In vitro toxicological characterisation of three arsenic-containing hydrocarbons.
Arsenic-containing hydrocarbons are one group of fat-soluble organic arsenic compounds (arsenolipids) found in marine fish and other seafood. A risk assessment of arsenolipids is urgently needed, but has not been possible because of the total lack of toxicological data. In this study the cellular toxicity of three arsenic-containing hydrocarbons was investigated in cultured human bladder (UROtsa) and liver (HepG2) cells. Cytotoxicity of the arsenic-containing hydrocarbons was comparable to that of arsenite, which was applied as the toxic reference arsenical. A large cellular accumulation of arsenic, as measured by ICP-MS/MS, was observed after incubation of both cell lines with the arsenolipids. Moreover, the toxic mode of action shown by the three arsenic-containing hydrocarbons seemed to differ from that observed for arsenite. Evidence suggests that the high cytotoxic potential of the lipophilic arsenicals results from a decrease in the cellular energy level. This first in vitro based risk assessment cannot exclude a risk to human health related to the presence of arsenolipids in seafood, and indicates the urgent need for further toxicity studies in experimental animals to fully assess this possible risk.


Nucleotide sequencing of the Proteus mirabilis calcium-independent hemolysin genes (hpmA and hpmB) reveals sequence similarity with the Serratia marcescens hemolysin genes (shlA and shlB).
We cloned a 13.5-kilobase EcoRI fragment containing the calcium-independent hemolysin determinant (pWPM110) from a clinical isolate of Proteus mirabilis (477-12). The DNA sequence of a 7,191-base-pair region of pWPM110 was determined. Two polypeptides are encoded in this region, HpmB and HpmA (in that transcriptional order), with predicted molecular masses of 63,204 and 165,868 daltons, respectively. A putative Fur-binding site was identified upstream of hpmB overlapping the -35 region of the proposed hpm promoter. In vitro transcription-translation of pWPM110 DNA and other subclones confirmed the assignment of molecular masses for the predicted polypeptides. These polypeptides are predicted to have NH2-terminal leader peptides of 17 and 29 amino acids, respectively. NH2-terminal amino acid sequence analysis of purified extracellular hemolysin (HpmA) confirmed the cleavage of the 29-amino-acid leader peptide in the secreted form of HpmA. Hemolysis assays and immunoblot analysis of Escherichia coli containing subclones expressing hpmA, hpmB, or both indicated that HpmB is necessary for the extracellular secretion and activation of HpmA. Significant nucleotide identity (52.1%) was seen between hpm and the shl hemolysin gene sequences of Serratia marcescens despite differences in the G+C contents of these genes (hpm, 38%; shl, 65%). The predicted amino acid sequences of HpmB and HpmA are also similar to those of ShlB and ShlA, the respective sequence identities being 55.4 and 46.7%. Predicted cysteine residues and major hydrophobic and amphipathic domains have been strongly conserved in both proteins. Thus, we have identified a new hemolysin gene family among gram-negative opportunistic pathogens.


The Pros and Cons of Getting Engaged in an Online Social Community Embedded Within Digital Cognitive Behavioral Therapy for Insomnia: Survey Among Users.
Sleepio is a proven digital sleep improvement program based on cognitive behavioral therapy techniques. Users have the option to join an online community that includes weekly expert discussions, peer-to-peer discussion forums, and personal message walls. The aim of this study was to conduct an online survey to (1) explore the reasons for deciding to engage with the Sleepio online community, (2) explore the potential benefits arising from engagement with the online community, and (3) identify and describe any problematic issues related to use of the online community. We developed an online survey and posted an invitation to the community discussion forum inviting users to participate. In addition, we sent an email invitation to 970 individuals who had previously or were currently working through the Sleepio program to participate in this study. In total, 100 respondents (70/100, 70% female; mean age 51 years, range 26-82 years) completed the online survey. Most respondents had started Sleepio with chronic sleep problems (59/100, 59% up to 10 years; 35/100, 35% >10 years) and had actively engaged with the online community (85/100, 85%) had made a discussion or wall post). At the time of the survey, respondents had used Sleepio for a median of 12 weeks (range from 3 weeks to 2 years). We analyzed responses to the open-ended questions using thematic analysis. This analysis revealed 5 initial drivers for engagement: (1) the desire to connect with people facing similar issues, (2) seeking personalized advice, (3) curiosity, (4) being invited by other members, and (5) wanting to use all available sleep improvement tools. Advantages of engagement included access to continuous support, a reduced sense of isolation, being part of a nonjudgmental community, personalized advice, positive comparisons with others, encouragement to keep going, and altruism. We found 5 potential disadvantages: design and navigation issues, uncertain quality of user-generated content, negative comparisons with others, excessive time commitments, and data privacy concerns. Participants related their community experiences to engagement with the Sleepio program, with many stating it had supported their efforts to improve their sleep, as well as helping with adherence and commitment to the program. Despite some concerns, members regarded the Sleepio community as a valuable resource. Online communities may be a useful means through which to support long-term engagement with Web-based therapy for insomnia.


Taking aim at a moving target: designing drugs to inhibit drug-resistant HIV-1 reverse transcriptases.
HIV undergoes rapid genetic variation; this variation is caused primarily by the enormous number of viruses produced daily in an infected individual. Because of this variation, HIV presents a moving target for drug and vaccine development. The variation within individuals has led to the generation of diverse HIV-1 subtypes, which further complicates the development of effective drugs and vaccines. In general, it is more difficult to hit a moving target than a stationary target. Two broad strategies for hitting a moving target (in this case, HIV replication) are to understand the movement and to aim at the portions that move the least. In the case of anti-HIV drug development, the first option can be addressed by understanding the mechanism(s) of drug resistance and developing drugs that effectively inhibit mutant viruses. The second can be addressed by designing drugs that interact with portions of the viral machinery that are evolutionarily conserved, such as enzyme active sites.


Challenges in the classification of fibrotic ILD: Patient case 2.
PATIENT PRESENTATION AND DIAGNOSTIC WORK-UP: The patient is a 72-year-old man presenting with dyspnoea on exertion. He is a former heavy smoker (40 pack-years) and his father, now deceased, had rheumatoid arthritis (RA). On physical examination he had mild bi-basilar crackles but no finger clubbing. Autoimmune serology was positive for rheumatoid factor (RF: 25 IU/ml). Lung function tests showed normal FVC, a FEV1/FVC ratio of 72% and a TLCO of 49% predicted. Lung CT showed centrilobular and paraseptal emphysema, subpleural fine reticulation and traction bronchiolectasis, with no clear basal predominance (Figure 1). BAL cytology showed 83% alveolar macrophages, 12% lymphocytes, 3% neutrophils, and 2% eosinophils.


An avrPto/avrPtoB mutant of Pseudomonas syringae pv. tomato DC3000 does not elicit Pto-mediated resistance and is less virulent on tomato.
AvrPto and AvrPtoB are type III effector proteins expressed by Pseudomonas syringae pv. tomato strain DC3000, a pathogen of both tomato and Arabidopsis spp. Each effector physically interacts with the tomato Pto kinase and elicits a hypersensitive response when expressed in tomato leaves containing Pto. An avrPto deletion mutant of DC3000 previously was shown to retain avirulence activity on Pto-expressing tomato plants. We developed an avrPtoB deletion mutant of DC3000 and found that it also retains Pto-specific avirulence on tomato. These observations suggested that avrPto and avrPtoB both contribute to avirulence. To test this hypothesis, we developed an deltaavrPtodeltaavrPtoB double mutant in DC3000. This double mutant was able to cause disease on a Pto-expressing tomato line. Thus, avrPto and avrPtoB are the only avirulence genes in DC3000 that elicit Pto-mediated defense responses in tomato. When inoculated onto susceptible tomato leaves and compared with wild-type DC3000, the mutants DC3000deltaavrPto and DC3000deltaavrPtoB each caused slightly less severe disease symptoms, although their growth rate was unaffected. However, DC3000deltaavr PtodeltaavrPtoB caused even less severe disease symptoms than the single mutants and grew more slowly than them on susceptible leaves. Our results indicate that AvrPto and AvrPtoB have phenotypically redundant avirulence activity on Pto-expressing tomato and additive virulence activities on susceptible tomato plants.


CLOCK/BMAL1 regulates circadian change of mouse hepatic insulin sensitivity by SIRT1.
The protein deacetylase, sirtuin 1 (SIRT1), involved in regulating hepatic insulin sensitivity, shows circadian oscillation and regulates the circadian clock. Recent studies show that circadian misalignment leads to insulin resistance (IR); however, the underlying mechanisms are largely unknown. Here, we show that CLOCK and brain and muscle ARNT-like protein 1 (BMAL1), two core circadian transcription factors, are correlated with hepatic insulin sensitivity. Knockdown of CLOCK or BMAL1 induces hepatic IR, whereas their ectopic expression attenuates hepatic IR. Moreover, circadian change of insulin sensitivity is impaired in Clock mutant, liver-specific Bmal1 knockout (KO) or Sirt1 KO mice, and CLOCK and BMAL1 are required for hepatic circadian expression of SIRT1. Further studies show that CLOCK/BMAL1 binds to the SIRT1 promoter to enhance its expression and regulates hepatic insulin sensitivity by SIRT1. In addition, constant darkness-induced circadian misalignment in mice decreases hepatic BMAL1 and SIRT1 levels and induces IR, which can be dramatically reversed by resveratrol. These findings offer new insights for coordination of the circadian clock and metabolism in hepatocytes by circadian regulation of hepatic insulin sensitivity via CLOCK/BMAL1-dependent SIRT1 expression and provide a potential application of resveratrol for combating circadian misalignment-induced metabolic disorders.


Coordinated Upregulation of Mitochondrial Biogenesis and Autophagy in Breast Cancer Cells: The Role of Dynamin Related Protein-1 and Implication for Breast Cancer Treatment.
Overactive mitochondrial fission was shown to promote cell transformation and tumor growth. It remains elusive how mitochondrial quality is regulated in such conditions. Here, we show that upregulation of mitochondrial fission protein, dynamin related protein-1 (Drp1), was accompanied with increased mitochondrial biogenesis markers (PGC1α, NRF1, and Tfam) in breast cancer cells. However, mitochondrial number was reduced, which was associated with lower mitochondrial oxidative capacity in breast cancer cells. This contrast might be owing to enhanced mitochondrial turnover through autophagy, because an increased population of autophagic vacuoles engulfing mitochondria was observed in the cancer cells. Consistently, BNIP3 (a mitochondrial autophagy marker) and autophagic flux were significantly upregulated, indicative of augmented mitochondrial autophagy (mitophagy). The upregulation of Drp1 and BNIP3 was also observed in vivo (human breast carcinomas). Importantly, inhibition of Drp1 significantly suppressed mitochondrial autophagy, metabolic reprogramming, and cancer cell viability. Together, this study reveals coordinated increase of mitochondrial biogenesis and mitophagy in which Drp1 plays a central role regulating breast cancer cell metabolism and survival. Given the emerging evidence of PGC1α contributing to tumor growth, it will be of critical importance to target both mitochondrial biogenesis and mitophagy for effective cancer therapeutics.


ERK1 and ERK2, two microtubule-associated protein 2 kinases, mediate the phosphorylation of tyrosine hydroxylase at serine-31 in situ.
Tyrosine hydroxylase (TH) is phosphorylated at four sites in situ and in vivo, and the protein kinases that phosphorylate three of these sites (Ser8,Ser19,Ser40) have been identified. In intact cells, the phosphorylation of the fourth site (Ser31) is increased in response to phorbol esters or nerve growth factor (NGF). Here, we show that Ser31 is phosphorylated by ERK1 and ERK2, two myelin basic protein and microtubule-associated protein kinases. Extracts of NGF- or bradykinin-treated PC12 rat pheochromocytoma cells were fractionated on Mono Q columns. Protein kinase activity toward Ser31 in TH was present in two peaks corresponding to myelin basic protein kinase activities previously identified as ERK1 and ERK2. Phosphorylation of purified TH in vitro by both kinases was selective for Ser31 up to at least 0.6 mol of phosphate per mol of TH subunit. Treatment of intact PC12 cells with bradykinin or NGF increased both the phosphorylation of TH-Ser31 in situ and the catalytic activity of ERKs (measured subsequently in vitro with myelin basic protein as substrate). Pretreatment of the cells with genistein (a protein-tyrosine kinase inhibitor) decreased the bradykinin- but not the NGF-induced changes in both TH-Ser31 phosphorylation and ERK activity. Genistein also inhibited the increases in Ser31 phosphorylation produced by phorbol dibutyrate, muscarine, and Ba2+. The data indicate that ERK activity is responsible for phosphorylating TH at Ser31 in intact cells and suggest that TH-Ser31 phosphorylation may be regulated by multiple signaling pathways that converge at or prior to the activation of the ERKs.


Interaction of Alu Polymorphisms and Novel Measures of Discrimination in Association with Blood Pressure in African Americans Living in Tallahassee, Florida.
African Americans are 40% more likely to be afflicted with hypertension than are non-Hispanic, white Americans, resulting in a 30% higher instance of mortality due to cardiovascular disease. There is debate about the relative contributions of genetic and sociocultural risk factors to the racial disparity in hypertension. We assayed three Alu insertion polymorphisms located in the ACE (angiotensin 1 converting enzyme), PLAT (plasminogen activator, tissue), and WNK1 (lysine deficient protein kinase 1) genes. We also estimated West African genetic ancestry and developed novel measures of perceived discrimination to create a biocultural model of blood pressure among African American adults in Tallahassee, Florida (n = 158). When tested separately, the ACE Alu noninsertion allele was significantly associated with higher systolic and diastolic blood pressure. In multiple regression analyses, West African genetic ancestry was not associated with blood pressure and reduced the strength of all blood pressure models tested. A gene × environment interaction was identified between the ACE Alu genotype and a new measure of unfair treatment that includes experiences by individuals close to the study participant. Inclusion of the WNK1 Alu genotype further improved this model of blood pressure variation. Our results suggest an association of the ACE and WNK1 genotypes with blood pressure that is consistent with their proposed gene functions. Measures of perceived unfair treatment of others show a threshold effect, with increased blood pressure occurring at higher values. The interaction between the ACE genotype and unfair treatment highlights the benefits of including both genetic and cultural data to investigate complex disease.


Effects of soy-derived isoflavones and a high-fat diet on spontaneous mammary tumor development in Tg.NK (MMTV/c-neu) mice.
Phytoestrogens such as isoflavonoids and lignans have been postulated as breast cancer protective constituents in soy and whole-grain cereals. We investigated the ability of isoflavones (IFs) and flaxseed to modulate spontaneous mammary tumor development in female heterozygous Tg.NK (MMTV/c-neu) mice. Two different exposure protocols were applied, either from 4 wk of age onward (postweaning) or during gestation and lactation (perinatal). In the postweaning exposure study, mice were fed IFs or flaxseed in a high-fat diet. In addition, flaxseed in a low-fat diet was tested. Postweaning exposure to IFs and flaxseed tended to accelerate the onset of mammary adenocarcinoma development, although tumor burden at necropsy was not changed significantly. Perinatal IF exposure resulted in enhanced mammary gland differentiation, but palpable mammary tumor onset was not affected. However, tumor burden at necropsy in the perinatal exposure study was significantly increased in the medium- and high-IF dose groups. Comparison of both exposure scenarios revealed a strongly accelerated onset of tumor growth after perinatal high-fat diet exposure compared with the low-fat diet. This study shows that breast cancer-modulating effects of phytoestrogens are dependent both on the background diet and on the timing of exposure in the life cycle.


Toxicity of statins on rat skeletal muscle mitochondria.
We investigated mitochondrial toxicity of four lipophilic stains (cerivastatin, fluvastatin, atorvastatin, simvastatin) and one hydrophilic statin (pravastatin). In L6 cells (rat skeletal muscle cell line), the four lipophilic statins (100 micromol/l) induced death in 27-49% of the cells. Pravastatin was not toxic up to 1 mmol/l. Cerivastatin, fluvastatin and atorvastatin (100 micromol/l) decreased the mitochondrial membrane potential by 49-65%, whereas simvastatin and pravastatin were less toxic. In isolated rat skeletal muscle mitochondria, all statins, except pravastatin, decreased glutamate-driven state 3 respiration and respiratory control ratio. Beta-oxidation was decreased by 88-96% in the presence of 100 micromol/l of the lipophilic statins, but only at higher concentrations by pravastatin. Mitochondrial swelling, cytochrome c release and DNA fragmentation was induced in L6 cells by the four lipophilic statins, but not by pravastatin. Lipophilic statins impair the function of skeletal muscle mitochondria, whereas the hydrophilic pravastatin is significantly less toxic.


Servicemembers and veterans with major traumatic limb loss from Vietnam war and OIF/OEF conflicts: survey methods, participants, and summary findings.
Care of veterans and servicemembers with major traumatic limb loss from combat theaters is one of the highest priorities of the Department of Veteran Affairs. We achieved a 62% response rate in our Survey for Prosthetic Use from 298 Vietnam war veterans and 283 servicemembers/veterans from Operation Iraqi Freedom/Operation Enduring Freedom (OIF/OEF) who sustained major traumatic limb loss. Participants reported their combat injuries; health status; quality of life; and prosthetic device use, function, rejection, and satisfaction. Despite the serious injuries experienced, health status was rated excellent, very good, or good by 70.7% of Vietnam war and 85.5% of OIF/OEF survey participants. However, many health issues persist for Vietnam war and OIF/OEF survey participants (respectively): phantom limb pain (72.2%/76.0%), chronic back pain (36.2%/42.1%), residual-limb pain (48.3%/62.9%), prosthesis-related skin problems (51.0%/58.0%), hearing loss (47.0%/47.0%), traumatic brain injury (3.4%/33.9%), depression (24.5%/24.0%), and posttraumatic stress disorder (37.6%/58.7%). Prosthetic devices are currently used by 78.2% of Vietnam war and 90.5% of OIF/OEF survey participants to improve function and mobility. On average, the annual rate for prosthetic device receipt is 10.7-fold higher for OIF/OEF than for Vietnam war survey participants. Findings from this cross-conflict survey identify many strengths in prosthetic rehabilitation for those with limb loss and several areas for future attention.


Prescriber and patient-oriented behavioural interventions to improve use of malaria rapid diagnostic tests in Tanzania: facility-based cluster randomised trial.
The increasing investment in malaria rapid diagnostic tests (RDTs) to differentiate malarial and non-malarial fevers, and an awareness of the need to improve case management of non-malarial fever, indicates an urgent need for high quality evidence on how best to improve prescribers' practices. A three-arm stratified cluster-randomised trial was conducted in 36 primary healthcare facilities from September 2010 to March 2012 within two rural districts in northeast Tanzania where malaria transmission has been declining. Interventions were guided by formative mixed-methods research and were introduced in phases. Prescribing staff from all facilities received standard Ministry of Health RDT training. Prescribers from facilities in the health worker (HW) and health worker-patient (HWP) arms further participated in small interactive peer-group training sessions with the HWP additionally receiving clinic posters and patient leaflets. Performance feedback and motivational mobile-phone text messaging (SMS) were added to the HW and HWP arms in later phases. The primary outcome was the proportion of patients with a non-severe, non-malarial illness incorrectly prescribed a (recommended) antimalarial. Secondary outcomes investigated RDT uptake, adherence to results, and antibiotic prescribing. Standard RDT training reduced pre-trial levels of antimalarial prescribing, which was sustained throughout the trial. Both interventions significantly lowered incorrect prescribing of recommended antimalarials from 8% (749/8,942) in the standard training arm to 2% (250/10,118) in the HW arm (adjusted RD (aRD) 4%; 95% confidence interval (CI) 1% to 6%; P = 0.008) and 2% (184/10,163) in the HWP arm (aRD 4%; 95% CI 1% to 6%; P = 0.005). Small group training and SMS were incrementally effective. There was also a significant reduction in the prescribing of antimalarials to RDT-negatives but no effect on RDT-positives receiving an ACT. Antibiotic prescribing was significantly lower in the HWP arm but had increased in all arms compared with pre-trial levels. Small group training with SMS was associated with an incremental and sustained improvement in prescriber adherence to RDT results and reducing over-prescribing of antimalarials to close to zero. These interventions may become increasingly important to cope with the wider range of diagnostic and treatment options for patients with acute febrile illness in Africa.


Oxygen therapeutics--current concepts.
In an effort to develop agents that enhance the oxygen-delivery capability of blood without the risks associated with allogeneic blood transfusions, several products are undergoing development and clinical trials. These oxygen transport agents can be divided into two main groups, perfluorocarbon (PFC) emulsions and modified hemoglobin solutions. Articles from the literature on the development and clinical trials of oxygen therapeutic agents were reviewed. PFCs are synthetic fluorinated hydrocarbons that increase dissolved oxygen in the fluid phase of the blood without binding the oxygen molecule. They enhance oxygen delivery significantly and may be used to augment the technique of intraoperative autologous donation. Two PFC products have been tested in Phase III clinical trials. Hemoglobin-based oxygen carriers (HBOCs) are either cross-linked or microencapsulated hemoglobin molecules. Modification of the human hemoglobin molecule with intra- and inter-molecular cross-linking eliminates renal toxicity and improves the oxygen dissociation characteristics of the molecule. These modifications are necessary because stroma-free hemoglobin (Hb) does not release oxygen in the physiologic range and dissociates into dimers which can be rapidly filtered by the kidney, leading to renal toxicity. In addition to human Hb, bovine hemoglobin is another source of raw material for HBOC products. Recombinant human Hb has also been produced, using an E. coli expression system, for HBOC manufacturing. Four cross-linked hemoglobin products have been tested in Phase III clinical trials. While no product has yet been approved for clinical use, preliminary studies with oxygen therapeutics show promising results, with effective oxygen carrying capacity and acceptable side effect profiles. In the future, the formation of a hybrid product which combines the best features from several of the products currently undergoing development may yield the ideal oxygen therapeutic agent.


Evidence for high-temperature in situ nifH transcription in an alkaline hot spring of Lower Geyser Basin, Yellowstone National Park.
Genes encoding nitrogenase (nifH) were amplified from sediment and photosynthetic mat samples collected in the outflow channel of Mound Spring, an alkaline thermal feature in Yellowstone National Park. Results indicate the genetic capacity for nitrogen fixation over the entire range of temperatures sampled (57.2°C to 80.2°C). Amplification of environmental nifH transcripts revealed in situ expression of nifH genes at temperatures up to 72.7°C. However, we were unable to amplify transcripts of nifH at the higher-temperature locations (> 72.7°C). These results indicate that microbes at the highest temperature sites contain the genetic capacity to fix nitrogen, yet either do not express nifH or do so only transiently. Field measurements of nitrate and ammonium show fixed nitrogen limitation as temperature decreases along the outflow channel, suggesting nifH expression in response to the downstream decrease in bioavailable nitrogen. Nitrogen stable isotope values of Mound Spring sediment communities further support geochemical and genetic data. DNA and cDNA nifH amplicons form several unique phylogenetic clades, some of which appear to represent novel nifH sequences in both photosynthetic and chemosynthetic microbial communities. This is the first report of in situ nifH expression in strictly chemosynthetic zones of terrestrial (non-marine) hydrothermal systems, and sets a new upper temperature limit (72.7°C) for nitrogen fixation in alkaline, terrestrial hydrothermal environments.


Improvements in Lower-Extremity Function Following a Rehabilitation Program With Patterned Electrical Neuromuscular Stimulation in Females With Patellofemoral Pain: A Randomized Controlled Trial.
Patellofemoral pain (PFP) is a challenging condition, with altered kinematics and muscle activity as 2 common impairments. Single applications of patterned electrical neuromuscular stimulation (PENS) have improved both kinematics and muscle activity in females with PFP; however, the use of PENS in conjunction with a rehabilitation program has not been evaluated. To determine the effects of a 4-week rehabilitation program with PENS on lower-extremity biomechanics and electromyography (EMG) during a single-leg squat (SLS) and a step-down task (SDT) in individuals with PFP. Double-blinded randomized controlled trial. Laboratory. Patients of Other Participants: Sixteen females with PFP (age 23.3 [4.9] y, mass 66.3 [13.5] kg, height 166.1 [5.9] cm). Patients completed a 4-week supervised rehabilitation program with or without PENS. Curve analyses for lower-extremity kinematics and EMG activity (gluteus maximus, gluteus medius, vastus medialis oblique, vastus lateralis, biceps femoris, and adductor longus) were constructed by plotting group means and 90% confidence intervals throughout 100% of each task, before and after the rehabilitation program. Mean differences (MDs) and SDs were calculated where statistical differences were identified. No differences at baseline in lower-extremity kinematics or EMG were found between groups. Following rehabilitation, the PENS group had significant reduction in hip adduction between 29% and 47% of the SLS (MD = 4.62° [3.85°]) and between 43% and 69% of the SDT (MD = 6.55° [0.77°]). Throughout the entire SDT, there was a decrease in trunk flexion in the PENS group (MD = 10.91° [1.73°]). A significant decrease in gluteus medius activity was seen during both the SLS (MD = 2.77 [3.58]) and SDT (MD = 4.36 [5.38]), and gluteus maximus during the SLS (MD = 1.49 [1.46]). No differences were seen in the Sham group lower-extremity kinematics for either task. Rehabilitation with PENS improved kinematics in both tasks and decreased EMG activity. This suggests that rehabilitation with PENS may improve muscle function during functional tasks.


Interleukin 1 beta propeptide is detected intracellularly and extracellularly when human monocytes are stimulated with LPS in vitro.
Human interleukin 1 beta (IL-1 beta) is synthesized as an inactive precursor that is cleaved by IL-1 converting enzyme (ICE) between Asp116 and Ala117 to form COOH-terminal mature IL-1 beta and NH2-terminal IL-1 beta propeptide. Little is known about the fate of the NH2-terminal cleavage product. In this study, human recombinant (hr)IL-1 beta propeptide (amino acids 2-116) was produced and used to prepare specific antibodies which do not recognize mature human IL-1 beta. These anti-propeptide antibodies were used for immunoprecipitation of biosynthetically labeled proteins from lipopolysaccharide-stimulated human monocytes. Analysis of immunoprecipitates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography revealed that these antibodies recognize precursor IL-1 beta and two unique proteins: one migrating at 17.5 kD and one at 14 kD. The larger of these two proteins has a migration nearly identical to that of the recombinant IL-1 beta propeptide, and most likely represents naturally derived propeptide. The protein migrating at 14 kD may result from a second cleavage by ICE, between Asp27 and Gly28. These proteins accumulate intracellularly and extracellularly during pulse-chase experiments, and therefore represent stable products of precursor IL-1 beta cleavage.


The Complexity of Making a Diagnosis.
Carl is a 12-year-old boy midline, with neurologic malformation, ataxia, bilateral strabismus with presumed residual visual impairment after surgical repair, and intellectual disability. He was referred to developmental-behavioral pediatrics for evaluation of possible autism spectrum disorder (ASD).Carl had a benign prenatal course and was born via spontaneous vaginal delivery at term. Bilateral strabismus was noted at birth. Despite surgery to address strabismus, Carl continues to be unable to raise his eyes above midline, and his visual status, including visual acuity, depth perception, color perception, and visual fields, is unclear. A recent auditory brainstem response evaluation was consistent with normal hearing. Multiple variants of undetermined significance were reported on chromosomal microarray. Magnetic resonance imaging of the brain demonstrated multiple malformations in the brainstem and posterior fossa. Electroencephalogram was without evidence of seizure activity. There is no family history of genetic disorders, brain malformation, or learning/cognitive disability.Carl was not able to sit independently until 2 years of age and did not walk until 5 years. Now, he can walk short distances by himself but is often unsteady on his feet. He uses a wheelchair but does not propel himself despite a physical therapy assessment indicating that he has adequate strength to do so. Carl spoke his first word at 5 years, and his parents report that he currently uses approximately 50 words spontaneously and appropriately. His speech is difficult to understand because of articulation errors. He can identify colors and numbers but has not mastered concepts of size and comparison. Standard scores for the conceptual, social, and practical domains and the General Adaptive Composite were in the mid-50s on the Adaptive Behavior Assessment System-3, as reported by the parents.At home, Carl spends much of his time watching videos on his iPad in his bedroom. At school, he tends to separate himself from others on the playground after lunch. Parents believe this to be due to communication difficulties and mobility limitations. Parents report that when he likes another classmate or family member, he will stay in the same room as that individual but will not try to interact. Teachers report that when a well-liked classmate sits near Carl, he will smile and say the classmate's name. He will not attempt to initiate or participate in conversation. A picture exchange communication system was introduced at school because of difficulties understanding his speech. Attempts have also been made to train him to use a keyboard to communicate. Carl has demonstrated limited interest in using either system. He has several repetitive behaviors such as hand flapping, body rocking, and rubbing his nose with a specific spoon. Repetitive, nonpurposeful vocalizations are reported at home and at school. Carl requires assistance with toileting and bathing because of refusal to complete the tasks otherwise. It is common for Carl to cover his ears in response to hearing specific sounds such as fire alarms and emergency sirens.The Autism Diagnostic Observation Schedule, Second Edition, Module 1 was administered to assess for behaviors consistent with a diagnosis of ASD. Carl scored within the severe range of symptoms and was diagnosed with ASD. Recommendations were made for completion of a functional vision assessment and implementation of home- and school-based applied behavior analysis programs. In addition, further testing with an assessment developed for children with low language abilities was recommended to clarify his nonverbal cognitive abilities. What else would you consider when making recommendations for Carl's care?


Dispersion-managed solitons in optical amplifier transmission systems with zero average dispersion.
Soliton propagation in a cascaded dispersion-managed optical amplifier system with zero net dispersion is examined. We present a qualitative physical explanation for the recently discovered fact that a soliton with finite energy can propagate down a fiber line with zero or normal average dispersion. We describe a specific practical system for the main properties of such a soliton, namely, the dependence of the soliton power on the pulse width at chirp-free points and the soliton average energy and width at chirp-free points as functions of the dispersion-allocation (strength of the map) parameter.


The effects of postactivation potentiation on sprint and jump performance of male academy soccer players.
The purpose of this investigation was to evaluate the postactivation potentiation (PAP) effects of both dynamic and isometric maximum voluntary contractions (MVCs) on sprint and jump performance and establish whether PAP methods could be used effectively in warm up protocols for soccer players. Twelve male soccer players performed 4 warm up protocols in a cross-over, randomized, and counterbalanced design. In addition to a control warm up, subjects performed deadlift (5 repetitions at 5 repetitions maximum), tuck jump (5 repetitions), and isometric MVC knee extensions (3 repetitions for 3 s) as PAP treatments in an otherwise identical warm up protocol. After each treatment, the subjects underwent 3 10 m and 20 m sprints 4, 5, and 6 minutes post-warm up and 3 vertical jumps (VJ) at 7, 8, and 9 minutes post-warm up. Repeated measures analysis of variance showed no significant differences in the first 10 m (p = 0.258) and 20 m (p = 0.253) sprint and VJ (p = 0.703) performance and the average 10 m (p = 0.215), 20 m (p = 0.388), and VJ (p = 0.529) performance between conditions. There were also no significant differences in performance responses between the strongest and weakest subjects, but large variations in individual responses were found between the subjects. The findings suggest that there was no significant group PAP effect on sprint and jump performance after dynamic and isometric MVCs compared with a control warm up protocol. However, the large variation in individual responses (-7.1% to +8.2%) suggests PAP should be considered on an individual basis. Factors such as method, volume, load, recovery, and interindividual variability of PAP must be considered in the practical application of PAP and the rigorous research design of future studies to evaluate the potential for performance enhancement.


Validity and reliability testing of the Quality of Life Scale, Swedish version in women with fibromyalgia -- statistical analyses.
One consequence of constant widespread pain is a low quality of life. The purpose of the study was to examine whether the Quality of Life Scale, Swedish version (QOLS-S), regarded as a generic quality-of-life instrument, is a reliable and valid instrument for use in women with fibromyalgia (FM). Women with FM (n = 113) contributed data on the QOLS-S and other standardized instrument at three points in time. Internal consistency reliability estimates ranged from 0.89 to 0.92. Convergent construct validity was indicated by moderate agreement with a global life satisfaction question. Discriminant construct validity was denoted by low correlations with the physical functioning subscale of the SF-36. In a factor analysis three factors emerged: 'personal and social well-being', 'relations with others' and 'active participation'. A few of the items have high cross-loadings, and the instrument could be improved by rewording those items to more closely reflect one specific factor. Overall, these results provide evidence that the QOLS-S has acceptable validity and reliability for use in women with FM.


Atypical antipsychotics rapidly and inappropriately switch peripheral fuel utilization to lipids, impairing metabolic flexibility in rodents.
Patients taking atypical antipsychotics are frequented by serious metabolic (eg, hyperglycemia, obesity, and diabetes) and cardiac effects. Surprisingly, chronic treatment also appears to lower free fatty acids (FFAs). This finding is paradoxical because insulin resistance is typically associated with elevated not lower FFAs. How atypical antipsychotics bring about these converse changes in plasma glucose and FFAs is unknown. Chronic treatment with olanzapine, a prototypical, side effect prone atypical antipsychotic, lowered FFA in Sprague-Dawley rats. Olanzapine also lowered plasma FFA acutely, concomitantly impairing in vivo lipolysis and robustly elevating whole-body lipid oxidation. Increased lipid oxidation was evident from accelerated losses of triglycerides after food deprivation or lipid challenge, elevated FFA uptake into most peripheral tissues (∼2-fold) except heart, rises in long-chain 3-hydroxylated acyl-carnitines observed in diabetes, and rapid suppression of the respiratory exchange ratio (RER) during the dark cycle. Normal rises in RER following refeeding, a sign of metabolic flexibility, were severely blunted by olanzapine. Increased lipid oxidation in muscle could be explained by ∼50% lower concentrations of the negative cytoplasmic regulator of carnitine palmitoyltransferase I, malonyl-CoA. This was associated with loss of anapleurotic metabolites and citric acid cycle precursors of malonyl-CoA synthesis rather than adenosine monophosphate-activated kinase activation or direct ACC1/2 inhibition. The ability of antipsychotics to lower dark cycle RER in mice corresponded to their propensities to cause metabolic side effects. Our studies indicate that lipocentric mechanisms or altered intermediary metabolism could underlie the FFA lowering and hyperglycemia (Randle cycle) as well as some of the other side effects of atypical antipsychotics, thereby suggesting strategies for alleviating them.


Post Flywheel Squat Potentiation of Vertical and Horizontal Ground Reaction Force Parameters during Jumps and Changes of Direction.
(1) Background: The aim of the study was to determine the post-activation performance enhancement (PAPE) of vertical and horizontal ground reaction force parameters during jumps and change of direction following flywheel squat exercise using two different flywheel inertias. (2) Methods: Eleven male athletes performed a countermovement jump (CMJ), standing broad jump (SBJ), and "modified 505" change of direction (COD) in a control condition and 6 minutes following three sets of six repetitions of flywheel half squats at one of two inertias (0.029 kg·m2 and 0.061 kg·m2). Peak directional ground reaction force, power, and rate of force development were calculated for each test. (3) Results: Higher inertia flywheel squats were able to acutely enhance CMJ peak vertical force (Bayes Factor (BF10) = 33.5, very strong; δ = 1.66; CI: 0.67, 2.70), whereas lower inertia flywheel squats were able to acutely enhance CMJ peak vertical power (BF10 = 3.65, moderate; δ = 0.93; CI: 0.11, 1.88). The vertical squat exercise induced no PAPE effect on resultant SBJ or horizontal COD ground reaction force parameters, nor were any differences observed between the inertias. (4) Conclusions: Researchers and practitioners should consider the kinetic and kinematic correspondence of a pre-load stimulus to the subsequent sport-specific activity (i.e., flywheel squat to CMJ).


Implementation and characterization of a quartz tuning fork based probe consisted of discrete resonators for dynamic mode atomic force microscopy.
The quartz tuning fork based probe {e.g., Akiyama et al. [Appl. Surf. Sci. 210, 18 (2003)]}, termed "A-Probe," is a self-sensing and self-actuating (exciting) probe for dynamic mode atomic force microscope (AFM) operation. It is an oscillatory force sensor consisting of the two discrete resonators. This paper presents the investigations on an improved A-Probe: its batch fabrication and assembly, mounting on an AFM head, electrical setup, characterization, and AFM imaging. The fundamental features of the A-Probe are electrically and optically characterized in "approach-withdraw" experiments. Further investigations include the frequency response of an A-Probe to small mechanical vibrations externally applied to the tip and the effective loading force yielding between the tip and the sample during the periodic contact. Imaging of an electronic chip, a compact disk stamper, carbon nanotubes, and Si beads is demonstrated with this probe at ambient conditions in the so-called frequency modulation mode. A special probe substrate, which can snap on a receptacle fixed on an AFM head, and a special holder including a preamplifier electronic are introduced. We hope that the implementation and characterization of the A-Probe described in this paper will provide hints for new scanning probe techniques.


The doctor as businessman: the changing politics of a cultural icon.
From just after the Civil War, when medicine began to professionalize, until the late 1970s, doctors and policy makers believed that clinical judgment should not be influenced by the financial interests of doctors. Physicians were highly entrepreneurial, and organized medicine fought to preserve their entrepreneurial interests, but the moral norm that justified their autonomy from state regulation was a strict separation of clinical judgment and pecuniary interests. Under managed care, the old norm is reversed. A good doctor takes financial considerations into account in making clinical decisions. Theoretically, doctors should consider measures of cost to society, but in practice, the payment systems of managed care plans induce doctors to consider the impact of each clinical decision on their own income. Because doctors share the risks of insuring patients with managed care plans, they have the same incentives as insurers to avoid patients who are expensively sick. The new cultural image of doctors as entrepreneurs masks their considerable loss of clinical autonomy under managed care. It also serves to persuade doctors to accept managed care arrangements and to persuade insurance consumers and patients to accept reduced benefits from employers and the government.


Determination of Spearman Correlation Coefficient (r) to Evaluate the Linear Association of Dermal Collagen and Elastic Fibers in the Perspectives of Skin Injury.
Difference in scar formation at different sites, in different directions at the same site, but with changes in the elasticity of skin with age, sex, and race or in some pathological conditions, is well known to clinicians. The inappropriate collagen syntheses and delayed or lack of epithelialization are known to induce scar formation with negligible elasticity at the site of damage. Changes in the elasticity of scars may be due to an unequal distribution of dermal collagen (C) and elastic (E) fibers. Spearman correlation coefficients (r) of collagen and elastic fibers in horizontal (H) and in vertical (V) directions (variables CV, CH, EV, and EH) were measured from the respective quantitative fraction data in 320 skin samples from 32 human cadavers collected at five selected sites over extremities. Spearman's correlation analysis revealed the statistically significant (p < 0.01) strong positive correlation between CH and CV in all the areas, that is, shoulder joint area (r = 0.66), wrist (r = 0.75), forearm (r = 0.75), and thigh (r = 0.80), except at the ankle (r = 0.26, p = 0.14) region. Similarly, positive correlation between EH and EV has been observed at the forearm (r = 0.65, moderate) and thigh (r = 0.42, low) regions. However, a significant moderate negative correlation was observed between CV and EV at the forearm (r = -0.51) and between CH and EH at the thigh region (r = -0.65). Significant differences of correlations of collagen and elastic fibers in different directions from different areas of extremities were noted. This may be one of the possible anatomical reasons of scar behavior in different areas and different directions of the same area.


Exercise with Blood Flow Restriction to Improve Quadriceps Function Long After ACL Reconstruction.
Quadriceps atrophy and weakness can persist for years after anterior cruciate ligament reconstruction (ACLR). We evaluated the effectiveness of a home-based blood flow restriction (BFR) exercise program to increase quadriceps size and strength several years after ACLR. Nine adults with ACLR (5±2 yrs post-surgery, ≤90% symmetry in quadriceps size and strength) and nine uninjured controls volunteered. ACLR participants exercised at home for 25 min, 5×/wk for 4 wks (single-leg knee extension, bodyweight half-squats, walking). Blood flow in only the involved leg was restricted using a thigh cuff inflated to 50% of limb occlusion pressure. Rectus femoris and vastus lateralis thickness and knee extensor strength were measured before and after training. Baseline and post-training symmetry (involved leg/uninvolved leg) indices were compared to uninjured controls. Rectus femoris and vastus lateralis thickness and knee extensor strength in the involved leg increased by 11±5%, 10±6%, and 20±14%, respectively (all P<0.01). Compared to baseline, post-training knee extensor strength symmetry increased from 88±4 to 99±5% (P<0.01) and did not differ from uninjured controls (99±5%, P=0.95). Implementation of BFR exercise at home was feasible, safe and effective. Results extend upon early post-operative application of BFR exercise for ACLR recovery and demonstrate that BFR can improve quadriceps function long after ACLR.


Androgens and dominance: sex-specific patterns in a highly social fish (Neolamprologus pulcher).
In most vertebrates, aggression and dominance are tightly linked to circulating testosterone. Fish, however, have two androgens (testosterone, T and 11-ketotestosterone, 11KT) that influence aggression and dominance. To date, few studies have compared the relationship between androgen levels and the outcome of aggressive contests in both females and males of the same species. To investigate sex differences in androgens we staged size-matched, limited-resource (territory) contests with 14 female-female and 10 male-male pairs of the highly social cichlid Neolamprologus pulcher. We then examined androgen levels in recently established dominants, who won the contest and subsequently acquired a territory (for 3h), and subordinates, who lost and did not acquire a territory. Newly dominant females had higher plasma T but similar 11KT levels to newly subordinate females. In contrast, newly dominant males had higher 11KT but similar T levels to subordinate males. The ratio of 11KT to T, which demonstrates physiological importance of T conversion to 11KT, was positively correlated with submissive behavior in female winners, and correlated weakly with aggressive behavior in male winners (p=0.05). These findings provide support for the hypothesis that different androgens play equivalent roles in female versus male dominance establishment, and suggest that relative levels of 11KT and T are implicated in female dominance behavior and perhaps behavior of both sexes.


Optimization of next-generation sequencing transcriptome annotation for species lacking sequenced genomes.
Next-generation sequencing methods, such as RNA-seq, have permitted the exploration of gene expression in a range of organisms which have been studied in ecological contexts but lack a sequenced genome. However, the efficacy and accuracy of RNA-seq annotation methods using reference genomes from related species have yet to be robustly characterized. Here we conduct a comprehensive power analysis employing RNA-seq data from Drosophila melanogaster in conjunction with 11 additional genomes from related Drosophila species to compare annotation methods and quantify the impact of evolutionary divergence between transcriptome and the reference genome. Our analyses demonstrate that, regardless of the level of sequence divergence, direct genome mapping (DGM), where transcript short reads are aligned directly to the reference genome, significantly outperforms the widely used de novo and guided assembly-based methods in both the quantity and accuracy of gene detection. Our analysis also reveals that DGM recovers a more representative profile of Gene Ontology functional categories, which are often used to interpret emergent patterns in genomewide expression analyses. Lastly, analysis of available primate RNA-seq data demonstrates the applicability of our observations across diverse taxa. Our quantification of annotation accuracy and reduced gene detection associated with sequence divergence thus provides empirically derived guidelines for the design of future gene expression studies in species without sequenced genomes.


Synaptogenesis in the giant-fibre system of Drosophila: interaction of the giant fibre and its major motorneuronal target.
The tergotrochanteral (jump) motorneuron is a major synaptic target of the Giant Fibre in Drosophila. These two neurons are major components of the fly's Giant-Fibre escape system. Our previous work has described the development of the Giant Fibre in early metamorphosis and the involvement of the shaking-B locus in the formation of its electrical synapses. In the present study, we have investigated the development of the tergotrochanteral motorneuron and its electrical synapses by transforming Drosophila with a Gal4 fusion construct containing sequences largely upstream of, but including, the shaking-B(lethal) promoter. This construct drives reporter gene expression in the tergotrochanteral motorneuron and some other neurons. Expression of green fluorescent protein in the motorneuron allows visualization of its cell body and its subsequent intracellular staining with Lucifer Yellow. These preparations provide high-resolution data on motorneuron morphogenesis during the first half of pupal development. Dye-coupling reveals onset of gap-junction formation between the tergotrochanteral motorneuron and other neurons of the Giant-Fibre System. The medial dendrite of the tergotrochanteral motorneuron becomes dye-coupled to the peripheral synapsing interneurons between 28 and 32 hours after puparium formation. Dye-coupling between tergotrochanteral motorneuron and Giant Fibre is first seen at 42 hours after puparium formation. All dye coupling is abolished in a shaking-B(neural) mutant. To investigate any interactions between the Giant Fibre and the tergotroachanteral motorneuron, we arrested the growth of the motorneuron's medial neurite by targeted expression of a constitutively active form of Dcdc42. This results in the Giant Fibre remaining stranded at the midline, unable to make its characteristic bend. We conclude that Giant Fibre morphogenesis normally relies on fasciculation with its major motorneuronal target.


Atmospheric transfer of a radio-frequency clock signal with a diode laser.
Remote transfer of a radio-frequency clock signal over a 60 m open atmospheric link has been experimentally investigated using a diode laser as the clock carrier. Phase-noise spectra and Allan deviations are both measured to characterize the excess clock instability incurred during the transfer process. Different detection schemes are used to assess the contributions from different noise sources. With an 80 MHz clock frequency, the total root-mean-square noise amplitude is measured to be about 5×10(-3) rad, with fractional frequency instability on the order of 1×10(-10) at 1 s. The majority of this excess noise is attributed to the transmitter noise, with the amplitude fluctuations of the diode laser identified as the main source. The excess phase noise caused by air turbulence is at the level of 10(-4) rad under the current experimental conditions. Our finding suggests that suppressing the transmitter noise is critical for improving the clock-transfer fidelity.


Murine mesenchymal progenitor cells from different tissues differentiated via mesenchymal microspheres into the mesodermal direction.
Because specific marker molecules for phenotypical identification of mesenchymal stem and progenitor cells are missing, the assessment of the in vitro-differentiation capacity is a prerequisite to characterize these cells. However, classical differentiation protocols are often cell-consuming and time intensive. Therefore, the establishment of novel strategies for differentiation is one topic of current efforts in stem cell biology. The goal of this study was to demonstrate the practicability of a new differentiation test using plastic adherent cell isolates from different tissues. We introduced the mesenchymal microsphere method as a feasible time- and cell saving screening method to analyse multilineage differentiation properties of adult progenitor cells in a three-dimensional system. For this purpose we isolated, characterized and analyzed new sources of adult murine mesenchymal progenitor cells from perirenal adipose tissue and mediastinal stromal tissue in comparison to bone marrow progenitor cells. The proliferation capacity of the cells was demonstrated by determination of the daily doubling index. Although the flow cytometry analysis of undifferentiated cells revealed differences in the expression of CD marker molecules, all isolates have the capacity for multilineage differentiation following the mesenchymal microsphere protocol as well as the classical "micro mass body" protocol for chondrogenic and the monolayer cultivation protocol for osteogenic and adipogenic differentiation. Differentiation was characterized using histochemical and immunhistochemical staining as well as RT-PCR. We were able to show that the mesenchymal microsphere method is an efficient test system for chondro-, osteo- and adipogenic differentiation of adult progenitor cells. The advantage of this system in comparison to classical protocols is that approximately 7 times lower cell numbers are necessary. Since classical culture procedures are time intensive because high cell numbers have to be obtained, the new differentiation method may also save cells and time in future clinical applications using human mesenchymal stromal cells.


Treatment of Zygomatic Complex Fractures with Surgical or Nonsurgical Intervention: A Retrospective Study.
Evaluate the 1-year treatment outcome of zygomatic complex fractures with surgical or nonsurgical intervention. One hundred and forty-two consecutive patients with a zygomatic complex fracture were enrolled. Sixty-eight patients underwent surgical intervention and 74 patients nonsurgical intervention. The 1-year examination evaluated cosmetic and functional outcome including malar symmetry, ocular motility, occlusion, mouth opening, neurosensory disturbances, and complications. Forty-six patients allocated to surgical intervention responded to the 1-year follow-up examination. Satisfying facial contour and malar alignment was observed in 45 patients. All patients presented with identical position of the eye globe without enophthalmos and normal ocular movement. A habitual occlusion was seen in all patients with a mean interincisal mouth opening without pain of 49 mm. One patient presented with minor ectropion. Wound infection occurred in five patients. Persistent infraorbital neurosensory disturbance was described by 19 patients. The 1-year radiographic examination showed adequate fracture alignment in all patients with satisfying facial contour. However, dissimilar position of the orbital floor was seen in three patients having orbital reconstruction. None of the patients were re-operated or needed secondary correction of the zygomatic complex or orbital floor. Surgical intervention is an effective treatment modality of depressed zygomatic complex fractures, whereas a nonsurgical approach is often used for nondisplaced fractures. Most zygomatic complex fractures can be treated solely by an intraoral approach and rigid fixation at the zygomaticomaxillary buttress. Further exposure of the zygomaticofrontal junction or inferior orbital rim is necessary for severely displaced fractures, which require additional fixation.


Antepartum fetal heart rate testing in preterm pregnancy.
The nonstress test (NST) and the contraction stress test (CST) have had wide application to term pregnancies, but little has been reported of use in the management of preterm fetuses. Seventy-two of 438 high-risk gravidas, receiving both tests between 25 and 34 weeks' gestation, delivered singleton infants before completion of their 34th week. On the basis of the last NST and CST preceding delivery, the 72 fetuses could be divided into reactive-negative and nonreactive-positive groups. Nonreactive-positive fetuses experienced greater perinatal mortality and significantly higher rates of intrapartum fetal distress, neonatal depression, respiratory distress syndrome (RDS), intrauterine growth retardation, and cesarean section. Pulmonary complications in nonreactive-positive fetuses, regardless of the lecithin: sphingomyelin ratio, were significantly increased if intrapartum fetal distress had preceded delivery. Most neonatal deaths stemmed from RDS-related complications, 86% preceding the 31st week of gestation; however, nearly half of the perinatal deaths before the 30th week followed reactive NSTs. The data suggest that for clinical management of preterm pregnancy, fetal heart rate testing should be initiated after the 29th week; earlier, test significance is less clear and infant survival less likely. Maternal estriol determinations aid little in the management of nonreactive-positive fetuses. Rather, the care of these selected pregnancies should be temporized through the 30th week; when delivery is then elected, intrapartum fetal distress should be avoided through liberal use of cesarean section in the nonreactive-positive group.


Numerical study of a confined slot impinging jet with nanofluids.
Heat transfer enhancement technology concerns with the aim of developing more efficient systems to satisfy the increasing demands of many applications in the fields of automotive, aerospace, electronic and process industry. A solution for obtaining efficient cooling systems is represented by the use of confined or unconfined impinging jets. Moreover, the possibility of increasing the thermal performances of the working fluids can be taken into account, and the introduction of nanoparticles in a base fluid can be considered. In this article, a numerical investigation on confined impinging slot jet working with a mixture of water and Al2O3 nanoparticles is described. The flow is turbulent and a constant temperature is applied on the impinging. A single-phase model approach has been adopted. Different geometric ratios, particle volume concentrations and Reynolds number have been considered to study the behavior of the system in terms of average and local Nusselt number, convective heat transfer coefficient and required pumping power profiles, temperature fields and stream function contours. The dimensionless stream function contours show that the intensity and size of the vortex structures depend on the confining effects, given by H/W ratio, Reynolds number and particle concentrations. Furthermore, for increasing concentrations, nanofluids realize increasing fluid bulk temperature, as a result of the elevated thermal conductivity of mixtures. The local Nusselt number profiles show the highest values at the stagnation point, and the lowest at the end of the heated plate. The average Nusselt number increases for increasing particle concentrations and Reynolds numbers; moreover, the highest values are observed for H/W = 10, and a maximum increase of 18% is detected at a concentration equal to 6%. The required pumping power as well as Reynolds number increases and particle concentrations grow, which is almost 4.8 times greater than the values calculated in the case of base fluid.List of symbols.


Microsaccades counteract perceptual filling-in.
Artificial scotomas positioned within peripheral dynamic noise fade perceptually during visual fixation (that is, the surrounding dynamic noise appears to fill-in the scotoma). Because the scotomas' edges are continuously refreshed by the dynamic noise background, this filling-in effect cannot be explained by low-level adaptation mechanisms (such as those that may underlie classical Troxler fading). We recently showed that microsaccades counteract Troxler fading and drive first-order visibility during fixation (S. Martinez-Conde, S. L. Macknik, X. G. Troncoso, & T. A. Dyar, 2006). Here we set out to determine whether microsaccades may counteract the perceptual filling-in of artificial scotomas and thus drive second-order visibility. If so, microsaccades may not only counteract low-level adaptation but also play a role in higher perceptual processes. We asked subjects to indicate, via button press/release, whether an artificial scotoma presented on a dynamic noise background was visible or invisible at any given time. The subjects' eye movements were simultaneously measured with a high precision video system. We found that increases in microsaccade production counteracted the perception of filling-in, driving the visibility of the artificial scotoma. Conversely, decreased microsaccades allowed perceptual filling-in to take place. Our results show that microsaccades do not solely overcome low-level adaptation mechanisms but they also contribute to maintaining second-order visibility during fixation.


Dynamic reweighting of visual and vestibular cues during self-motion perception.
The perception of self-motion direction, or heading, relies on integration of multiple sensory cues, especially from the visual and vestibular systems. However, the reliability of sensory information can vary rapidly and unpredictably, and it remains unclear how the brain integrates multiple sensory signals given this dynamic uncertainty. Human psychophysical studies have shown that observers combine cues by weighting them in proportion to their reliability, consistent with statistically optimal integration schemes derived from Bayesian probability theory. Remarkably, because cue reliability is varied randomly across trials, the perceptual weight assigned to each cue must change from trial to trial. Dynamic cue reweighting has not been examined for combinations of visual and vestibular cues, nor has the Bayesian cue integration approach been applied to laboratory animals, an important step toward understanding the neural basis of cue integration. To address these issues, we tested human and monkey subjects in a heading discrimination task involving visual (optic flow) and vestibular (translational motion) cues. The cues were placed in conflict on a subset of trials, and their relative reliability was varied to assess the weights that subjects gave to each cue in their heading judgments. We found that monkeys can rapidly reweight visual and vestibular cues according to their reliability, the first such demonstration in a nonhuman species. However, some monkeys and humans tended to over-weight vestibular cues, inconsistent with simple predictions of a Bayesian model. Nonetheless, our findings establish a robust model system for studying the neural mechanisms of dynamic cue reweighting in multisensory perception.


Broad-band near-infrared plasmonic nanoantennas for higher harmonic generation.
We propose a broad-band near-infrared trapezoidal plasmonic nanoantenna, analyze it numerically using finite integration and difference time domain techniques, and explain qualitatively its performance via a multidipolar scenario as well as a conformal transformation. The plasmonic nanoantenna reported here intercepts the incoming light as if it were of cross-sectional area larger than double its actual physical size for a 1500 nm bandwidth expanding from the near-infrared to the visible spectrum. Within this bandwidth, it also confines the incoming light to its center with more than 1 order of magnitude field enhancement. This wide-band operation is achieved due to the overlapping of the different dipole resonances excited across the nanoantenna. We further demonstrate that the broad-band field enhancement leads to efficient third harmonic generation in a simplified wire trapezoidal geometry when a Kerr medium is introduced, due to the lightning rod effect at the fundamental and the Purcell effect at the induced third harmonic.


Adenosine receptor transcriptomic profile in cardiac tissue of a Zucker rat model.
To evaluate the possible variations of adenosine receptor (AR) profile together with TNF-α and IL-6 mRNA in cardiac tissue of obese Zucker rats (OZR) during fasting conditions (fc) and during the induction of acute hyperglycemia (AH). OZR (O, n=21) and age-matched lean control rats (CO, n=18) were studied during fc (COfc, n=8; Ofc, n=13) and during the induction of AH (COAH, n=10; OAH, n=8). The histopathologic analysis performed on O and CO heart samples did not show abnormalities of myocardial structure. The AR transcriptomic profile was analyzed in O and CO by real-time polymerase chain reaction (PCR) and a significantly lower mRNA expression was observed for A2AR in O with respect to CO (p=0.047), while a significant upregulation was observed for A3R in O with respect to CO (p=0.002). No significant differences between O and CO were observed for TNF-α or IL-6. Correlations were found between glycemia and A1R (p=0.03) and A2BR (p=0.002); total cholesterol and A2BR (p=0.02) and A3R (p=0.0002), as well as between IL-6 and A1R (p=0.05) and TNF-α and A2AR (p<0.0001). The modulation of ARs in these settings could represent a promising approach to pharmacological treatment, which must be supported by diet restrictions and physical exercise.


Role of attention and perceptual grouping in visual statistical learning.
Statistical learning has been widely proposed as a mechanism by which observers learn to decompose complex sensory scenes. To determine how robust statistical learning is, we investigated the impact of attention and perceptual grouping on statistical learning of visual shapes. Observers were presented with stimuli containing two shapes that were either connected by a bar or unconnected. When observers were required to attend to both locations at which shapes were presented, the degree of statistical learning was unaffected by whether the shapes were connected or not. However, when observers were required to attend to just one of the shapes' locations, statistical learning was observed only when the shapes were connected. These results demonstrate that visual statistical learning is not just a passive process. It can be modulated by both attention and connectedness, and in natural scenes these factors may constrain the role of stimulus statistics in learning.


Sepsis following burns, trauma, and intra-abdominal infections.
In the last ten years anaerobic organisms have emerged as the major infecting agent in surgical patients. While these groups of organisms including Bacteroides fragilis, clostridia, and anaerobic cocci persist, there has, in addition, developed in the last few years a virulent group of nosocomial infections, and modern management of sepsis is primarily directed at gram-negative and anaerobic infections, which include nosocomial infections, for example, those caused by the Serratia group. Much has been learned about control of infections from the patient who has sustained thermal injury. While topical water-soluble antibiotics have been a remarkable advance in the care of the burn patient, systemic and subeschar antibiotics have proved essential in the management of severe burn injury. There is increasing evidence that there is remarkable interference with host defense mechanisms in patients who have sustained burns or significant trauma or intraabdominal infection. The patient sustaining nonthermal traumatic injury also sustains reduction in host resistance. Because of this and the additional initial contamination, in the traumatized patient antibiotic therapy should be started early and as a therapeutic measure. Newer localization techniques, including sonography and computed axial tomography scanning, have helped localize abdominal infections early. Specific antimicrobial therapy may be begun as an adjunct to the surgical therapy of intra-abdominal infection.


Central representation of dental structures in the kitten, including projections to the mesencephalic trigeminal nucleus.
Horseradish peroxidase (HRP) was injected into either a single maxillary or a single mandibular primary (deciduous) cuspid tooth of 8- to 10-week-old kittens. The large apex of the primary cuspid allowed for some leakage of the HRP from the pulpal chamber to the periodontal ligament (PDL). Thus, the injection procedure resulted in the application of HRP to the PDL as well as to the pulpal tissues. The transganglionic transport of HRP resulted in discrete terminal fields within the spinal trigeminal nucleus (STN) and the main sensory nucleus (MSN). These projections were clearly somatotopically organized within the STN, but less so within MSN. Within pars oralis (PO) and pars interpolaris (PI), mandibular cuspid dental structures (MdCDS) were represented in a dorsal position relative to the maxillary cuspid dental structures (MxCDS), whereas within pars caudalis (PC) and the adjacent reticular formation the somatotopic representation was not dorsoventral, but rather mediolateral, with the MdCDS represented more medially than the MxCDS. Areas of overlap between MxCDS and MdCDS were found within MSN and to a lesser degree within the superficial laminae of PC. In addition, the fiber pathway leading to labeled somata in the mesencephalic trigeminal (Mes V) nucleus was clearly identified. The majority of the fibers traced to the Mes V nucleus exited the spinal trigeminal tract at the level of the transition from PO to the MSN and traversed the nuclear region in a position dorsal to and separate from the trigeminal motor tract. As in STN, fibers within the caudal Mes V tract appeared to be somatotopically organized, with the fibers from the MdCDS generally more dorsal than the ones from the MxCDS. Labeled fibers, some with terminal arbors, were also identified in close association with the trigeminal motor tract. The findings show a complex pattern of central representation in the immature feline central nervous system for deciduous dental structures.


Impact of efalizumab on patient-reported outcomes in high-need psoriasis patients: results of the international, randomized, placebo-controlled Phase III Clinical Experience Acquired with Raptiva (CLEAR) trial [NCT00256139].
Chronic psoriasis can negatively affect patients' lives. Assessing the impact of treatment on different aspects of a patient's health-related quality of life (HRQOL) is therefore important and relevant in trials of anti-psoriasis agents. The recombinant humanized IgG1 monoclonal antibody efalizumab targets multiple T-cell-dependent steps in the immunopathogenesis of psoriasis. Efalizumab has demonstrated safety and efficacy in several clinical trials, and improves patients' quality of life. To evaluate the impact of efalizumab on HRQOL and other patient-reported outcomes in patients with moderate to severe plaque psoriasis, including a large cohort of High-Need patients for whom at least 2 other systemic therapies were unsuitable because of lack of efficacy, intolerance, or contraindication. A total of 793 patients were randomized in a 2:1 ratio to receive efalizumab 1 mg/kg/wk (n = 529) or placebo (n = 264) for 12 weeks. The study population included 526 High-Need patients (342 efalizumab, 184 placebo). The treatment was evaluated by patients using the HRQOL assessment tools Short Form-36 (SF-36) and Dermatology Life Quality Index (DLQI). Other patient-reported assessments included the Psoriasis Symptom Assessment (PSA), a visual analog scale (VAS) for itching, and the Patient's Global Psoriasis Assessment (PGPA). Efalizumab was associated with improvements at Week 12 from baseline in patient-reported outcomes, both in the total study population and in the High-Need cohort. Among all efalizumab-treated patients, the DLQI improved by 5.7 points from baseline to Week 12, relative to an improvement of 2.3 points for placebo patients (P < .001). Corresponding improvements in DLQI in the High-Need cohort were 5.4 points for efalizumab compared to 2.3 for placebo (P < .001). Improvements from baseline on the SF-36, PSA, PGPA, and itching VAS at Week 12 were also significantly greater in efalizumab-treated patients than for placebo. A 12-week course of efalizumab improved HRQOL and other patient-reported outcomes in patients with moderate to severe plaque psoriasis. The benefits of efalizumab therapy in High-Need patients were similar to those observed in the total study population, indicating that the beneficial impact of efalizumab on QOL is consistent regardless of disease severity, prior therapy, or contraindications to previous therapies.


Self-rated health showed a consistent association with serum HDL-cholesterol in the cross-sectional Oslo Health Study.
To examine the association between serum HDL-cholesterol concentration (HDL-C) and self rated health (SRH) in several age groups of men and women. The study had a cross-sectional design and included 18,770 men and women of the Oslo Health Study aged 30; 40 and 45; 69-60; 75-76 years. In both sexes and all age groups, SRH (3 categories: poor, good, very good) was positively correlated with HDL-C. Logistic regression analysis on dichotomized values of SRH (i.e. poor vs. good health) in each age group of men and women showed that increasing HDL-C values were associated with increasing odds for reporting good health; the odds ratio (OR) was highest in young men, and was generally lower in women than in men. Odds ratios in the 4 age groups of men were 4.94 (2.63-9.29), 2.25 (1.63-3.09), 2.12 (1.58-2.86), 1.87 (1.37-2.54); and in women: 3.58 (2.46-5.21), 2.81 (2.23-3.53), 2.28 (1.84-2.82), 1.61 (1.31-1.99). In the whole material, 1 mmol/L increase in HDL-C increased the odds for reporting good health by 2.27 (2.06-2.50; p<0.001), when adjusting for sex, age group, time since food intake and use of cholesterol lowering drugs. Chronic diseases, pain, psychological distress, smoking, alcohol, length of education, and dietary items did not have any major influence on the pattern of the HDL-C vs. SRH association. There was a consistent positive association between HDL-C and SRH, in both men and women in four different age groups, with the strongest association in young people.


Identification of sequence variants and analysis of the role of the glycogen synthase kinase 3 beta gene and promoter in late onset Alzheimer's disease.
Alzheimer's disease (AD) is a disorder characterised by a progressive deterioration in memory and other cognitive functions. Glycogen synthase kinase 3 beta (GSK3 beta) phosphorylates the microtubule associated protein tau at sites that are aberrantly phosphorylated in AD. GSK3 beta binds to presenilin 1 and plays a role in wnt and insulin signalling cascades, both of which have been proposed to be linked to AD. Moreover GSK3 beta activity may be altered in AD brain. These observations suggest a central role for GSK3 beta in AD and led us to investigate GSK3 beta as a candidate gene for AD. We sought to identify sequence variations in the gene and its promoter, as these could have an effect on activity and expression leading to abnormal function. Sequencing over 3000 bp of the GSK3 beta putative promoter revealed there to be five sequence variations, two of which were common (>10%). However on further examination none of these, either alone or in synergy, had any association with late onset AD. Stratification of the data by APOE epsilon 4 status also produced no significant association. Sequencing of the GSK3 beta coding region revealed no variations. This would suggest that the aberrant phosphorylation of tau by GSK3 beta in AD is not due to sequence variations in the gene or its promoter.


A Framework of a Lower Limb Musculoskeletal Model With Implemented Natural Proprioceptive Feedback and Its Progressive Evaluation.
Proprioceptive senses play an important role in human body reflex and movement, so far implementing physiological mathematical models of proprioceptors in the musculoskeletal model and investigating their effects have not been sufficiently investigated. The purpose of the present study was to establish a compact framework for a lower limb musculoskeletal model by considering both ascending signals from central nervous system and descending feedback neural signal from physiologically realistic proprioceptors and evaluate it with progressive experimental data as well as investigating the effects of the proprioceptive feedback on the human movement. The simulation framework was established by combining a lower limb musculoskeletal model, the forward dynamic tool from OpenSim codes, and an executive program based on Python codes. The physiological mathematical models of the muscle spindle and Golgi tendon organs were included in the feedback control loop for the model. The model was evaluated through both previous literature data and currently implemented volunteer reflex experiments from the neural organ level to the monosynaptic reflex loop, and finally the complicated movement, such as the firing rate of the proprioceptors, the knee-jerk reflex, and the normal gait. Simultaneously, the effects of the proprioceptors on human normal gait were initially investigated. The reliability of the framework was properly evaluated by comparing the experimental data of neural firing rate, electromyography signals, and joint kinematics. The gait analysis indicated that the introduction of the proprioceptors in the motor control loop can substantially resist the external disturbance. The established framework has been evaluated at different levels, and it can be extended and applied to different musculoskeletal models for human movement analysis and evaluate the effects of the proprioceptors on them.


Comparative performance of species richness estimation methods.
In most real-world contexts the sampling effort needed to attain an accurate estimate of total species richness is excessive. Therefore, methods to estimate total species richness from incomplete collections need to be developed and tested. Using real and computer-simulated parasite data sets, the performances of 9 species richness estimation methods were compared. For all data sets, each estimation method was used to calculate the projected species richness at increasing levels of sampling effort. The performance of each method was evaluated by calculating the bias and precision of its estimates against the known total species richness. Performance was evaluated with increasing sampling effort and across different model communities. For the real data sets, the Chao2 and first-order jackknife estimators performed best. For the simulated data sets, the first-order jackknife estimator performed best at low sampling effort but, with increasing sampling effort, the bootstrap estimator outperformed all other estimators. Estimator performance increased with increasing species richness, aggregation level of individuals among samples and overall population size. Overall, the Chao2 and the first-order jackknife estimation methods performed best and should be used to control for the confounding effects of sampling effort in studies of parasite species richness. Potential uses of and practical problems with species richness estimation methods are discussed.


The mutagenic effect of ultraviolet-A1 on human skin demonstrated by sequencing the p53 gene in single keratinocytes.
Sun exposure is accepted as the major risk factor for developing skin cancer, the most common cancer in the western world. Ultraviolet-B (UV-B) radiation is considered the causative agent, but recently several findings suggest a role also for ultraviolet-A (UV-A) radiation. Repeated suberythemal doses of ultraviolet-A1 (UV-A1) on healthy human skin induce an increase of p53 immunoreactive cells in epidermis, which may indicate cell cycle arrest and/or occurrence of p53 mutations. We have investigated the possible mutagenic effect of UV-A1 on skin by sequencing exons 4-11 and adjacent intron sequence of the p53 gene in immunoreactive single cells from three healthy individuals. Previously unexposed buttock skin was irradiated three times a week for 2 weeks with physiological fluences (40 J/cm2) of UV-A1. Punch biopsies were taken before and at different time-points after the exposure, and from these single p53 immunoreactive cells were isolated by using laser-assisted microdissection. Three mutations--all being indicative of oxidative damage and most likely related to UV-A exposure--were found among the 37 single cells from exposed skin, whereas no mutations were found in the 22 single cells taken before exposure. The findings indicate a mutagenic effect of low-dose UV-A1 on healthy human skin, which further demonstrates the importance of considering UV-A when taking protective measures against skin cancer.


Grasping the concept of personal property.
The concept of property is integral to personal and societal development, yet understanding of the cognitive basis of ownership is limited. Objects are the most basic form of property, so our physical interactions with owned objects may elucidate nuanced aspects of ownership. We gave participants a coffee mug to decorate, use and keep. The experimenter also designed a mug of her own. In Experiment 1, participants performed natural lifting actions with each mug. Participants lifted the Experimenter's mug with greater care, and moved it slightly more towards the Experimenter, while they lifted their own mug more forcefully and drew it closer to their own body. In Experiment 2, participants responded to stimuli presented on the mug handles in a computer-based stimulus-response compatibility task. Overall, participants were faster to respond in trials in which the handles were facing in the same direction as the response location compared to when the handles were facing away. The compatibility effect was abolished, however, for the Experimenter's mug--as if the action system is blind to the potential for action towards another person's property. These findings demonstrate that knowledge of the ownership status of objects influences visuomotor processing in subtle and revealing ways.


Lymphoepithelioma-like carcinoma of the breast: is it a distinct entity? Clinicopathological evaluation of two cases and review of the literature.
Lymphoepithelioma-like carcinomas (LELCs) are tumors with morphologic features identical to those of undifferentiated nasopharyngeal carcinoma. They are characterized by a pronounced lymphocytic infiltrate, often obscuring the neoplastic epithelial component. LELCs have been described in several organs, but are extremely rare in the breast. In this report, two cases of LELC of the breast are presented with their histological and immunohistochemical features. In situ hybridization for Epstein-Barr viral genome was negative in both cases. We discuss the differential diagnosis of LELC of the breast and review the reports of this entity that have appeared in the literature.


Effects of Depth of Propofol and Sevoflurane Anesthesia on Upper Airway Collapsibility, Respiratory Genioglossus Activation, and Breathing in Healthy Volunteers.
Volatile anesthetics and propofol impair upper airway stability and possibly respiratory upper airway dilator muscle activity. The magnitudes of these effects have not been compared at equivalent anesthetic doses. We hypothesized that upper airway closing pressure is less negative and genioglossus activity is lower during deep compared with shallow anesthesia. In a randomized controlled crossover study of 12 volunteers, anesthesia with propofol or sevoflurane was titrated using a pain stimulus to identify the threshold for suppression of motor response to electrical stimulation. Measurements included bispectral index, genioglossus electromyography, ventilation, hypopharyngeal pressure, upper airway closing pressure, and change in end-expiratory lung volume during mask pressure drops. A total of 393 attempted breaths during occlusion maneuvers were analyzed. Upper airway closing pressure was significantly less negative at deep versus shallow anesthesia (-10.8 ± 4.5 vs. -11.3 ± 4.4 cm H2O, respectively [mean ± SD]) and correlated with the bispectral index (P < 0.001), indicating a more collapsible airway at deep anesthesia. Respiratory genioglossus activity during airway occlusion was significantly lower at deep compared with light anesthesia (26 ± 21 vs. 35 ± 24% of maximal genioglossus activation, respectively; P < 0.001) and correlated with bispectral index (P < 0.001). Upper airway closing pressure and genioglossus activity during airway occlusion did not differ between sevoflurane and propofol anesthesia. Propofol and sevoflurane anesthesia increased upper airway collapsibility in a dose-dependent fashion with no difference at equivalent anesthetic concentrations. These effects can in part be explained by a dose-dependent inhibiting effect of anesthetics on respiratory genioglossus activity.


Probing the multifactorial basis of Plasmodium falciparum quinine resistance: evidence for a strain-specific contribution of the sodium-proton exchanger PfNHE.
Quinine (QN) continues to be an important treatment option for severe malaria, however resistance to this drug has emerged in field isolates of the etiologic agent Plasmodium falciparum. Quantitative trait loci investigations of QN resistance have mapped three loci of this complex trait. Two coincide with pfcrt and pfmdr1, involved in resistance to chloroquine (CQ) and other quinoline-based antimalarials. A third locus on chromosome 13 contains the sodium-proton exchanger (pfnhe) gene. Previous studies have associated pfnhe polymorphisms with reduced QN sensitivity in culture-adapted field isolates. Here, we provide direct evidence supporting the hypothesis that pfnhe contributes to QN resistance. Using allelic exchange, we reduced pfnhe expression by introducing a truncated 3' untranslated region (UTR) from pfcrt into the endogenous pfnhe 3'UTR. Transfections were performed with 1BB5 and 3BA6 (both CQ- and QN-resistant) as well as GC03 (CQ- and QN-sensitive), all progenies of the HB3xDd2 genetic cross. RNA and protein analyses of the ensuing recombinant clones demonstrated a approximately 50% decrease in pfnhe expression levels. A statistically significant 30% decrease in QN IC(50) values was associated with these decreased expression levels in 1BB5 and 3BA6 but not in GC03. CQ, mefloquine and lumefantrine IC(50) values were unaltered. Cytosolic pH values were similar in all parental lines and recombinant clones. Our observations support a role for pfnhe in QN resistance in a strain-dependent manner, which might be contingent on pre-existing resistance to CQ and/or QN. These data bolster observations that QN resistance is a complex trait requiring the contribution of multiple transporter proteins.


Transforming Growth Factor Beta Gene Signatures are Spatially Enriched in Keloid Tissue Biopsies and Ex vivo-Cultured Keloid Fibroblasts.
The keloid lesion is recognised as a spatially heterogeneous mass both in cellular and acellular composition and biological activity. Here, we have utilised a bioinformatic approach to determine whether this spatial heterogeneity is also evident at the molecular level and to identify key upstream regulators of signalling pathways enriched in the lesion in a spatially-restricted manner. Differentially expressed genes (20% change, p < 0.05) obtained from microarray datasets derived from whole keloid biopsies and ex vivo-cultured keloid fibroblasts, both from distinct regions of the keloid lesion (leading edge, centre, and top) have been analysed to show that the TGFβ family plays a significant but spatially dependent role in regulation of keloid gene expression. Furthermore, we have identified additional upstream signalling molecules involved in driving keloid biology and provide information on therapeutic targets whose modulation might be expected to lead to significant therapeutic efficacy.


The effect of concentrated pomegranate juice consumption on risk factors of cardiovascular diseases in women with polycystic ovary syndrome: A randomized controlled trial.
Polycystic ovary syndrome (PCOS) is associated with insulin resistance and dyslipidemia. Pomegranate juice is a rich source of polyphenols that may improve blood pressure, lipid profile, and other risk factors of cardiovascular diseases (CVDs). No study has examined the effects of pomegranate juice consumption on the clinical outcomes of PCOS. This study aimed to examine the effects of concentrated pomegranate juice (CPJ) consumption on risk factors of CVDs in women with PCOS. In this randomized clinical trial, 44 PCOS women with an age range of 18-40 years and body mass index (BMI) of ≥25 kg/m2 were recruited. They were randomly allocated to the CPJ (n = 22) and control (n = 22) groups, and followed for 8 weeks. Anthropometric measures, blood pressure, and biochemical parameters were measured at baseline and at the end of the trial. Compared with the control group, CPJ consumption significantly reduced systolic (-9.77 ± 1.46 vs. -1.18 ± 1.46 mmHg, p < .001) and diastolic (-3.87 ± 1.36 vs. 0.30 ± 1.36 mmHg, p = .05) blood pressure, serum triglyceride (TG) levels (-7.82 ± 6.32 vs. 16.63 ± 6.32 mg/dl, p = .01), TG/high-density lipoprotein-cholesterol (HDL-C) ratio (-0.39 ± 0.24 vs. 0.66 ± 0.24, p = .007), and increased serum concentrations of low-density lipoprotein-cholesterol (LDL-C) (6.03 ± 2.47 vs. -1.98 ± 2.47 mg/dl, p = .04) and HDL-C (1.93 ± 1.10 vs. -1.78 ± 1.10 mg/dl, p = .03). These findings were obtained after taking baseline measurements and potential confounders into account. No adverse event or complication was reported due to CPJ consumption throughout the study. We found that CPJ consumption has a beneficial effect on blood pressure, serum concentrations of TG and HDL-C, and TG/HDL-C ratio in women with PCOS. However, this juice could increase serum LDL-C levels in these patients. This trial was registered at www.irct.ir as IRCT20191109045383N1.


Genomic and transcriptomic analyses of Agrobacterium tumefaciens S33 reveal the molecular mechanism of a novel hybrid nicotine-degrading pathway.
Agrobacterium tumefaciens S33 is able to degrade nicotine via a novel hybrid of the pyridine and pyrrolidine pathways. It can be utilized to remove nicotine from tobacco wastes and transform nicotine into important functionalized pyridine precursors for some valuable drugs and insecticides. However, the molecular mechanism of the hybrid pathway is still not completely clear. Here we report the genome analysis of strain S33 and its transcriptomes grown in glucose-ammonium medium and nicotine medium. The complete gene cluster involved in nicotine catabolism was found to be located on a genomic island composed of genes functionally similar but not in sequences to those of the pyridine and pyrrolidine pathways, as well as genes encoding plasmid partitioning and replication initiation proteins, conjugal transfer proteins and transposases. This suggests that the evolution of this hybrid pathway is not a simple fusion of the genes involved in the two pathways, but the result of a complicated lateral gene transfer. In addition, other genes potentially involved in the hybrid pathway could include those responsible for substrate sensing and transport, transcription regulation and electron transfer during nicotine degradation. This study provides new insights into the molecular mechanism of the novel hybrid pathway for nicotine degradation.


Urinary kallikrein in normotensive subjects and in patients with essential hypertension.
1. Excretion of urinary kallikrein was normal in 13 out of 15 patients with uncomplicated essential hypertension. 2. Frusemide increased urinary kallikrein excretion in normotensive subjects and in patients with essential hypertension. The stimulating effect of frusemide on urinary kallikrein was significantly diminished in patients with essential hypertension. 3. No correlations of urinary kallikrein with sodium, potassium, and aldosterone excretion were found. 4. The results do not support the idea that urinary kallikrein plays a primary role in the pathogenesis of essential hypertension.


Can rapid integrated polymerase chain reaction-based diagnostics for gastrointestinal pathogens improve routine hospital infection control practice? A diagnostic study.
Every year approximately 5000-9000 patients are admitted to a hospital with diarrhoea, which in up to 90% of cases has a non-infectious cause. As a result, single rooms are 'blocked' by patients with non-infectious diarrhoea, while patients with infectious diarrhoea are still in open bays because of a lack of free side rooms. A rapid test for differentiating infectious from non-infectious diarrhoea could be very beneficial for patients. To evaluate MassCode multiplex polymerase chain reaction (PCR) for the simultaneous diagnosis of multiple enteropathogens directly from stool, in terms of sensitivity/specificity to detect four common important enteropathogens: Clostridium difficile, Campylobacter spp., Salmonella spp. and norovirus. A retrospective study of fixed numbers of samples positive for C. difficile (n = 200), Campylobacter spp. (n = 200), Salmonella spp. (n = 100) and norovirus (n = 200) plus samples negative for all these pathogens (n = 300). Samples were sourced from NHS microbiology laboratories in Oxford and Leeds where initial diagnostic testing was performed according to Public Health England methodology. Researchers carrying out MassCode assays were blind to this information. A questionnaire survey, examining current practice for infection control teams and microbiology laboratories managing infectious diarrhoea, was also carried out. MassCode assays were carried out at Oxford University Hospitals NHS Trust. Further multiplex assays, carried out using Luminex, were run on the same set of samples at Leeds Teaching Hospitals NHS Trust. The questionnaire was completed by various NHS trusts. Sensitivity and specificity to detect C. difficile, Campylobacter spp., Salmonella spp., and norovirus. Nucleic acids were extracted from 948 clinical samples using an optimised protocol (200 Campylobacter spp., 199 C. difficile, 60 S. enterica, 199 norovirus and 295 negative samples; some samples contained more than one pathogen). Using the MassCode assay, sensitivities for each organism compared with standard microbiological testing ranged from 43% to 94% and specificities from 95% to 98%, with particularly poor performance for S. enterica. Relatively large numbers of unexpected positives not confirmed with quantitative PCR were also observed, particularly for S. enterica, Giardia lamblia and Cryptosporidium spp. As the results indicated that S. enterica detection might provide generic challenges to other multiplex assays for gastrointestinal pathogens, the Luminex xTag(®) gastrointestinal assay was also run blinded on the same extracts (937/948 remaining) and on re-extracted samples (839/948 with sufficient material). For Campylobacter spp., C. difficile and norovirus, high sensitivities (> 92%) and specificities (> 96%) were observed. For S. enterica, on the original MassCode/Oxford extracts, Luminex sensitivity compared with standard microbiological testing was 84% [95% confidence interval (CI) 73% to 93%], but this dropped to 46% on a fresh extract, very similar to MassCode, with a corresponding increase in specificity from 92% to 99%. Overall agreement on the per-sample diagnosis compared with combined microbiology plus PCR for the main four/all pathogens was 85.6%/64.7%, 87.0%/82.9% and 89.8%/86.8% for the MassCode assay, Luminex assay/MassCode extract and Luminex assay/fresh extract, respectively. Luminex assay results from fresh extracts implied that 5% of samples did not represent infectious diarrhoea, even though enteropathogens were genuinely present. Managing infectious diarrhoea was a significant burden for infection control teams (taking 21% of their time) and better diagnostics were identified as having major potential benefits for patients. Overall, the Luminex xTag gastrointestinal panel showed similar or superior sensitivity and specificity to the MassCode assay. However, on fresh extracts, this test had low sensitivity to detect a key enteric pathogen, S. enterica; making it an unrealistic option for most microbiology laboratories. Extraction efficiency appears to be a major obstacle for nucleic acid-based tests for this organism, and possibly the whole Enterobacteriaceae family. To improve workflows in service microbiology laboratories, to reduce workload for infection control practitioners, and to improve outcomes for NHS patients, further research on deoxyribonucleic acid-based multiplex gastrointestinal diagnostics is urgently needed. The Health Technology Assessment programme of the National Institute for Health Research.