl)-1H-imidazoles Represent a Common Pharmacophore for Substrate Selective Inhibition of Linoleate Oxygenase Activity of ALOX15?
Mammalian 15-lipoxygenases (ALOX15) are lipid peroxidizing enzymes that exhibit variable functionality in different cancer and inflammation models. The pathophysiological role of linoleic acid- and arachidonic acid-derived ALOX15 metabolites rendered this enzyme a target for pharmacological research. Several indole and imidazole derivatives inhibit the catalytic activity of rabbit ALOX15 in a substrate-specific manner, but the molecular basis for this allosteric inhibition remains unclear. Here, we attempt to define a common pharmacophore, which is critical for this allosteric inhibition. We found that substituted imidazoles induce weaker inhibitory effects when compared with the indole derivatives. In silico docking studies and molecular dynamics simulations using a dimeric allosteric enzyme model, in which the inhibitor occupies the substrate-binding pocket of one monomer, whereas the substrate fatty acid is bound at the catalytic center of another monomer within the ALOX15 dimer, indicated that chemical modification of the core pharmacophore alters the enzyme-inhibitor interactions, inducing a reduced inhibitory potency. In our dimeric ALOX15 model, the structural differences induced by inhibitor binding are translated to the hydrophobic dimerization cluster and affect the structures of enzyme-substrate complexes. These data are of particular importance since substrate-specific inhibition may contribute to elucidation of the putative roles of ALOX15 metabolites derived from different polyunsaturated fatty acids in mammalian pathophysiology.


Some consequences of capillary permeability to macromolecules: Starling's hypothesis reconsidered.
Starling's hypothesis ascribes fluid movements across capillary walls to the interaction of hydrostatic and colloid osmotic forces. For 90 years it has been recognized as the basis of plasma-to-interstitial fluid balance. Its original statement was based on the notion of capillary impermeability to plasma proteins. However, as knowledge of transcapillary exchange of plasma proteins developed, its formulation was progressively modified to allow for protein transport and for interaction of protein transport with volume flow. The most important aspects of the conceptual evolution of Starling's hypothesis are reviewed in the text of this lecture.


Spontaneous apoptosis in lymphocytes from patients with Wiskott-Aldrich syndrome: correlation of accelerated cell death and attenuated bcl-2 expression.
Wiskott-Aldrich syndrome (WAS) is an X-linked recessive disorder characterized by thrombocytopenia, eczema, and a progressive deterioration of immune function. WAS is caused by mutations in an intracellular protein, WASP, that is involved in signal transduction and regulation of actin cytoskeleton rearrangement. Because immune dysfunction in WAS may be due to an accelerated destruction of lymphocytes, we examined the susceptibility to apoptosis of resting primary lymphocytes isolated from WAS patients in the absence of exogenous apoptogenic stimulation. We found that unstimulated WAS lymphocytes underwent spontaneous apoptosis at a greater frequency than unstimulated normal lymphocytes. Coincident with increased apoptotic susceptibility, WAS lymphocytes had markedly attenuated Bcl-2 expression, whereas Bax expression did not differ. A negative correlation between the frequency of spontaneous apoptosis and the level of Bcl-2 expression was demonstrated. These data indicate that accelerated lymphocyte destruction by spontaneous induction of apoptosis may be one pathogenic mechanism by which the progressive immunodeficiency in WAS patients develops.


Assessment of lingual frenulum lengths in skeletal malocclusion.
The orofacial musculature plays a pivotal role in maintaining a balance in positioning of the teeth and any imbalance which occurs in this , results in malocclusion. Lingual frenum is a soft tissue structure which tethers the ventral surface of the tongue to the floor of the mouth. This study was performed to analyze the lingual frenal lengths in skeletal class I, class II and Class III malocclusion and to correlate relationship between both. This study comprised of 30 subjects, with 10 in each group and an impression was made with the maximum mouth opening position and the tip of tongue touching the incisive papilla. The length of the lingual frenum was then measured from the casts. The maximum mouth opening position was also determined by measuring the interincisal distance with the maximum mouth opening position. Statistical analysis was done to analyze the relationship between both. The lingual frenum was found to be longest in class III malocclusion, with a statistical significant value of p<0.01. The class II and class I malocclusion did not show much difference. The maximum mouth opening position was also increased in class III malocclusion, followed by class II and class I malocclusion, in a descending order. The lingual frenum exerts erratic forces and a long lingual frenum pushes the mandibular anteriors forwards, resulting in malocclusion. Hence, a relationship between the lingual frenum and malocclusion is essential, so that the erratic forces can be eliminated and excellent results can be achieved, following the correction of malocclusion.


Overexpression of Arabidopsis AGD7 causes relocation of Golgi-localized proteins to the endoplasmic reticulum and inhibits protein trafficking in plant cells.
ADP ribosylation factor (Arf) GTPase-activating proteins (GAPs) promote the hydrolysis of GTP bound to Arfs to GDP, which plays a pivotal role in regulating Arfs by converting the active GTP-bound forms of these proteins into their inactive GDP-bound forms. Here, we investigated the biological role of AGD7, an Arf GAP homolog, in Arabidopsis (Arabidopsis thaliana). We show that AGD7 bears a highly conserved N-terminal region and a unique C-terminal region, interacts with Arf1 both in vitro and in vivo, and stimulates Arf1 GTPase activity in a phosphatidic acid-dependent manner in vitro. In plant cells, AGD7 localized to the Golgi complex, where its overexpression was found to inhibit the Golgi localization of gamma-subunit of coat proteins and promote the relocation of Golgi proteins into the endoplasmic reticulum in both protoplasts and transgenic plants. Furthermore, overexpression of AGD7 inhibited anterograde trafficking of proteins from the endoplasmic reticulum. We propose that AGD7 functions as a GAP for Arf1 in the Golgi complex and plays a critical role in protein trafficking by controlling Arf1 activity.


Daily energy expenditure is related to plasma leptin concentrations in older African-American women but not men.
Recent research suggests that leptin may control body weight by regulating energy expenditure and energy intake in mice. To explore the possible role of leptin in the regulation of energy expenditure in humans, we used doubly-labeled water methodology to determine whether fasting plasma leptin concentrations were related to total daily energy expenditure (TEE) and its components, resting energy expenditure (REE) and physical activity energy expenditure (PAEE), in free-living older African-American men (n = 21) and women (n = 25). Plasma leptin concentrations were higher in women than men, even after the adjustment for differences in fat mass (28 +/- 3 ng/ml for women vs. 17 +/- 3 ng/ml for men; P < 0.01). The logarithm of plasma leptin concentrations correlated with fat mass in both women (r = 0.80) and men (r = 0.78) (P < 0.0001). After statistical adjustment for sex differences in fat-free mass and fat mass, women had lower TEE (22%) and REE (15%) (P < 0.01) and a trend (P = 0.08) toward lower PAEE, compared with men. After controlling for the effects of fat-free mass on energy expenditure, plasma leptin concentrations were related to REE (r = 0.68, P < 0.001) and tended to be related to TEE (r = 0.37, P = 0.07) in African-American women but not men (r = 0.18 and -0.03, respectively). Plasma leptin concentrations were not related to PAEE in either men or women. These results suggest that leptin may contribute to the regulation of TEE in older African-American women through its effects on resting energy metabolism, but the role of leptin in the regulation of energy expenditure is less apparent in older African-American men.


Protists in Arctic drift and land-fast sea ice.
Global climate change is having profound impacts on polar ice with changes in the duration and extent of both land-fast ice and drift ice, which is part of the polar ice pack. Sea ice is a distinct habitat and the morphologically identifiable sympagic community living within sea ice can be readily distinguished from pelagic species. Sympagic metazoa and diatoms have been studied extensively since they can be identified using microscopy techniques. However, non-diatom eukaryotic cells living in ice have received much less attention despite taxa such as the dinoflagellate Polarella and the cercozoan Cryothecomonas being isolated from sea ice. Other small flagellates have also been reported, suggesting complex microbial food webs. Since smaller flagellates are fragile, often poorly preserved, and are difficult for non-experts to identify, we applied high throughput tag sequencing of the V4 region of the 18S rRNA gene to investigate the eukaryotic microbiome within the ice. The sea ice communities were diverse (190 taxa) and included many heterotrophic and mixotrophic species. Dinoflagellates (43 taxa), diatoms (29 taxa) and cercozoans (12 taxa) accounted for ~80% of the sequences. The sympagic communities living within drift ice and land-fast ice harbored taxonomically distinct communities and we highlight specific taxa of dinoflagellates and diatoms that may be indicators of land-fast and drift ice.


Magnetic resonance imaging findings in 84 patients with early rheumatoid arthritis: bone marrow oedema predicts erosive progression.
To examine the spectrum and severity of magnetic resonance imaging (MRI) findings in patients with early rheumatoid arthritis (RA), and to investigate the predictive value of MRI findings for subsequent development of conventional radiographic (CR) damage and MRI erosions. 84 consecutive patients with RA with disease duration <1 year were enrolled. Patients were treated according to standard clinical practice, and evaluated at baseline, 3, 6 and 12 months by core measures of disease activity, conventional radiographs of both hands and wrists and MRI of the dominant wrist. MR images were scored according to the OMERACT rheumatoid arthritis magnetic resonance imaging score (RAMRIS), and conventional radiographs according to the van der Heijde modified Sharp score. MRI findings reflecting inflammation (synovitis, bone marrow oedema and tenosynovitis) decreased during follow-up, while there was a small increase in MRI erosion score and CR damage. The proportion of patients with erosive progression at 1 year was 48% for conventional radiography and 66% for MRI. Baseline MRI bone marrow oedema (score >2 RAMRIS units) was identified as an independent predictor of both CR (odds ratio = 2.77 (95% confidence interval (CI) 1.06 to 7.21)) and MRI erosive progression (B = 0.21 (95% CI 0.08 to 0.34)). MRI findings were common in early RA, and MRI bone marrow oedema was an independent predictor of radiographic damage. These results suggest that MRI scans of the dominant wrist may help clinicians to determine which patients need early and aggressive treatment to avoid subsequent joint damage.


Metabolic assessment of a novel chronic myelogenous leukemic cell line and an imatinib resistant subline by H NMR spectroscopy.
The goal of this study was to examine metabolic differences between a novel chronic myelogenous leukemic (CML) cell line, MyL, and a sub-clone, MyL-R, which displays enhanced resistance to the targeted Bcr-Abl tyrosine kinase inhibitor imatinib. (1)H nuclear magnetic resonance (NMR) spectroscopy was carried out on cell extracts and conditioned media from each cell type. Both principal component analysis (PCA) and specific metabolite identification and quantification were used to examine metabolic differences between the cell types. MyL cells showed enhanced glucose removal from the media compared to MyL-R cells with significant differences in production rates of the glycolytic end-products, lactate and alanine. Interestingly, the total intracellular creatine pool (creatine + phosphocreatine) was significantly elevated in MyL-R compared to MyL cells. We further demonstrated that the MyL-R cells converted the creatine to phosphocreatine using non-invasive monitoring of perfused alginate-encapsulated MyL-R and MyL cells by in vivo (31)P NMR spectroscopy and subsequent HPLC analysis of extracts. Our data demonstrated a clear difference in the metabolite profiles of drug-resistant and sensitive cells, with the biggest difference being an elevation of creatine metabolites in the imatinib-resistant MyL-R cells.


Bone status in nonambulant, epileptic, institutionalized youth. Improvement with vitamin D therapy.
This study was conducted to determine if the adverse effects of anticonvulsant drug therapy and nonambulancy on bone status could be overcome with vitamin D therapy in severely handicapped individuals. Six male and five female gastrostomy fed, nonambulant, epileptic, profoundly mentally retarded individuals ranging in age from 7 to 17 years were given vitamin D therapy at a dosage of 4,000 IU/m2 body surface area/day for 6 months. Photon absorptiometry and biochemical indices of bone status were measured to follow the effects of therapy. Bone mineral content expressed as a percentage of normal improved by 11 percent (p less than 0.01), from 59.6 to 66.1 percent. Tartrate-resistant acid phosphatase, total alkaline phosphatase, and the bone isoenzyme activities declined 11 percent, 18 percent, and 11 percent respectively. These reductions were not statistically significant but they were consistent with the improvements observed by photon absorptiometry. The results of our study suggest that a conservative supplement of vitamin D will improve the bone status of severely disabled youths.


The hospital as an enterprise: management strategies.
After introducing the context in which health systems exist in developing as well as in industrialized countries, the author describes the role that hospitals are expected to play as 'enterprises'. The rationale behind such a role is explored and the experience so far available discussed. Evidence from several developing countries suggests caution and trends to discourage radical and indiscriminate introduction of market-oriented reforms, which international donors seem to be spearheading in Africa's health systems. It is likely however, that a limited, gradual introduction of selected elements of these reforms (such as some aspects of contracting out, managerial decentralization, the split between purchasing and providing functions) may find its place and eventually have a positive impact on the performance of the health systems. But 'mystique of markets' must not be allowed to deprive public hospitals of the sense of social utility and of the caring ethos which are essential requisites of a meaningful and effective public health system.


[Changes in the serum marker of cardiac fibrosis and inflammation associated with atrial fibrillation].
This work was designed to study characteristics of cardiac fibrosis and inflammation associated with idiopathic atrial fibrillation (AF) and AF with concomitant arterial hypertension (AH). The same parameters were investigated in connection with different types of arrhythmia. It is shown that patients with AF have increased levels of serum markers of fibrosis and inflammation (C-terminalpropeptide of type I procollagen (PICP) and IL-6); they are especially high in case of combination ofAF and AH. Characteristics of fibrosis are related to the form of AF. PICP levels increase in patients with chronic and recurrent AF. Activity of matrix metalloproteinase-9 (MMP-9) does not depend on the type of AF whereas concentration of tissue metalloproteinase inhibitor-1 (TMPI-1) tends to decrease in case of permanent AF. Patients with recurrent and especially permanent AF have higher IL-6 levels than practically healthy subjects. Permanent AF is associated with a slightly elevated level of tumour necrosis factor-alpha.


Dietary Azomite, a natural trace mineral complex, improved the growth, immunity response, intestine health and resistance against bacterial infection in largemouth bass (Micropterus salmoides).
Azomite is a hydrated calcium sodium aluminosilicat rich in rare earth elements. To investigate the dietary effects of Azomite on growth, intestine microbiota and morphology, immunohematological changes and disease resistance, seven diets with Azomite supplementation of 0 (the control), 1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 g/kg (A0, A1, A2, A3, A4, A5, A6), were prepared and fed to largemouth bass, Micropterus salmoides (7.96 ± 0.19) for 60 days. The results revealed that the weight gain (WG) increased first and then decreased with the increasing dietary Azomite, and the A2 group presented the highest WG and lowest feed conversion ratio among all the groups. The supplementation of 2.0 g/kg Azomite significantly increased the intestine protease activity, the crude protein of whole body and protein retention (P < 0.05), and high inclusion of Azomite (6.0 g/kg) significantly reduced the lipid retention (P < 0.05). The amounts of red blood cells in A5, A6 groups, white blood cells in A3, A5, A6 groups and lymphocyte in A2-A6 groups were all significantly higher than those in the control group (P < 0.05). In addition, serum superoxide dismutase and catalase activities in A5, A6 groups, and serum alkaline phosphatase and lysozyme activities in A2-A4 groups showed significantly higher values than the control group (P < 0.05). Intestinal microbiota analysis indicated that the Tenericutes abundance was increased, whereas Proteobacteria abundance was decreased in all Azomite supplemented groups. The villus height in A2-A4 groups, and the villus width in A2 group were significantly higher than those of the control group (P < 0.05). The cumulative mortality was reduced by the addition of 2.0-5.0 g/kg Azomite after challenging with A. hydrophila (P < 0.05). In conclusion, proper addition of Azomite in diets improved the growth, intestine morphology, immune response and disease resistance in largemouth bass, and the optimal inclusion was estimated to be 2.0-3.0 g/kg diet.


Absence of the lectin activation pathway of complement does not increase susceptibility to Pseudomonas aeruginosa infections.
Pseudomonas aeruginosa remains one of the major clinical pathogens that burden immuno-compromised patients and patients with cystic fibrosis. The present study aimed to define the role of the lectin pathway of complement in the immune-defence against P. aeruginosa in a mouse model of invasive pneumonia. Using in vitro assays specific for each of the three complement pathways, we demonstrate that some strains of P. aeruginosa bind lectin pathway recognition sub-components and initiate complement activation in a lectin pathway-specific mode. All of the tested strains activated complement via classical and alternative pathways. We assessed the importance of lectin pathway activation in fighting P. aeruginosa infections by testing a lectin pathway activating strain in a mouse model of intra-nasal infection. MASP-2 (mannan binding lectin associated serine protease-2) deficient mice, which have no lectin pathway activity, had no significant survival disadvantage compared to wild type littermates (72.7% and 81.8% survival, respectively, p=0.48). Likewise, no difference in opsonising activity was seen between MASP-2 sufficient and MASP-2 deficient mouse sera. Moreover, cytokine expression profiles in the lungs of WT mice and MASP-2-/- mice were similar throughout the course of P. aeruginosa infection. We conclude that the lectin pathway does not play an essential role in fighting P. aeruginosa infection in mice.


Why the fair innings argument is not persuasive.
The fair innings argument (FIA) is frequently put forward as a justification for denying elderly patients treatment when they are in competition with younger patients and resources are scarce. In this paper I will examine some arguments that are used to support the FIA. My conclusion will be that they do not stand up to scrutiny and therefore, the FIA should not be used to justify the denial of treatment to elderly patients, or to support rationing of health care by age. There are six issues arising out of the FIA which are to be addressed. First, the implication that there is such a thing as a fair share of life. Second, whether it makes sense to talk of a fair share of resources in the context of health care and the FIA. Third, that 'fairness' is usually only mentioned with regard to the length of a person's life, and not to any other aspect of it. Fourth, if it is sensible to discuss the merits of the FIA without taking account of the 'all other things being equal' argument. Fifth, the difference between what is unfair and what is unfortunate. Finally, that it is tragic if a young person dies, but only unfortunate if an elderly person does.


Aging and the representation of spatial situations in narrative understanding.
Age differences in the construction of the situation model during text understanding were investigated. Situation model processing was measured in terms of the distance effect, the tendency for readers to process information about objects in a narrative more quickly when the objects are spatially closer to the protagonist than when they are farther away. To examine readers' ability to construct the model directly from the text, the distance effect was contrasted for objects that were either presented in a layout of the narrative setting prior to reading (learned objects) or introduced in the narrative itself (new objects). Both younger and older readers showed strong distance effects for learned objects. When objects had not been learned but were only introduced in the text, however, younger adults did not show a reliable distance effect. Older adults with high levels of comprehension did, lending support to the position that older readers differentially rely on the situation model for effective narrative understanding.


Immunity to syphilis: passive transfer in rabbits using serial doses of immune serum.
The influence of prolonged treatment with serial doses of immune serum initiated either 2 or 48 h before intracutaneous challenge with Treponema pallidum on the subsequent appearance and development of lesions was studied. Whereas treatment with large doses of immune serum initiated 48 h before treponemal challenge caused a delay of 9 to 20 days in the onset of lesions and marked suppression of lesion development. Treatment initiated 2 h before treponemal challenge failed to delay the onset of lesions and only moderately suppressed their development. Lesion development was suppressed as long as treatment was continued. The significance of the results is discussed.


The ACT domain in chloroplast precursor-phosphorylating STY kinases binds metabolites and allosterically regulates kinase activity.
Protein import of nucleus-encoded proteins into plant chloroplasts is a highly regulated process, requiring fine-tuning mechanisms especially during chloroplast differentiation. One way of altering import efficiency is phosphorylation of chloroplast transit peptides in the cytosol. We recently investigated the role of three serine/threonine/tyrosine (STY) kinases, STY8, STY17, and STY46, in precursor phosphorylation. These three kinases have a high degree of similarity and harbor a conserved aspartate kinase-chorismate mutase-tyrA (prephenate dehydrogenase) (ACT) domain upstream of the kinase domain. The ACT domain is a widely distributed structural motif known to be important for allosteric regulation of many enzymes. In this work, using biochemical and biophysical techniques in vitro and in planta, including kinase assays, microscale thermophoresis, size exclusion chromatography, as well as site-directed mutagenesis approaches, we show that the ACT domain regulates autophosphorylation and substrate phosphorylation of the STY kinases. We found that isoleucine and S-adenosylmethionine bind to the ACT domain, negatively influencing its autophosphorylation ability. Moreover, we investigated the role of the ACT domain in planta and confirmed its involvement in chloroplast differentiation in vivo Our results provide detailed insights into the regulation of enzyme activity by ACT domains and establish that it has a role in binding amino acid ligands during chloroplast biogenesis.


Developmental patterns of glucagon-like peptide-1-(7-36) amide and peptide-YY in rat pancreas and gut.
Glucagon-like peptide-(17-36) amide [GLP-1-(7-36) amide] and peptide tyrosine tyrosine (PYY) are both products of the enteroglucagon cell. To examine the changes occurring during development, we analyzed by RIA the pancreatic concentrations of these two peptides in fetal and neonatal rats. The levels obtained were compared with those of the classical islet hormones, insulin, somatostatin, and glucagon. The total hormone content of the pancreas increased with age for insulin, glucagon, and somatostatin. The amounts of GLP-1-(7-36) amide immunoreactivity (IR) and PYY, however, remained approximately constant in the 3-, 8-, and 30-day-old and adult pancreas. GLP-1-(7-36) amide IR showed only a single peak by gel chromatography, but further analysis by anion exchange chromatography showed that during the fetal period (-18 days) and 3 days postpartum GLP-1-(7-36) amide was the main product, whereas 30 days postpartum and in adult pancreas mainly GLP-1 and an intermediate form were found. Similar analyses of gut extracts revealed that only GLP-1-(7-36) amide is produced during intestinal development. The gut content of GLP-1-(7-36) amide IR and PYY IR increased approximately 100 times between the fetus and the 30-day-old rat. This study reveals a complex and specific regulation of posttranslational processing during maturation for these two peptides.


Activation of electrogenic Na+/K+ exchange by extracellular K+ in canine cardiac Purkinje fibers.
Transient increments in sodium pump current were elicited in small voltage-clamped Purkinje fibers suspended in a fast flow system by briefly exposing them to K+-free fluid, to temporarily inhibit the pump, and then suddenly returning them to K+-containing fluid. The exponential time course of decay of the current increment provides a measure of the pump rate constant for Na+ extrusion. The dependence of that rate constant, and of the peak amplitude of the increment in pump current, on the extracellular K+ concentration was determined. The results indicate: that in cardiac Purkinje cells, as in many other cells, the pump is half-maximally activated by about 1 mM K+; that the coupling ratio for Na+/K+ exchange is independent of either intracellular Na+ concentration or external K+ concentration; and that a simple model in which intracellular Na+ concentration is determined by a passive "leak," and an active extrusion of Na+, seems sufficient to account for moderate changes in cellular Na+ concentration.


Development and application of a method for identification of isothiocyanate-targeted molecules in colon cancer cells.
In this study, we have developed a novel method to identify isothiocyanate (ITC)-targeted molecules using two well-studied ITCs: benzyl ITC (BITC) and phenethyl ITC (PEITC). The principle of this method is based on identifying a pattern of differences between BITC and PEITC given that they show similar chemical and biological behaviors. For method validation, dithiothreitol-reduced bovine insulin as a model molecule was incubated with either BITC or PEITC, and digested peptides were analyzed by ultra-performance liquid chromatography time-of-flight mass spectrometry (UPLC-TOF-MS) and liquid chromatography quadrupole TOF-MS (LC-Q-TOF-MS). Three peptides-NYCN, FVNQHLCGSHLVE, and ALYLVCGE-were identified as being adducted with BITC or PEITC on their cysteine residues. Each set of peptides adducted with either BITC or PEITC showed retention times (RT(BITC)<RT(PEITC)) by reverse-phase column chromatography with a difference of molecular mass (Δ14.01565). On the basis of these findings, computational mathematical schemes were constructed to extract sets of MS ions satisfying the above criteria. Application of the developed method to an extract of ITC-treated human colon cancer HCT116 cells, thiocarbamoylation of cysteine residues of glutathione, and the N-terminal proline residues of PMFIVNTNVPR from macrophage migration inhibitory factor were successfully identified as one of the intracellular targets of ITCs. Moreover, the method also detected the thiocarbamoylated conjugates of ITCs with intracellular free cysteines and lysines.


Comparison of sterol import under aerobic and anaerobic conditions in three fungal species, Candida albicans, Candida glabrata, and Saccharomyces cerevisiae.
Sterol import has been characterized under various conditions in three distinct fungal species, the model organism Saccharomyces cerevisiae and two human fungal pathogens Candida glabrata and Candida albicans, employing cholesterol, the sterol of higher eukaryotes, as well as its fungal equivalent, ergosterol. Import was confirmed by the detection of esterified cholesterol within the cells. Comparing the three fungal species, we observe sterol import under three different conditions. First, as previously well characterized, we observe sterol import under low oxygen levels in S. cerevisiae and C. glabrata, which is dependent on the transcription factor Upc2 and/or its orthologs or paralogs. Second, we observe sterol import under aerobic conditions exclusively in the two pathogenic fungi C. glabrata and C. albicans. Uptake emerges during post-exponential-growth phases, is independent of the characterized Upc2-pathway and is slower compared to the anaerobic uptake in S. cerevisiae and C. glabrata. Third, we observe under normoxic conditions in C. glabrata that Upc2-dependent sterol import can be induced in the presence of fetal bovine serum together with fluconazole. In summary, C. glabrata imports sterols both in aerobic and anaerobic conditions, and the limited aerobic uptake can be further stimulated by the presence of serum together with fluconazole. S. cerevisiae imports sterols only in anaerobic conditions, demonstrating aerobic sterol exclusion. Finally, C. albicans imports sterols exclusively aerobically in post-exponential-growth phases, independent of Upc2. For the first time, we provide direct evidence of sterol import into the human fungal pathogen C. albicans, which until now was believed to be incapable of active sterol import.


Acute respiratory failure in severe blunt chest trauma.
From 1973 through 1980 144 patients with chest trauma were treated. Concomitant acute respiratory failure was considered severe in 125 (83%). Morbidity and mortality were found to be related to the presence of shock, head injury Glasgow score 3-4, and size of the flail segment, but not by the extent of the thoracic or intrathoracic injuries. Mortality for the whole group and for the flail chest group were 8.3 and 9.5%, respectively. Treatment was analyzed in two historical periods: In the first, 1973 through 1976, controlled mandatory ventilation and tracheostomy were used in 83 and 70% of the cases, respectively. In the second period, 1977 through 1980, intermittent mandatory ventilation plus soft-cuff endotracheal tube were used in 77% of the cases. Ventilator time did not vary in these two periods but the lung oxygen transport was better in the group treated with intermittent mandatory ventilation.


Lipolytic Activity and the Utilization of Fatty Acids Associated with Bat Sebum by Pseudogymnoascus destructans.
Pseudogymnoascus destructans is the causative agent of a fungal infection of bats known as white-nose syndrome (WNS). Since its discovery in 2006, it has been responsible for precipitous declines of several species of cave-dwelling North American bats. While numerous advancements in the understanding of the disease processes underlying WNS have been made in recent years, there are still many aspects of WNS, particularly with respect to pathogen virulence, that remain unknown. In this preliminary investigation, we sought to further elucidate the disease cycle by concentrating on the pathogen, with specific focus on its ability to utilize lipids that compose bat wing sebum and are found in wing membranes, as a substrate for energy and growth. In vitro growth experiments were conducted with the three most common fatty acids that comprise bat sebum: oleic, palmitic, and stearic acids. None of the fatty acids were observed to contribute a significant difference in mean growth from controls grown on SDA, although morphological differences were observed in several instances. Additionally, as an accompaniment to the growth experiments, bat wing explants from Perimyotis subflavus and Eptesicus fuscus were fluorescently stained to visualize the difference in distribution of 16- and 18-carbon chain fatty acids in the wing membrane. Which substrates contribute to the growth of P. destructans is important to understanding the progressive impact P. destructans has on bat health through the course of the disease cycle.


The evolutionary position of nematodes.
The complete genomes of three animals have been sequenced by global research efforts: a nematode worm (Caenorhabditis elegans), an insect (Drosophila melanogaster), and a vertebrate (Homo sapiens). Remarkably, their relationships have yet to be clarified. The confusion concerns the enigmatic position of nematodes. Traditionally, nematodes have occupied a basal position, in part because they lack a true body cavity. However, the leading hypothesis now joins nematodes with arthropods in a molting clade, Ecdysozoa, based on data from several genes. We tested the Ecdysozoa hypothesis with analyses of more than 100 nuclear protein alignments, under conditions that would expose biases, and found that it was not supported. Instead, we found significant support for the traditional hypothesis, Coelomata. Our result is robust to different rates of sequence change among genes and lineages, different numbers of taxa, and different species of nematodes. We conclude that insects (arthropods) are genetically and evolutionarily closer to humans than to nematode worms.


Study of Methylene Blue Ototoxicity in the Guinea Pig.
Methylene blue is widely used in the medical field, especially as a blue dye for staining. It is also used as a photosensitizing agent in antimicrobial photodynamic therapy, which once photoactivated is effective for the eradication of several multi-resistant bacteria. The objective of this study was to investigate the ototoxic potential of methylene blue and precise its use in otology. It was a prospective animal study performed on guinea pigs in our tertiary medical center. We divided the animals into two groups: an experimental group and a control group, who underwent a series of three intratympanic (IT) injections. In the control group (n = 10), they received injections of gentamicin in one ear (positive control) and normal saline in the contralateral ear (negative control). The experimental group (n = 10) received injections of methylene blue in one ear, compared to injections of normal saline in the contralateral ear. We conducted auditory-evoked brainstem response (ABR) before and 1 week after the injection series. Once this is completed, the cochlea was dissected and caspase-3 was analyzed by immunohistochemistry. The mean difference of hearing loss in the methylene blue group compared to normal saline was 1.50 dB, and it was not shown to be statistically significant (P = 0.688). For the positive control group, which received IT injections of gentamicin, the mean threshold of hearing loss difference for all the frequencies combined was 66.25 dB (P < 0.001). Furthermore, uptake of caspase-3 by immunohistochemistry (apoptotic marker) was negative in our group, which received injections of methylene blue. In light of our results, IT injections of methylene blue did not demonstrate an ototoxic potential. We recommend further studies to precise its use in the otologic field.


Identification and Management of Children and Adolescents with Obesity Referred to a General Paediatric Outpatient Department.
Aims To identify all children and adolescents with overweight or obesity attending the outpatient department and audit our processes in their identification and management against NICE standards. Methods A retrospective chart review was performed. BMI charts were used to identify children and adolescents with overweight/obesity. The patient journey was audited to ascertain if overweight/obesity was identified by the clinician, whether this was communicated to the child or adolescent/their carer and whether intervention was offered. Results There were 669 scheduled appointments and 27.3%(n=127) of children 2 years and adolescents were identified with overweight/obesity. Children and adolescents referred for reasons not primarily related to obesity management were identified (90.6% (n=115)) and this group was analysed. Height and weight and/or BMI were communicated in 13.9% (n=16) of referral letters. A record of discussing growth was observed in 15.7% (n=18) of cases. Growth measurements were included in the post-clinic correspondence to the primary care physician in 56.8% (n=63) of letters. Discussion Further research is required to ascertain what barriers exist to the discussion of growth. Additional education of healthcare providers is necessary to develop standardised procedures around processes related to child and adolescent growth.


Low temperature solution synthesis of reduced two dimensional Ti3C2 MXenes with paramagnetic behaviour.
MXenes - two dimensional, 2D, early transition metal, M, carbides and nitrides, X - are the latest addition to the 2D materials' world. Herein, we report on a facile low temperature solution chemical synthesis method to reduce Ti3C2Tx multilayered, ML, MXenes. Using X-ray photoelectron spectroscopy, electron spin resonance, magnetization measurements and other techniques, we concluded that immersing Ti3C2Tx MLs in the reducing agent Li-ethylenediamine (Li-EDA) - held at temperatures varying from room to 120 °C - reduces the 2D layers creating Ti3+ ions and oxygen vacancies. Above a temperature (T) of ≈10 K, the magnetic susceptibilities, χ, are temperature independent, implying that the resulting powders are Pauli paramagnetic. The loss of the magnetic signal upon intercalation of Li+ or EDA, together with a Curie-like increase in χ at T < 10 K, is consistent with that of a disordered metal that is close to a metallic to insulator transition and proves that the magnetism is associated with the 2D flakes. This result is the first evidence of any magnetism of any MXene.


The effect of appropriate and inappropriate stimulus color on odor discrimination.
Color can strongly affect participants' self-report of an odor's qualities. In Experiment 1, we examined whether color influences a more objective measure of odor quality, discrimination. Odor pairs, presented in their appropriate color (e.g., strawberry and cherry in red water), an inappropriate color (e.g., strawberry and cherry in green water), or uncolored water were presented for discrimination. Participants made significantly more errors when odors were discriminated in an inappropriate color. In Experiment 2, the same design was utilized, but with an articulatory suppression task (AST), to examine whether the effect of color was mediated by identification or by a more direct effect on the percept. Here, the AST significantly improved discrimination for the inappropriate color condition, relative to Experiment 1. Although color does affect a more objective measure of odor quality, this is mediated by conceptual, rather than perceptual, means.


Environmental DNA (eDNA) metabarcoding reveals strong discrimination among diverse marine habitats connected by water movement.
While in recent years environmental DNA (eDNA) metabarcoding surveys have shown great promise as an alternative monitoring method, the integration into existing marine monitoring programs may be confounded by the dispersal of the eDNA signal. Currents and tidal influences could transport eDNA over great distances, inducing false-positive species detection, leading to inaccurate biodiversity assessments and, ultimately, mismanagement of marine environments. In this study, we determined the ability of eDNA metabarcoding surveys to distinguish localized signals obtained from four marine habitats within a small spatial scale (<5 km) subject to significant tidal and along-shore water flow. Our eDNA metabarcoding survey detected 86 genera, within 77 families and across 11 phyla using three established metabarcoding assays targeting fish (16S rRNA gene), crustacean (16S rRNA gene) and eukaryotic (cytochrome oxidase subunit 1) diversity. Ordination and cluster analyses for both taxonomic and OTU data sets show distinct eDNA signals between the sampled habitats, suggesting dispersal of eDNA among habitats was limited. Individual taxa with strong habitat preferences displayed localized eDNA signals in accordance with their respective habitat, whereas taxa known to be less habitat-specific generated more ubiquitous signals. Our data add to evidence that eDNA metabarcoding surveys in marine environments detect a broad range of taxa that are spatially discrete. Our work also highlights that refinement of assay choice is essential to realize the full potential of eDNA metabarcoding surveys in marine biodiversity monitoring programs.


Albuminuria, not only a cardiovascular/renal risk marker, but also a target for treatment?
Albuminuria has been identified as a marker for predicting both cardiovascular and renal risk. From normal to overt proteinuria levels, albuminuria shows a continuous marked increase in risk. This is independent of other well-known cardiovascular and renal risk markers and factors, such as blood pressure, cholesterol, smoking, overweight, and others. The predictive power is not only present in already diseased populations with either nondiabetic or diabetic renal disease, but also in hypertensive and even in otherwise healthy populations. New antihypertensive intervention strategies, such as angiotensin-converting enzyme (ACE) inhibitors and angiotensin II (Ang II) receptor-antagonists are claimed to have cardioprotective and renoprotective benefits that go beyond blood pressure control. Interestingly, these new therapeutic classes share the ability to lower urinary albumin excretion by an average of 40%, a characteristic that is not observed with the other antihypertensive drug classes. This short-term-induced antiproteinuric effect appears to predict the long-term cardiovascular and renal protection: the more albuminuria is lowered, the more that individual (or group) is protected. These data suggest that albumin is not only a risk marker for cardiovascular and or renal disease, but it may also be a useful target for therapy. Monitoring of albuminuria should be daily practice in subjects at risk for cardiovascular and renal disease. In addition to new clinical trials that prove that albumin can be targeted to obtain cardiovascular protection, guidelines should be made to help the physician in deciding how to measure albumin in the urine, what are normal levels, how to target "abnormal" levels, and how low we should go.


Pseudomonas aeruginosa rash associated with a whirlpool.
A generalized pruritic pustular rash was reported by 32 of 61 (53%) persons who had used the swimming pool and whirlpool at a Minnesota motel in March 1975. A questionnaire survey indicated that attack rates were highest in periods of heaviest bather load. The rash appeared 8 to 48 hours after exposure and resolved within seven days. No rash was reported by 37 motel guests who did not use the pool. Pseudomonas aeruginosa, serogroup 11, was isolated from the skin lesions of two patients and from the water from both pools. Circumstantial evidence implicated the whirlpool as the most probable source of infection. Deficiencies in disinfecting equipment and technique were identified and corrected.


Unveiling Candida albicans intestinal carriage in healthy volunteers: the role of micro- and mycobiota, diet, host genetics and immune response.
Candida albicans is a commensal yeast present in the gut of most healthy individuals but with highly variable concentrations. However, little is known about the host factors that influence colonization densities. We investigated how microbiota, host lifestyle factors, and genetics could shape C. albicans intestinal carriage in 695 healthy individuals from the Milieu Intérieur cohort. C. albicans intestinal carriage was detected in 82.9% of the subjects using quantitative PCR. Using linear mixed models and multiway-ANOVA, we explored C. albicans intestinal levels with regard to gut microbiota composition and lifestyle factors including diet. By analyzing shotgun metagenomics data and C. albicans qPCR data, we showed that Intestinimonas butyriciproducens was the only gut microbiota species whose relative abundance was negatively correlated with C. albicans concentration. Diet is also linked to C. albicans growth, with eating between meals and a low-sodium diet being associated with higher C. albicans levels. Furthermore, by Genome-Wide Association Study, we identified 26 single nucleotide polymorphisms suggestively associated with C. albicans colonization. In addition, we found that the intestinal levels of C. albicans might influence the host immune response, specifically in response to fungal challenge. We analyzed the transcriptional levels of 546 immune genes and the concentration of 13 cytokines after whole blood stimulation with C. albicans cells and showed positive associations between the extent of C. albicans intestinal levels and NLRP3 expression, as well as secreted IL-2 and CXCL5 concentrations. Taken together, these findings open the way for potential new interventional strategies to curb C. albicans intestinal overgrowth.


Targeting Cytoprotective Autophagy to Enhance Anticancer Therapies.
Autophagy is a highly regulated multi-step process that occurs at the basal level in almost all cells. Although the deregulation of the autophagy process has been described in several pathologies, the role of autophagy in cancer as a cytoprotective mechanism is currently well established and supported by experimental and clinical evidence. Our understanding of the molecular mechanism of the autophagy process has largely contributed to defining how we can harness this process to improve the benefit of cancer therapies. While the role of autophagy in tumor resistance to chemotherapy is extensively documented, emerging data point toward autophagy as a mechanism of cancer resistance to radiotherapy, targeted therapy, and immunotherapy. Therefore, manipulating autophagy has emerged as a promising strategy to overcome tumor resistance to various anti-cancer therapies, and autophagy modulators are currently evaluated in combination therapies in several clinical trials. In this review, we will summarize our current knowledge of the impact of genetically and pharmacologically modulating autophagy genes and proteins, involved in the different steps of the autophagy process, on the therapeutic benefit of various cancer therapies. We will also briefly discuss the challenges and limitations to developing potent and selective autophagy inhibitors that could be used in ongoing clinical trials.


Characterization of β-lactoglobulin from buffalo (Bubalus bubalis) colostrum and its possible interaction with erythrocyte lipocalin-interacting membrane receptor.
Lipocalins form a widespread class of proteins involved in the transport of weakly soluble vitamins, hormones or hydrophobic molecules. β-lactoglobulin (BLG-col), a major lipocalin present in whey was purified and characterized from buffalo colostrum. The molecular weight of BLG-col as determined by Liquid chromatography -electrospray ionization mass spectrometry (LC-ESI-MS) was 18.257 kDa and the peptide mass fingerprint of the purified protein revealed 67% sequence homology to buffalo milk β-lg. The N-terminal-IIVTQ and LC-ESI-collision-induced dissociation-Electron transfer dissociation mass spectrometry/mass spectrometry analyses of doubly (m/z 1156(+2)) and triply (m/z 546(+3)) charged ion pairs corresponding to VYVEELKPTPEGDLEILLQK (41-60) and TPEVDDEALEKFDK (125-138) sequences confirmed the identity of BLG-col. Using these peptide sequences, the location of a gene encoding for BLG-col was identified on chromosome 11 at 11q28 loci of bovine genome. The unique property of the BLG-col isolated from buffalo colostrum was its strong and specific haemagglutinating activity with 'O' blood of human erythrocytes with 10,309 HAU/mg protein. The cell surface localization of BLG-col on human erythrocytes was confirmed by immunocytochemistry and the specificity of interaction was established by immunoblot analysis of human erythrocyte membrane proteins. Based on these observations, we suggest the presence of lipocalin receptor (70 kDa) on human erythrocyte membrane and the multiple sequence alignment supported structural diversity among lipocalin receptors.


Effects of small bowel transection, resection, or bypass in 1,2-dimethylhydrazine-induced rat intestinal neoplasia.
The incidence, distribution, size, and histopathology of rat small and large bowel tumors induced by sequential administration of 1,2-dimethylhydrazine followed by either small bowel resection, 50% jejunoileal resection, or 50% jejunoileal bypass were examined in addition to measurements of transit times and beta-glucuronidase activities in large bowel contents. The results indicate that even limited small bowel resection or bypass promotes intestinal neoplasia, particularly in the large bowel, and this effect seems independent of the chronic injury imposed by suture lines. Although no differences in transit times were observed, increased beta-glucuronidase activities in both cecum and distal colon of resected but not bypassed rats was detected. Moreover, an apparent subsite redistribution of small bowel tumors to ileum and large bowel tumors to more proximal colon in bypassed rats suggests that the mechanisms involved for tumor enhancement differ substantially from those in resected rats.


EasyEyes - A new method for accurate fixation in online vision testing.
Online methods allow testing of larger, more diverse populations, with much less effort than in-lab testing. However, many psychophysical measurements, including visual crowding, require accurate eye fixation, which is classically achieved by testing only experienced observers who have learned to fixate reliably, or by using a gaze tracker to restrict testing to moments when fixation is accurate. Alas, both approaches are impractical online as online observers tend to be inexperienced, and online gaze tracking, using the built-in webcam, has a low precision (±4 deg). EasyEyes open-source software reliably measures peripheral thresholds online with accurate fixation achieved in a novel way, without gaze tracking. It tells observers to use the cursor to track a moving crosshair. At a random time during successful tracking, a brief target is presented in the periphery. The observer responds by identifying the target. To evaluate EasyEyes fixation accuracy and thresholds, we tested 12 naive observers in three ways in a counterbalanced order: first, in the laboratory, using gaze-contingent stimulus presentation; second, in the laboratory, using EasyEyes while independently monitoring gaze using EyeLink 1000; third, online at home, using EasyEyes. We find that crowding thresholds are consistent and individual differences are conserved. The small root mean square (RMS) fixation error (0.6 deg) during target presentation eliminates the need for gaze tracking. Thus, this method enables fixation-dependent measurements online, for easy testing of larger and more diverse populations.


Glucuronidation of PhIP and N-OH-PhIP by UDP-glucuronosyltransferase 1A10.
The UDP-glucuronosyltransferase (UGT) 1A10 is an extra-hepatic enzyme that plays an important role in the glucuronidation of a variety of endogenous and exogenous substances and is expressed throughout the aerodigestive and digestive tracts. Two classes of carcinogens that target the colon, heterocyclic amines (HCAs) and polycyclic aromatic hydrocarbons, are known to be detoxified by the UGT family of enzymes. Recently, our laboratory demonstrated that UGT1A10 has considerably more activity against polycyclic aromatic hydrocarbons in vitro than any other UGT family member. In this study, we focused on the glucuronidation of the HCA, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), and its bioactivated metabolite, N-hydroxy-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (N-OH-PhIP). We demonstrated that UGT1A10 exhibited a significantly higher glucuronidation rate against PhIP and N-OH-PhIP than any other UGT family member in vitro using whole-cell homogenates of HEK293 cells over-expressing individual UGTs. Kinetic analysis revealed a 9- and 22-fold higher level of activity for UGT1A10 homogenates as compared with the next most active UGT, UGT1A1, against N-OH-PhIP as determined by maximum rate/apparent Michaelis constant (V(max)/K(M)) at the N3 and N2 positions, respectively. The polymorphic UGT1A10(139Lys) variant exhibited a 2- to 16-fold decrease in glucuronidation activity against PhIP and N-OH-PhIP, as compared with the wild-type UGT1A10(139Glu) isoform. These data suggest that UGT1A10 is the most active UGT against PhIP and N-OH-PhIP and that UGT1A10 may play an important role in susceptibility to HCA-induced colon cancer.


Fabry disease in a large Nova Scotia kindred: carrier detection using leucocyte alpha-galactosidase activity and an NcoI polymorphism detected by an alpha-galactosidase cDNA clone.
Fabry disease is an X linked recessive disorder of glycosphingolipid metabolism resulting from a deficiency of the lysosomal hydrolase alpha-galactosidase (alpha-gal). Measurement of the enzyme activity, however, is not an accurate method for identification of female carriers among at risk relatives of affected males. The alpha-gal cDNA and gene have been cloned previously and found to provide useful probes for the molecular analysis of affected families but these clones have not been available to us. Thus, to analyse Fabry disease in Nova Scotia, especially within a large kindred known to contain 30 affected males and 50 possible carrier females, we isolated an independent cDNA for alpha-gal. Using this clone as a probe, the mutation in the Nova Scotia kindred was shown not to be a major DNA alteration, but was found to be linked to the rarer allele (frequency 0.20) of the polymorphic NcoI site located 3' to the gene. Affected males from two Nova Scotia families who cannot be associated with the kindred by history were also found to have the rarer NcoI allele, which suggests they are, in fact, part of the kindred. The coupling of the mutation to an infrequent marker also helped carrier identification in the kindred where all of 17 obligate carriers examined, including six who were not identified as carriers by enzyme assays, were found to be heterozygous for the RFLP. Thus, DNA analysis can be used for presymptomatic and prenatal diagnosis in most portions of the Nova Scotia kindred affected with Fabry disease.


Differentiation of Micrococcus luteus and Micrococcus varians on the basis of catalase isoenzymes.
Crude extracts prepared from four Micrococcus varians strains, 11 M. luteus strains and four laboratory isolates subsequently classified with M. luteus were assayed for catalase activity following electrophoresis on polyacrylamide gels. The enzyme patterns produced from the M. varians strains exhibited three catalase isoenzymes which were distinguished into two types of patterns depending upon the location of the major band. The extracts from all the M. luteus strains produced the same pattern, composed of two catalase isoenzymes of similar electrophoretic mobility. For both species the isoenzyme patterns agreed with the differentiation based on biochemical properties. The catalase activity activity staining method was shown to be a restricted yet reliable assay in the intrageneric but not intraspecies differentiation of yellow-pigmented micrococci.


Steroids in the treatment of clinical septic shock.
A prospective (Part I) and a retrospective (Part II) study were used to determine the safety and efficacy of corticosteroids in the treatment of septic shock. In Part I, 172 consecutive patients in septic shock admitted over an 8-year period were treated with either steroid or saline: 43 received dexamethasone (DMP), 43 received methylprednisolone (MPS), and 86 received saline. The study was double-blind and randomized, and the three groups were compared for age, severity of shock, presence of underlying disease, and year of study. In the 86 saline-treated patients, the mortality rate was 38.4% (33/86); in the steroid-treated patients, it was 10.4% (9/86). With MPS the mortality rate was 11.6% (5/43), and with DMP it was 9.3% (4/43). Thus, overall mortality was significantly less in the steroid-treated group than in the control group. Further, there was no significant difference in mortality rate between the DMP- and the MPS-treated patients. In Part II, 328 patients were studied retrospectively. One-hundred sixty were treated without steroid, and 168 were treated with either DMP or MPS. Again, the two groups of patients were compared for severity of shock, underlying disease, age, and year of study. Mortality among patients treated without steroid was 42.5% (68/160) and among patients treated with steroid was 14% (24/168); there was no significant difference in mortality rate between DMP- and MPS-treated patients. In Parts I and II combined, complications occurred in 6% of steroid-treated patients with no significant difference between DMP- and MPS-treated groups.


TAAR6 variation effect on clinic presentation and outcome in a sample of schizophrenic in-patients: an open label study.
We recently reported an association between TAAR6 (trace amine associated receptor 6 gene) variations and schizophrenia (SZ). We now report an association of a set of TAAR6 variations and clinical presentation and outcome in a sample of 240 SZ Korean patients. Patients were selected by a Structured Clinical Interview, DSM-IV Axis I disorders - Clinical Version (SCID-CV). Other psychiatric or neurologic disorders, as well as medical diseases, were exclusion criteria. To assess symptom severity, patients were administered the CGI scale and the PANSS at baseline and at the moment of discharge, 1 month later on average. TAAR6 variations rs6903874, rs7452939, rs8192625 and rs4305745 were investigated; rs6903874, rs7452939 and rs8192625 entered the statistical investigation after LD analysis. Rs8192625 G/G homozygosis was found to be significantly associated both with a worse clinical presentation at PANSS total and positive scores and with a shorter period of illness before hospitalization. No haplotype significant findings were found. The present study stands for a role of the TAAR6 in the clinical presentation of SZ. Moreover, our results show that this genetic effect may be counteracted by a correct treatment. Haplotype analysis was not informative in our sample, probably also because of the incomplete SNPs' coverage of the gene we performed. Further studies in this direction are warranted.


Efficacy of eye movement desensitization and reprocessing on the phantom limb pain of patients with amputations within a 24-month follow-up.
The aim of this study was to evaluate the efficacy of eye movement desensitization and reprocessing (EMDR) on the phantom limb pain (PLP) of patients with amputations within a 24-month follow-up. This study was a randomized-controlled trial. A total of 60 patients with amputations were selected by a purposive sampling and patients were divided randomly into two experimental and control groups. Samples were assigned through randomized allocation. EMDR therapy was administered individually to the experimental group participants in 12 one-hour sessions over a 1-month period In each session, the patient completed the Subjective Units of Distress Scale and a pain-rating scale before and after the intervention. Follow-up measures were obtained 24 months later for the experimental group. The participants in the control group were measured on the two scales at an initial session and again after 1- and 24-month follow-up. The mean PLP decreased in the experimental group between the first and last sessions and remained so at a 24-month follow-up. No decrease occurred for the control group over the 1- and 24-month period. The differences were statistically significant (P<0.001) according to a repeated-measures analysis of variance. EMDR therapy proved to be a successful treatment for PLP. Because of its efficacy and the fact that the positive effects were maintained at the 24-month follow-up, this therapy is recommended for the treatment of PLP.


Macronutrient intake, physical activity, serum lipids and increased body weight in primary schoolchildren in Istanbul.
The aim of this cross-sectional study was to record the prevalence of underweight, overweight and obesity in primary school children living in Istanbul and to examine the relationship between increased body weight and certain cardiovascular disease (CVD) risk factors. A total of 510 randomly selected children aged 12 and 13 years of age (257 boys, 253 girls) were examined. Information regarding anthropometrical indices, energy and macronutrient intake, physical activity, physical fitness and lipid profile were collected. Classification of children in overweight and obese subgroups was based on the cut-off points proposed by Cole et al. The prevalence of underweight, overweight and obesity was found to be 15.3%, 10.6% and 1.6%, respectively. Both overweight boys and girls were found to have lower physical fitness compared to their normal-weight counterparts, but no difference was observed for energy and macronutrient intake. Overweight boys were found to have higher total cholesterol (P < 0.001), low-density lipoprotein cholesterol (P < 0.01), triglycerides (P < 0.01) and total cholesterol/high-density lipoprotein cholesterol (HDL-C) ratio (P < 0.05) compared to their normal-weight counterparts, while overweight girls were found to have lower HDL-C (P < 0.05) compared to their normal-weight peers. Underweight and overweight coexisted in the current population. Increased body weight was accompanied by unfavorable lipid profiles and lower fitness levels. Consequently, there is an emergent need for early identification and understanding of behavioral and physiological variables related to obesity and CVD, so that appropriate interventions can be targeted to children who are at risk for adult onset of these diseases.


Reactions of patients to video recording of consultations in general practice.
The potential value of video recording for examining medical consultations depends on the extent to which recordings are representative of unfilmed consultations. This paper examines the views of 295 patients in two general practices whose consultations were filmed and compares them with the views of a control group of 185 patients. Most of those who were filmed reported that the consultation was not directly affected, and no overall effect of filming was discovered when patients rated their stress after the consultations, their rapport with the doctor, or other aspects of the consultation. At one practice, however, filming was significantly associated with lower ratings of rapport between doctor and patient among those patients who reported some direct effect of filming. Patient refusal rates from other studies are also examined and shown to vary systematically--the more opportunity patients are given to decline the more likely they are to take it. Consideration of doctors' responses to being filmed would usefully complement the emphasis on the views of patients.


Turnover of strain-level diversity modulates functional traits in the honeybee gut microbiome between nurses and foragers.
Strain-level diversity is widespread among bacterial species and can expand the functional potential of natural microbial communities. However, to what extent communities undergo consistent shifts in strain composition in response to environmental/host changes is less well understood. Here, we used shotgun metagenomics to compare the gut microbiota of two behavioral states of the Western honeybee (Apis mellifera), namely nurse and forager bees. While their gut microbiota is composed of the same bacterial species, we detect consistent changes in strain-level composition between nurses and foragers. Single nucleotide variant profiles of predominant bacterial species cluster by behavioral state. Moreover, we identify strain-specific gene content related to nutrient utilization, vitamin biosynthesis, and cell-cell interactions specifically associated with the two behavioral states. Our findings show that strain-level diversity in host-associated communities can undergo consistent changes in response to host behavioral changes modulating the functional potential of the community.


Trends for inflammatory breast cancer: is survival improving?
The purpose of this study was to evaluate whether the survival of women with inflammatory breast cancer (IBC) treated at our institution has improved over the past 30 years. Three-hundred ninety-eight patients with IBC were treated between 1974 and 2005. Patient characteristics and outcomes were tabulated and compared among decades of diagnosis. Survival outcomes were estimated with the Kaplan-Meier product limit method and compared among groups with the log-rank statistic. Cox proportional hazards models were fit to determine the association between year of diagnosis and survival outcomes after adjustment for patient and disease characteristics and treatments received. The median follow-up was 5.8 years (range, 0.3-23.8 years). There were 238 recurrences and 236 deaths. The median recurrence-free survival (RFS) duration was 2.3 years and the median overall survival (OS) time was 4.2 years. In the models for RFS and OS, after adjustment for patient and disease characteristics, increasing year of diagnosis was not associated with a decrease in the risk for recurrence (hazard ratio, [HR], 1.00; 95% confidence interval [CI], 0.97-1.04) or death (HR, 0.97; 95% CI, 0.94-1.01). Our data show that there has not been an important change in the prognosis of patients with IBC in the last 30 years. Clinical trials focusing on the management of this aggressive disease are warranted. Disclosure of potential conflicts of interest is found at the end of this article.


Terminal alkene formation by the thioesterase of curacin A biosynthesis: structure of a decarboxylating thioesterase.
Curacin A is a polyketide synthase (PKS)-non-ribosomal peptide synthetase-derived natural product with potent anticancer properties generated by the marine cyanobacterium Lyngbya majuscula. Type I modular PKS assembly lines typically employ a thioesterase (TE) domain to off-load carboxylic acid or macrolactone products from an adjacent acyl carrier protein (ACP) domain. In a striking departure from this scheme the curacin A PKS employs tandem sulfotransferase and TE domains to form a terminal alkene moiety. Sulfotransferase sulfonation of β-hydroxy-acyl-ACP is followed by TE hydrolysis, decarboxylation, and sulfate elimination (Gu, L., Wang, B., Kulkarni, A., Gehret, J. J., Lloyd, K. R., Gerwick, L., Gerwick, W. H., Wipf, P., Håkansson, K., Smith, J. L., and Sherman, D. H. (2009) J. Am. Chem. Soc. 131, 16033-16035). With low sequence identity to other PKS TEs (<15%), the curacin TE represents a new thioesterase subfamily. The 1.7-Å curacin TE crystal structure reveals how the familiar α/β-hydrolase architecture is adapted to specificity for β-sulfated substrates. A Ser-His-Glu catalytic triad is centered in an open active site cleft between the core domain and a lid subdomain. Unlike TEs from other PKSs, the lid is fixed in an open conformation on one side by dimer contacts of a protruding helix and on the other side by an arginine anchor from the lid into the core. Adjacent to the catalytic triad, another arginine residue is positioned to recognize the substrate β-sulfate group. The essential features of the curacin TE are conserved in sequences of five other putative bacterial ACP-ST-TE tridomains. Formation of a sulfate leaving group as a biosynthetic strategy to facilitate acyl chain decarboxylation is of potential value as a route to hydrocarbon biofuels.


Probing the permeability of polyelectrolyte multilayer capsules via a molecular beacon approach.
Application of polyelectrolyte multilayer (PEM) capsules as vehicles for the controlled delivery of substances, such as drugs, genes, pesticides, cosmetics, and foodstuffs, requires a sound understanding of the permeability of the capsules. We report the results of a detailed investigation into probing capsule permeability via a molecular beacon (MB) approach. This method involves preparing MB-functionalized bimodal mesoporous silica (BMSMB) particles, encapsulating the BMSMB particles within the PEM film to be probed, and then incubating the encapsulated BMSMB particles with DNA target sequences of different lengths. Permeation of the DNA targets through the capsule shell causes the immobilized MBs to open due to hybridization of the DNA targets with the complementary loop region of the MBs, resulting in an increase in the MB fluorescence. The assay conditions (BMSMB particle concentration, MB loading within the BMS particles, DNA target concentration, DNA target size, pH, sodium chloride concentration) where the MB-DNA sensing process is effective were first examined. The permeability of DNA through poly(sodium 4-styrenesulfonate) (PSS)/poly(allylamine hydrochloride) (PAH) multilayer films, with and without a poly(ethyleneimine) (PEI) precursor layer, was then investigated. The permeation of the DNA targets decreases considerably as the thickness of the PEM film encapsulating the BMSMB particles increases. Furthermore, the presence of a PEI precursor layer gives rise to less permeable PSS/PAH multilayers. The diffusion coefficients calculated for the DNA targets through the PEM capsules range from 10-19 to 10-18 m2 s-1. This investigation demonstrates that the MB approach to measuring permeability is an important new tool for the characterization of PEM capsules and is expected to be applicable for probing the permeability of other systems, such as membranes, liposomes, and emulsions.


Effects of Ag and Cu ions on the microbial corrosion of 316L stainless steel in the presence of Desulfovibrio sp.
The utilization of Ag and Cu ions to prevent both microbial corrosion and biofilm formation has recently increased. The emphasis of this study lies on the effects of Ag and Cu ions on the microbial corrosion of 316L stainless steel (SS) induced by Desulfovibrio sp. Electrochemical impedance spectroscopy (EIS) and potentiodynamic polarization were used to analyze the corrosion behavior. The biofilm formation, corrosion products and Ag and Cu ions on the surfaces were investigated using scanning electron microscopy (SEM), energy dispersive X-ray spectrometry (EDS) and elemental mapping. Through circuit modeling, EIS results were used to interpret the physicoelectric interactions between the electrode, biofilm and culture interfaces. EIS results indicated that the metabolic activity of Desulfovibrio sp. accelerated the corrosion rate of SS in both conditions with and without ions. However, due to the retardation in the growth of Desulfovibrio sp. in the presence of Ag and Cu ions, significant decrease in corrosion rate was observed in the culture with the ions. In addition, SEM and EIS analyses revealed that the presence of the ions leads to the formation on the SS of a biofilm with different structure and morphology. Elemental analysis with EDS detected mainly sulfide- and phosphorous-based corrosion products on the surfaces.