CD13 as a new tumor target for antibody-drug conjugates: validation with the conjugate MI130110.
In the search for novel antibody-drug conjugates (ADCs) with therapeutic potential, it is imperative to identify novel targets to direct the antibody moiety. CD13 seems an attractive ADC target as it shows a differential pattern of expression in a variety of tumors and cell lines and it is internalized upon engagement with a suitable monoclonal antibody. PM050489 is a marine cytotoxic compound tightly binding tubulin and impairing microtubule dynamics which is currently undergoing clinical trials for solid tumors. Anti-CD13 monoclonal antibody (mAb) TEA1/8 has been used to prepare a novel ADC, MI130110, by conjugation to the marine compound PM050489. In vitro and in vivo experiments have been carried out to demonstrate the activity and specificity of MI130110. CD13 is readily internalized upon TEA1/8 mAb binding, and the conjugation with PM050489 did not have any effect on the binding or the internalization of the antibody. MI130110 showed remarkable activity and selectivity in vitro on CD13-expressing tumor cells causing the same effects than those described for PM050489, including cell cycle arrest at G2, mitosis with disarrayed and often multipolar spindles consistent with an arrest at metaphase, and induction of cell death. In contrast, none of these toxic effects were observed in CD13-null cell lines incubated with MI130110. Furthermore, in vivo studies showed that MI130110 exhibited excellent antitumor activity in a CD13-positive fibrosarcoma xenograft murine model, with total remissions in a significant number of the treated animals. Mitotic catastrophes, typical of the payload mechanism of action, were also observed in the tumor cells isolated from mice treated with MI130110. In contrast, MI130110 failed to show any activity in a xenograft mouse model of myeloma cells not expressing CD13, thereby corroborating the selectivity of the ADC to its target and its stability in circulation. Our results show that MI130110 ADC combines the antitumor potential of the PM050489 payload with the selectivity of the TEA1/8 monoclonal anti-CD13 antibody and confirm the correct intracellular processing of the ADC. These results demonstrate the suitability of CD13 as a novel ADC target and the effectiveness of MI130110 as a promising antitumor therapeutic agent.


Activation of the TLR4 signaling pathway and abnormal cholesterol efflux lead to emphysema in ApoE-deficient mice.
Smokers with airflow obstruction have an increased risk of atherosclerosis, but the relationship between the pathogenesis of these diseases is not well understood. To determine whether hypercholesterolemia alters lung inflammation and emphysema formation, we examined the lung phenotype of two hypercholesterolemic murine models of atherosclerosis at baseline and on a high-fat diet. Airspace enlargement developed in the lungs of apolipoprotein E-deficient (Apoe(-/-)) mice exposed to a Western-type diet for 10 wk. An elevated number of macrophages and lymphocytes accompanied by an increase in matrix metalloproteinase-9 (MMP-9) activity and MMP-12 expression was observed in the lungs of Apoe(-/-) mice on a Western-type diet. In contrast, low-density lipoprotein receptor-deficient (Ldlr(-/-)) mice did not exhibit lung destruction or inflammatory changes. Most importantly, we revealed augmented expression of the downstream targets of the Toll-like receptor (TLR) pathway, interleukin-1 receptor-associated kinase 1, and granulocyte colony-stimulating factor, in the lungs of Apoe(-/-) mice fed with a Western-type diet. In addition, we demonstrated overexpression of MMP-9 in Apoe(-/-) macrophages treated with TLR4 ligand, augmented with the addition of oxidized LDL, suggesting that emphysema in these mice results from the activation of the TLR pathway secondary to known abnormal cholesterol efflux. Our findings indicate that, in Apoe(-/-) mice fed with an atherogenic diet, abnormal cholesterol efflux leads to increased systemic inflammation with subsequent lung damage and emphysema formation.


The human pro-opiomelanocortin gene: organization, sequence, and interspersion with repetitive DNA.
The human pro-opiomelanocortin (POMC) gene has been characterized by molecular cloning and DNA sequence analysis. Although this gene codes for several different polypeptide hormones, only a single intron interrupts the protein coding region. The DNA in this intron, and in a second intron found in the region of the gene homologous to the mRNA 5'-untranslated sequence, contains repetitive DNA sequences. At least some of these sequences belong to the Alu family of transcribed middle repetitive DNA. The determination of the complete nucleotide sequence of the coding regions of the gene demonstrates that the pattern of homologous and variable regions seen in the POMC protein between different species is reflected at the DNA level. DNA sequences encoding the highly conserved regions of POMC are 90-95% homologous between species while the coding sequences for the variable regions of the protein are approximately 70% homologous. The very high degree of homology in the amino terminal portion of POMC is consistent with an important physiological role for this peptide.


[Advances in the treatment of castration-resistant prostate cancer: emphasis in new hormonal therapies].
Prostate cancer represents the second cancer-related cause of death in North American and Chilean men. The main treatment for incurable stages of disease is surgical or pharmacological castration. However, with time and despite the addition of anti-androgens, the disease progresses to a clinical state that has been commonly referred to as "hormone refractory". In recent years, the concept of hormone refractoriness has been challenged and replaced by "castration resistance", acknowledging that further and optimal hormonal manipulation can be attained, beyond achieving testosterone levels at castration range. The purpose of this review is to summarize the recent therapeutic breakthroughs in the management of metastatic castrate resistant prostate cancer (mCRPC), with greater emphasis in the newer hormonal therapy agents such as Abiraterone and Enzalutamide. Future combination and sequential treatment strategies are contextualized in the current era of personalized cancer medicine and genomic characterization of prostate cancer.


HIT'nDRIVE: patient-specific multidriver gene prioritization for precision oncology.
Prioritizing molecular alterations that act as drivers of cancer remains a crucial bottleneck in therapeutic development. Here we introduce HIT'nDRIVE, a computational method that integrates genomic and transcriptomic data to identify a set of patient-specific, sequence-altered genes, with sufficient collective influence over dysregulated transcripts. HIT'nDRIVE aims to solve the "random walk facility location" (RWFL) problem in a gene (or protein) interaction network, which differs from the standard facility location problem by its use of an alternative distance measure: "multihitting time," the expected length of the shortest random walk from any one of the set of sequence-altered genes to an expression-altered target gene. When applied to 2200 tumors from four major cancer types, HIT'nDRIVE revealed many potentially clinically actionable driver genes. We also demonstrated that it is possible to perform accurate phenotype prediction for tumor samples by only using HIT'nDRIVE-seeded driver gene modules from gene interaction networks. In addition, we identified a number of breast cancer subtype-specific driver modules that are associated with patients' survival outcome. Furthermore, HIT'nDRIVE, when applied to a large panel of pan-cancer cell lines, accurately predicted drug efficacy using the driver genes and their seeded gene modules. Overall, HIT'nDRIVE may help clinicians contextualize massive multiomics data in therapeutic decision making, enabling widespread implementation of precision oncology.


Analysis of the FGFR3 gene in Japanese patients with achondroplasia and hypochondroplasia.
It has been reported that mutations in the FGFR3 gene cause autosomal dominant forms of dwarfism, achondroplasia (ACH) and hypochondroplasia (HCH). In the present study, we analyzed the FGFR3 gene in 26 Japanese patients with ACH and 14 with HCH. Genomic DNAs of the patients were isolated from whole blood. For the ACH patients, a 164-bp fragment of the FGFR3 gene that spans the entire transmembrane domain was amplified by polymerase chain reaction (PCR), and the PCR products were analyzed by direct sequencing of the PCR products and by digestion of the PCR products with restriction enzymes. For the HCH patients, a 206-bp fragment of the FGFR3 gene which encodes a part of the TK1 domain was amplified, and the PCR products were directly sequenced. The heterozygous G380R mutations were identified in all of the 26 ACH patients, whereas the heterozygous N540K mutations were identified in 8 out of 14 HCH patients. These results were consistent with previous reports from abroad.


Mesenchymal stromal cells isolated from gestationally diabetic human placenta exhibit insulin resistance, decreased clonogenicity and angiogenesis.
Pregnancy is known to be a diabetogenic state. With sedentary lifestyle and wrong dietary choices, gestational diabetes mellitus is on the rise. This raises a concern as placenta is becoming an acceptable choice, as a source of Mesenchymal Stromal Cells (MSCs). In our current study we questioned whether there exists a difference between MSCs isolated from normal and diabetic (Gd-P-MSCs) placenta, as the health of the cells used in therapy is of prime importance. We isolated and verified the Gd-P-MSCs based on their surface markers and differentiation potential. We looked at viability and proliferation and did not see a difference between the two. We analysed the glucose uptake potential of these cells by assessing the remnant glucose in the media, glucose within the cells by 2-NBDG and by glycogen storage. Despite only a slight downregulation of mRNA expression levels of glucose transporters, Gd-P-MSCs exhibited decreased glucose uptake even upon insulin stimulation and decreased glycogen storage, indicative of an insulin resistant state. We then assessed the colony forming ability of the cells and found a decreased clonogenicity in Gd-P-MSCs. We also examined the angiogenic potential of the cells by tube formation. Gd-P-MSCs showed decreased angiogenic potential when compared to normal cells. Thus we show for the first time, the effect of gestational diabetes on cells isolated from the chorionic villi of term placenta. Gd-P-MSCs are indeed insulin resistant, exhibit decreased clonogenicity and angiogenic potential. The present investigation is of relevance to the choice of sample for MSC isolation for therapeutic purposes.


Successful application of hyperbranched multidisplacement genomic amplification to detect HIV-1 sequences in single neurons removed from autopsy brain sections by laser capture microdissection.
To confirm studies suggesting that HIV-1 infects neurons and to determine whether CD8(+) T lymphocytes traffic to HIV-1-infected neurons, we used laser capture microdissection to remove hippocampal neurons with and without perineuronal CD8(+) T cells from AIDS patients with HIV-1 encephalitis (HIVE) or without HIVE and from normal controls. We used hyperbranched multidisplacement amplification for whole gene amplification (MDA-WGA) plus two rounds of PCR to amplify housekeeping sequences (HK(+)) and, in HK(+) samples, to amplify HIV-1 gag, nef, and pol sequences. Sample size and, in single neurons, MDA-WGA correlated with housekeeping gene amplification (P < 0.05), whereas patient group and postmortem interval did not (P > 0.05). Neuronal viral sequences correlated with HIVE (43% vs. 13% and 0 in non-HIVE and controls, respectively) and, in HIVE cases, with perineuronal CD8(+) T lymphocytes (70% in CD8(+) samples vs. 37% of CD8(-) samples). Our results suggest that MDA-WGA is a useful technique when analyzing DNA from single cells from autopsy brains, supporting prior studies that show that neurons may contain HIV-1 neuronal sequences in vivo. The association between neuronal infection and perineuronal CD8(+) T cells supports our hypothesis that these cells specifically traffic to infected neurons but raises the possibility that CD8(+) T cells, if infected, could transmit virus to neurons.


MT-SP1 proteolysis and regulation of cell-microenvironment interactions.
MT-SP1 is a type II transmembrane serine protease implicated in a range of human cancers including those of the breast, cervix, ovaries, prostate, colon and gastrointestinal tract. Mouse models have shown it to be critical for proper epidermal development and postnatal survival. However, the role of this enzyme in normal and malignant biology has not yet been fully elucidated. Several groups have identified putative substrates of MT-SP1 in an effort to understand the possible biological processes in which this protease may be involved. Methods for substrate identification include comparing known protein cleavage sequences with MT-SP1 specificity data, in vitro cleavage assays, examining genetic microarrays for enzyme/substrate coexpression, immunohistochemistry for colocalization, and a variety of phenotypic observations using cell culture and mouse models. Given the inherent limitations of each individual method, substrate plausibility is best substantiated using a combination of experimental approaches. Here we review MT-SP1 substrates identified to date and the possible physiological implications of substrate cleavage in cell-microenvironment interactions. This data indicates that MT-SP1 is capable of playing roles in growth factor activation, receptor activation and inactivation, protease activation, and ectodomain shedding. We also present for the first time vascular endothelial growth factor receptor 2 (VEGFR-2) as a putative substrate for MT-SP1.


Acid-sensing ion channels detect moderate acidifications to induce ocular pain.
Sensory nerve fibers innervating the ocular anterior surface detect external stimuli producing innocuous and painful sensations. Protons are among the first mediators released by damaged cells during inflammation, tissue injury, or other chronic ophthalmic conditions. We studied whether acid-sensing ion channels (ASICs) are expressed in corneal sensory neurons and their roles in the response to moderate acidifications of the ocular surface and in pathologies producing ocular surface inflammation. Moderate acidic pH (6.6) activated ASIC-like currents in corneal sensory neurons, which were blocked by ASIC1- or ASIC3-specific toxins. Acidic pH depolarizes corneal sensory neurons to fire action potentials, an effect blocked by the ASIC3 inhibitor APETx2. 2-Guanidino-4-methylquinazoline, an ASIC3 agonist, activated a population of corneal polymodal sensory nerve fibers and significantly increased the blinking and tearing rate. The nocifensive behaviors produced by application of either a moderate acidic stimulus or ophthalmic drugs formulated in acidic solution were abolished by ASIC blockers. In a model of allergic keratoconjunctivitis, nocifensive behavior was greatly reduced by ASIC3 blockade, presumably by reducing nociceptor sensitization during the inflammatory process. Our results show that, in addition to the established role of TRPV1, ASICs play a significant role in the detection of acidic insults at the ocular surface. The identification of ASICs in corneal neurons and their alterations during different diseases is critical for the understanding of sensory ocular pathophysiology. They are likely to mediate some of the discomfort sensations accompanying several ophthalmic formulations and may represent novel targets for the development of new therapeutics for ocular pathologies.


Automated image analysis of alveolar expansion patterns in immature newborn rabbits treated with natural or artificial surfactant.
Automated image analysis of histological lung sections was used to compare the efficacy of an artificial surfactant (dipalmitoylphosphatidylcholine + high-density lipoprotein, 10:1) and a natural surfactant (the phospholipid fraction of porcine surfactant, isolated by liquid-gel chromatography in ventilated immature newborn rabbits delivered after 27 days' gestation. Tidal volumes were significantly improved in each group treated with surfactant when compared with controls, but natural surfactant-treated rabbits had significantly higher tidal volumes than those receiving artificial surfactant. There were no statistically significant differences in alveolar expansion between the artificial surfactant group and the controls, but alveolar volume density and a shape factor (assessing the 'circularity' of terminal airspaces) were significantly higher in animals receiving natural surfactant. These animals also had a lower coefficient of variation of alveolar volume density and a lower alveolar average integral mean surface curvature, indicating a uniform pattern of alveoli with a smooth profile. We conclude that automated image analysis is useful for the quantitation of alveolar expansion patterns in immature neonatal lungs and that natural surfactant is superior to the artificial surfactant tested in the present study.


Biocide-Induced Emergence of Antibiotic Resistance in Escherichia coli.
Biocide use is essential and ubiquitous, exposing microbes to sub-inhibitory concentrations of antiseptics, disinfectants, and preservatives. This can lead to the emergence of biocide resistance, and more importantly, potential cross-resistance to antibiotics, although the degree, frequency, and mechanisms that give rise to this phenomenon are still unclear. Here, we systematically performed adaptive laboratory evolution of the gut bacteria Escherichia coli in the presence of sub-inhibitory, constant concentrations of ten widespread biocides. Our results show that 17 out of 40 evolved strains (43%) also decreased the susceptibility to medically relevant antibiotics. Through whole-genome sequencing, we identified mutations related to multidrug efflux proteins (mdfA and acrR), porins (envZ and ompR), and RNA polymerase (rpoA and rpoBC), as mechanisms behind the resulting (cross)resistance. We also report an association of several genes (yeaW, pyrE, yqhC, aes, pgpA, and yeeP-isrC) and specific mutations that induce cross-resistance, verified through mutation repairs. A greater capacity for biofilm formation with respect to the parent strain was also a common feature in 11 out of 17 (65%) cross-resistant strains. Evolution in the biocides chlorophene, benzalkonium chloride, glutaraldehyde, and chlorhexidine had the most impact in antibiotic susceptibility, while hydrogen peroxide and povidone-iodine the least. No cross-resistance to antibiotics was observed for isopropanol, ethanol, sodium hypochlorite, and peracetic acid. This work reinforces the link between exposure to biocides and the potential for cross-resistance to antibiotics, presents evidence on the underlying mechanisms of action, and provides a prioritized list of biocides that are of greater concern for public safety from the perspective of antibiotic resistance. Bacterial resistance and decreased susceptibility to antimicrobials is of utmost concern. There is evidence that improper biocide (antiseptic and disinfectant) use and discard may select for bacteria cross-resistant to antibiotics. Understanding the cross-resistance emergence and the risks associated with each of those chemicals is relevant for proper applications and recommendations. Our work establishes that not all biocides are equal when it comes to their risk of inducing antibiotic resistance; it provides evidence on the mechanisms of cross-resistance and a risk assessment of the biocides concerning antibiotic resistance under residual sub-inhibitory concentrations.


A prospective, randomized, multicenter, double-blind study of early corticosteroid cessation versus long-term maintenance of corticosteroid therapy with tacrolimus and mycophenolate mofetil in primary renal transplant recipients: one year report.
This report summarizes year 1 blinded data from a 5-year prospective, randomized (1:1, stratified by race and donor type), multicenter, double-blind study comparing treatment failure rates between early cessation of corticosteroid therapy and long-term corticosteroid maintenance therapy in 397 primary renal transplant recipients receiving tacrolimus and mycophenolate mofetil with antibody induction. Sixteen (4.1%) patients died or experienced graft loss during year 1 (9 deaths, 6 graft losses, and 1 death with a graft loss). Fifty-nine (15.3%) patients experienced rejection; 40 (10.4%) of these cases were biopsy-confirmed. Values for 10-year predictive risk of coronary heart disease (CHD) estimated from the Framingham Heart Study scores showed a shift from baseline toward a more favorable risk profile. Renal transplant recipients treated with a tacrolimus-based immunosuppressive regimen and either early cessation or maintenance of corticosteroid therapy experienced excellent patient and graft survival, low acute rejection rates, low rate of posttransplantation diabetes mellitus (9.0%), and improved CHD risk profile at 1 year posttransplantation.


Characterisation of a homogeneous plant aminoaldehyde dehydrogenase.
According to our knowledge, this is the first purification method developed, enabling isolation of a homogeneous aminoaldehyde dehydrogenase (AMADH) from etiolated pea seedlings. The procedure involved initial purification with precipitants followed by three low pressure chromatographic steps. Partially purified enzyme was further subjected to fast protein liquid chromatography on a Mono Q column and to affinity-interaction chromatography on 5'-AMP Sepharose. Purity of the final enzyme preparation was checked by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and chromatofocusing. Pea AMADH exists as a tetramer of 230 kDa in the native state, a molecular mass of one subunit was determined as 57 kDa. The enzyme was found to be an acidic protein with pI 5.4. AMADH showed a broad substrate specificity utilising various aminoaldehydes (C3-C6) as substrates. The best substrate of pea AMADH was 3-aminopropionaldehyde, the enzyme also efficiently oxidised 4-aminobutyraldehyde and omega-guanidinoanalogues of the aminoaldehydes. Pea AMADH was inhibited by SH reagents, several elementary aldehydes and metal-binding agents. Although AMADH did not oxidise betaine aldehyde at all, the N-terminal amino acid sequence of the enzyme shows a high degree of homology with those of plant betaine aldehyde dehydrogenases (BADHs) of spinach, sugar beet and amaranth. Several conserved amino acids were found in comparison with BADH from cod liver of known crystal structure.


Astrocytes protect neurons against methylmercury via ATP/P2Y(1) receptor-mediated pathways in astrocytes.
Methylmercury (MeHg) is a well known environmental pollutant that induces serious neuronal damage. Although MeHg readily crosses the blood-brain barrier, and should affect both neurons and glial cells, how it affects glia or neuron-to-glia interactions has received only limited attention. Here, we report that MeHg triggers ATP/P2Y1 receptor signals in astrocytes, thereby protecting neurons against MeHg via interleukin-6 (IL-6)-mediated pathways. MeHg increased several mRNAs in astrocytes, among which IL-6 was the highest. For this, ATP/P2Y1 receptor-mediated mechanisms were required because the IL-6 production was (i) inhibited by a P2Y1 receptor antagonist, MRS2179, (ii) abolished in astrocytes obtained from P2Y1 receptor-knockout mice, and (iii) mimicked by exogenously applied ATP. In addition, (iv) MeHg released ATP by exocytosis from astrocytes. As for the intracellular mechanisms responsible for IL-6 production, p38 MAP kinase was involved. MeHg-treated astrocyte-conditioned medium (ACM) showed neuro-protective effects against MeHg, which was blocked by anti-IL-6 antibody and was mimicked by the application of recombinant IL-6. As for the mechanism of neuro-protection by IL-6, an adenosine A1 receptor-mediated pathway in neurons seems to be involved. Taken together, when astrocytes sense MeHg, they release ATP that autostimulates P2Y1 receptors to upregulate IL-6, thereby leading to A1 receptor-mediated neuro-protection against MeHg.


Activin A targets extrasynaptic NMDA receptors to ameliorate neuronal and behavioral deficits in a mouse model of Huntington disease.
Cortical-striatal synaptic dysfunction, including enhanced toxic signaling by extrasynaptic N-methyl-d-aspartate receptors (eNMDARs), precedes neurodegeneration in Huntington disease (HD). A previous study showed Activin A, whose transcription is upregulated by calcium influx via synaptic NMDARs, suppresses eNMDAR signaling. Therefore, we examined the role of Activin A in the YAC128 HD mouse model, comparing it to wild-type controls. We found decreased Activin A secretion in YAC128 cortical-striatal co-cultures, while Activin A overexpression in this model rescued altered eNMDAR expression. Striatal overexpression of Activin A in vivo improved motor learning on the rotarod task, and normalized striatal neuronal eNMDAR-mediated currents, membrane capacitance and spontaneous excitatory postsynaptic current frequency in the YAC128 mice. These results support the therapeutic potential of Activin A signaling and targeting eNMDARs to restore striatal neuronal health and ameliorate behavioral deficits in HD.


Swim performance following creatine supplementation in Division III athletes.
Creatine (Cr) supplementation has yielded inconsistent results when applied to competitive swimming. To further define the role of Cr, we tested the hypothesis that a Cr supplementation group of Division III swimmers would demonstrate enhanced performance when compared with placebo. In order to test this hypothesis, 8 male and 7 female collegiate Division III swimmers were assigned in a random, double-blind manner into either a Cr supplementation group (0.3 g Cr.kg(-1) body mass) or a placebo group. Loading was maintained for 5 days followed by a 9-day period where Cr-supplemented subjects consumed 2.25 g Cr regardless of body weight. A 50- and 100-yd sprint was performed prior to and following the supplementation regimens. The Cr supplementation group decreased their finish times in both the 50- and 100-yd sprints. Support of the hypothesis suggests that Cr supplementation for swimming events is effective for singular effort sprints of 50 and 100 yd in Division III athletes.


Elevated baseline soluble FcεRI may be linked to early response to omalizumab treatment in chronic spontaneous urticaria.
Omalizumab, an anti-IgE monoclonal antibody, is an effective treatment in chronic spontaneous urticaria (CSU). Predictors of fast and good response for omalizumab treatment have not yet been identified and characterized. To evaluate whether soluble FcεRI (sFcεRI), a marker of IgE-mediated mast cell activation, predicts the time of response to omalizumab in CSU. Sera of 67 CSU patients were obtained before omalizumab treatment and analysed for sFcεRI levels by ELISA (2 ng/mL was used as cut-off for elevated sFcɛRI). Treatment response during the first 4 weeks was assessed with the urticaria activity score (UAS7), urticaria control test (UCT) and the rolling UAS7 (rUAS7). Elevated pre-treatment sFcɛRI levels were detected in more than 70% of patients with completely controlled disease (UCT = 16) and well-controlled disease (UCT = 12-15) and were significantly associated with disease control (χ2 = 4.94, p < 0.05). More than half of the patients (14/25) with low levels had poor disease control (UCT < 12). Of the patients who achieved complete and marked UAS7 response, respectively, 75% and 63% had elevated baseline sFcɛRI levels. Post-treatment UAS7 scores were lower in patients with elevated sFcɛRI levels reaching statistical significance at Week 3 (p < 0.05). Patients with elevated baseline sFcɛRI levels achieved rUAS7 ≤ 6 and = 0 earlier than those with lower levels (Days 9 vs. 13 and Days 12 vs. 14, respectively). Elevated sFcεRI serum levels predict early and good response to treatment with omalizumab, which may help to better design treatment options for CSU patients.


Anesthetic efficacy of combinations of 0.5 m mannitol and lidocaine with epinephrine in inferior alveolar nerve blocks: a prospective randomized, single-blind study.
The purpose of this prospective, randomized, single-blind study was to determine the anesthetic efficacy of lidocaine with epinephrine compared to lidocaine with epinephrine plus 0.5 M mannitol in inferior alveolar nerve (IAN) blocks. Forty subjects randomly received an IAN block in 3 separate appointments spaced at least 1 week apart using the following formulations: a 1.8 mL solution of 36 mg lidocaine with 18 µg epinephrine (control solution); a 2.84 mL solution of 36 mg lidocaine with 18 µg epinephrine (1.80 mL) plus 0.5 M mannitol (1.04 mL); and a 5 mL solution of 63.6 mg lidocaine with 32 µg epinephrine (3.18 mL) plus 0.5 M mannitol (1.82 mL). Mandibular teeth were blindly electric pulp tested at 4-minute cycles for 60 minutes postinjection. No response from the subject to the maximum output (80 reading) of the pulp tester was used as the criterion for pulpal anesthesia. Mean percent total pulpal anesthesia was defined as the total of all the times of pulpal anesthesia (80 readings) over the 60 minutes. Pain of solution deposition and postoperative pain were also measured. The results demonstrated that 2.84 mL of lidocaine with epinephrine plus 0.5 M mannitol was significantly better than 1.8 mL of lidocaine with epinephrine for the molars and premolars. The 5 mL of lidocaine with epinephrine plus 0.5 M mannitol was statistically better than 1.8 mL of lidocaine with epinephrine and 2.84 mL of lidocaine with epinephrine plus 0.5 M mannitol for all teeth except the central incisor. Solution deposition pain and postoperative pain were not statistically different among the mannitol formulations and the lidocaine formulation without mannitol. We concluded that adding 0.5 M mannitol to lidocaine with epinephrine formulations significantly improved effectiveness in achieving a greater percentage of total pulpal anesthesia compared with a lidocaine formulation without mannitol for IAN block.


Fertility factors in Pseudomonas putida: selection and properties of high-frequency transfer and chromosome donors.
The octane plasmid (OCT) in Pseudomonas putida strains has been shown to be transferred at low frequency. However, bacteria which had newly received this plasmid showed a transient increase in donor ability. Using Octane+ P. putida as the donor, the transfer of most chromosomal markers was shown to be independent of OCT transfer, whereas the mobilization of the octanoate catabolism genes (octanoic and acetate) was dependent on OCT plasmid transfer. The presence of a fertility factor termed FPo has been postulated to explain these results. Strains carrying only this fertility factor have been obtained from strains carrying both OCT and FPo plasmids. Strains in which the OCT plasmid was transferred at high frequencies have also been isolated, and chromosome mobilization by OCT and FPo has been compared. A different gradient of transmission by OCT and FPo has been observed. It has also been shown that chromosome transfer by OCT was dependent on the bacterial recombination system, whereas the chromosome transfer by FPo was unaffected by the presence of a rec mutation in the donor strain.


280 cases of hyperplasia of mammary gland treated by ru he chong ji.
280 patients with hyperplasia of mammary gland were treated by Ru He Chong Ji, a breast nodule granular dissolvable agent. Thirty-one were cured, 117 markedly improved, 119 improved, and 13 showed no change. The rate of cure and marked effectiveness was 52.86%, with a total effective rate of 95.36%. The agent is also effective for treatment of depression of liver-energy and menoxenia.


Acute DeBakey Type I Dissection Repair Using Frozen Elephant Trunk: The Cleveland Clinic Technique.
Over the past 3 decades, the standard surgical management for acute DeBakey type I aortic dissection has not significantly changed. Most patients undergo ascending aortic replacement with an interposition graft under hypothermic circulatory arrest with selective root replacement. Nevertheless, with the improvement in overall patient care, acute surgical outcomes continue to improve. The frozen elephant trunk (FET) technique was introduced as a modification to conventional elephant trunk to treat extensive degenerative and chronic dissecting aneurysms involving the aortic arch and the proximal descending aorta. The FET technique converts the conventional elephant trunk procedure-an inherently 2-stage operation-into a 1-stage repair in selected patients and greatly facilitates more distal aortic repair, for others. The surgical strategy is tailored to address individual pathology, based on each patient's aortic anatomy and disease. FET technique in acute DeBakey type I aortic dissection extends the surgical repair into and beyond the arch with the potential to address some of the limitations of the conventional type I aortic dissection repair, but it has not been widely adopted yet. Only a limited number of institutions have embraced this modality during the acute phase of aortic dissection out of concern that it may be too complicated during an emergency situation. We have developed a simplified technique that has evolved over the past several years and the purpose of this report is to describe it.


Catestatin Regulates Epithelial Cell Dynamics to Improve Intestinal Inflammation.
Ulcerative colitis (UC) is characterized by aberrant regulation of tight junctions (TJ), signal transducer and activator of transcription 3 (STAT3), and interleukin (IL)-8/18, which lead to intestinal barrier defects. Catestatin (CST), an enterochromaffin-derived peptide, regulates immune communication and STAT-3 in the inflamed intestine. Here, we investigated the effects of CST during the development of inflammation using human biopsies from patients with active UC, human colonic epithelial cells (Caco2), and an experimental model of UC (dextran sulfate sodium [DSS]-colitis). In UC patients, the protein and mRNA level of CST was significantly decreased. Colonic expression of CST showed a strong positive linear relationship with TJ proteins and STAT3, and a strong negative correlation with IL-8 and IL-18. Intra-rectal administration of CST reduced the severity of experimental colitis, IL-18 colonic levels, maintained TJ proteins and enhanced the phosphorylation of STAT3. CST administration increased proliferation, viability, migration, TJ proteins, and p-STAT3 levels, and reduced IL-8 & IL-18 in LPS- & DSS-induced Caco2 cell epithelial injury, and the presence of STAT-3 inhibitor abolished the beneficial effect of CST. In inflammatory conditions, we conclude that CST could regulate intestinal mucosal dynamic via a potential STAT3-dependent pathway that needs to be further defined. Targeting CST in intestinal epithelial cells (IECs) should be a promising therapeutic approach such as when intestinal epithelial cell homeostasis is compromised in UC patients.


Primary bone lymphoma: report of a case with multifocal skeletal involvement.
Primary bone lymphoma is an uncommon tumour accounting for approximately 4-5% of extra nodal lymphoma and less than 1% of all Non-Hodgkin's lymphoma. The radiographic appearance of primary bone lymphoma is variable. As lesions frequently resemble other disease processes namely chronic osteomyelitis and metastases, further imaging evaluation and histopathological examination allow early identification for appropriate treatment. The authors present a case of anaplastic large cell lymphoma of bone presenting with multifocal osseus involvement.


Jingzhaotoxin-XII, a gating modifier specific for Kv4.1 channels.
Jingzhaotoxin-XII (JZTX-XII), a 29-residue polypeptide, was purified from the venom of the Chinese tarantula Chilobrachys jingzhao. Electrophysiological recordings carried out in Xenopus laevis oocytes showed that JZTX-XII is specific for Kv4.1 channels, with the IC50 value of 0.363 microM. It interacts with the channels by modifying the gating behavior. JZTX-XII shares 80% sequence identity with phrixotoxin1, a potent inhibitor for Kv4.2 and Kv4.3 channels. Structure analysis indicates that the difference of the charge distribution in the interactive surface perhaps influences the specific pharmacology of the toxins. JZTX-XII should be a valuable tool for the investigation of the Kv4.1 channels.


Temporal trend of leprosy among the elderly in Brazil, 2001 - 2018.
To describe the temporal trends of leprosy indicators among the elderly in Brazil in 2001 - 2018. This was an ecological time-series study of new leprosy cases in the elderly reported to the Notifiable Diseases Information System. Prais-Winsten generalized linear regression was used to estimate temporal variations. There were 687 317 new leprosy cases in Brazil from 2001 - 2018, of which 129 214 (18.8%) were among elderly people. Overall detection rates in the elderly and of new cases with grade-2 disability showed a falling trend, with an annual percent change of -4.6% (95%CI = -5.1 to -4.0) and -3.9% (95%CI = -4.6 to -3.2). New case and new multibacillary case proportions showed an increasing trend, with an annual percent change of 2.9% (95%CI = 2.6 to 3.3) and 1.4% (95%CI = 1.0 to 1.7), respectively. Detection rates for new leprosy cases in elderly people in Brazil are decreasing, but the proportion of new cases and multibacillary cases are trending upwards. New cases are shifting to older age groups, and demographic transition and immunosenescence are an influence. Inadequate reduction of grade-2 disability indicates a high risk of physical disability persists. Improved contact tracing and more effective action are needed in this age group.


Contributing factors to suicide: Political, social, cultural and economic.
This review summarizes recent research in four environmental areas affecting risk of deaths by suicide. Politically, the weight of the evidence suggests that laws increasing social welfare expenditures and other policies assisting persons with low incomes (e.g., minimum wage) tend to lower suicide rates. Other legal changes such as those restricting firearms and alcohol availability can also prevent suicides. The social institutions of marriage, as well as parenting, continue to serve as protective factors against suicide, although the degree of protection is often gendered. Religiousness tends to be inversely associated with suicide deaths at the individual level of analysis, but the mediators need exploration to determine what accounts for the association: social support, better mental health, better physical health, less divorce, or other covariates. Cultural definitions of the traditional male role (e.g., breadwinner culture) continue to help explain the high male to female suicide ratio. New work on the "culture of suicide" shows promise. The degree of approval of suicide is sometimes the single most important factor in predicting suicide. At the individual level of analysis, two of the strongest predictors of suicide are economic ones: unemployment and low socio-economic status. Attention is drawn to enhancing the minimum wage as a policy known to lower state suicide rates. Limitations of research include model mis-specification, conflicting results especially when ecological data are employed, and a need for more research exploring moderators of established patterns such as that between religiousness and suicide.


Coding for spatial goals in the prelimbic/infralimbic area of the rat frontal cortex.
Finding one's way in space requires a distributed neural network to support accurate spatial navigation. In the rat, this network likely includes the hippocampus and its place cells. Although such cells allow the organism to locate itself in the environment, an additional mechanism is required to specify the animal's goal. Here, we show that firing activity of neurons in medial prefrontal cortex (mPFC) reflects the motivational salience of places. We recorded mPFC neurons from rats performing a place navigation task, and found that a substantial proportion of cells in the prelimbic/infralimbic area had place fields. A much smaller proportion of cells with such properties was found in the dorsal anterior cingulate area. Furthermore, the distribution of place fields in prelimbic/infralimbic cells was not homogeneous: goal locations were overrepresented. Because such locations were spatially dissociated from rewards, we suggest that mPFC neurons might be responsible for encoding the rat's goals, a process necessary for path planning.


Towards improving the co-ordination of supportive cancer care services in the community.
In this paper an evaluation approach to assess the co-ordination of supportive community cancer care is presented. The aim of the study was to identify current gaps in co-ordination of services in a selected region in the province of Ontario, Canada, determine how consistent these gaps were across the province of Ontario, and develop service design considerations for improving the co-ordination of supportive cancer care services in the province of Ontario. The study addressed services required by two populations -clients who had been recently diagnosed and those in the palliative stages of cancer. The evaluation was theory-driven and incorporated evidence from three methods: a systematic literature review, a community case study and a provincial scan. The results revealed the absence of a formal supportive cancer care system and a complex community care system. Supportive cancer care was shown to be delivered by a range of generalist programs that lacked specialisation in addressing the unique needs of cancer clients. In addition, there was no clear evidence of leadership for co-ordinating supportive cancer care, where client care was most often provided by multiple programs at any given point in time. The study generated recommendations to improve co-ordination of supportive cancer care at both the administrative as well as direct care level.


Polymerase chain reaction for the detection of Bordetella pertussis in clinical nasopharyngeal aspirates.
A PCR procedure for the detection of Bordetella pertussis in nasopharyngeal aspirates (NPAs) was developed with primers derived from the pertussis toxin promoter region. The amplification resulted in a 191-bp PCR product specific for B. pertussis. A total of 681 NPAs collected from children with cough lasting >7 days was evaluated by PCR and culture; 104 aspirates were positive by PCR and 93 by culture. Sixteen cases were positive only by PCR and five culture positive aspirates were negative by PCR. An internal control was included in the assay to monitor the performance of the PCR and to identify possible inhibitory components in clinical samples. The PCR method was more efficient than culture in detecting B. pertussis in samples collected late in the disease, in antibiotic-treated children and in patients with mild disease.


Critical period of weed control in winter canola (Brassica napus L.) in a semi-arid region.
In order to determine the critical period of weed control in winter canola (Brassica napus L. cv. Okapi) an experiment was carried out at research field of Tarbiat Modarres University, Tehran, Iran on 2004-2005 growing season. Fourteen experimental treatments which divided into two sets were arranged in Randomized complete blocks design with four replications. In the first set, the crop was kept weed-free from emergence time to two-leaf stage (V2), four-leaf stage (V4), six-leaf stage (V6), eight-leaf stage (V8), early flowering (IF), 50% of silique set (50% SS) and final harvest (H). In the second set, weeds where permitted to grow with the crop until above mentioned stages. In this study critical period of weed control was determined according to evaluate seed bank emerged weed biomass effect on canola grain yield loss using Gompertz and logistic equations. Result showed a critical time of weed control about 25 days after emergence (between four to six-leaf stages) with 5% accepted yield loss. Therefore, weed control in this time could provide the best result and avoid yield loss and damage to agroecosystem.


A narrative review of the empirical evidence on public attitudes on brain death and vital organ transplantation: the need for better data to inform policy.
Vital organ transplantation is premised on 'the dead donor rule': donors must be declared dead according to medical and legal criteria prior to donation. However, it is controversial whether individuals diagnosed as 'brain dead' are really dead in accordance with the established biological conception of death-the irreversible cessation of the functioning of the organism as a whole. A basic understanding of brain death is also relevant for giving valid, informed consent to serve as an organ donor. There is therefore a need for reliable empirical data on public understanding of brain death and vital organ transplantation. We conducted a review of the empirical literature that identified 43 articles with approximately 18,603 study participants. These data demonstrate that participants generally do not understand three key issues: (1) uncontested biological facts about brain death, (2) the legal status of brain death and (3) that organs are procured from brain dead patients while their hearts are still beating and before their removal from ventilators. These data suggest that, despite scholarly claims of widespread public support for organ donation from brain dead patients, the existing data on public attitudes regarding brain death and organ transplantation reflect substantial public confusion. Our review raises questions about the validity of consent for vital organ transplantation and suggests that existing data are of little assistance in developing policy proposals for organ transplantation from brain dead patients. New approaches to rigorous empirical research with educational components and evaluations of understanding are urgently needed.


Interaction of Co+ and Co2+ with glycine. A theoretical study.
Cobalt cations are open shell systems with several possible electronic states arising from the different occupations of the 3d and 4s orbitals. The influence of these occupations on the relative stability of the coordination modes of the metal cation to glycine has been studied by means of theoretical methods. The structure and vibrational frequencies have been determined using the B3LYP method. Single-point calculations have also been carried out at the CCSD(T) level. The most stable structure of Co(+)-glycine is bidentate, with the Co(+) cation interacting with the amino group and the carbonyl oxygen of neutral glycine, and the ground electronic state being (3)A. For Co(2+)-glycine, the lowest energy structure corresponds to the interaction of the metal cation with the carboxylate group of the zwitterionic glycine, the ground electronic state being (4)A''.


Convenient and Live Movement (CALM) for women undergoing breast cancer treatment: Challenges and recommendations for internet-based yoga research.
To conduct a pilot trial of internet-based, cancer-adapted yoga for women receiving breast cancer treatment. Women undergoing radiation or chemotherapy for breast cancer were recruited for 12, 75-min, biweekly, cancer-adapted yoga classes delivered via internet-based, multipoint videoconferencing. Data were collected on feasibility and acceptability, including qualitative feedback from participants and the yoga instructor. Among 42 women approached, 13 declined eligibility screening, and 23 were ineligible. All 6 women who were eligible provided consent, but 2 withdrew prior to beginning yoga classes. The remaining 4 participants attended 1-11 of 12 online yoga classes. In post-intervention interviews, participants and the instructor agreed that internet-based yoga classes hold great potential for increasing access and improving psychological outcomes in adults with cancer. Qualitative feedback from participants revealed suggestions for future trials of internet-based, cancer-adapted yoga classes, including: continued use of group format; offering more varied class times to accommodate patients' demanding schedules and fluctuating symptoms; enrolling patients after they have acclimated to or completed cancer treatment; streamlining the technology interface; and careful attention to participant burden when designing surveys/forms. The instructor recommended closed session courses, as opposed to rolling enrollment; teaching the same modified poses for all participants, rather than individual tailoring; and using a large screen to allow closer monitoring of students' class experience. Internet delivery may increase patients' access to cancer-adapted yoga classes, but cancer-related and technological barriers remain. This study informs how to optimally design yoga classes, technology, and research procedures to maximize feasibility and acceptability in future trials.


[Association between socioeconomic status and all-cause mortality after breast cancer surgery: nationwide retrospective cohort study].
This study aims to evaluate and explain the socioeconomic inequalities of all-cause mortality after breast cancer surgery in South Korea. This population based study included all 8868 females who underwent radical mastectomy for breast cancer between January 2002 and June 2003. Follow-up for mortality continued from January 2002 to June 2006. The patients were divided into 4 socioeconomic classes according to their socioeconomic status as defined by the National Health Insurance contribution rate. The relationship between socioeconomic status and all-cause mortality after breast cancer surgery was assessed using the Cox proportional hazards model with adjusting for age, the Charlson's index score, emergency hospitalization, the type of hospital and the hospital ownership. Those in the lowest socioeconomic status group had a significantly higher hazard ratio of 2.09 (95% CI =1.50 - 2.91) compared with those in the highest socioeconomic group after controlling for all the identifiable confounding variables. For all-cause mortality after radical mastectomy, all the other income groups showed significantly higher 3-year mortality rates than did the highest income group. The socioeconomic status of breast cancer patients should be considered as an independent prognostic factor that affects all-cause mortality after radical mastectomy, and this is possibly due to a delayed diagnosis, limited access or minimal treatment leading to higher mortality. This study may provide tangible support to intensify surveillance and treatment for breast cancer among low socioeconomic class women.


Predictive Modeling Of Alzheimer's Disease Prognosis Using Anatomical & Diffusion MRI.
Mild cognitive impairment (MCI) is an intermediate stage between healthy aging and Alzheimer's disease (AD), and AD is a progressive neurodegenerative disorder that affects around 50 million people worldwide. As new AD treatments begin to be developed, one key goal of AD research is to predict which individuals with MCI are most likely to progress to AD over a given interval (such as 2 years); if successful, these individuals could be preferentially enrolled in drug trials that aim to slow AD progression. Here we benchmarked a range of MCI-to-AD predictive models including linear regressions, support vector machines, and random forests, using predictors from anatomical and diffusion-weighted brain MRI, age, sex, APOE genotype and standardized clinical scores. In evaluations on 2,448 subjects (1,132 MCI, 883 healthy controls, 433 with dementia) from the ADNI study, models including PCA-compacted features achieved a balanced accuracy of 75.3% (using cortical features) and 89.7% using diffusion MRI measures on test set, suggesting the added prognostic value of microstructural metrics obtainable with diffusion MRI.


Clinical significance of variceal hemorrhage in recent years in patients with liver cirrhosis and esophageal varices.
Recent progress in the treatment of variceal bleeding might have reduced the impact of variceal bleeding on survival in patients with esophageal varices. We conducted a retrospective cohort study in an attempt to re-evaluate the clinical significance of variceal bleeding. A cohort of 304 patients with liver cirrhosis and esophageal varices, who had no previous history of variceal bleeding and no prophylactic therapy, was studied. During a median follow-up period of 32 months, 55 patients (18%) bled from varices and 111 (37%) died. Variceal hemorrhages accounted for 15% of total deaths. The mortality of first variceal bleeding was 25% in the whole group, but was remarkably different depending on liver function at the time of bleeding (0% in grade Child A vs 55% in grade C; P < 0.05). Among the survivors of first bleeding, 30% experienced rebleeding. Form of varix, red color sign and heavy drinking were the independent risk factors for first variceal bleeding. Multivariate analysis revealed that variceal bleeding still had a significant (P < 0.001) impact on death in the whole cohort, when other independent prognostic factors such as age, ascites, encephalopathy, platelet count, serum albumin level and hepatocellular carcinoma were adjusted. Furthermore, in subgroup analyses, variceal bleeding was more strongly (P < 0.001) linked to death in patients with alcoholic cirrhosis than in those with non-alcoholic cirrhosis, and showed a significant association with survival only for the patients in Child grade B. Variceal bleeding has various prognostic impacts depending on the etiology of cirrhosis or on the degree of liver dysfunction, and this needs to be taken into account in the prophylaxis against first variceal bleeding.


Mountain biodiversity patterns at low and high latitudes.
This paper presents an overview of mountain biodiversity at a multitude of scales in space, time, and function. Even though species richness is usually the focal component in nature conservation, genetic diversity within species is equally important. The small-scale distribution of species in the tropical Andes, as exemplified by the plant genera Calceolaria and Bartsia, contrasts against the situation in high-latitude mountains, e.g., the Scandes, where species have wide ranges and many are circumpolar. Recent studies on alpine plants based on molecular methods show that the intraspecific genetic diversity tends to increase with latitude, a situation brought about by the glaciation history with repeated contraction-expansion episodes of species' distributions. In tropical mountains, species distributions are geographically much narrower, often as a result of relatively recent, local speciation. Thus, whereas species richness in mountains decreases from the Equator towards the poles, genetic diversity shows the opposite trend. Finally, a comparison of ecosystem diversity in low- and high-latitude mountain ranges (tropical Andes vs. Scandes) shows that the landscapes differ profoundly with regard to timberline ecotones, snow distribution, and climate variables, and are subject to widely different impacts of global change


Impaired HLA Class I Antigen Processing and Presentation as a Mechanism of Acquired Resistance to Immune Checkpoint Inhibitors in Lung Cancer.
Mechanisms of acquired resistance to immune checkpoint inhibitors (ICI) are poorly understood. We leveraged a collection of 14 ICI-resistant lung cancer samples to investigate whether alterations in genes encoding HLA Class I antigen processing and presentation machinery (APM) components or interferon signaling play a role in acquired resistance to PD-1 or PD-L1 antagonistic antibodies. Recurrent mutations or copy-number changes were not detected in our cohort. In one case, we found acquired homozygous loss of B2M that caused lack of cell-surface HLA Class I expression in the tumor and a matched patient-derived xenograft (PDX). Downregulation of B2M was also found in two additional PDXs established from ICI-resistant tumors. CRISPR-mediated knockout of B2m in an immunocompetent lung cancer mouse model conferred resistance to PD-1 blockade in vivo, proving its role in resistance to ICIs. These results indicate that HLA Class I APM disruption can mediate escape from ICIs in lung cancer.Significance: As programmed death 1 axis inhibitors are becoming more established in standard treatment algorithms for diverse malignancies, acquired resistance to these therapies is increasingly being encountered. Here, we found that defective antigen processing and presentation can serve as a mechanism of such resistance in lung cancer. Cancer Discov; 7(12); 1420-35. ©2017 AACR.This article is highlighted in the In This Issue feature, p. 1355.


The key importance of soy isoflavone bioavailability to understanding health benefits.
Research over the past two decades has provided significant epidemiological and other evidence for the health benefits of the consumption of soy-based foods. A large number of dietary intervention studies have examined the effects of soy isoflavones on risk factors for cardiovascular disease and hormone-dependent cancers. However, these report large variability in outcome measures, very limited reproducibility between studies, and in some cases, controversy between the results of clinical trials using dietary soy or soy protein and isoflavone supplementation. This highlights a major gap in our understanding of soy isoflavone uptake, metabolism, distribution, and overall bioavailability. There are many potential factors that may influence bioavailability and a better knowledge is necessary to rationalize the inconsistencies in the intervention and clinical studies. This review focuses attention on our current state of knowledge in this area and highlights the importance of metabolism of the parent soy isoflavones and the critical role of gut microbiota on the bioavailability of these compounds and their metabolites.


Predictability in the evolution of Orthopteran cardenolide insensitivity.
The repeated evolutionary specialization of distantly related insects to cardenolide-containing host plants provides a stunning example of parallel adaptation. Hundreds of herbivorous insect species have independently evolved insensitivity to cardenolides, which are potent inhibitors of the alpha-subunit of Na+,K+-ATPase (ATPα). Previous studies investigating ATPα-mediated cardenolide insensitivity in five insect orders have revealed remarkably high levels of parallelism in the evolution of this trait, including the frequent occurrence of parallel amino acid substitutions at two sites and recurrent episodes of duplication followed by neo-functionalization. Here we add data for a sixth insect order, Orthoptera, which includes an ancient group of highly aposematic cardenolide-sequestering grasshoppers in the family Pyrgomorphidae. We find that Orthopterans exhibit largely predictable patterns of evolution of insensitivity established by sampling other insect orders. Taken together the data lend further support to the proposal that negative pleiotropic constraints are a key determinant in the evolution of cardenolide insensitivity in insects. Furthermore, analysis of our expanded taxonomic survey implicates positive selection acting on site 111 of cardenolide-sequestering species with a single-copy of ATPα, and sites 115, 118 and 122 in lineages with neo-functionalized duplicate copies, all of which are sites of frequent parallel amino acid substitution. This article is part of the theme issue 'Convergent evolution in the genomics era: new insights and directions'.


Video head impulse and suppression head impulse test in vestibular migraine.
Vestibular Migraine (VM) is a frequent cause of recurrent spontaneous vertigo. While some report a normal Video Head Impulse Test (vHIT) in VM, others observed abnormal results on this test. Whether or not methodological discrepancies could be the cause of these differences is not known. There are 2 vHIT methods: subjects fixating an earth-fixed target (HIMP paradigm) or a head-fixed target, the suppression head impulse test (SHIMP paradigm). The present study aimed to compare VM patients against healthy controls on both HIMP and SHIMP in order to unravel any differences between them. Forty-eight VM patients and 27 healthy controls tested with both the HIMP and SHIMP paradigm. Results: Our 48 VM patients showed mean VOR normal range gains in both the HIMP and SHIMP paradigms, although there were some VOR impairments in individual semicircular SCCs. VM patients with motion sickness had lower horizontal VOR gain than those without motion sickness, with the HIMP, but not the SHIMP paradigm. VM patients have normal VOR gain with either vHIT paradigm. The clinical significance of this observation is that a definitely abnormal vHIT with either method is unlikely to be due to vestibular migraine and an alternative diagnosis should be sought.


Carrier rates of the ancestral Indian W24X mutation in GJB2 in the general Gypsy population and individual subisolates.
Mutations in the GJB2 gene are the most common cause of autosomal recessive nonsyndromic hearing loss and occur in approximately 20% of all cases of prelingual deafness. Previous studies of Roma/Gypsies in Slovakia, the Czech Republic, and Spain have shown that W24X, the most common GJB2 mutation in India, is also the prevalent molecular defect in the Gypsy population. The reported W24X frequencies vary broadly from 23% to 93% of Gypsy mutant alleles, likely reflecting local founder effects, drift, and differential admixture in the subisolates of this genetically structured population. Our goal was to provide more representative data on W24X carrier rates in European Gypsies, which can inform individual diagnostic investigations and public health initiatives across countries. Mutation testing in 603 control subjects of Gypsy ethnicity, representing 8 traditional subisolates in southeastern Europe and 4 additional European regions revealed that W24X is spread across subisolates, as expected for an ancestral founder mutation. While variation between subisolates does exist, the average carrier rates, overall and in the major linguistic/migrational categories of Balkan Gypsies, Vlax Roma, and west European Gypsies, are consistently in the 4%-5% range. The results place W24X among the three most common founder mutations in the Gypsies, and classify them as one of the high-risk populations for prelingual deafness. Higher demands on language acquisition in this bilingual population, together with poorer quality of health care compared to autochthonous Europeans, make the consequences of congenital deafness even more damaging than is usually the case. Neonatal screening for W24X among Gypsies would be a justified and cost-effective public health intervention.


Individual differences in the development of children's arithmetic fluency from grades 2 to 3.
In the present research, we provide empirical evidence for the process of symbolic integration of number associations, focusing on the development of simple addition (e.g., 5 + 3 = 8), subtraction (e.g., 5 - 3 = 2), and multiplication (e.g., 5 × 3 = 15). Canadian children were assessed twice, in Grade 2 and Grade 3 (N = 244; 55% girls). All families were English-speaking, and parent education levels ranged from high school to postgraduate, with a median of community college. In Grade 2, children completed general cognitive tasks (i.e., receptive vocabulary, working memory, nonverbal reasoning, and inhibitory control). In both grades, children completed single-digit addition and complementary subtraction problems. In Grade 3, they completed single-digit multiplication problems and measures of applied mathematics, specifically, word-problem solving, algebra, and measurement. We found that addition and subtraction were reciprocally related (controlling for cognitive skills). Subtraction fluency predicted multiplication in Grade 3, whereas addition fluency did not. In Grade 3, both subtraction and multiplication fluency were predictors of applied mathematics, with multiplication partially mediating the relation between subtraction and applied mathematics performance. These findings support the view that learning arithmetic associations is a hierarchical process. As students practice each new skill, individual differences reflect the integration of the novel component into the developing associative network. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Periampullary diverticula: consequences of failed ERCP.
Periampullary diverticula (PAD) are associated with biliary disease and contribute to failure of endoscopic retrograde cholangiopancreatography (ERCP), especially in elderly patients. The presence of PAD and causes of failure to cannulate the ampulla were noted in 1211 consecutive patients undergoing ERCP. Case notes of 100 consecutive patients with PAD were reviewed retrospectively. Overall prevalence of PAD was 9%. Prevalence was higher in patients > or = 75 years when compared with those < 75 years (19.2% vs 4.8%, P < 0.0001). Ampullary cannulation was successful in 62.4% of patients with PAD and 92.7% without PAD (P < 0.0001). Success rates were lower in patients with intradiverticular papillae than in those with juxtapapillary diverticula (38.1% vs 77.6%; P < 0.0001). Of 19 patients with PAD who did not have any imaging other than ultrasound, 16 were asymptomatic over a median follow-up of 20 months. Biliary surgery was performed on 35 patients, with no major complication. PAD are a major cause of failed ERCP. Failure rates are higher in patients with intradiverticular papillae than juxtapapillary diverticula. Though a large proportion of patients not imaged remain asymptomatic on follow-up, it is difficult to predict which patients may form this group. Surgery, when indicated, is safe and effective in elderly patients in whom ERCP has failed.


Cortical reflex myoclonus responds to intravenous lisuride.
Six patients with myoclonus were given 0.1-0.15 mg lisuride i.v. All patients had stimulus-sensitive myoclonus and an increased size of somatosensory evoked potentials, and in three there was electrophysiological evidence of a cortical event time-locked to the jerks. All subjects showed a considerable diminution of spontaneous, action- and stimulus-evoked jerking. Lisuride has potent central dopaminergic and serotonergic actions. Administration of the dopamine agonists levodopa or apomorphine had no effect on myoclonic jerking in any of the six patients. Detailed pharmacological analysis of the myoclonus in one patient showed that levodopa, apomorphine, and haloperidol had no effect, and that haloperidol did not prevent the therapeutic action of lisuride. 5-Hydroxytryptophan abolished the myoclonus, and methysergide prevented the beneficial effect of lisuride, although it did not alter spontaneous myoclonus. These observations suggest that lisuride improves some types of reflex, stimulus-sensitive cortical myoclonus by a serotonin agonist action.


A new training approach in endoscopic percutaneous tracheostomy using a simulation model based on biological tissue.
Ciaglia et al. originally proposed the percutaneous tracheostomy in 1985 as an alternative to the surgical approach. Several variants have been developed in the years with a convincing evidence that, compared to surgical tracheostomy, the percutaneous approach is more cost-effective without compromising safety. However the procedure is not immune from complications and requires procedural skills and adequate level of expertise with a steep learning curve. In this contest, to optimize safety and efficacy of percutaneous tracheostomy in different clinical scenarios, physicians should be familiar with the different available percutaneous techniques. Traditionally, the training has been performed using manikins or animal models. However, by definition the manikin is not able to replicate the complex anatomy and does not provide a realist model for learning. A sheep model was implemented and was tested in a simulation-based course for percutaneous dilational tracheostomy and cricothyrotomy organized by the Department of Surgical Sciences of University of Turin at the Special Educational Section of the Veterinary Sciences Department (University of Turin). From September 2012 to December 2014, 2 sessions of the training course were performed. Thirty-two anesthesiologists (16 men and 16 women, mean (SD) age 40.4 (SD 8.6) attended the course. A quality assessment of the course was then collected by participants. The biological model we implemented proved to be adequate and effective. Future studies are required to compare its efficacy with previously proposed training methods.


Somatosensory cortical remodelling after rehabilitation and clinical benefit of in writer's cramp.
In order to explore the pathophysiological basis of a new rehabilitation therapy in writer's cramp (WC), healthy controls, untreated WC patients and WC patients who recovered a legible handwriting after rehabilitation were explored using magnetoencephalography, and the somatosensory evoked fields of fingers I, II, III and V in the sensory cortex were studied. In the cortex controlling the dystonic limb, the size of the hand representation in the trained patients was similar to that of healthy controls, and significantly different from that of untrained patients. Trained patients exhibited 'super-normal' reorganisation of the finger maps. In the cortex controlling the non-dystonic limb, there was little difference between trained and untrained patients, and the hand representation was enlarged and disorganised. The authors hypothesise that prolonged tailored rehabilitation in WC may induce long-term plasticity phenomena, lateralised to the cortex controlling the dystonic hand.


Effect of mild irritant on gastric mucosal offensive and defensive factors.
The effect of hypotonic medium (Distilled water: DW) and hypertonic saline (HS: 5% NaCl) compared to control normal saline (NS) was studied on gastric ulcer induced by aspirin, 6 h cold restraint stress, ethanol, and pylorus ligation in rats. DW did not afford any protection while HS showed significant ulcer protective effects in all gastric ulcer models studied. The cytoprotective effect of HS seemed to be not only due to its effect on gastric acid secretion but also its effect on mucosal defensive factors like enhanced mucin secretion and decreased cell shedding. As determined by radioimmunoassay, DW did not produce any change in the accumulation of PGE and PGI2, while HS increased them significantly in the human gastric mucosal incubates compared to NS. However, in the incubates of human colonic mucosa, both DW and HS showed a significant increase in PGE with a tendency to increase in PGI2 accumulation.


Neuromuscular training improves performance and lower-extremity biomechanics in female athletes.
The purpose of this study was to examine the effects of a comprehensive neuromuscular training program on measures of performance and lower-extremity movement biomechanics in female athletes. The hypothesis was that significant improvements in measures of performance would be demonstrated concomitant with improved biomechanical measures related to anterior cruciate ligament injury risk. Forty-one female basketball, soccer, and volleyball players (age, 15.3 +/- 0.9 years; weight, 64.8 +/- 9.96 kg; height, 171.2 +/- 7.21 cm) underwent 6 weeks of training that included 4 main components (plyometric and movement, core strengthening and balance, resistance training, and speed training). Twelve age-, height-, and weight-matched controls underwent the same testing protocol twice 6 weeks apart. Trained athletes demonstrated increased predicted 1 repetition maximum squat (92%) and bench press (20%). Right and left single-leg hop distance increased 10.39 cm and 8.53 cm, respectively, and vertical jump also increased from 39.9 +/- 0.9 cm to 43.2 +/- 1.1 cm with training. Speed in a 9.1-m sprint improved from 1.80 +/- 0.02 seconds to 1.73 +/- 0.01 seconds. Pre- and posttest 3-dimensional motion analysis demonstrated increased knee flexion-extension range of motion during the landing phase of a vertical jump (right, 71.9 +/- 1.4 degrees to 76.9 +/- 1.4 degrees ; left, 71.3 +/- 1.5 degrees to 77.3 +/- 1.4 degrees ). Training decreased knee valgus (28%) and varus (38%) torques. Control subjects did not demonstrate significant alterations during the 6-week interval. The results of this study support the hypothesis that the combination of multiple-injury prevention-training components into a comprehensive program improves measures of performance and movement biomechanics.


Simultaneously cooperative, but serially antagonistic: a neuropsychological study of diagonistic dyspraxia in a case of Marchiafava-Bignami disease.
We describe a patient with Marchiafava-Bignami disease who showed, in addition to signs of callosal interruption, a peculiar form of diagonistic dyspraxia. Unlike the typical diagonistic dyspraxia, both of the patient's hands could simultaneously cooperate in a sequence of bimanual actions. More specifically, his right hand could start a commanded action with the cooperation of his left hand. However, once the action was completed, his left hand started an antagonistic action, undoing the result, with the cooperation of his right hand. Once this countermanding action was completed, the original action started again. These antagonistic actions repeated themselves alternately unless he was restrained. The patient's diagonistic dyspraxia was apparent in only some bimanual actions, and he showed no diagonistic dyspraxia when performing voluntary actions; the antagonistic actions occurred in response to oral commands or by imitation. Magnetic resonance imaging showed symmetrical demyelination with partial necrosis in the genu, body, and anterior splenium of the corpus callosum. We speculate that the bimanual coordination is possible because part of the corpus callosum is intact, whereas the antagonistic actions may be caused by conflict between the two hemispheres due to interhemispheric disinhibition elicited by the demyelinated part of the corpus callosum.


Phenomenology and neurobiology of cocaine withdrawal: are they related?
1. The presence of mood disturbances and platelet 3H-imipramine binding, a putative peripheral serotonergic marker, were evaluated in a group of 27 cocaine users three days after drug withdrawal. 2. Parameters of cocaine use and the linkage between cocaine withdrawal and "post-cocaine depression" were also investigated. In a subgroup of 10 patients, both psychopathological and biological measurements were repeated after 5 or 6 weeks. 3. Interpretation of the data by Pearson's analysis showed a statistically significant and positive correlation between Hamilton Rating Scale for Depression (HAM-D) scores and period of use. A trend towards a negative correlation, which however did not reach the statistical significance, was found between 3H-Imipramine binding and period of cocaine use, number of days of abstinence and HAM-D scores 4. When compared with normal volunteers at baseline, patients had significantly lower Bmax and Kd values which returned towards normal values after 5 or 6 weeks of cocaine withdrawal. 5. These results indicate the presence of a decreased platelet imipramine binding during cocaine withdrawal which may be due to the effect of the drug or alternatively, a result of concomitant depression which may be primary or secondary in origin. The decreased imipramine binding is a reversible phenomenon, since it increases with the time, in parallel with the improvement of depressive symptoms.


Association between major surgical admissions and the cognitive trajectory: 19 year follow-up of Whitehall II cohort study.
To quantify the association between major surgery and the age related cognitive trajectory. Prospective longitudinal cohort study. United Kingdom. 7532 adults with as many as five cognitive assessments between 1997 and 2016 in the Whitehall II study, with linkage to hospital episode statistics. Exposures of interest included any major hospital admission, defined as requiring more than one overnight stay during follow-up. The primary outcome was the global cognitive score established from a battery of cognitive tests encompassing reasoning, memory, and phonemic and semantic fluency. Bayesian linear mixed effects models were used to calculate the change in the age related cognitive trajectory after hospital admission. The odds of substantial cognitive decline induced by surgery defined as more than 1.96 standard deviations from a predicted trajectory (based on the first three cognitive waves of data) was also calculated. After accounting for the age related cognitive trajectory, major surgery was associated with a small additional cognitive decline, equivalent on average to less than five months of aging (95% credible interval 0.01 to 0.73 years). In comparison, admissions for medical conditions and stroke were associated with 1.4 (1.0 to 1.8) and 13 (9.6 to 16) years of aging, respectively. Substantial cognitive decline occurred in 2.5% of participants with no admissions, 5.5% of surgical admissions, and 12.7% of medical admissions. Compared with participants with no major hospital admissions, those with surgical or medical events were more likely to have substantial decline from their predicted trajectory (surgical admissions odds ratio 2.3, 95% credible interval 1.4 to 3.9; medical admissions 6.2, 3.4 to 11.0). Major surgery is associated with a small, long term change in the average cognitive trajectory that is less profound than for major medical admissions. The odds of substantial cognitive decline after surgery was about doubled, though lower than for medical admissions. During informed consent, this information should be weighed against the potential health benefits of surgery.


Effect of in-line drinking water chlorination at the point of collection on child diarrhoea in urban Bangladesh: a double-blind, cluster-randomised controlled trial.
Previous blinded trials of household water treatment interventions in low-income settings have failed to detect a reduction in child diarrhoea. Technological advances have enabled the development of automated in-line chlorine dosers that can disinfect drinking water without electricity, while also allowing users to continue their typical water collection practices. We aimed to evaluate the effect of installing novel passive chlorination devices at shared water points on child diarrhoea prevalence in low-income, densely populated communities in urban Bangladesh. In this double-blind cluster-randomised controlled trial, 100 shared water points (clusters) in two low-income urban communities in Bangladesh were randomly assigned (1:1) to have their drinking water automatically chlorinated at the point of collection by a solid tablet chlorine doser (intervention group) or to be treated by a visually identical doser that supplied vitamin C (active control group). The trial followed an open cohort design; all children younger than 5 years residing in households accessing enrolled water points were measured every 2-3 months during a 14-month follow-up period (children could migrate into or out of the cluster). The primary outcome was caregiver-reported child diarrhoea (≥3 loose or watery stools in a 24-h period [WHO criteria]) with a 1-week recall, including all available childhood observations in the analyses. This trial is registered with ClinicalTrials.gov, number NCT02606981, and is completed. Between July 5, 2015, and Nov 11, 2015, 100 water points with 920 eligible households were enrolled into the study and randomly assigned to the treatment (50 water points; 517 children at baseline; 2073 child observations included in the primary analysis) or control groups (50; 519; 2154). Children in the treatment group had less WHO-defined diarrhoea than did children in the control group (control 216 [10·0%] of 2154; treatment 156 [7·5%] of 2073; prevalence ratio 0·77, 95% CI 0·65-0·91). Drinking water at the point of collection at treatment taps had detectable free chlorine residual 83% (mean 0·37 ppm) of the time compared with 0% at control taps (0·00 ppm). Passive chlorination at the point of collection could be an effective and scalable strategy in low-income urban settings for reducing child diarrhoea and for achieving global progress towards Sustainable Development Goal 6.1 to attain universal access to safe and affordable drinking water. Targeting a low chlorine residual (<0·5 ppm) in treated water can increase taste acceptability of chlorinated drinking water while still reducing the risk of diarrhoea. The World Bank.


Value of internal mammary artery bypass.
Coronary artery bypass in 150 patients with angina has been accomplished with 225 IMA grafts and 85 saphenous vein grafts. There were 7 (4.7%) hospital deaths. Posterative catheterization in 85% of patients revealed that 9 (4.7%) IMA grafts and 11 (15%) vein graft had thrombosed. One-year catheterization in 32 patients (38 IMA grafts and 13 vein grafts) revealed no additional graft failures. The better patency of the IMA grafts appears to be related to their smaller size and therfore greater velocity of blood flow and freedom from intimal proliferation.


Dominant alleles of Saccharomyces cerevisiae CDC20 reveal its role in promoting anaphase.
We identified an allele of Saccharomyces cerevisiae CDC20 that exhibits a spindle-assembly checkpoint defect. Previous studies indicated that loss of CDC20 function caused cell cycle arrest prior to the onset of anaphase. In contrast, CDC20-50 caused inappropriate cell cycle progression through M phase in the absence of mitotic spindle function. This effect of CDC20-50 was dominant over wild type and was eliminated by a second mutation causing loss of function, suggesting that it encodes an overactive form of Cdc20p. Overexpression of CDC20 was found to cause a similar checkpoint defect, causing bypass of the preanaphase arrest produced by either microtubule-depolymerizing compounds or MPS1 overexpression. CDC20 overexpression was also able to overcome the anaphase delay caused by high levels of the anaphase inhibitor Pds1p, but not a mutant form immune to anaphase-promoting complex- (APC-)mediated proteolysis. CDC20 overexpression was unable to promote anaphase in cells deficient in APC function. These findings suggest that Cdc20p is a limiting factor that promotes anaphase entry by antagonizing Pds1p. Cdc20p may promote the APC-dependent proteolytic degradation of Pds1p and other factors that act to inhibit cell cycle progression through mitosis.


Race/Ethnicity and Treatment Outcome in a Randomized Controlled Trial for Trichotillomania (Hair-Pulling Disorder).
Treatment outcome was compared among non-Hispanic White and racial/ethnic minority participants with trichotillomania (TTM), or hair-pulling disorder. Symptom severity, quality of life, and TTM-related disability were compared in a behavior therapy trial with a stepped care approach: web-based self-help and then individual behavior therapy. The sample comprised 72% (n = 38) non-Hispanic White participants and 28% (n = 15) minority participants. The ethnic groups responded differently to treatment, with fewer minority participants showing improvement during web-based self-help. Response rates were equivalent between ethnic groups during the in-person behavior therapy. These results should be interpreted with caution because of the small sample size of minorities in the study and consequent inability to analyze results for each racial/ethnic group individually. Future studies should focus on the investigation of factors that may enable or hinder racial and ethnic minority participants to benefit from online and/or self-help behavior therapy for TTM.


Antimicrobial effect of 2% sodium hypochlorite and 2% chlorhexidine tested by different methods.
The objective of this study was to analyze the antimicrobial effect of 2% sodium hypochlorite (NaOCl) and 2% chlorhexidine (CHX) by agar diffusion test and by direct exposure test. Five microorganisms: Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aernginosa, Bacillus subtilis, Candida albicans, and one mixture of these were used. These strains were inoculated in brain heart infusion (BHI) and incubated at 37 degrees C for 24 h. For the agar diffusion test (ADT), 18 Petri plates with 20 ml of BHI agar were inoculated with 0.1 ml of the microbial suspensions, using sterile swabs that were spread on the medium, obtaining growth injunction. Fifty-four paper disks (9 mm in diameter) were immersed in the experimental solutions for 1 min. Subsequently, three papers disks containing one of the substances were placed on the BHI agar surface in each agar plate. The plates were maintained for 1 h at room temperature, and then incubated at 37 degrees C for 48 h. The diameter of microbial inhibition was measured around the papers disks containing the substances. For the direct exposure test, 162#50 sterile absorbent paper points were immersed in the experimental suspensions for 5 min, and were then placed on Petri plates and covered with one of the irrigant solutions, or with sterile distilled water (control group). After intervals of 5, 1 0 and 30 min, the paper points were removed from contact with the solutions and individually immersed in 7 ml of Letheen Broth, followed by incubation at 37 degrees C for 48 h. Microbial growth was evaluated by turbidity of the culture medium. A 0.1 ml inoculum obtained from the Letheen Broth was transferred to 7 ml of BHI, and incubated at 37 degrees C for 48 h. Bacterial growth was again evaluated by turbidity of the culture medium. Gram stain of BHI cultures was used for verification of contamination and growth was determined by macroscopic and microscopic examination. The best performance of antimicrobial effectiveness of NaOCI was observed in the direct exposure test, and of CHX was observed in the agar diffusion test. The magnitude of antimicrobial effect was influenced by the experimental methods, biological indicators and exposure time.


Selective autophagy: the fulcrum of plant-virus interaction.
Selective autophagy receptors play both proviral and antiviral roles during plant-virus interaction. However, little is known about the balance between such contradictory dual roles of these receptors. Tong et al. have deciphered the temporal regulation of antiviral and antiplant roles of a selective autophagy receptor, a virus-induced small peptide 1 (VISP1).


IRF4 Regulates the Ratio of T-Bet to Eomesodermin in CD8+ T Cells Responding to Persistent LCMV Infection.
CD8+ T cell exhaustion commonly occurs in chronic infections and cancers. During T cell exhaustion there is a progressive and hierarchical loss of effector cytokine production, up-regulation of inhibitory co-stimulatory molecules, and eventual deletion of antigen specific cells by apoptosis. A key factor that regulates T cell exhaustion is persistent TCR stimulation. Loss of this interaction results in restoration of CD8+ T cell effector functions in previously exhausted CD8+ T cells. TCR stimulation is also important for the differentiation of Eomeshi anti-viral CD8+ effector T cells from T-bethi precursors, both of which are required for optimal viral control. However, the molecular mechanisms regulating the differentiation of these two cell subsets and the relative ratios required for viral clearance have not been described. We show that TCR signal strength regulates the relative expression of T-bet and Eomes in antigen-specific CD8+ T cells by modulating levels of IRF4. Reduced IRF4 expression results in skewing of this ratio in the favor of Eomes, leading to lower proportions and numbers of T-bet+ Eomes- precursors and poor control of LCMV-clone 13 infection. Manipulation of this ratio in the favor of T-bet restores the differentiation of T-bet+ Eomes- precursors and the protective balance of T-bet to Eomes required for efficient viral control. These data highlight a critical role for IRF4 in regulating protective anti-viral CD8+ T cell responses by ensuring a balanced ratio of T-bet to Eomes, leading to the ultimate control of this chronic viral infection.


Intact-protein analysis system for discovery of serum-based disease biomarkers.
Profiling of serum and plasma proteins has substantial relevance to the discovery of circulating disease biomarkers. However, the extreme complexity and vast dynamic range of protein abundance in serum and plasma present a formidable challenge for protein analysis. Thus, integration of multiple technologies is required to achieve high-resolution and high-sensitivity proteomic analysis of serum or plasma. In this chapter, we describe an orthogonal multidimensional intact-protein analysis system (IPAS) (Wang et al., Mol Cell Proteomics 4:618-625, 2005) coupled with protein tagging (Faca et al., J Proteome Res 5:2009-2018, 2006) to profile the serum and plasma proteomes quantitatively, which we have applied in our biomarker discovery studies (Katayama et al., Genome Med 1:47, 2009; Faca et al., PLoS Med 5:e123, 2008; Zhang et al. Genome Biol 9:R93, 2008).


Ultraviolet radiation exposure of children and adolescents in Durban, South Africa.
The solar ultraviolet radiation (UVR) exposure of 30 children and adolescents in three age groups (4-6 years, 7-9 years and 13-14 years) was measured for 1 week in late summer (February-March) in Durban, South Africa, using UVR-sensitive polysulfone film badges (PSFB) attached to the lapel region of the body. The mean and median values for all ages over the study period were 2.0 and 1.2 standard erythemal dose (SED) units, respectively, where 1 SED = 100 J x m(-2). Individual PSFB doses were analyzed as a function of age, gender and behavior. No significant statistical differences were found between different age groups; however, there was a statistical difference between males and females, with males generally receiving higher PSFB doses. Subjects completed UVR exposure journals documenting their time outdoors, shade versus sun conditions, nature of their activities, clothing worn and their use of sunscreen for each day of the study. Activity patterns were noted as the most important factor influencing individual UVR dose. Ambient erythemal UVR was measured by a Yankee Environmental Systems UVB pyranometer, and a relationship between ambient UVR and individual UVR dose was derived. On average, subjects received a dose of 4.6% of the total daily erythemal UVR. Based on this factor, the potential dose of an individual over a full annual cycle was estimated. Accordingly, there were 139 days during the year when an individual with skin type I (light skin) would be likely to experience minimal erythema and 97 and 32 days for individuals with skin types II and III, respectively.


Impacts of human-induced environmental change in wetlands on aquatic animals.
Many wetlands harbour highly diverse biological communities and provide extensive ecosystem services; however, these important ecological features are being altered, degraded and destroyed around the world. Despite a wealth of research on how animals respond to anthropogenic changes to natural wetlands and how they use created wetlands, we lack a broad synthesis of these data. While some altered wetlands may provide vital habitat, others could pose a considerable risk to wildlife. This risk will be heightened if such wetlands are ecological traps - preferred habitats that confer lower fitness than another available habitat. Wetlands functioning as ecological traps could decrease both local and regional population persistence, and ultimately lead to extinctions. Most studies have examined how animals respond to changes in environmental conditions by measuring responses at the community and population levels, but studying ecological traps requires information on fitness and habitat preferences. Our current lack of knowledge of individual-level responses may therefore limit our capacity to manage wetland ecosystems effectively since ecological traps require different management practices to mitigate potential consequences. We conducted a global meta-analysis to characterise how animals respond to four key drivers of wetland alteration: agriculture, mining, restoration and urbanisation. Our overarching goal was to evaluate the ecological impacts of human alterations to wetland ecosystems, as well as identify current knowledge gaps that limit both the current understanding of these responses and effective wetland management. We extracted 1799 taxon-specific response ratios from 271 studies across 29 countries. Community- (e.g. richness) and population-level (e.g. density) measures within altered wetlands were largely comparable to those within reference wetlands. By contrast, individual fitness measures (e.g. survival) were often lower, highlighting the potential limitations of using only community- and population-level measures to assess habitat quality. Only four studies provided habitat-preference data, preventing investigation of the potential for altered wetlands to function as ecological traps. This is concerning because attempts to identify ecological traps may detect previously unidentified conservation risks. Although there was considerable variability amongst taxa, amphibians were typically the most sensitive taxon, and thus, may be a valuable bio-indicator of wetland quality. Despite suffering reduced survival and reproduction, measures such as time to and mass at metamorphosis were similar between altered and reference wetlands, suggesting that quantifying metamorphosis-related measures in isolation may not provide accurate information on habitat quality. Our review provides the most detailed evaluation to date of the ecological impacts of human alterations to wetland ecosystems. We emphasise that the role of wetlands in human-altered ecosystems can be complex, as they may represent important habitat but also pose potential risks to animals. Reduced availability of natural wetlands is increasing the importance of altered wetlands for aquatic animals. Consequently, we need to define what represents habitat quality from the perspective of animals, and gain a greater understanding of the underlying mechanisms of habitat selection and how these factors could be manipulated. Furthermore, strategies to enhance the quality of these wetlands should be implemented to maximise their conservation potential.


Divisions of general practice in Australia: how do they measure up in the international context?
Since the late 1980s, there has been evidence of an international trend towards more organised primary care. This has taken a number of forms including the emergence of primary care organisations. Underpinning such developments is an inherent belief in evidence that suggests that well-developed primary care is associated with improved health outcomes and greater cost-effectiveness within health systems. In Australia, primary care organisations have emerged as divisions of general practice. These are professionally-led, regionally-based, and largely government-funded voluntary associations of general practitioners that seek to co-ordinate local primary care services, and improve the quality of care and health outcomes for local communities. In this paper, we examine and debate the development of divisions in the international context, using six roles of primary care organisations outlined in published research. The six roles that are used as the basis for the critique are the ability of primary care organisations to: improve health outcomes; manage demand and control costs; engage primary care physicians; enable greater integration of health services; develop more accessible services in community and primary care settings; and enable greater scrutiny and assurance of quality of primary care services. We conclude that there has been an evolutionary approach to divisions' development and they now appear embedded as geographically-based planning and development organisations within the Australian primary health care system. The Australian Government has to date been cautious in its approach to intervention in divisions' direction and performance. However, options for the next phase include: making greater use of contracts between government and divisions; introducing and extending proposed national quality targets for divisions, linked with financial or other incentives for performance; government sub-contracting with state-based organisations to act as purchasers of care; pursuing a fund-holding approach within divisions; and developing divisions as a form of health maintenance organisation. The challenge for the Australian Government, should it wish to see divisions' role expand, is to find mechanisms to enable this without compromising the relatively strong GP engagement that increasingly distinguishes divisions of general practice within the international experience of primary care organisations.


The Epstein-Barr Virus Lytic Protein BZLF1 as a Candidate Target Antigen for Vaccine Development.
The Epstein-Barr virus (EBV) is an oncogenic, γ-herpesvirus associated with a broad spectrum of disease. Although most immune-competent individuals can effectivley develop efficient adaptive immune responses to EBV, immunocompromised individuals are at serious risk for developing life-threatening diseases, such as Hodgkin lymphoma and posttransplant lymphoproliferative disorder (PTLD). Given the significant morbidity associated with EBV infection in high-risk populations, there is a need to develop vaccine strategies that restore or enhance EBV-specific immune responses. Here, we identify the EBV immediate-early protein BZLF1 as a potential target antigen for vaccine development. Primary tumors from patients with PTLD and a chimeric human-murine model of EBV-driven lymphoproliferative disorder (EBV-LPD) express BZLF1 protein. Pulsing human dendritic cells (DC) with recombinant BZLF1 followed by incubation with autologous mononuclear cells led to expansion of BZLF1-specific CD8(+) T cells in vitro and primed BZLF1-specific T-cell responses in vivo. In addition, vaccination of hu-PBL-SCID mice with BZLF1-transduced DCs induced specific cellular immunity and significantly prolonged survival from fatal EBV-LPD. These findings identify BZLF1 as a candidate target protein in the immunosurveillance of EBV and provide a rationale for considering BZLF1 in vaccine strategies to enhance primary and recall immune responses and potentially prevent EBV-associated diseases.