Free radical initiation in proteins and amino acids by ionizing and ultraviolet radiations and lipid oxidation--Part I: ionizing radiation.
Parallels and similarities in chemical and functional damage to proteins by ionizing and UV radiations and oxidizing lipids have been recognized for some time. However, only recently have oxidizing lipids been shown directly by electron spin resonance to be radiomimetic also in their capacity for protein free radical production. Free radicals play a key role in the transformation of energy to molecular and cellular damage. It is thus of critical importance to elucidate the general mechanisms of free radical formation and reactions in proteins in order to understand protein involvement in various pathological conditions and in food deterioration. Accordingly, this review is a detailed comparison of gamma radiation, UV radiation, and lipid oxidation for what is presently known concerning (1) the specific modes of energy deposition and free radical formation, (2) the free radicals formed in proteins and amino acids, and (3) the typical damage correlating with these radicals.


Genetic and epigenetic alterations in primary-progressive paired oligodendroglial tumors.
The aim of the present study was to identify genetic and epigenetic alterations involved in the progression of oligodendroglial tumors. We characterized 21 paired, World Health Organization (WHO) grade II and III oligodendroglial tumors from patients who received craniotomies for the partial or complete resection of primary and secondary oligodendroglial tumors. Tumor DNA was analyzed for alterations in selected genetic loci (1p36, 9p22, 10q23-24, 17p13, 19q13, 22q12), isocitrate dehydrogenase 1 (IDH1), isocitrate dehydrogenase 2 (IDH2) and the CpG island methylation status of critical tumor-related genes (MGMT, P16, DAPK, PTEN, RASSF1A, Rb1). Alterations of these markers were common early in the tumorigenesis. In the primary tumors we identified 12 patients (57.1%) with 1p36 deletions, 17 (81.0%) with 19q13 deletions, 9 (42.9%) with 1p36/19q13 codeletions, 11 (52.3%) with 9p22 deletions, and 12 (57.1%) with IDH1 mutation. Epigenetic analysis detected promoter methylation of the MGMT, P16, DAPK, PTEN, RASSF1A, and Rb1 genes in 38.1%, 19.0%, 38.1%, 33.3%, 66.7%, and 14.3% of primary tumors, respectively. After progression, additional losses of 1p, 9p, 10q, 17p, 19q and 22q were observed in 3 (14.3%), 1 (4.8%), 3 (14.3%), 2 (9.5%), 1 (4.8%) and 3 (14.3%) cases, respectively. Additional methylations of the MGMT, P16, DAPK, PTEN, RASSF1A, and RB1 promoters was observed in 4 (19.0%), 2 (9.5%), 0 (0%), 6 (28.6%), 2(9.5%) and 3 (14.3%) cases, respectively. The status of IDH1 mutation remained unchanged in all tumors after progression. The primary tumors of three patients with subsequent progression to high-grade astrocytomas, all had 9p deletion, intact 1p, intact 10q and unmethylated MGMT. Whether this may represent a molecular signature of patients at-risk for the development of aggressive astrocytomas needs further investigation.


Lifetime occupation is not associated with radiographic osteoarthritis of the first metatarsophalangeal joint in a cohort study of UK women.
The study aim was to determine whether lifetime occupation was associated with the presence of radiographic osteoarthritis (ROA) of the first metatarsophalangeal joint (MTPJ) in women. Data were collected from the prospective, population-based Chingford 1000 Women study. This cohort of women, aged 45-64 years at inception, was established in 1989 from a single general practice in Chingford, UK. Data has subsequently been collected repeatedly. Data from baseline, year six and year ten was used for the purposes of this cross-sectional study. The primary outcome was the presence of dorsal view ROA of the first MTPJ. The main exposure was lifetime occupation, categorised according to levels of occupation previously defined via international consensus: 1. Sedentary, 2. Light, 3. Light manual, 4. Heavy manual. Logistic regression analyses were conducted to quantify the relationship between lifetime occupation type and the presence of ROA of the first MTPJ, adjusting for age, body mass index and lifetime high-heeled footwear use as potential interactive variables for each decade. Data for 209 women were included within this study. The mean (SD) age was 57 (±5.2) years. Predominant lifetime occupation was reported as sedentary by 51.7%, as light by 0%, as light manual by 33.5% and as heavy manual by 14.8% of participants. There were no statistical associations between lifetime occupation type and the presence of ROA of the first MTPJ in either the unadjusted (OR = 0.99, CI = 0.78-1.26,P = 0.96) partially adjusted (for age and BMI; OR = 1.00, CI = 0.78-1.29, P = 0.99) or fully adjusted models (for age, BMI and lifetime high heel footwear use for each decade of working life (OR = 1.02, CI = 0.79-1.31, P = 0.91); high-heel footwear use up to 20s (OR = 0.83, CI = 0.71-1.31, P = 0.83); high-heel footwear use in 20-30s (OR = 1.00, CI = 0.75-1.3, P = 0.98); high-heel footwear use in 30-40s (OR = 1.00, CI = 0.70-1.42, P = 0.99); high-heel footwear use in 40-50s (OR = 0.90, CI = 0.58-1.40, P = 0.65); high-heel footwear use in 50s (OR = 0.63,CI = 0.36-1.09, P = 0.10). The findings suggest that lifetime occupation is not associated with the presence of ROA of the fist metatarsophalangeal joint. There does not appear to be any interactive effect between lifetime occupation, lifetime high-heel footwear use, age or BMI and ROA of the first MTPJ. In later life a positive trend towards increased ROA in those who reported lifetime high-heel footwear use was noted and this may be worthy of further research.


Where did the chili get its spice? Biogeography of capsaicinoid production in ancestral wild chili species.
The biogeography of pungency in three species of wild chili in the chaco and surrounding highland habitats of southeastern Bolivia is described. We report that Capsicum chacoense, C. baccatum, and C. eximium are polymorphic for production of capsaicin and its analogs, such that completely pungent and completely nonpungent individuals co-occur in some populations. In C. chacoense, the density of plants and the proportion of pungent plants increased with elevation. Above 900 m, all individuals in all populations except two were pungent; nonpungent individuals in at least one of the two polymorphic populations were likely a result of spreading by humans. The occurrence of pungent and nonpungent individuals in three species of ancestral Capsicum and the geographic variation of pungency within species suggest that production of capsaicin and its analogs entails both costs and benefits, which shift from one locality to another. Determining the selection pressures behind such shifts is necessary to understand the evolution of pungency in chilies.


Cancer survivorship monitoring systems for the collection of patient-reported outcomes: a systematic narrative review of international approaches.
This systematic narrative review describes and compares the development and operational approaches of monitoring systems without a clinical care component that collect patient-reported outcome (PRO) data from cancer survivors. Searches were conducted using Medline, PubMed, PsycINFO, the Cochrane Library, CINAHL, Scopus, Joanna Briggs Institute EBP Database and Google Scholar (Advanced). Sources of grey literature and websites of relevant organisations were also searched for relevant published and unpublished material. Articles were included if they described the development (including piloting) of monitoring systems with ongoing recruitment that collect PRO at more than one time point, from 6 months post-diagnosis onward. The initial searches returned 7290 unique citations. After screening titles and abstracts, 39 full-text articles were retrieved for more detailed examination. Eleven articles were included in the review, representing seven international monitoring systems. Systems varied in their scope, implementation process, governance and administration, recruitment and data collection, consent rates, PRO collection, use of PRO and translation strategies. The most suitable approach for setting-up and implementing a monitoring system for ongoing surveillance will differ depending on the unique requirements, aims and level of resourcing available within a particular context. Better specification and consideration of how PRO data will be used, for what purpose, and by whom, is required to inform effective translational strategies to improve outcomes for cancer survivors. The findings from this review may inform the future development of survivorship monitoring systems in varied environments, which in turn may improve practices that lead to better outcomes for survivors.


The effect of peanut and grain bar preloads on postmeal satiety, glycemia, and weight loss in healthy individuals: an acute and a chronic randomized intervention trial.
Peanut consumption favorably influences satiety. This study examined the acute effect of peanut versus grain bar preloads on postmeal satiety and glycemia in healthy adults and the long-term effect of these meal preloads on body mass in healthy overweight adults. In the acute crossover trial (n = 15; 28.4 ± 2.9 y; 23.1 ± 0.9 kg/m2), the preload (isoenergetic peanut or grain bar with water, or water alone) was followed after 60 min with ingestion of a standardized glycemic test meal. Satiety and blood glucose were assessed immediately prior to the preload and to the test meal, and for two hours postmeal at 30-min intervals. In the parallel-arm, randomized trial (n = 44; 40.5 ± 1.6 y, 31.8 ± 0.9 kg/m2), the peanut or grain bar preload was consumed one hour prior to the evening meal for eight weeks. Body mass was measured at 2-week intervals, and secondary endpoints included blood hemoglobin A1c and energy intake as assessed by 3-d diet records collected at pre-trial and trial weeks 1 and 8. Satiety was elevated in the postprandial period following grain bar ingestion in comparison to peanut or water ingestion (p = 0.001, repeated-measures ANOVA). Blood glucose was elevated one hour after ingestion of the grain bar as compared to the peanut or water treatments; yet, total glycemia did not vary between treatments in the two hour postprandial period. In the 8-week trial, body mass was reduced for the grain bar versus peanut groups after eight weeks (-1.3 ± 0.4 kg versus -0.2 ± 0.3 kg, p = 0.033, analysis of covariance). Energy intake was reduced by 458 kcal/d in the first week of the trial for the grain bar group as compared to the peanut group (p = 0.118). Hemoglobin A1c changed significantly between groups during the trial (-0.25 ± 0.07% and -0.18 ± 0.12% for the grain bar and peanut groups respectively, p = 0.001). Compared to an isoenergetic peanut preload, consumption of a grain bar preload one hour prior to a standardized meal significantly raised postmeal satiety. Moreover, consumption of the grain bar prior to the evening meal was associated with significant weight loss over time suggesting that glycemic carbohydrate ingestion prior to meals may be a weight management strategy.


Structure of the human neutrophil elastase gene.
The gene for human neutrophil elastase (NE), a powerful serine protease carried by blood neutrophils and capable of destroying most connective tissue proteins, was cloned from a genomic DNA library of a normal individual. The NE gene consists of 5 exons and 4 introns included in a single copy 4-kilobase segment of chromosome 11 at q14. The coding exons of the NE gene predict a primary translation product of 267 residues including a 29-residue N-terminal precursor peptide and a 20-residue C-terminal precursor peptide. Analysis of the N-terminal peptide sequence suggests it contains a 27-residue "pre" signal peptide followed by a "proN" dipeptide, similar to that of other blood cell lysosomal proteases. The sequences for the mature 218-residue NE protein are included in exons II-V. The 5'-flanking region of the gene includes typical TATA, CAAT, and GC sequences within 61 base pairs (bp) of the cap site. The sequence 1.5 kilobases 5' to exon I contains several interesting repetitive sequences including six tandem repeats of unique 52- or 53-bp sequences. The 5'-flanking region also contains a 19-bp segment with 90% homology to a segment of the 5'-flanking region of the human myeloperoxidase (MPO) gene, a gene also expressed in bone marrow precursor cells and a protein stored in the same neutrophil granules as NE. In addition, like the MPO gene, the NE 5'-flanking region has several regions with greater than or equal to 75% homology to sequences 5' to c-myc, but there is no overlap between the NE-c-myc and MPO-c-myc homologous sequences.


Ocular damage induced by near-ultraviolet laser radiation.
A quantitative study was conducted of ocular damage thresholds in the rhesus monkey eye from krypton, argon, and nitrogen laser radiation. Corneal and lenticular thresholds are reported for various laser beam parameters. Corneal damage was found to occur following incident energy doses of approximately 60 to 70 Joules per square centimeter (J./cm.2) for pulsewidths ranging from 250 musec to 120 sec. The results are consistent with a photochemical damage mechanism. With certain exposure parameters, cataracts were induced with lower energy doses than required to cause corneal damage. The lenticular thresholds, however, appear to be consistent with a thermal rather than a photochemical mechanism. Corneal and lenticular hazards of near-ultraviolet (near-UV) lasers are discussed in terms of existing safety standards for laser radiation.


Living Materials Herald a New Era in Soft Robotics.
Living beings have an unsurpassed range of ways to manipulate objects and interact with them. They can make autonomous decisions and can heal themselves. So far, a conventional robot cannot mimic this complexity even remotely. Classical robots are often used to help with lifting and gripping and thus to alleviate the effects of menial tasks. Sensors can render robots responsive, and artificial intelligence aims at enabling autonomous responses. Inanimate soft robots are a step in this direction, but it will only be in combination with living systems that full complexity will be achievable. The field of biohybrid soft robotics provides entirely new concepts to address current challenges, for example the ability to self-heal, enable a soft touch, or to show situational versatility. Therefore, "living materials" are at the heart of this review. Similarly to biological taxonomy, there is a recent effort for taxonomy of biohybrid soft robotics. Here, an expansion is proposed to take into account not only function and origin of biohybrid soft robotic components, but also the materials. This materials taxonomy key demonstrates visually that materials science will drive the development of the field of soft biohybrid robotics.


An in vivo model of postinflammatory hyperpigmentation and erythema: clinical, colorimetric and molecular characteristics.
Postinflammatory hyperpigmentation (PIH) is a common, acquired pigmentary disorder of the skin associated with significant quality-of-life impairment, especially in individuals with skin of colour. Current treatment for PIH is limited, largely due to a poor understanding of disease pathogenesis and the lack of a representative disease model. This study is intended to further develop, update and validate our previously designed in vivo model of acne-induced PIH/postinflammatory erythema (PIE) using different concentrations of trichloroacetic acid (TCA), a medium-depth chemical peel. Twenty-nine patients with skin types II-VI and clinician-confirmed presence of two or more truncal acne pustules and PIH/PIE were included. On the basis of Investigator's Global Assessment (IGA), clinical polarized photography (CPP), colorimetry and Skindex, we experimentally determined an optimum TCA concentration and assessed our model's ability to exhibit a dose-response relationship between degree of inciting insult and severity of resulting pigmentation. We also performed differential microRNA profiling and pathway analysis to explore the potential of microRNAs as molecular adjuncts to our model. Application of TCA 30% produced lesions indistinguishable from acne-induced PIH and PIE lesions on the basis of colorimetry data without causing epidermal necrosis. Application of progressively increasing TCA doses from 20% to 30% resulted in concentration-dependent increases in CPP, IGA and colorimetry scores at all timepoints during the study. miRNA-31 and miRNA-23b may play a role in PIH pathogenesis, although further validation is required. Our TCA-based in vivo model, using TCA concentrations between 20% and 30% with an optimum of 30%, enables the quantitative assessment of the pigmentary response to varying degrees of cutaneous inflammation in a fashion that mirrors natural acne-induced PIH and PIE.


Bone Broth Unlikely to Provide Reliable Concentrations of Collagen Precursors Compared With Supplemental Sources of Collagen Used in Collagen Research.
Intake of dietary sources of collagen may support the synthesis of collagen in varying tissues, with the availability of key amino acids being a likely contributor to its effectiveness. This study analyzed commonly consumed preparations of bone broth (BB) to assess the amount and consistency of its amino acid content. Commercial and laboratory-prepared samples, made with standardized and variable (nonstandardized) protocols, were analyzed for key amino acids (glycine, lysine, proline, leucine, hydroxyproline, and hydroxylysine). The main finding of this study was that amino acid concentrations in BB made to a standardized recipe were significantly lower for hydroxyproline, glycine, and proline (p = .003) and hydroxylysine, leucine, and lysine (p = .004) than those provided by a potentially therapeutic dose (20 g) of reference collagen supplements (p > .05). There was a large variability in the amino acid content of BB made to nonstandardized recipes, with the highest levels of all amino acids found within the café-prepared varieties. For standardized preparations, commercial BBs were lower in all amino acids than the self-prepared varieties. There were no differences (p > .05) in the amino acid content of different batches of BB when prepared according to a standardized recipe. If the intake of collagen precursors is proven to support the synthesis of new collagen in vivo, it is unlikely that BB can provide a consistently reliable source of key amino acids. Research on the provision of key amino acids from dietary sources should continue to focus on the standard sources currently being researched.


Crystallographic studies of the catalytic and a second site in fumarase C from Escherichia coli.
Fumarase C catalyzes the stereospecific interconversion of fumarate to L-malate as part of the metabolic citric acid or Kreb's cycle. The recent three-dimensional structure of fumarase C from Escherichia coli has identified a binding site for anions which is generated by side chains from three of the four subunits within the tetramer (Weaver et al., 1995). These same side chains are found in the three most highly conserved regions within the class II fumarase superfamily. The site was initially characterized by crystallographic studies through the binding of a heavy atom derivative, tungstate. A number of additional crystallographic structures using fumarase crystals with bound inhibitors and poor substrates have now been studied. The new structures have both confirmed the originally proposed active site, site A, and led to the discovery of a novel second binding site that is structurally nearby, site B. Site A utilizes a combination of residues, including H188, T187, K324, N326, T100, N141, S139, and S140, to form direct hydrogen bonds to each of the inhibitors. The A-site has been demonstrated by studying crystalline fumarase with the bound competitive inhibitors-citrate and 1,2,4,5-benzenetetracarboxylic acid. The crystal structure of fumarase C with beta-(trimethylsilyl)maleate, a cis substrate for fumarase, has led to the discovery of the second site or B-site. Sites A and B have different properties in terms of their three-dimensional structures. Site B, for example, is formed by atoms from only one of the subunits within the tetramer and mainly by atoms from a pi-helix between residues H129 through N135. The crystal structures show that the two locations are separated by approximately 12 A. A highly coordinated buried water molecule is also found at the active or A-site. The high-resolution crystal structures describe both sites, and atoms near the A-site are used to propose a likely enzyme/substrate complex.


First Report of Bark Cracking of Koelreuteria bipinnata var integrifoliola Caused by Lasiodiplodia theobromae in China.
Koelreuteria bipinnata var integrifoliola is becoming a popular urban green tree in Ningbo City, Zhejiang Province, China, because of its adaptation ability to local conditions, fast growth, and beautiful appearance. A survey conducted from 2007 to 2010 revealed serious bark cracking on greenbelt trees approximately 15 to 16 years old that had been transplanted 5 to 6 years ago. Bark cracks increased in size over time, extending into the phloem and leading to extensive areas of bark loss with discoloration of the underlying xylem. Symptomatic trees had fewer new shoots in spring; many wilted and died in summer. Root rot was not observed in the withered trees but large light brown lesions were observed on cross sections of the main stem, each with a dark brown outer margin. In a September 2009 survey, 95% of symptomatic trees had stem lesions more than 50 cm long. Pieces of xylem (2 × 2 × 1 mm thick) were obtained from the margin of lesions surface sterilized using 0.1% mercuric chloride for 30 s, washed in sterile distilled water, and placed on 2% potato dextrose agar (PDA) at 28°C for 2 days. The fungus was then isolated and 12 colonies were obtrained. Three isolates KL-1-2, KL-3-2, and KL-4-3 were incubated on 2% PDA at 28°C for 30 days to produce spores. On PDA, the colonies were circular or near circular with irregular gray edges turning black green or black. The fungus also produced abundant aerial hyphae that were villous, septate, and irregular branched. Conidia were elliptical (or rounded) and hyaline when immature, becoming dark brown and septate longitudinally when mature and ranged from 23.2 to 27.0 × 10.8 to 16.2 μm (average 25.3 × 13.6 μm), similar to Lasiodiplodia theobromae (Patouillard) Griffon =Botryodiplodia theobromae Pa.t, Botryosphaeria rhodina (Berkeley & Curtis) von Arx (2). DNA extraction directly from the mycelium of KL-1-2, KL-3-2, and KL-4-3 was performed after 10 days' growth on PDA (1). The identities of the three isolates were confirmed by ITS1-5.8S-ITS2 rDNA sequence (GenBank Accession Nos. JN681172, JQ894322, and JQ894323, respectively) analysis that showed 99%, 100%, and 100% sequence similarity to L. theobromae xsd08006 (Accession No. FJ478102), L. theobromae PD20 (Accession No. GU251120), and L. theobromae xsd08008 (Accession No. EU918707), respectively. Pathogenicity tests were performed on 20 five-year-old K. bipinnata var integrifoliola plants by placing mycelia plugs of isolate KL-1-2 (10 × 10 mm) on the main trunk after wounding with a metal needle. Control plants received PDA plugs without mycelium. After inoculation, humidity was maintained using wet absorbent cotton and PE wrap film. Stem bark and phloem cracking was observed after 60 days on 85% of inoculated plants; 30% of those trees also had xylem discoloration. Symptoms were similar to those with natural infection. Control plants remained symptomless. The same fungus was reisolated from the brown xylem of inoculated plants. To our knowledge, this is the first report of bark cracking of K. bipinnata var integrifoliola caused by L. theobromae in China. References: (1) M.-J. Côté et al. Plant Dis. 88:1219, 2004. (2) G. Fu et al. Australas. Plant Dis. Notes 2:75, 2007.


Evidence for hepatic formation, export and covalent binding of reactive naphthalene metabolites in extrahepatic tissues in vivo.
Previous studies have shown that cytochrome P-450-mediated metabolism of naphthalene results in dose-dependent bronchiolar necrosis in mice and in the formation of reactive metabolites which deplete reduced glutathione and become bound covalently to tissue macromolecules. The finding that pulmonary glutathione levels were nearly totally depleted after large doses of naphthalene suggested that hepatic formation of reactive metabolites may contribute substantially to glutathione depletion and covalent binding in extrahepatic tissues. This possibility has been supported by several new lines of evidence: 1) similar levels of covalent binding were observed in lung, liver and kidney in vivo, yet the rate of kidney microsomal metabolic activation of naphthalene was much lower than in liver or lung; 2) phenobarbital pretreatment markedly increased in vivo covalent binding in lung, liver and kidney and increased hepatic but decreased pulmonary microsomal covalent binding; 3) 3-methylcholanthrene pretreatment resulted in slightly increased levels of covalent binding in lung, liver and kidney yet decreased pulmonary microsomal covalent binding; 4) administration of p-xylene, at doses which selectively decreased pulmonary microsomal metabolism of biphenyl (4-hydroxylation) and naphthalene (to reactive metabolites), decreased in vivo covalent binding in liver and kidney to the same extent as lung after [14C]naphthalene; and 5) pretreatment with buthionine sulfoximine preferentially depleted hepatic and renal but not pulmonary glutathione levels and markedly increased covalent binding in all three tissues. The severity of naphthalene-induced bronchiolar damage was unaffected by pretreatment with phenobarbital, 3-methylcholanthrene or p-xylene but was increased by prior administration of buthionine sulfoximine. These studies suggest that a portion of the reactive metabolites which deplete glutathione and bind covalently in extrahepatic tissues originate in the liver. Whether these circulating metabolites play a role in naphthalene-induced pulmonary bronchiolar damage is not clear.


Transsexualism: a review of etiology, diagnosis and treatment.
Transsexualism is considered to be the extreme end of the spectrum of gender identity disorders characterized by, among other things, a pursuit of sex reassignment surgery (SRS). The origins of transsexualism are still largely unclear. A first indication of anatomic brain differences between transsexuals and nontranssexuals has been found. Also, certain parental (rearing) factors seem to be associated with transsexualism. Some contradictory findings regarding etiology, psychopathology and success of SRS seem to be related to the fact that certain subtypes of transsexuals follow different developmental routes. The observations that psychotherapy is not helpful in altering a crystallized cross-gender identity and that certain transsexuals do not show severe psychopathology has led clinicians to adopt sex reassignment as a treatment option. In many countries, transsexuals are now treated according to the Standards of Care of the Harry Benjamin International Gender Dysphoria Association, a professional organization in the field of transsexualism. Research on postoperative functioning of transsexuals does not allow for unequivocal conclusions, but there is little doubt that sex reassignment substantially alleviates the suffering of transsexuals. However, SRS is no panacea. Psychotherapy may be needed to help transsexuals in adapting to the new situation or in dealing with issues that could not be addressed before treatment.


Non-invasive blood pressure measurement in mice.
Hypertension is a leading cause of heart attack, stroke, and kidney failure and represents a serious medical issue worldwide. The genetic basis of hypertension is well-established, but few causal genes have been identified thus far. Non-invasive blood pressure measurements are a critical component of high-throughput genetic studies to identify genes controlling blood pressure. Whereas this technique is fairly routine for blood pressure measurements in rats, non-invasive blood pressure measurement in mice has proven to be more challenging. This chapter describes an experimental protocol measuring blood pressure in mice using a CODA non-invasive blood pressure monitoring system. This method enables accurate blood pressure phenotyping in mice for linkage or mutagenesis studies, as well as for other experiments requiring high-throughput blood pressure measurement.


Biologic brachytherapy: ex vivo transduction of microvascular beds for efficient, targeted gene therapy.
Gene therapy for cancer holds enormous therapeutic promise, but its clinical application has been limited by the inability to achieve targeted, high-level transgene expression with limited systemic toxicity. The authors have developed a novel method for delivering genes to microvascular free flaps (commonly used during reconstructive surgery) to avoid these problems. During the finite period in which a free flap is separated from the host (ex vivo), it can be perfused with extremely high titers of genetic material through the afferent artery, resulting in efficient transduction of the tissue. Before reanastomosis, unincorporated genetic material is flushed from the flap, minimizing systemic toxicity. In a rodent model using an adenoviral vector containing the lacZ reporter gene, high regional expression of beta-galactosidase was achieved in all the different cells in a microvascular free flap. Moreover, no beta-galactosidase staining was observed outside of the transduced flap, and viral sequence was undetectable by polymerase chain reaction analysis in other tissues. Further analysis confirmed that high-level transgene expression was precisely localized to the explanted tissue, with no collateral transduction. Targeting gene delivery with minimal systemic toxicity is essential for successful gene therapy. This form of "biological brachytherapy" provides a new opportunity to deliver targeted therapeutic transgenes to patients undergoing reconstructive surgery and allows microvascular free flaps to perform therapeutic and reconstructive functions.


Pharmacokinetics of cefazolin applied topically to the surgical wound.
Topical application of antibiotics is used in the prophylaxis of postoperative surgical infections. However, whether topically applied antibiotics remain primarily in the surgical wound fluid or are systemically absorbed is uncertain. The pharmacokinetics of topically applied cefazolin were studied in a canine model that allowed simultaneous determination of serum and wound fluid antibiotic concentrations. Topical administration of cefazolin resulted in high antibiotic concentrations in the wound fluid for prolonged periods and rapid systemic absorption. Bioavailability after topical administration was 95%. Within 1 hour, the serum concentrations after topical administration equaled the serum concentrations after intravenous administration, and the concentration time curves declined in parallel. In wound fluid, the mean time above the susceptibility break point minimum inhibitory concentration after topical administration of cefazolin was 5.76 hours compared with the estimated time above the minimum inhibitory concentration of 2.55 hours after intravenous administration.


Symptom severity moderates the outcome of attention bias modification for depression: An exploratory study.
A recent meta-analysis has questioned the relevance of attention bias modification (ABM) for depression outcomes. However, there might be patient characteristics not yet accounted for, that are relevant to the outcome. In the context of personalized treatment, the lack of moderator studies have limited the potential for matching ABM-treatment to individual patient characteristics. Subjects (N = 301) were randomly assigned 1:1 to receive either active or placebo Attention Bias Modification (ABM) twice daily for 14 days in a double-blind design (placebo n = 148; ABM n = 153). The outcome was change in symptoms based on the Hamilton Depression Rating Scale (HDRS). Moderator variables were self-reported depression (Beck Depression Inventory-II; BDI-II), anxiety (Beck Anxiety Inventory; BAI) and attentional bias (AB) assessed at baseline. This trial was registered with ClinicalTrials.gov, number NCT02658682. Only BAI (p for interaction = .01, Bootstrap 95% CI [0.046, 0.337]) moderated the effects of ABM on change in clinician rated depressive symptoms. Interactions were significant for BAI scores ≥8. The relative effect of the intervention increased with the highest symptom load. ABM was not effective in patients with the lowest symptom load. Future research should validate this finding and continue investigating moderators of the ABM-intervention to further enhance personalization of treatment to individual symptom characteristics.


Microbial community transcriptomes reveal microbes and metabolic pathways associated with dissolved organic matter turnover in the sea.
Marine dissolved organic matter (DOM) contains as much carbon as the Earth's atmosphere, and represents a critical component of the global carbon cycle. To better define microbial processes and activities associated with marine DOM cycling, we analyzed genomic and transcriptional responses of microbial communities to high-molecular-weight DOM (HMWDOM) addition. The cell density in the unamended control remained constant, with very few transcript categories exhibiting significant differences over time. In contrast, the DOM-amended microcosm doubled in cell numbers over 27 h, and a variety of HMWDOM-stimulated transcripts from different taxa were observed at all time points measured relative to the control. Transcripts significantly enriched in the HMWDOM treatment included those associated with two-component sensor systems, phosphate and nitrogen assimilation, chemotaxis, and motility. Transcripts from Idiomarina and Alteromonas spp., the most highly represented taxa at the early time points, included those encoding TonB-associated transporters, nitrogen assimilation genes, fatty acid catabolism genes, and TCA cycle enzymes. At the final time point, Methylophaga rRNA and non-rRNA transcripts dominated the HMWDOM-amended microcosm, and included gene transcripts associated with both assimilatory and dissimilatory single-carbon compound utilization. The data indicated specific resource partitioning of DOM by different bacterial species, which results in a temporal succession of taxa, metabolic pathways, and chemical transformations associated with HMWDOM turnover. These findings suggest that coordinated, cooperative activities of a variety of bacterial "specialists" may be critical in the cycling of marine DOM, emphasizing the importance of microbial community dynamics in the global carbon cycle.


Echidna venom gland transcriptome provides insights into the evolution of monotreme venom.
Monotremes (echidna and platypus) are egg-laying mammals. One of their most unique characteristic is that males have venom/crural glands that are seasonally active. Male platypuses produce venom during the breeding season, delivered via spurs, to aid in competition against other males. Echidnas are not able to erect their spurs, but a milky secretion is produced by the gland during the breeding season. The function and molecular composition of echidna venom is as yet unknown. Hence, we compared the deeply sequenced transcriptome of an in-season echidna crural gland to that of a platypus and searched for putative venom genes to provide clues into the function of echidna venom and the evolutionary history of monotreme venom. We found that the echidna venom gland transcriptome was markedly different from the platypus with no correlation between the top 50 most highly expressed genes. Four peptides found in the venom of the platypus were detected in the echidna transcriptome. However, these genes were not highly expressed in echidna, suggesting that they are the remnants of the evolutionary history of the ancestral venom gland. Gene ontology terms associated with the top 100 most highly expressed genes in echidna, showed functional terms associated with steroidal and fatty acid production, suggesting that echidna "venom" may play a role in scent communication during the breeding season. The loss of the ability to erect the spur and other unknown evolutionary forces acting in the echidna lineage resulted in the gradual decay of venom components and the evolution of a new role for the crural gland.


Adaptation reveals that facial expression averaging occurs during rapid serial presentation.
How do we interpret the rapidly changing visual stimuli we encounter? How does our past visual experience shape our perception? Recent work has suggested that our visual system is able to interpret multiple faces presented temporally via integration or ensemble coding. Visual adaptation is widely used to probe such short term plasticity. Here we use an adaptation paradigm to investigate whether integration or averaging of emotional faces occurs during a rapid serial visual presentation (RSVP). In four experiments, we tested whether the RSVP of distinct emotional faces could induce adaptation aftereffects and whether these aftereffects were of similar magnitudes as their statistically averaged face. Experiment 1 showed that the RSVP faces could generate significant facial expression aftereffects (FEAs) across happy and sad emotions. Experiment 2 revealed that the magnitudes of the FEAs from RSVP faces and their paired average faces were comparable and significantly correlated. Experiment 3 showed that the FEAs depended on the mean emotion of the face stream, regardless of variations in emotion or the temporal frequency of the stream. Experiment 4 further indicated that the emotion of the average face of the stream, but not the emotion of individual faces matched for identity to the test faces, determined the FEAs. Together, our results suggest that the visual system interprets rapidly presented faces by ensemble coding, and thus implies the formation of a facial expression norm in face space.


Can overnight portable pulse oximetry be used to stratify obstructive sleep apnea risk in infants? A correlation analysis.
There is limited evidence on the accuracy of oximetry in the evaluation of infant obstructive sleep apnea (OSA). We aimed to determine the utility of overnight oximetry to stratify infants at risk for OSA, to determine urgency for definitive screening with an overnight in-laboratory polysomnogram (PSG). Retrospective single-institution cohort study of infants undergoing PSG and a separate overnight oximetry over an 8-year period. Correlations, using oximetry in both in-hospital (attended) or at-home (unattended) settings, for ODI410 (decrease in oxygen saturation ≥ 4% from baseline, duration ≥ 10 seconds) and ODI40 (duration > 0 second) with the obstructive apnea-hypopnea index (AHIo) were obtained. The area under the curve was calculated, and sensitivity and specificity values have been presented as receiver operating characteristic curves. Thirty-eight infants were included. The mean (SD) age (months) was 5.7 (3.9) at diagnostic PSG and 5.5 (3.7) at the time of oximetry. The mean AHIo for the entire cohort was 6.7 (6.2). The mean (SD) ODI40 was 8.6 (9.0) and the mean (SD) ODI410 was 5.4 (5.1).The correlation between ODI and AHIo was statistically significant for the cohort (ODI40 vs. AHIo [r = .59, P < .001] and ODI410 vs AHIo [r = .55, P = .0003]). Using an ODI40 cutoff of 3, the sensitivity, specificity, negative predictive value and positive predictive value for diagnosing OSA was: 86%, 40%, 50%, and 80% respectively for an AHIo greater than 2, and 100%, 35%, 100%, and 58% respectively for an AHIo greater than or equal to 5. There is a significant positive correlation between the ODI4 obtained from oximetry and the AHIo obtained from PSG in infants at risk for OSA. An ODI40 greater than 3 may be useful to stratify infants at risk for moderate to severe OSA when used in attended (in-hospital) or unattended (in-home) settings.


One- or two-stage bilateral total hip replacement.
It is not clear whether bilateral hip replacement should be done in 1 or 2 stages. The total number of total hip replacements (THRs) done in our center between 1989 and 1995 was approximately 4,000. The number of hips that were bilateral was 404, or 9% of the total number of THRs performed during this time period. Of these bilateral hip replacements, 190 (95 patients) were done as a 1-stage procedure, whereas 214 (107 patients) were done in 2 stages with 2 to 24 months in between the operations. In contrast to previous studies, there were no significant preoperative differences between the 2 groups of patients having 1-stage or 2-stage THRs, and, in particular, the comorbidity assessed by the American Society of Anesthesiologists (ASA) grade was not significantly different. Our results demonstrate that, in our patient population, bilateral THR was equally safe whether performed as a 1-stage or 2-stage procedure. This was the case in the low-risk (ASA 1 and 2) and high-risk (ASA 3 and 4) patient subgroups. One-stage bilateral THR is cheaper and involves less time in the hospital.


Expanding the Malaria Antibody Toolkit: Development and Characterisation of Plasmodium falciparum RH5, CyRPA, and CSP Recombinant Human Monoclonal Antibodies.
Malaria, an infection caused by apicomplexan parasites of the genus Plasmodium, continues to exact a significant toll on public health with over 200 million cases world-wide, and annual deaths in excess of 600,000. Considerable progress has been made to reduce malaria burden in endemic countries in the last two decades. However, parasite and mosquito resistance to frontline chemotherapies and insecticides, respectively, highlights the continuing need for the development of safe and effective vaccines. Here we describe the development of recombinant human antibodies to three target proteins from Plasmodium falciparum: reticulocyte binding protein homologue 5 (PfRH5), cysteine-rich protective antigen (PfCyRPA), and circumsporozoite protein (PfCSP). All three proteins are key targets in the development of vaccines for blood-stage or pre-erythrocytic stage infections. We have developed potent anti-PfRH5, PfCyRPA and PfCSP monoclonal antibodies that will prove useful tools for the standardisation of assays in preclinical research and the assessment of these antigens in clinical trials. We have generated some very potent anti-PfRH5 and anti-PfCyRPA antibodies with some clones >200 times more potent than the polyclonal anti-AMA-1 antibodies used for the evaluation of blood stage antigens. While the monoclonal and polyclonal antibodies are not directly comparable, the data provide evidence that these new antibodies are very good at blocking invasion. These antibodies will therefore provide a valuable resource and have potential as biological standards to help harmonise pre-clinical malaria research.


Risk Assessment Tools and Data-Driven Approaches for Predicting and Preventing Suicidal Behavior.
Risk assessment of suicidal behavior is a time-consuming but notoriously inaccurate activity for mental health services globally. In the last 50 years a large number of tools have been designed for suicide risk assessment, and tested in a wide variety of populations, but studies show that these tools suffer from low positive predictive values. More recently, advances in research fields such as machine learning and natural language processing applied on large datasets have shown promising results for health care, and may enable an important shift in advancing precision medicine. In this conceptual review, we discuss established risk assessment tools and examples of novel data-driven approaches that have been used for identification of suicidal behavior and risk. We provide a perspective on the strengths and weaknesses of these applications to mental health-related data, and suggest research directions to enable improvement in clinical practice.


The sexual behaviour of bisexual men in relation to HIV transmission.
Relatively little is known about the behaviour of bisexual men which may help in assessing their role in HIV transmission. A sample of 60 behaviourally bisexual men were asked about their sexual behaviour with male and female partners and their perceptions of risk of HIV infection. Only a minority of men engaged in unprotected anal sex with their male partners while two thirds had unprotected vaginal sex with their female partners. This asymmetrical pattern of sexual behaviour reflects a differential perception of risk of HIV infection with male and female partners. A quarter of the men had unprotected penetrative sex with both male and female partners in the previous year. The pattern of risk behaviour varied amongst men living in gay, heterosexual or bisexual contexts.


Reflections on a methodology for analyzing professional practice and its possible use in nursing.
In order for nursing work not to become mechanistic, the team needs to develop a critical eye to its knowledge, its know-how and its know-to-be. The training of nursing professionals from a critical and reflective perspective motivated this study, which was based on the theoretical reference and the experience of the authors with use of the analytical methodology of the Groupe Entraînement de Analyze de Situations Éducatives (Training Group to Analyze Educational Situations) professional practice. This methodology aims to analyze professional situations in a group with the presence of a facilitator. The objective is to analyze the contributions of the Groupe Entraînement de Analyze de Situations Éducatives to train reflective professionals in nursing. The steps, rules of operation and the role of the participants are explained in order to promote a safe environment for all, enabling group members to develop hypotheses to understand the situation and develop analysis and reflection skills. It is concluded that this methodology offers the opportunity to open a space for reflective thinking, respecting the different roles and times intended for the speech of each participant, building their professional identity.


Physically active children with epilepsy have good objective sleep duration and efficiency despite subjective reports of fatigue and sleep problems.
The aim of this study was to longitudinally characterize in children with epilepsy the objective and subjective sleep quality and the relationship between increased physical activity and sleep as well as measures of psychosocial well-being. Baseline physical activity and sleep were established in children with epilepsy over four weeks, prior to a 12-week exercise intervention (weekly meeting with exercise counselor). Participants continuously wore a wrist pedometer (Fitbit Flex®) to capture daily number of steps, sleep efficiency, and total sleep time. The Early Childhood Epilepsy Severity Scale (E-Chess) assessed baseline epilepsy severity. Subjective sleep quality (Children's Sleep Habits Questionnaire, CSHQ), quality of life (KIDSCREEN-27; Pediatric Quality of Life Inventory, PedsQL™, 4.0 Core), fatigue (PedsQL™ Multidimensional Fatigue Scale), depression (Children's Depression Inventory-Short), and anxiety (Multidimensional Anxiety Scale for Children) were assessed pre- and post-interventions. Our cohort of 22 children with epilepsy aged 8-14 years was similarly active to peers (11,271 ± 3189 mean steps per day) and displayed normal sleeping patterns (mean sleep efficiency: 87.4% ± 3.08 and mean total sleep time: 521 ± 30.4). Epilepsy severity assessed by E-Chess was low to moderate (median baseline E-Chess score of 6, interquartile range: 5-7). Study outcomes did not change with the intervention. Older children and those with lower baseline activity were more likely to increase their activity during the intervention. Changes in physical activity were not associated with changes in sleep outcomes when accounting for age, sex, and baseline E-Chess score. Subjective sleep quality marginally improved with the intervention (CSHQ total score: 44.5 ± 5.8 at baseline and 41.6 ± 7.2 at the end of study, p = 0.05). Quality of life, fatigue, depression, and anxiety did not change with the intervention (p = 0.55, 0.60, 0.12, and 0.69, respectively). Children with epilepsy who are as active as peers without epilepsy have good objective measures of sleep despite self-reported fatigue and parent-reported sleep problems. The physical activity of initially less active and older children with epilepsy may benefit from an exercise counseling intervention.


Genome-wide copy number analysis of Hodgkin Reed-Sternberg cells identifies recurrent imbalances with correlations to treatment outcome.
In classical Hodgkin lymphoma (cHL) the mechanisms underlying primary refractory disease and relapse remain unknown. To gain further insight into cHL pathogenesis and genomic changes linked to treatment response, we studied 53 cHL patients by array comparative genomic hybridization, including 23 patients whose primary treatment failed, using DNA from microdissected HRS cells. Copy number alterations found in more than 20% of cases included gains of 2p, 9p, 16p, 17q, 19q, 20q, and losses of 6q, 11q, and 13q. We identified at high resolution recurrent changes defining minimally gained and lost regions harboring genes involved in nuclear factor kappaB signaling, such as REL, IKBKB, CD40, and MAP3K14. Gains of chromosome 16p11.2-13.3 were significantly more frequent in pretreatment and relapse biopsies of unresponsive patients and were associated with shortened disease-specific survival (P = .028). In the therapy-resistant HL cell line KMH2, we found genomic gains and overexpression of the multidrug resistance gene ABCC1 mapping to cytoband 16p13.11. We show that doxorubicin exposure to KMH2 induces ABCC1 expression and that siRNA silencing of ABCC1 sensitizes KMH2 cells to doxorubicin toxicity in vitro, suggesting that overexpression of ABCC1 contributes to the drug resistance phenotype found in KMH2.


[Epidemiology of overweight and obesity and its determinants in Belo Horizonte (MG), Brazil: a cross-sectional population-based study].
To determine the prevalence of overweight and obesity and to study potential risk factors for these conditions in persons over 18 years of age in the metropolitan area of Belo Horizonte, Brazil. Data were obtained from a population-based study conducted in 1996 and 1997 by the Brazilian Institute of Geography and Statistics (Instituto Brasileiro de Geografia e Estatística, IBGE) with a random sample of 1 105 individuals. Height and weight were measured by trained personnel. Overweight (body mass index [BMI] > or = 25.0 kg/m2) and obesity (BMI > or = 30.0 kg/m2) were the dependent variables. Bivariate analysis and multivariate logistic regression were used to identify potential risk factors for overweight and obesity. The prevalence of overweight and obesity was 31.1% (95% confidence interval 195% CI]: 27.3 to 35.2) and 5.7% (95% CI: 4.0 to 8.1), respectively, in men, and 25.9% (95% CI: 22.4 to 29.8) and 14.7% (95% CI: 11.9 to 17.9), respectively, in women. Multivariate analysis revealed that age and marital status were independent risk factors for overweight, whereas age, sex, and education were independent risk factors for obesity. Being female and well-educated showed a protective effect against overweight (odds ratio [OR] = 0.52; 95% CI: 0.33 to 0.83), but not against obesity (OR = 3.01; IC 95%: 1.14 to 7.94). Women with low education had a significantly greater risk (OR = 5.95; 95%CI: 2.51 to 14.12) of developing obesity than men having a high educational level. These results suggest that overweight and obesity may be serious public health problems in the metropolitan area of Belo Horizonte, Brazil. It is important to carry out further studies in order to explore the potential relationship between overweight and obesity on the one hand, and behavioral variables, such as smoking and alcohol consumption, on the other.


Heterogeneity of long-history migration predicts smiling, laughter and positive emotion across the globe and within the United States.
Recent findings demonstrate that heterogeneity of long-history migration predicts present-day emotion behaviors and norms. Residents of countries characterized by high ancestral diversity display emotion expressions that are easier to decode by observers, endorse norms of higher emotion expressivity, and smile more in response to certain stimuli than residents of countries that lack ancestral diversity. We build on the extant findings and investigate historical heterogeneity as a predictor of daily smiling, laughter, and positive emotion across the world's countries and the states of the United States. Study 1 finds that historical heterogeneity is positively associated with self-reports of smiling, laughter, and positive emotions in the Gallup World Poll when controlling for GDP and present-day population diversity. Study 2 extends the findings to effects of long-history migration within the United States. We estimated the average percentage of foreign-born citizens in each state between 1850 and 2010 based on US Census information as an indicator of historical heterogeneity. Consistent with the world findings of Study 1, historical heterogeneity predicted smiling, laughter, and positive, but not negative, emotion. The relationships remained significant when controlling for per capita income and present-day population diversity of each state. Together, the findings further demonstrate the important role of long-history migration in shaping emotion cultures of countries and states, which persist beyond the original socio-ecological conditions, and open promising avenues for cross-cultural research.


Postmarket surveillance of Menicon Z rigid gas-permeable contact lenses for up to 30 days continuous wear in the United States.
To evaluate the incidence of microbial keratitis associated with the use of the Menicon Z rigid gas-permeable (RGP) contact lenses (CLs) for up to 30 days continuous wear (CW). For this study, CW was defined as wearing CLs in an extended-wear modality with an average wearing time of 22 days or more. Microbial keratitis was the sole endpoint of the study safety evaluation. Subjects who were interested in RGP lens CW were recruited for the study through 33 investigational practices. To be eligible for the study, subjects must have worn the Menicon Z lens for CW for at least 1.5 months with an average CW time of 22 days or more. Eligible wearers were then contacted at 6-month intervals for up to 24 months after enrollment to determine their typical wearing schedules and continuance of wear and to detect any problems that would be indicative of microbial keratitis. Discontinuation from the study was defined as a discontinuation of contact with or CL wear by the study participants. Study volunteers could be discontinued from the study at 6, 12, 18, or 24 months if they could not be contacted (lost to follow-up); if they were not wearing their CL for CW; or at their own request. Wearing time data collected for individuals discontinued from the study was included in the wearing time analysis. The study population included 507 participants, of whom 173 had discontinued by the end of study at 24 months. Lack of 22 days or more of CW was the most frequently cited reason for discontinuation and accounted for 121 of the 173 subject discontinuations. Lost to follow-up, study participant choice, and other reasons accounted for the remaining 52 subject discontinuations. Of the reported wearing times over the duration of the study, 1,275 of the 1,527 contacts resulted in reported wearing times of 22 days or more of CW. There were no findings of microbial keratitis during the study. Overall, there was a low reported rate of complications for the enrolled study participants. On the basis of this study, CW of the Menicon Z hyper-Dk/t RGP lens provides a safe and full-time vision correction alternative for patients who can adapt to RGP lens wear.


Brief report: Under-representation of African americans in autism genetic research: a rationale for inclusion of subjects representing diverse family structures.
African American children with autism are seriously under-represented in existing genetic registries and biomedical research studies of autism. We estimated the number of African American children with autism in the St. Louis region using CDC surveillance data and present the outcomes of a concerted effort to enroll approximately one-third of that population into either of two large national genetic autism registries. The results revealed that even after traditional barriers to research participation were addressed and all contacted families expressed a willingness to participate, 67% of the reachable families were disqualified from participation because of family structure alone. Comprehensive efforts-including expansion of eligibility to families of diverse structure-are warranted to facilitate the inclusion of African American children in biomedical research.


Ser/Thr-rich domains are associated with genetic variation and morphogenesis in Saccharomyces cerevisiae.
Proteins containing regions of amino acid bias are often found in eukaryotes and are associated with particular functional groups. We have carried out a genomic analysis of yeast proteins containing regions with a significant bias of Ser and Thr residues. Our findings reveal that a high number are cell surface proteins or regulatory proteins involved in many aspects of cell differentiation. Furthermore, in Saccharomyces-related species, a highly significant correlation exists between the frequency of Ser-rich regions and DNA repeats, indicating that their generation may rely on similar factors. Cluster analysis shows that Ser/Thr-rich regions, located within the tandem repeats of cell surface proteins, are encoded to an increased frequency by UCU (Ser) and ACU/ACC (Thr), implying that mutational events that generate iterations could involve these codons. Replication slippage is proposed to be a contributing factor, as mounting evidence suggests that repeat generation in cell surface proteins can occur independently of meiosis. To reinforce this argument, we have discovered a premeiotic association between Mre11p, a nuclease involved in DNA repair, and ORFs encoding Ser/Thr-rich regions. Several macromolecules involved in the glycosylation and phosphorylation of proteins require Ser and Thr residues as binding sites. Ser/Thr-rich regions, through polymorphisms, are associated with the evolution of functional sites, particularly in providing motifs for glycosylation and phosphorylation. These results point to a Ser/Thr-biased somatic mutation mechanism that contributes to rapid evolution in yeast.


Evidence for adaptive diet-induced thermogenesis in man during intravenous nutrition with hypertonic glucose.
This study was designed to investigate the thermogenic effect of intravenously administered nutrition with glucose (given a fixed nitrogen intake of 12.5 g daily as amino acids) as the principal source of energy. The protocol was designed so that each patient received their energy intake in five consecutive periods of 3 days with intakes ranging from 6650 to 17,100 kJ/day with increments or decrements of 2600 kJ. Thermogenesis from administered glucose was evident between levels of energy supply of 6650 kJ/day and 17,100 kJ/day. The progressive rise in oxygen consumption and carbon dioxide production accounted for a total of 31% of the additional glucose which was administered. The net rate of fat synthesis from glucose reached a maximum 147 g/day at an energy supply of 14,500 kJ/day. This study suggests that both fat synthesis and the associated obligatory thermogenesis is the main component of diet-induced thermogenesis in response to glucose intakes in excess of 150 kJ day-1 kg-1. If the energy cost of fat synthesis (fat associated obligatory thermogenesis) is taken to be 22% of the total energy of the increase in glucose supplied, then only 9% (31-22%) of the glucose can be accounted for by adaptive thermogenesis.


Measuring pain intensity in patients with neck pain: does it matter how you do it?
The aim of this study was to investigate whether variations in the way that pain intensity is measured in patients with neck pain influences the magnitude of pain ratings. The study uses data from 3 longitudinal studies (n = 361 at baseline) on people with neck pain due to whiplash injuries. Pain measures included verbal rating scales, numerical rating scales and a visual analog scale. Different measures asked patient to rate current pain, average pain over 24 hours, over 1 week, or over 4 weeks. Scores were converted to a 0-100 scale and tracked over time, correlations between measures were calculated. Mixed models regression was used to explore the factors which influenced the differences between scores on the measures. Scores on the different measures were significantly different from each other in each dataset (P < 0.02). The effect of recall period was significant in all datasets and the effect of number of response options was significant in 2 of 3 datasets. Pain intensity ratings appear to be sensitive to method of measurement. It is likely the length of recall time (eg, pain today vs. average pain over 4 weeks) has a significant influence on pain ratings. The influence of number of response options is less certain. Systematic reviewers should not uncritically rescale and pool absolute pain scores from instruments with varying scale descriptors or recall periods.


Dysphagia in Friedreich Ataxia.
The objective of the study was to comprehensively characterise dysphagia in Friedreich ataxia (FRDA) and identify predictors of penetration/aspiration during swallowing. We also investigated the psychosocial impact of dysphagia on individuals with FRDA. Sixty participants with FRDA were screened for dysphagia using a swallowing quality of life questionnaire (Swal-QOL) and case history. Individuals reporting dysphagia underwent a standardised oromotor assessment (Frenchay Dysarthria Assessment, 2, FDA-2) and videofluoroscopic study of swallowing (VFSS). Data were correlated with disease parameters (age at symptom onset, age at assessment, disease duration, FXN intron 1 GAA repeat sizes, and Friedreich Ataxia Rating Scale (FARS) score). Predictors of airway penetration/aspiration were explored using logistic regression analysis. Ninety-eight percent (59/60) of participants reported dysphagia, of whom 35 (58.3%) underwent FDA-2 assessment, and 38 (63.3%) underwent VFSS. Laryngeal, respiratory, and tongue dysfunction was observed on the FDA-2. A Penetration-Aspiration Scale score above 3 (deemed significant airway compromise based on non-clinical groups) was observed on at least one consistency in 13/38 (34.2%) participants. All of those who aspirated (10/38, 26.3%) did so silently, with no overt signs of airway entry such as reflexive cough. Significant correlations were observed between dysphagic symptoms and disease duration and severity. No reliable predictors of penetration or aspiration were identified. Oropharyngeal dysphagia is commonly present in individuals with FRDA and worsens with disease duration and severity. Individuals with FRDA are at risk of aspiration at any stage of the disease and should be reviewed regularly. Instrumental analysis remains the only reliable method to detect aspiration in this population. Dysphagia significantly affects the quality of life of individuals with FRDA.


The chemokinetic inhibitory factor (CIF) in serum of CLL patients: correlation with infection propensity and disease activity.
We have recently described the partial purification and characterization of a neutrophil migration inhibitory activity present in serum from patients with chronic lymphocytic leukaemia (CLL). This new lymphokine, the chemokinetic inhibitory factor (CIF), is produced by B-CLL cells. It is a heat-labile glycoprotein of an approximate molecular weight (m. w.) of 30000. In this extended investigation 64/89 CLL-patients had CIF in their serum. CLL serum diluted to a concentration of 0.02% gave significantly decreased chemokinetic activity, suggesting that CIF is potent at very low concentrations. 31/89 patients had increased infection propensity. Significantly more patients with CIF in serum had infections compared to the group with normal susceptibility to infections. The combination of low Ig levels and CIF in serum discriminated even better between the infection-prone and non-infection-prone patients. CIF in serum was not correlated to tumour cell mass - estimated by Rai clinical staging - tumour progression or deoxythymidine kinase, S-TK, an enzyme that may reflect proliferating cells. The existence of this new lymphokine in serum seems to contribute to the increased susceptibility to infections seen in CLL patients.


Effects of citalopram infusion on the serotonin transporter binding of [11C]DASB in healthy controls.
The positron emission tomography (PET) ligand [(11)C]DASB is currently the most widely used imaging agent for quantitative studies of the serotonin transporter (SERT) in human brain. The aim of this work was to assess the effects of an intravenous infusion of 10 mg citalopram, a selective serotonin reuptake inhibitor (SSRI), before the PET scan on the kinetics of [(11)C]DASB in arterial plasma and in selected brain regions. Four healthy male volunteers underwent two PET scans with a mean of 523 MBq injected activity after either placebo or citalopram infusion in a randomised design. The citalopram infusion led to a substantial increase of the area under the curve of the metabolite-corrected arterial plasma input function. Total volumes of distribution V(T) were estimated applying the Logan plot to regional time-activity curves or by generating parametric maps with spectral analysis. A mean reduction of the cerebellar V(T) of 19% with Logan analysis and of 24% with spectral analysis was observed after citalopram infusion, which confirms previous findings of displaceable SERT ligand binding in cerebellar grey matter. The SERT occupancy for six target regions with moderate to high binding was 60% derived from BP(ND) and 69% derived from BP(P).


A pilot study of oral calcitriol (1,25-dihydroxyvitamin D3) for relapsing-remitting multiple sclerosis.
Epidemiological and ecological studies suggest links between vitamin D deficiency and increased multiple sclerosis (MS) prevalence. To evaluate the safety and tolerability of oral calcitriol therapy in an open label pilot study. 15 ambulatory patients with relapsing-remitting MS and at least one clinical relapse within the previous 12 months received oral calcitriol (target dose: 2.5 microg/d) for 48 weeks. Dietary calcium was restricted to 800 mg/d. Patients were monitored using frequent clinical and laboratory examinations, the expanded disability status scale (EDSS), and brain magnetic resonance imaging (MRI). Two patients withdrew because of symptomatic hypercalcaemia (serum calcium >3.35 mmol/l in each case) resulting from persistent dietary indiscretion. Two diet compliant patients required temporary dose adjustments for mild asymptomatic hypercalcaemia. Diet compliant patients experienced mild adverse effects. The on-study exacerbation rate (27%) was less than baseline. Four patients experienced five clinical relapses but only one patient worsened by >1 EDSS point. Brain MRI revealed enhancing lesions in five patients at baseline (33%) and in four (29%) at both 24 and 48 weeks. Oral calcitriol is safe and well tolerated for up to one year by diet compliant relapsing-remitting MS patients. Further study of vitamin D related mechanisms is warranted in MS.


Virulence properties and clonal structures of strains of Escherichia coli O119 serotypes.
A total of 110 Escherichia coli strains of serogroup O119 were examined for the presence of virulence properties characteristic of enteropathogenic E. coli (EPEC). Three virulence patterns were distinguished based on the detection of a chromosomal gene mediating intimate attachment (eaeA) and plasmid DNA involved in localized adherence (EAF and bfpA). The first pattern, represented by strains which hybridized with three gene probes, was the most common (68%) and, with a single exception, included only O119:H6 strains. Of these strains, 90% showed a typical localized adherence (LA) pattern in HEp-2 cells and 96% were positive for intimate attachment in a fluorescent-actin staining test with a 3-h incubation period. The second pattern was represented by strains which hybridized with the eaeA gene only. Most (89.5%) of these strains showed the LA phenotype but only after 6 h of incubation (LA-like phenotype). The third pattern consisted of strains which were positive for eaeA and bfpA but did not hybridize with the EAF probe. Most (80%) of these strains exhibited the LA-like phenotype. Analysis of several eaeA+ bfpA+ strains for the expression of the pilin subunit (BfpA) of the bundle-forming pili demonstrated that all LA strains expressed BfpA whereas the LA-like strains did not. The study of the clonal relationships, carried out by multilocus enzyme electrophoresis in 79 representative strains, defined 11 distinct electrophoretic types (ETs). ET1 included 66% of the strains, most of which displayed the eaeA+ bfpA+ EAF+ pattern and were serotyped as O119:H6 or O119:H-. The remaining 10 ETs were each represented by no more than five strains and, with the exception of ET8, included strains of a single serotype. The genetic relatedness of the ETs revealed two main clusters, with most strains in cluster A having the eaeA+ bfpA+ EAF+ combination and a O119:H6 serotype. Cluster B was represented by atypical EPEC strains with only the eaeA+ and the eaeA+ bfpA+ virulence pattern.


Identification of RUNX3 as a component of the MST/Hpo signaling pathway.
Recent genetic screens of fly mutants and molecular analysis have revealed that the Hippo (Hpo) pathway controls both cell proliferation and cell death. Deregulation of its human counterpart (the MST pathway) has been implicated in human cancers. However, how this pathway is linked with the known tumor suppressor network remains to be established. RUNX3 functions as a tumor suppressor of gastric cancer, lung cancer, bladder cancer, and colon cancer. Here, we show that RUNX3 is a principal and evolutionarily conserved component of the MST pathway. SAV1/WW45 facilitates the close association between MST2 and RUNX3. MST2, in turn, stimulates the SAV1-RUNX3 interaction. In addition, we show that siRNA-mediated RUNX3 knockdown abolishes MST/Hpo-mediated cell death. By establishing that RUNX3 is an endpoint effector of the MST pathway and that RUNX3 is capable of inducing cell death in cooperation with MST and SAV1, we define an evolutionarily conserved novel regulatory mechanism loop for tumor suppression in human cancers.


Breastfeeding duration predicts greater maternal sensitivity over the next decade.
The current study represents the first longitudinal investigation of the potential effects of breastfeeding duration on maternal sensitivity over the following decade. This study also examined whether infant attachment security at 24 months would mediate longitudinal relations between breastfeeding duration and changes in maternal sensitivity over time. Using data from 1,272 families from the National Institute of Child Health and Human Development's Study of Early Child Care and Youth Development, we found that longer breastfeeding duration (assessed up to age 3) predicted increases in observed maternal sensitivity up to child age 11, after accounting for maternal neuroticism, parenting attitudes, ethnicity, maternal years of education, and presence of a romantic partner. Additionally, secure attachment at 24 months was predicted by breastfeeding duration, but it did not act as a mediator of the link from breastfeeding duration to maternal sensitivity in this study. Generating a more specific understanding of how breastfeeding impacts the mother-child dyad beyond infancy will inform recommendations for best practices regarding breastfeeding. (PsycINFO Database Record


Dying cells expose a nuclear antigen cross-reacting with anti-PD-1 monoclonal antibodies.
Checkpoint molecules such as programmed death 1 (PD-1) dampen excessive T cell activation to preserve immune homeostasis. PD-1-specific monoclonal antibodies have revolutionized cancer therapy, as they reverse tumour-induced T cell exhaustion and restore CTL activity. Based on this success, deciphering underlying mechanisms of PD-1-mediated immune functions has become an important field of immunological research. Initially described for T cells, there is emerging evidence of unconventional PD-1 expression by myeloid as well as tumor cells, yet, with cell-intrinsic functions in various animal tumor models. Here, we describe positive PD-1 antibody staining of various murine immune and tumour cells that is, unlike for T cells, not the PD-1 receptor and restricted to cells with low forward scatter characteristics. Based on flow cytometry and various approaches, including two established murine anti-PD-1 antibody clones, CRISPR/Cas9 genome editing and confocal imaging, we describe a staining pattern assigned to a nuclear antigen cross-reacting with anti-PD-1 monoclonal antibodies. Lack of PD-1 expression was further underlined by the analysis of PD-1 expression from B16-F10-derived 3D cultures and ex vivo tumours. Thus, our data provide multiple lines of evidence that PD-1 expression by non-T cells is unlikely to be the case and, taking recent data of PD-1 tumour cell-intrinsic functions into account, suggest that other antibody-mediated pathways might apply.


C57BL/6 and 129 inbred mouse strains differ in Gbp2 and Gbp2b expression in response to inflammatory stimuli in vivo.
Background: Infections cause the production of inflammatory cytokines such as Interferon gamma (IFNγ). IFNγ in turn prompts the upregulation of a range of host defence proteins including members of the family of guanylate binding proteins (Gbps). In humans and mice alike, GBPs restrict the intracellular replication of invasive microbes and promote inflammation. To study the physiological functions of Gbp family members, the most commonly chosen in vivo models are mice harbouring loss-of-function mutations in either individual Gbp genes or the entire Gbp gene cluster on mouse chromosome 3. Individual Gbp deletion strains differ in their design, as some strains exist on a pure C57BL/6 genetic background, while other strains contain a 129-derived genetic interval encompassing the Gbp gene cluster on an otherwise C57BL/6 genetic background. Methods: To determine whether the presence of 129 alleles of paralogous Gbps could influence the phenotypes of 129-congenic Gbp-deficient strains, we studied the expression of Gbps in both C57BL/6J and 129/Sv mice following in vivo stimulation with adjuvants and after infection with either Toxoplasma gondii or Shigella flexneri. Results: We show that C57BL/6J relative to 129/Sv mice display moderately elevated expression of Gbp2, but more prominently, are also defective for Gbp2b (formerly Gbp1) mRNA induction upon immune priming. Notably, Toxoplasma infections induce robust Gbp2b protein expression in both strains of mice, suggestive of a Toxoplasma-activated mechanism driving Gbp2b protein translation. We further find that the higher expression of Gbp2b mRNA in 129/Sv mice correlates with a gene duplication event at the Gbp2b locus resulting in two copies of the Gbp2b gene on the haploid genome of the 129/Sv strain. Conclusions: Our findings demonstrate functional differences between 129 and C57BL/6 Gbp alleles which need to be considered in the design and interpretation of studies utilizing mouse models, particularly for phenotypes influenced by Gbp2 or Gbp2b expression.


Loss of heterozygosity for genes on 11p and the clinical course of patients with lung carcinoma.
Forty-five primary human lung carcinomas were evaluated for the loss of heterozygosity for genes on the short end of chromosome 11. Of 40 evaluable heterozygous cases, loss of the 11p genes c-H-ras and insulin was documented in nine cases (22%). The clinical parameters investigated for each patient included the disease stage at presentation, the presence of metastatic disease in either bronchial or mediastinal lymph nodes, and the presence of positive parietal pleural margins in the surgically resected specimen. There were no differences found with respect to these indicators when patients exhibiting the loss of heterozygosity were compared with those who did not have such genetic loss. In addition, when the clinical courses of the two patient groups were compared, there was no difference in survival. We conclude that the loss of heterozygosity for c-H-ras and insulin on 11p is a common finding in primary non-small cell human lung carcinomas but does not confer a more aggressive phenotype on these tumors. Although this genetic lesion may be important in the initial transformation of the cells to carcinoma, the available data for lung carcinoma are insufficient to prove causality.


Outcomes after arthroscopic repair of type-II SLAP lesions.
To our knowledge, there has been no prospective study on the results of arthroscopic repair of superior labrum-biceps anchor complex (SLAP) tears with use of modern techniques. The purpose of the present study was to prospectively evaluate the minimum two-year results for patients with type-II SLAP tears that were treated with arthroscopic suture anchor fixation. Forty-seven patients with symptomatic type-II SLAP tears were evaluated preoperatively and at least two years postoperatively with use of the American Shoulder and Elbow Surgeons (ASES) and L'Insalata outcomes instruments and physical examination. The study group included thirty-nine male and eight female patients with a mean age of thirty-six years; thirty-four of the forty-seven patients were athletes. Patients with rotator cuff tears requiring repair or concomitant shoulder instability were excluded. At an average of 2.7 years, the median ASES and L'Insalata scores were 97 and 93, respectively, compared with baseline scores of 62 and 65 (p < 0.05). The median patient-reported satisfaction rating was 9 (of 10); forty-one patients (87%) rated the outcome as good or excellent. The median patient-reported satisfaction rating was significantly higher for patients with a discrete traumatic etiology than for those with an atraumatic etiology (9 compared with 7); however, there was no significant difference between these groups in terms of the ASES or L'Insalata outcome scores. Overall, twenty-five (74%) of the thirty-four athletes were able to return to their preinjury level of competition, whereas eleven (92%) of the twelve athletes who reported a discrete traumatic event were able to return to their previous level of competition. There were five complications, including four cases of refractory postoperative stiffness. Our findings indicate that favorable outcomes can be anticipated in the majority of patients after arthroscopic SLAP lesion repair. While only three of four patients overall may be capable of returning fully to their previous level of competition, patients with a distinct traumatic etiology have a greater likelihood of a successful return to sports.


GABAB Receptor Agonist R-Baclofen Reverses Social Deficits and Reduces Repetitive Behavior in Two Mouse Models of Autism.
Autism spectrum disorder (ASD) is diagnosed by two core behavioral criteria, unusual reciprocal social interactions and communication, and stereotyped, repetitive behaviors with restricted interests. Excitatory/inhibitory imbalance is a prominent hypothesis for the etiology of autism. The selective GABAB receptor agonist R-baclofen previously reversed social deficits and reduced repetitive behaviors in a mouse model of Fragile X syndrome, and Arbaclofen improved some clinical symptoms in some Fragile X and ASD patients. To evaluate R-baclofen in a broader range of mouse models of ASD, we tested both the R-baclofen enantiomer and the less potent S-baclofen enantiomer in two inbred strains of mice that display low sociability and/or high repetitive or stereotyped behaviors. R-baclofen treatment reversed social approach deficits in BTBR T+ Itpr3tf/J (BTBR), reduced repetitive self-grooming and high marble burying scores in BTBR, and reduced stereotyped jumping in C58/J (C58), at nonsedating doses. S-baclofen produced minimal effects at the same doses. These findings encourage investigations of R-baclofen in other preclinical model systems. Additional clinical studies may be warranted to further evaluate the hypothesis that the GABAB receptor represents a promising pharmacological target for treating appropriately stratified subsets of individuals with ASD.


Krill phosphatidylserine improves learning and memory in Morris water maze in aged rats.
The ameliorating effect of phosphatidylserine (PS) isolated from krill (KR-PS) on the learning and memory deficits associated with normal aging in rats was investigated, as compared with soybean PS (SOY-PS). Rats were orally administered with KR-PS (20, 50 mg kg-1) and SOY-PS (50 mg kg-1) daily, for 7 days, 30 min before behavioral assessment using the Morris water maze (MWM). Changes in the cholinergic system were examined by measuring choline acetyltransferase (ChAT) and acetylcholinesterase (AchE) immunoreactivity in the hippocampus. The daily administration of KR-PS produced a significant improvement in the escape latency for finding the platform in the MWM, as compared with SOY-PS. Consistent with the behavioral results, KR-PS treatments significantly alleviated age-associated losses of cholinergic immunoreactivity, and muscarinic acetylcholine receptor type 1 (mAChR-M1) and choline transporter (CHT) mRNA expression in the hippocampus. These findings demonstrate that KR-PS showed significant neuroprotective activity against the neuronal and cognitive impairments that occur with normal aging in rats; comparable results were obtained with SOY-PS. These data indicate that oral administration of PS derived from marine life could substitute for bovine cerebral cortex PS (BC-PS) as therapy for the improvement of diminished memory function in elderly people.


Tensile bond strength of veneering resins to PEEK: impact of different adhesives.
This study tested tensile bond strength (TBS) between veneering resins and polyetheretherketone (PEEK) after pre-treatment with adhesive systems. Five-hundred-seventy-six PEEK disks were fabricated, air-abraded and divided into six pre-treatment groups (n=96/group): Z-Prime Plus, Ambarino P60, Monobond Plus, Visio.link, Signum PEEK Bond, and control group without pre-treatment. Each group was divided into three subgroups of different veneering resins (n=32): Sinfony, GC Gradia and VITA VM LC. After specimen preparation with a bond area of 6.6 mm(2), half of each subgroup (n=16) was tested initially, and the other half was thermo-cycled. TBS measurements were analysed by three-way and one-way ANOVA, t-test and Weibull statistics. Groups without pre-treatment and groups pre-treated by Z-Prime Plus and Ambarino P60 showed no TBS. Pre-treatment with Monobond Plus increased the TBS values. The highest TBS before and after thermo-cycling between PEEK and all tested veneering resins was observed for groups pre-treated with Visio.link and Signum PEEK Bond.


The effects of temperature and body size on immunological development and responsiveness in juvenile shortnose sturgeon (Acipenser brevirostrum).
Sturgeon are an important evolutionary taxa of which little is known regarding their responses to environmental factors. Water temperature strongly influences growth in fish; however, its effect on sturgeon immune responses is unknown. The objective of this study was to assess how 2 different temperatures affect immune responses in shortnose sturgeon (Acipenser brevirostrum) relevant immune organs such as the meningeal myeloid tissue, spleen, thymus and skin. These responses were studied in 2 different sizes of same age juvenile sturgeon kept at either 11 °C or 20 °C (4 treatment groups), before and after exposure to an ectoparasitic copepod (Dichelesthium oblongum). Based on a differential cell count, temperature was found to strongly influence immune cell production in the meningeal myeloid tissue, regardless of the fish sizes considered. Morphometric analysis of splenic white pulp showed a transient response to temperature. There were no differences between the groups in the morphometric analysis of thymus size. Splenic IRF-1 and IRF-2 had similar expression profiles, significantly higher in fish kept at 20 °C for the first 6 weeks of the study but not by 14 weeks. In the skin, IRF-1 was significantly higher in the fish kept at 11 °C over the first 6 weeks of the study. IRF-2 had a similar profile but there were no differences between the groups by the end of the trial. In conclusion, higher water temperatures (up to 20 °C) may have beneficial effects in maximizing growth and improving immunological capacity, regardless of the fish sizes considered in this study.


Is the canine fossa puncture approach really necessary for the severely diseased maxillary sinus during endoscopic sinus surgery?
To evaluate the necessity of canine fossa puncture (CFP) by comparing the symptom scores and postoperative computed tomography (CT) findings between patients with severe maxillary sinus disease who underwent CFP and those who underwent maxillary sinus clearance through a middle meatal antrostomy (MMA). A prospective, randomized study. Fourteen patients met the inclusion criteria for each of the CFP and MMA groups. In both groups, all diseased sinuses were addressed in the same manner using the same surgical techniques; the only difference was management of the maxillary sinus. Most of the patients completed the Sinonasal Outcome Test 20 (SNOT-20) and visual analogue scales (VAS) for the six main symptoms preoperatively and 3, 6, and 12 months postoperatively. The Lund-Mackay scores for the maxillary sinus and for all sinuses were calculated from the preoperative CT scan and another scan taken 12 months postoperatively. The mucosal thickening as a percentage of the total volume of the maxillary sinus was also evaluated on the postoperative CT scans, and complications related to both procedures were investigated. Twenty-four patients completed the follow-up, questionnaires, and postoperative CT scans and were included in the analysis: 11 CFP patients and 13 MMA patients. All of the patients had chronic rhinosinusitis with nasal polyposis (NP). There were no significantdifferences in polyp extent or Lund-Mackay score for the maxillary sinus and for all sinuses on the pre- and postoperative CT scans between the groups. The volume of mucosal thickening also did not differ significantly between the groups on the postoperative CT scans. The SNOT-20 and VAS scores improved significantly 3, 6, and 12 months after the procedure in both groups. However, there were no significant differences between the two groups except for the VAS for postnasal drip 3 months postoperatively, which was better in the CFP group. Six of 11 patients in the CFP group experienced one or more complications after the procedure, although all of the symptoms resolved spontaneously within 3 months. In the MMA group, three patients had bleeding from the branches of the sphenopalatine artery during widening of the ostium, which was controlled intraoperatively with suction cauterization. We could not find any benefits of the CFP procedure over the conventional MMA method in the present study. Although CFP is a useful method for removing severe mucosal disease that cannot be reached through the MMA, it does not guarantee a better subjective or objective surgical outcome in patients who have accompanying NP.


The spectrum of beta thalassaemia in Burma.
The molecular defects causing beta thalassaemia have been analysed in 85 unrelated Burmese patients. The patients included 14 with homozygous beta thalassaemia, 70 with HbE/beta thalassaemia and one with HbS/beta thalassaemia. Using a combination of allele-specific oligoprobe hybridization and direct sequencing of genomic DNA amplified by the polymerase chain reaction, 95/99 of the beta-thalassaemia alleles have been characterized. Six mutations have been identified of which three, the G-T at IVS-1 position 1, the G-C at IVS-1 position 5 and the deletion of TCTT in codons 41/42, accounted for 85% of the alleles. Despite the diversity of ethnic groups in Burma, the number of beta-thalassaemia alleles in Burma is relatively small. Thus, diagnosis of the majority of the beta thalassaemias would be possible using a limited number of oligonucleotide probes.


Role of the terminal complement pathway in experimental membranous nephropathy in the rabbit.
Our recent observations of a complement-mediated, cell-independent mechanism of altered glomerular permeability in rat membranous nephropathy suggested a possible role for the terminal complement pathway in the mediation of proteinuria in certain forms of glomerular disease. To directly determine whether the membranolytic terminal complement components (C5b-C9) are involved in glomerular injury, we studied the development of proteinuria in normal and C6-deficient (C6D) rabbits, in both of which a membranous nephropathy-like lesion develops early in the course of immunization with cationized bovine serum albumin (cBSA) (pI 8.9-9.2). C6 hemolytic activity of C6D was 0.01% that of control rabbits. After 1 wk of daily intravenous injections of cBSA, proteinuria developed in 71% of controls (median 154, range 1-3,010 mg/24 h, n = 24), whereas none of C6D were proteinuric (median 6, range 2-12 mg/24 h, n = 12, P less than 0.01). After 1 wk of cBSA, both groups had qualitatively identical glomerular deposits of BSA, rabbit IgG, and C3 on immunofluorescence microscopy, predominantly subepithelial electron-dense deposits on electron microscopy, and minimal glomerular inflammatory cell infiltration of glomeruli. Glomeruli were isolated from individual animals after 1 wk of cBSA and deposits of rabbit IgG antibody were quantitated by a standardized in vitro assay using anti-rabbit IgG-125I. Rabbit IgG deposits were found to be similar in control (29.8 +/- 13.2, range 12.7-48.6 micrograms anti-IgG/2,000 glomeruli, n = 6) and C6D rabbits (32.6 +/- 13.8, range 16.8-48.8 micrograms anti-IgG/2,000 glomeruli, n = 5, P greater than 0.05). After 2 wk, coincident with a prominent influx of mononuclear cells and neutrophils, proteinuria developed in C6D rabbits. These results document, for the first time, a requirement for a terminal complement component in the development of immunologic glomerular injury. Since the only known action of C6 is in the assembly of the membrane attack complex, these observations suggest that the membranolytic properties of complement may contribute to glomerular damage.


JH Zero: New Naturally Occurring Insect Juvenile Hormone from Developing Embryos of the Tobacco Hornworm.
A new insect juvenile hormone was isolated from developing embryos of the tobacco hornworm moth, Manduca sexta. The new hormone was found with juvenile hormone I and is a 1-carbon homolog of this substance. The assigned structure is methyl (2E,6E,10-cis)-10,11-epoxy-3,7-diethyl-11-methyl-2,6-tridecadienoate, which constitutes a trishomosesquiterpenoid skeleton. This is the first chemical idetification of any juvenile hormone from insect eggs.


Long-term survival after acute kidney injury following ruptured abdominal aortic aneurysm repair.
Acute kidney injury (AKI) is a major complication of ruptured abdominal aortic aneurysm (RAAA). Severe AKI is associated with high morbidity and mortality in the short term. The objective of this study was to determine the association between AKI after RAAA repair and long-term survival. We conducted a retrospective cohort study of all patients undergoing RAAA repair in three hospitals between 2004 and 2011. Outcomes were long-term survival after RAAA repair, incidence of postoperative AKI, and chronic dialysis rates. Survival rates were compared between different AKI groups (no AKI, Risk, Injury, Failure) with Kaplan-Meier survival analyses and log-rank tests. Univariable and multivariable Cox regression analyses were carried out to assess the association of survival with AKI, preoperative shock, postoperative shock, and sex. The main analysis focused on the group of patients surviving initial hospital stay. Our study encompassed 362 patients with RAAA. AKI occurred in 267 of 362 patients (74%). At discharge, 267 patients were alive (74%). Median survival in this group was 7.2 years. Survival was not significantly different between the four AKI groups (P = .07). However, the univariable Cox regression analysis demonstrated a significant association between Failure and reduced long-term survival compared with having no AKI (hazard ratio, 1.85; 95% confidence interval, 1.15-2.97). This association did not remain significant after multivariable adjustment. Four patients were discharged with chronic dialysis, and four other patients needed chronic dialysis later after discharge. This study demonstrates no significant independent association between AKI after RAAA repair and long-term survival. Only a small proportion of patients developed end-stage renal disease at a later stage in life.


In silico approach to predict candidate R proteins and to define their domain architecture.
Plant resistance genes, which encode R-proteins, constitute one of the most important and widely investigated gene families. Thanks to the use of both genetic and molecular approaches, more than 100 R genes have been cloned so far. Analysis of resistance proteins and investigation of domain properties may afford insights into their role and function. Moreover, genomic experiments and availability of high-throughput sequence data are very useful for discovering new R genes and establish hypotheses about R-genes architecture. We surveyed the PRGdb dataset to provide valuable information about hidden R-protein features. Through an in silico approach 4409 putative R-proteins belonging to 33 plant organisms were analysed for domain associations frequency. The proteins showed common domain associations as well as previously unknown classes. Interestingly, the number of proteins falling into each class was found inversely related to domain arrangement complexity. Out of 31 possible theoretical domain combinations, only 22 were found. Proteins retrieved were filtered to highlight, through the visualization of a Venn diagram, candidate classes able to exert resistance function. Detailed analyses performed on conserved profiles of those strong putative R proteins revealed interesting domain features. Finally, several atypical domain associations were identified. The effort made in this study allowed us to approach the R-domains arrangement issue from a different point of view, sorting through the vast diversity of R proteins. Overall, many protein features were revealed and interesting new domain associations were found. In addition, insights on domain associations meaning and R domains modelling were provided.


Direct radiocarbon dating and genetic analyses on the purported Neanderthal mandible from the Monti Lessini (Italy).
Anatomically modern humans replaced Neanderthals in Europe around 40,000 years ago. The demise of the Neanderthals and the nature of the possible relationship with anatomically modern humans has captured our imagination and stimulated research for more than a century now. Recent chronological studies suggest a possible overlap between Neanderthals and anatomically modern humans of more than 5,000 years. Analyses of ancient genome sequences from both groups have shown that they interbred multiple times, including in Europe. A potential place of interbreeding is the notable Palaeolithic site of Riparo Mezzena in Northern Italy. In order to improve our understanding of prehistoric occupation at Mezzena, we analysed the human mandible and several cranial fragments from the site using radiocarbon dating, ancient DNA, ZooMS and isotope analyses. We also performed a more detailed investigation of the lithic assemblage of layer I. Surprisingly we found that the Riparo Mezzena mandible is not from a Neanderthal but belonged to an anatomically modern human. Furthermore, we found no evidence for the presence of Neanderthal remains among 11 of the 13 cranial and post-cranial fragments re-investigated in this study.


Computed tomographic adrenal gland quantification in canine adrenocorticotroph hormone-dependent hyperadrenocorticism.
We conducted a retrospective study to determine whether multidetector computed tomography (CT) could be of value for adrenal gland assessment in dogs with pituitary-dependent hyperadrenocorticism. Adrenal gland attenuation and volume values of 49 dogs with hyperadrenocorticism were recorded and age, body weight, and gender were examined to determine if a relationship existed between these variables and adrenal gland morphology. There was not a statistically significant difference in mean X-ray attenuation of the left vs. right adrenal gland in normal dogs (35.3 +/- 6.1 HU), or in dogs with hyperadrenocorticism. The mean adrenal X-ray attenuation (+/- standard deviation [SD]) in dogs with microadenoma was 33.1 +/- 6.8 vs. 31.8 +/- 12.7 HU for dogs with macroadenoma, and these values were not statistically different. The mean volume of the left adrenal gland in normal dogs (0.59 +/- 0.17 cm3) was greater than that of the right adrenal gland (0.54 +/- 0.19 cm3) (P < 0.05). The mean CT volume (+/- SD) of the adrenal glands in dogs with microadenoma vs. macroadenoma were 1.60 +/- 1.25 vs. 2.88 +/- 1.60 cm3, respectively. There was no effect of age or gender on adrenal gland morphology or X-ray attenuation. The weight effect was the most important source of variation for the volume measurement in dogs with hyperadrenocorticism.