Identification of putative parasitism genes expressed in the esophageal gland cells of the soybean cyst nematode Heterodera glycines.
Cloning parasitism genes encoding secretory proteins expressed in the esophageal gland cells is the key to understanding the molecular basis of nematode parasitism of plants. Suppression subtractive hybridization (SSH) with the microaspirated contents from Heterodera glycines esophageal gland cells and intestinal region was used to isolate genes expressed preferentially in the gland cells of parasitic stages. Twenty-three unique cDNA sequences from a SSH cDNA library were identified and hybridized to the genomic DNA of H. glycines in Southern blots. Full-length cDNAs of 21 clones were obtained by screening a gland-cell long-distance polymerase chain reaction cDNA library. Deduced proteins of 10 clones were preceded by a signal peptide for secretion, and PSORT II computer analysis predicted eight proteins as extracellular, one as nuclear, and one as plasmalemma localized. In situ hybridization showed that four of the predicted extracellular clones were expressed specifically in the dorsal gland cell, one in the subventral gland cells, and three in the intestine in H. glycines. The predicted nuclear clone and the plasmalemma-localized clone were expressed in the subventral gland cells and the dorsal gland cell, respectively. SSH is an efficient method for cloning putative parasitism genes encoding esophageal gland cell secretory proteins that may have a role in H. glycines parasitism of soybean.


Brain metabolism is abnormal in the mdx model of Duchenne muscular dystrophy.
Duchenne muscular dystrophy (DMD) is an X-linked genetic disorder primarily affecting young boys, often causing mental retardation in addition to the well-known progressive muscular weakness. Normal dystrophin expression is lacking in skeletal muscle and the CNS of both DMD children and the mdx mouse model. To date, 31P-magnetic resonance spectroscopy (MRS) has shown in vivo several abnormalities within skeletal muscle of mdx mice and DMD boys. In this study, we determined whether similar abnormalities occur in mdx brain in vivo by using 31P-MRS in addition to metabolite and enzyme analysis to study cerebral metabolism. An increased inorganic phosphate (P(i))/phosphocreatine (PCr) and pH was found in vivo for mdx brain compared with controls, and biochemical analysis showed a reduction in total creatine, an increased extracellular and decreased intracellular volume in mdx brain. No differences were found in any glycolytic or mitochondrial maximal enzyme activities. These changes are discussed with respect to the biochemical changes found in muscle from DMD patients and mdx mice. It is proposed that these biochemical changes may be a factor in the reduced cognitive capacity of mdx mice and some DMD children.


Giant vertebral notochordal rest: magnetic resonance and diffusion weighted imaging findings.
A giant vertebral notochordal rest is a newly described, benign entity that is easily confused with a vertebral chordoma. As microscopic notochordal rests are rarely found in adult autopsies, the finding of a macroscopic vertebral lesion is a new entity with only seven previously presented cases. We report here radiological findings, including diffusion weighted images, of a patient with a giant notochordal remnant confined to the L5 vertebra, with an emphasis on its distinction from a chordoma.


Numerically correcting the joint misplacement of the sub-holograms in spatial synthetic aperture digital Fresnel holography.
We propose an effective reconstruction method for correcting the joint misplacement of the sub-holograms caused by the displacement error of CCD in spatial synthetic aperture digital Fresnel holography. For every two adjacent sub-holograms along the motion path of CCD, we reconstruct the corresponding holographic images under different joint distances between the sub-holograms and then find out the accurate joint distance by evaluating the quality of the corresponding synthetic reconstructed images. Then the accurate relative position relationships of the sub-holograms can be confirmed according to all of the identified joint distances, with which the accurate synthetic reconstructed image can be obtained by superposing the reconstruction results of the sub-holograms. The numerical reconstruction results are in agreement with the theoretical analysis. Compared with the traditional reconstruction method, this method could be used to not only correct the joint misplacement of the sub-holograms without the limitation of the actually overlapping circumstances of the adjacent sub-holograms, but also make the joint precision of the sub-holograms reach sub-pixel accuracy.


[Epidemiologic study of head injuries in Taipei City, Taiwan].
This study was designed to examine the descriptive epidemiology of head injuries in an urban population in the Taiwan area, Taipei City, during the period from July 1, 1987 to June 30, 1988. Clinical records reviewed included emergency room (ER) charts, inpatient charts of 19 major hospitals, death registration forms and medical examiners' reports in this city. A formulated definition was used to identify patients with head injuries. A total of 4,692 cases were collected, which included 4,319 hospital inpatients and 373 non-hospital deaths. The average incidence rate for head injury was 180/100,000 per year, 246/100,000 for males and 111/100,000 for females. The highest incidence rate was observed in the elderly group followed by the age group of 20-29 years. Sixty eight percent of the head injuries were caused by traffic accidents, 59.7 percent of which were involved in motorcycle rides. This rate was higher than those in any western reports. Among the 10-39 age group, the head injuries caused by traffic accidents were mainly due to motorcycle accidents. However, most of the children and the elderly who were injured were either pedestrians or bicycle riders. The average mortality rate was 23/100,000 per year, 34/100,000 for males and 12/100,000 for females. Initially the Glasgow Coma Scale was used in assessing the severity of head injuries. Seventy-two percent of the cases were considered mild, and 28 percent moderate to severe in degree, including 609 deaths.(ABSTRACT TRUNCATED AT 250 WORDS)


Responses of thoracic spinal neurons to activation and desensitization of cardiac TRPV1-containing afferents in rats.
The purpose of this study was to examine how upper thoracic spinal neurons responded to activation and desensitization of cardiac transient receptor potential vanilloid-1 (TRPV1)-containing afferent fibers. Extracellular potentials of single T3 spinal neurons were recorded in pentobarbital-anesthetized, paralyzed, and ventilated male rats. To activate cardiac nociceptive receptors, a catheter was placed in the pericardial sac to administer various chemicals: bradykinin (BK; 10 microg/ml, 0.2 ml), capsaicin (CAP, 10 microg/ml, 0.2 ml), or a mixture of algesic chemicals (AC; 0.2 ml) containing adenosine 10(-3) M, BK, serotonin, histamine, and PGE(2), 10(-5) M for each. Spinal neurons that responded to intrapericardial BK and/or CAP were used in this study. Results showed that 81% (35/43) of the neurons had excitatory responses to both intrapericardial BK and CAP, and the remainder responded to either BK or CAP. Intrapericardial resiniferatoxin (RTX) (0.2 microg/ml, 0.2 ml, 1 min), which desensitizes TRPV1-containing nerve endings, abolished excitatory responses to both BK (n = 8) and CAP (n = 7), and to AC (n = 5) but not to somatic stimuli. Intrapericardial capsazepine (1 mg/ml, 0.2 ml, 3 min), a specific antagonist of TRPV1, sharply attenuated excitatory responses to CAP in 5/5 neurons, but responses to BK in 5/5 neurons was maintained. Additionally, intrapericardial capsazepine had no significant effect on excitatory responses to AC in 3/3 neurons. These data indicated that intrapericardial BK-initiated spinal neuronal responses were linked to cardiac TRPV1-containing afferent fibers, but were not dependent on TRPV1. Intraspinal signaling for cardiac nociception was mediated through CAP-sensitive afferent fibers innervating the heart.


Antibody-onconase conjugates: cytotoxicity and intracellular routing.
Onconase, a member of the pancreatic ribonuclease A superfamily, is currently in Phase III clinical trials for treatment of unresectable malignant mesothelioma. The anticancer effect of onconase may relate to its intracellular target, a non-coding RNA. Some non-coding RNAs are aberrantly expressed in cancer cells. This discovery is creating new interest in drugs that target RNA. Conjugating onconase to agents that recognize tumor associated molecules further increases its potency and specificity. Analysis of onconase activity when directed to two different internalizing and one non-internalizing receptor reveals that the ideal targeting agents would rapidly enter lysosomal compartments before onconase escaped to the cytosol. Antibody-onconase conjugates are effective in preclinical models, cause little non-specific toxicities in mice and have favorable formulation properties. Understanding the reason for their potency coupled with understanding novel RNA-based mechanisms of tumor cell death will lead to improved variations of targeted onconase.


Emerging trends in regenerative medicine: a scientometric analysis in CiteSpace.
Regenerative medicine involves research in a number of fields and disciplines such as stem cell research, tissue engineering and biological therapy in general. As research in these areas advances rapidly, it is critical to keep abreast of emerging trends and critical turns of the development of the collective knowledge. A progressively synthesized network is derived from 35,963 original research and review articles that cite 3875 articles obtained from an initial topic search on regenerative medicine between 2000 and 2011. CiteSpace is used to facilitate the analysis of the intellectual structure and emerging trends. A major ongoing research trend is concerned with finding alternative reprogramming techniques as well as refining existing ones for induced pluripotent stem cells (iPSCs). A more recent emerging trend focuses on the structural and functional equivalence between iPSCs and human embryonic stem cells and potential clinical and therapeutic implications on regenerative medicine in a long run. The two trends overlap in terms of what they cite, but they are distinct and have different implications on future research. Visual analytics of the literature provides a valuable, timely, repeatable and flexible approach in addition to traditional systematic reviews so as to track the development of new emerging trends and identify critical evidence.


Recent advancements in stem cell and gene therapies for neurological disorders and intractable epilepsy.
The potential applications of stem cell therapies for treating neurological disorders are enormous. Many laboratories are focusing on stem cell treatments for CNS diseases, including spinal cord injury, Amyotrophic lateral sclerosis, Parkinson's disease, Huntington's disease, multiple sclerosis, stroke, traumatic brain injury, and epilepsy. Among the many stem cell types under testing for neurological treatments, the most common are fetal and adult brain stem cells, embryonic stem cells, induced pluripotent stem cells, and mesenchymal stem cells. An expanding toolbox of molecular probes is now available to allow analyses of neural stem cell fates prior to and after transplantation. Concomitantly, protocols are being developed to direct the fates of stem cell-derived neural progenitors, and also to screen stem cells for tumorigenicity and aneuploidy. The rapid progress in the field suggests that novel stem cell and gene therapies for neurological disorders are in the pipeline.


Validated age, growth and maturity of the bonnethead Sphyrna tiburo in the western North Atlantic Ocean.
The age, growth and maturity of bonnetheads Sphyrna tiburo inhabiting the estuarine and coastal waters of the western North Atlantic Ocean (WNA) from Onslow Bay, North Carolina, south to West Palm Beach, Florida, were examined. Vertebrae were collected and aged from 329 females and 217 males ranging in size from 262 to 1043 mm and 245 to 825 mm fork length, LF , respectively. Sex-specific von Bertalanffy growth curves were fitted to length-at-age data. Female von Bertalanffy parameters were L∞ = 1036 mm LF , k = 0·18, t0 = -1·64 and L0 = 272 mm LF . Males reached a smaller theoretical asymptotic length and had a higher growth coefficient (L∞ = 782 mm LF , k = 0·29, t0 = -1·43 and L0 = 266 mm LF ). Maximum observed age was 17·9 years for females and 16·0 years for males. Annual deposition of growth increments was verified by marginal increment analysis and validated for age classes 2·5+ to 10·5+ years through recapture of 13 oxytetracycline-injected specimens at liberty in the wild for 1-4 years. Length (LF50 ) and age (A50 ) at 50% maturity were 819 mm and 6·7 years for females, and 618 mm and 3·9 years for males. Both female and male S. tiburo in the WNA had a significantly higher maximum observed age, LF50 , A50 and L∞ , and a significantly lower k and estimated L0 than evident in the Gulf of Mexico (GOM). These significant differences in life-history parameters, as well as evidence from tagging and genetic studies, suggest that S. tiburo in the WNA and GOM should be considered separate stocks.


Analytic gradients, geometry optimization and excited state potential energy surfaces from the particle-particle random phase approximation.
The energy gradient for electronic excited states is of immense interest not only for spectroscopy but also for the theoretical study of photochemical reactions. We present the analytic excited state energy gradient of the particle-particle random phase approximation (pp-RPA). The analytic gradient formula is developed from an approach similar to that of time-dependent density-functional theory (TDDFT). The formula is verified for both the Hartree-Fock and (Generalized) Kohn-Sham reference states via comparison with finite difference results. The excited state potential energy surfaces and optimized geometries of some small molecules are investigated, yielding results of similar or better quality compared to adiabatic TDDFT. The singlet-to-triplet instability in TDDFT resulting in underestimated energies of the lowest triplet states is eliminated by pp-RPA. Charge transfer excitations and double excitations, which are challenging for most adiabatic TDDFT methods, can be reasonably well captured by pp-RPA. Within this framework, ground state potential energy surfaces of stretched single bonds can also be described well.


Ascorbate prophylaxis with mitomycin-C for corneal haze after laser-assisted sub-epithelial keratectomy.
Corneal haze is a significant complication of photorefractive keratectomy (PRK) and laser-assisted subepithelial keratectomy (LASEK). To evaluate the effect of ascorbic acid supplementation in addition to perioperative topical mitomycin-C for the prevention of haze after LASEK. We performed a retrospective, non-randomized case series study of two groups of 48 consecutive patients (96 myopic eyes) who had LASEK surgery. The treatment group was given ascorbic acid (vitamin C) orally, 500 mg, twice daily from 1 week before to 2 weeks after surgery. The control group was not offered any additional treatment. Ascorbate supplementation was the only difference in the postoperative treatment protocol between the treatment and control groups. Haze was assessed on a scale from 0 to 4 at the 1 year visit. Overall, 33.3% and 37.5% of the patients in the treatment and control groups respectively developed corneal haze. The trend of increased haze severity in the control group did not reach statistical significance. Our results showed that systemic ascorbate supplementation does not have an additional effect on the prevention of haze after LASEK compared to the effect of topical mitomycin-C alone.


Postnatal administration of 2-oxoglutaric acid improves articular and growth plate cartilages and bone tissue morphology in pigs prenatally treated with dexamethasone.
The potential effects of prenatal administration of dexamethasone (DEX) and postnatal treatment with 2-oxoglutaric acid (2-Ox) on postnatal development of connective tissue of farm animals were not examined experimentally. The aim of this study was to establish changes in morphological parameters of bone and articular and growth plate cartilages damaged by the prenatal action of DEX in piglets supplemented with 2-Ox. The 3 mg of DEX was administered by intramuscular route every second day from day 70 of pregnancy to parturition and then piglets were supplemented with 2-Ox during 35 days of postnatal life (0.4 g/kg body weight). The mechanical properties, BMD and BMC of bones, and histomorphometry of articular and growth plate cartilages were determined. Maternal treatment with DEX decreased the weight by 48%, BMD by 50% and BMC by 61% of the tibia in male piglets while such action of DEX in female piglets was not observed. DEX led to thinning of articular and growth plate cartilages and trabeculae thickness and reduced the serum GH concentration in male piglets. The administration of 2-Ox prevented the reduction of trabeculae thickness, the width of articular and growth plate cartilages in male piglets connected with higher growth hormone concentration compared with non-supplemented male piglets. The result showed that the presence of 2-Ox in the diet had a positive effect on the development of connective tissue in pigs during suckling and induced a complete recovery from bone and cartilage damage caused by prenatal DEX action.


Genome-Wide Association Study Reveals Novel Genomic Regions Associated with 10 Grain Minerals in Synthetic Hexaploid Wheat.
Synthetic hexaploid wheat (SHW; Triticum durum L. × Aegilops tauschii Coss.) is a means of introducing novel genes/genomic regions into bread wheat (T. aestivum L.) and a potential genetic resource for improving grain mineral concentrations. We quantified 10 grain minerals (Ca, Cd, Cu, Co, Fe, Li, Mg, Mn, Ni, and Zn) using an inductively coupled mass spectrometer in 123 SHWs for a genome-wide association study (GWAS). A GWAS with 35,648 single nucleotide polymorphism (SNP) markers identified 92 marker-trait associations (MTAs), of which 60 were novel and 40 were within genes, and the genes underlying 20 MTAs had annotations suggesting a potential role in grain mineral concentration. Twenty-four MTAs on the D-genome were novel and showed the potential of Ae. tauschii for improving grain mineral concentrations such as Ca, Co, Cu, Li, Mg, Mn, and Ni. Interestingly, the large number of novel MTAs (36) identified on the AB genome of these SHWs indicated that there is a lot of variation yet to be explored and to be used in the A and B genome along with the D-genome. Regression analysis identified a positive correlation between a cumulative number of favorable alleles at MTA loci in a genotype and grain mineral concentration. Additionally, we identified multi-traits and stable MTAs and recommended 13 top 10% SHWs with a higher concentration of beneficial grain minerals (Cu, Fe, Mg, Mn, Ni, and Zn), a large number of favorable alleles compared to low ranking genotypes and checks that could be utilized in the breeding program for the genetic biofortification. This study will further enhance our understanding of the genetic architecture of grain minerals in wheat and related cereals.


Postservice lower limb amputation in Scottish military veterans.
Recent attention has focused on veterans who have lost limbs in conflict, but the number of UK veterans who lose limbs to disease is unknown. We used data from the Trends in Scottish Veterans' Health study to explore postservice lower limb amputation. We carried out a retrospective cohort study of 78 000 veterans and 253 000 non-veterans born between 1945 and 1995, matched for age, sex and area of residence. We used survival analysis to examine the risk of amputation in veterans compared with non-veterans, and explored associations with antecedent disease. We found no difference between veterans and non-veterans in the risk of lower limb amputation, which was recorded in 145 (0.19%) veterans and 464 (0.18%) non-veterans (Cox proportional hazard ratio (HR) 1.00, 95% CIs 0.82 to 1.20, p=0.961). Peripheral arterial disease was recorded in two-thirds of both veteran and non-veteran amputees, and type 2 diabetes in 41% of veterans and 33% of non-veterans, with a dual diagnosis in 32% of veterans and 26% of non-veterans. Trauma was an infrequent cause of amputation. Although in later life veterans are no more likely to lose a limb to disease than non-veterans, the number so affected greatly outweighs those who have lost limbs in conflict. The high public profile of conflict-related limb loss risks eclipsing the needs of veterans with disease-related loss. Support for ageing veterans who have lost limbs due to disease will require planning with the same care as that afforded to the victims of conflict if inequalities are to be avoided.


Exertional compartment syndrome of the thigh: a rare diagnosis and literature review.
Exercise-induced acute compartment syndrome of the thigh is an uncommon entity. We present a rare case of bilateral exercise-induced three-compartment syndrome of the thighs that required fasciotomies. The objective of this study was to understand the history, physical examination, signs, symptoms, pathophysiology, diagnosis, and treatment of compartment syndrome and rhabdomyolysis. A 42-year-old man presented to the Emergency Department (ED) complaining of worsening pain and swelling in both thighs 45 h after performing a lower extremity exercise regimen. The patient's thighs were tender and swollen, but there was no ecchymosis or evidence of trauma. Admitting serum creatinine kinase (CK) was 106,289 U/L. Treatment for rhabdomyolysis was initiated. The next day, he complained of escalating bilateral thigh pain. Repeat serum CK was 346,580 U/L. The patient was diagnosed with bilateral thigh compartment syndrome and immediately taken to the operating room for fasciotomies. Postoperatively, the patient's symptoms improved rapidly and his serum CK quickly returned to normal. His incisions were closed and he returned to normal activities of daily living. Because exercise-induced compartment syndrome is an extremely rare diagnosis with a high risk of poor outcome, this article serves to emphasize the importance of considering this diagnosis during the work-up of patients presenting to the ED with rhabdomyolysis.


Vision loss in older persons.
Family physicians have an essential role in assessing, identifying, treating, and preventing or delaying vision loss in the aging population. Approximately one in 28 U.S. adults older than 40 years is visually impaired. Vision loss is associated with depression, social isolation, falls, and medication errors, and it can cause disturbing hallucinations. Adults older than 65 years should be screened for vision problems every one to two years, with attention to specific disorders, such as diabetic retinopathy, refractive error, cataracts, glaucoma, and age-related macular degeneration. Vision-related adverse effects of commonly used medications, such as amiodarone or phosphodiesterase inhibitors, should be considered when evaluating vision problems. Prompt recognition and management of sudden vision loss can be vision saving, as can treatment of diabetic retinopathy, refractive error, cataracts, glaucoma, and age-related macular degeneration. Aggressive medical management of diabetes, hypertension, and hyperlipidemia; encouraging smoking cessation; reducing ultraviolet light exposure; and appropriate response to medication adverse effects can preserve and protect vision in many older persons. Antioxidant and mineral supplements do not prevent age-related macular degeneration, but may play a role in slowing progression in those with advanced disease.


Comparison of power water--assisted and traditional liposuction: a prospective randomized trial of postoperative pain.
Postoperative pain has always been underestimated by the majority of plastic surgeons. A prospective randomized trial compared power water-assisted liposuction with the traditional tumescent technique. All patients with a body mass index (BMI) of 25 to 30 and excessive localized pathologic fat were recruited. Exclusion criteria specified a BMI greater than 30 or redundant anelastic skin. Patients were randomly assigned to power water-assisted or traditional liposuction. From September 2005 to December 2005, 60 patients were recruited and analyzed. For the study, 28 patients were randomized to traditional liposuction and 32 to power water-assisted liposuction. A significant difference in postoperative pain was observed (p < 0.05). After 4 days, 87% of the patients who underwent power water-assisted liposuction were completely pain free, as compared with 3.6% of those treated with traditional liposuction. Furthermore, ecchymosis was significantly less for the patients who underwent power-assisted liposuction (p < 0.05). The study findings demonstrate that power water-assisted liposuction is an almost painless procedure that produces less tissue trauma than traditional liposuction.


[Physical activity reference for health promotion 2013 and the prevalence of metabolic syndrome: A cross-sectional study].
This study aimed to examine the association between meeting the reference value for physical activity (PA) defined in the "Physical activity reference for health promotion 2013" recommendations or incremental increases in PA (add 10-minute per day) and the prevalence of metabolic syndrome (MS). A total of 906 adults aged 23-64 years participated in this cross-sectional study. The amount of moderate to vigorous PA was measured with a tri-axial accelerometer. The participants were classified into 2 groups-those with a PA level greater than or equal to the reference value of 23 metabolic equivalents (METs)-hours/week versus those with a PA level less than the reference value. Pre-MS and MS were determined based on the diagnostic criteria used in Japan. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for the prevalence of pre-MS and MS by adherence with the PA reference value were analyzed using a multivariable logistic regression model. Furthermore, adjusted ORs were calculated for each 3.5 METs-hours/week increase in PA, which is comparable to 10-minute increments of PA such as walking. The prevalence rates of pre-MS and MS were 10.3% (n=93) and 4.4% (n=40), respectively. The adjusted OR (95% CI) for the prevalence of pre-MS/MS among the participants meeting the PA reference compared with those not meeting the reference was 0.49 (0.33-0.74). Each 3.5 METs-hours/week increase in PA was associated with a lower adjusted OR for the prevalence of pre-MS/MS (OR 0.92, 95% CI 0.87-0.98). Our results suggest that meeting the reference value for PA (as defined in the "Physical activity reference for health promotion 2013" recommendations) and an increase in PA (add 10-minute) are both associated with a lower prevalence of pre-MS/MS.


The Arabic Version of the Mobile App Rating Scale: Development and Validation Study.
With thousands of health apps in app stores globally, it is crucial to systemically and thoroughly evaluate the quality of these apps due to their potential influence on health decisions and outcomes. The Mobile App Rating Scale (MARS) is the only currently available tool that provides a comprehensive, multidimensional evaluation of app quality, which has been used to compare medical apps from American and European app stores in various areas, available in English, Italian, Spanish, and German. However, this tool is not available in Arabic. This study aimed to translate and adapt MARS to Arabic and validate the tool with a sample of health apps aimed at managing or preventing obesity and associated disorders. We followed a well-established and defined "universalist" process of cross-cultural adaptation using a mixed methods approach. Early translations of the tool, accompanied by confirmation of the contents by two rounds of separate discussions, were included and culminated in a final version, which was then back-translated into English. Two trained researchers piloted the MARS in Arabic (MARS-Ar) with a sample of 10 weight management apps obtained from Google Play and the App Store. Interrater reliability was established using intraclass correlation coefficients (ICCs). After reliability was ascertained, the two researchers independently evaluated a set of additional 56 apps. MARS-Ar was highly aligned with the original English version. The ICCs for MARS-Ar (0.836, 95% CI 0.817-0.853) and MARS English (0.838, 95% CI 0.819-0.855) were good. The MARS-Ar subscales were highly correlated with the original counterparts (P<.001). The lowest correlation was observed in the area of usability (r=0.685), followed by aesthetics (r=0.827), information quality (r=0.854), engagement (r=0.894), and total app quality (r=0.897). Subjective quality was also highly correlated (r=0.820). MARS-Ar is a valid instrument to assess app quality among trained Arabic-speaking users of health and fitness apps. Researchers and public health professionals in the Arab world can use the overall MARS score and its subscales to reliably evaluate the quality of weight management apps. Further research is necessary to test the MARS-Ar on apps addressing various health issues, such as attention or anxiety prevention, or sexual and reproductive health.


Omega-3 Fatty Acids and Skeletal Muscle Health.
Skeletal muscle is a plastic tissue capable of adapting and mal-adapting to physical activity and diet. The response of skeletal muscle to adaptive stimuli, such as exercise, can be modified by the prior nutritional status of the muscle. The influence of nutrition on skeletal muscle has the potential to substantially impact physical function and whole body metabolism. Animal and cell based models show that omega-3 fatty acids, in particular those of marine origin, can influence skeletal muscle metabolism. Furthermore, recent human studies demonstrate that omega-3 fatty acids of marine origin can influence the exercise and nutritional response of skeletal muscle. These studies show that the prior omega-3 status influences not only the metabolic response of muscle to nutrition, but also the functional response to a period of exercise training. Omega-3 fatty acids of marine origin therefore have the potential to alter the trajectory of a number of human diseases including the physical decline associated with aging. We explore the potential molecular mechanisms by which omega-3 fatty acids may act in skeletal muscle, considering the n-3/n-6 ratio, inflammation and lipidomic remodelling as possible mechanisms of action. Finally, we suggest some avenues for further research to clarify how omega-3 fatty acids may be exerting their biological action in skeletal muscle.


Well-differentiated papillary mesothelioma: A 17-year single institution experience with a series of 75 cases.
We present our experience with 75 cases of well-differentiated papillary mesothelioma (WDPM) that were diagnosed at our institution between 2000 and 2017. The patients included 58 females and 17 males with age ranging from 18 to 69 years (mean, 42 years). Clinically, the vast majority of WDPMs were incidental findings during laparotomy or laparoscopic surgery for a variety of benign or malignant disease. The lesion manifested as either a small solitary nodule or multiple miliary nodules on the peritoneum or serosal surfaces of internal organs. Histologically, 67 cases were consistent with a classical WDPM, of which 6 cases contained microinvasive foci and 1 case had malignant transformation. Eight cases were hybrid tumors with variable combined component of adenomatoid tumor (n = 4), multicystic mesothelioma (n = 2), and both (n = 2). By immunohistochemistry, besides calretinin, D2-40, CK5/6 and WT1, 94% (29/31) of cases also showed immunostaining for PAX8. In comparison, PAX8 staining was only present in 12% (6/50) of epithelioid malignant mesothelioma selected as control cases. Follow-up information available in 46 cases revealed no signs of tumor progression or local recurrence except for the case that showed transformation to a fully malignant mesothelioma after a period of 15 years. Our comprehensive study further expanded the clinical and histopathological spectrum of WDPM. Compared with epithelioid malignant mesothelioma, PAX8 staining is highly sensitive and specific for WDPM (P < 0.001).


Divergent trends for gastric cancer incidence by anatomical subsite in US adults.
Age-specific analyses of non-cardia gastric cancer incidence reveal divergent trends among US whites: rates are declining in individuals aged 40 years and older but rising in younger persons. To investigate this heterogeneity further, incidence trends were evaluated by anatomical subsite. Gastric cancer incidence data for 1976-2007 were obtained from the US National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) Program and the US Centers for Disease Control and Prevention's National Program of Cancer Registries (NPCR). Incidence rates and estimated annual percentage change were calculated by age group (25-39, 40-59 and 60-84 years), race/ethnicity and subsite. Based on data from the nine oldest SEER registries (covering ~10% of the US population), rates for all non-cardia subsites decreased in whites and blacks, except for corpus cancer, which increased between 1976 and 2007 with estimated annual percentage changes of 1.0% (95% CI 0.1% to 1.9%) for whites and 3.5% (95% CI 1.8% to 5.2%) for blacks. In contrast, rates for all non-cardia subsites including corpus cancer declined among other races. In combined data from NPCR and SEER registries (covering 89% of the US population), corpus cancer significantly increased between 1999 and 2007 among younger and middle-aged whites; in ethnic-specific analyses, rates significantly increased among the same age groups in non-Hispanic whites and were stable among Hispanic whites. Age-specific rates for all subsites declined or were stable in this period among blacks and other races. Long- and short-term incidence trends for gastric cancers indicate a shifting distribution by anatomical subsite. Corpus cancer may have distinctive aetiology and changing risk factor exposures, warranting further investigation.


Effect of essential oils, tylosin, and monensin on finishing steer performance, carcass characteristics, liver abscesses, ruminal fermentation, and digestibility.
A feedlot (Exp. 1) experiment was conducted to evaluate the effects of an essential oil mixture (EOM), experimental essential oil mixture (EXP), tylosin, and monensin (MON) on performance, carcass characteristics, and liver abscesses. A metabolism experiment (Exp. 2) was conducted to evaluate the effects of EOM, EXP, and MON on ruminal fermentation and digestibility in finishing steers. In Exp. 1, 468 yearling steers (398 +/- 34 kg initial BW) were used in 50 pens (10 pens/treatment) and received their respective dietary treatments for 115 d. Five dietary treatments were compared in Exp. 1: 1) control, no additives (CON); 2) EOM, 1.0 g/steer daily; 3) EXP, 1.0 g/steer daily; 4) EOM, 1.0 g/steer daily plus tylosin, 90 mg/steer daily (EOM+T); and 5) monensin, 300 mg/steer daily plus tylosin, 90 mg/steer daily (MON+T). Compared with CON, steers fed MON+T had decreased DMI (P < 0.01), and steers fed EOM+T and MON+T had improved G:F (P </= 0.02). Average daily gain was not different among treatments (P > 0.58). There was a trend (P = 0.09) for a treatment effect on 12th-rib fat thickness, which resulted in a significant increase in calculated yield grade for the EOM+T treatment. No other carcass characteristics were affected by treatment (P >/= 0.10). Prevalence of total liver abscesses was reduced for steers fed tylosin compared with no tylosin (P < 0.05). In Exp. 2, 8 ruminally fistulated steers (399 +/- 49 kg initial BW) were assigned randomly to 1 of 4 treatments in a replicated 4 x 4 Latin square designed experiment. Treatments were 1) CON, 2) EOM, 3) EXP, and 4) MON with feeding rates similar to Exp. 1. There were no differences in DMI, OM intake, and apparent total tract DM or OM digestibilities among treatments (P > 0.30). Feed intake patterns were similar among feed additive treatments (P > 0.13). Total VFA (P = 0.10) and acetate (P = 0.06) concentrations tended to be affected by treatment with EOM numerically greater than CON. Average ruminal pH ranged from 5.59 to 5.72 and did not differ among treatments. Addition of a EOM or monensin to a diet containing tylosin improves G:F, but little difference was observed in metabolism or digestibility.


MicroRNAs, miR-23a-3p and miR-151-3p, Are Regulated in Dentate Gyrus Neuropil following Induction of Long-Term Potentiation In Vivo.
Translation of synaptic mRNA contributes to alterations in the proteome necessary to consolidate long-term potentiation (LTP), a model of memory processes. Yet, how this process is controlled is not fully resolved. MicroRNAs are non-coding RNAs that negatively regulate gene expression by suppressing translation or promoting mRNA degradation. As specific microRNAs are synaptically located, we hypothesized that they are ideally suited to couple synaptic activation, translational regulation, and LTP persistence. The aim of this study was to identify LTP-regulated microRNAs at or near synapses. Accordingly, LTP was induced unilaterally at perforant path-dentate gyrus synapses in awake adult Sprague-Dawley rats. Five hours later, dentate gyrus middle molecular layer neuropil, containing potentiated synapses, was laser-microdissected. MicroRNA expression profiling, using TaqMan Low Density MicroRNA Microarrays (n = 4), identified eight regulated microRNAs. Subsequent individual TaqMan assays confirmed upregulation of miR-23a-3p (1.30 ± 0.10; p = 0.015) and miR-151-3p (1.17 ± 0.19; p = 0.045) in a second cohort (n = 7). Interestingly, bioinformatic analysis indicated that miR-151-3p and miR-23a-3p regulate synaptic reorganisation and transcription, respectively. In summary, we have demonstrated for the first time that microRNAs are regulated in isolated neuropil following LTP induction in vivo, supporting the hypothesis that synaptic, LTP-responsive microRNAs contribute to LTP persistence via regulation of the synaptic proteome.


Prenatal stress-induced programming of genome-wide promoter DNA methylation in 5-HTT-deficient mice.
The serotonin transporter gene (5-HTT/SLC6A4)-linked polymorphic region has been suggested to have a modulatory role in mediating effects of early-life stress exposure on psychopathology rendering carriers of the low-expression short (s)-variant more vulnerable to environmental adversity in later life. The underlying molecular mechanisms of this gene-by-environment interaction are not well understood, but epigenetic regulation including differential DNA methylation has been postulated to have a critical role. Recently, we used a maternal restraint stress paradigm of prenatal stress (PS) in 5-HTT-deficient mice and showed that the effects on behavior and gene expression were particularly marked in the hippocampus of female 5-Htt+/- offspring. Here, we examined to which extent these effects are mediated by differential methylation of DNA. For this purpose, we performed a genome-wide hippocampal DNA methylation screening using methylated-DNA immunoprecipitation (MeDIP) on Affymetrix GeneChip Mouse Promoter 1.0 R arrays. Using hippocampal DNA from the same mice as assessed before enabled us to correlate gene-specific DNA methylation, mRNA expression and behavior. We found that 5-Htt genotype, PS and their interaction differentially affected the DNA methylation signature of numerous genes, a subset of which showed overlap with the expression profiles of the corresponding transcripts. For example, a differentially methylated region in the gene encoding myelin basic protein (Mbp) was associated with its expression in a 5-Htt-, PS- and 5-Htt × PS-dependent manner. Subsequent fine-mapping of this Mbp locus linked the methylation status of two specific CpG sites to Mbp expression and anxiety-related behavior. In conclusion, hippocampal DNA methylation patterns and expression profiles of female prenatally stressed 5-Htt+/- mice suggest that distinct molecular mechanisms, some of which are promoter methylation-dependent, contribute to the behavioral effects of the 5-Htt genotype, PS exposure and their interaction.


Detection of erythromycin-resistant determinants by PCR.
Erythromycin resistance determinants include Erm methylases, efflux pumps, and inactivating enzymes. To distinguish the different mechanisms of resistance in clinical isolates, PCR primers were designed so that amplification of the partial gene products could be detected in multiplex PCRs. This methodology enables the direct sequencing of amplified PCR products that can be used to compare resistance determinants in clinical strains. Further, this methodology could be useful in surveillance studies of erythromycin-resistant determinants.


Acrylamide in human diet, its metabolism, toxicity, inactivation and the associated European Union legal regulations in food industry.
Nowadays acrylamide is known not only as synthetic material used in industry, but also as carcinogenic, cyto- and genotoxic compound which forms during heat-induced process (due to Maillard reaction) mostly in foodstuff such as potato, bakery, plant derivatives products and coffee. The International Agency for Research on Cancer in 1994 declared acrylamide as a probable carcinogenic agent in humans. After metabolic process, acrylamide is distributed to all organs and tissues in human body. Acrylamide is classified as human neurotoxin, because this effect was observed in humans occupationally exposed to this compound. Acrylamide was found to cause apoptosis by mitochondrial dysfunction. Methods of acrylamide inactivation by microorganisms and bioactive diet compounds have also been reviewed. Moreover, there is still deficit of the European Union legal regulation concerning acrylamide mitigation strategies in food. Regulation 2017/2158 from 20 November 2017 is a step in the right direction when it comes to ensuring food safety and maximum levels of acrylamide in foodstuffs, however when exceeding those, it should result in elimination of such food from the market.


Type IIx myosin heavy chain transcripts are expressed in type IIb fibers of human skeletal muscle.
Several members of the sarcomeric myosin heavy chain (MHC) gene family have been mapped in the human genome but many of them have not yet been identified. In this study we report the identification of two human skeletal MHC genes as fast IIa and IIx MHC based on pattern of expression and sequence homology with the corresponding rat genes in the 3'-translated and untranslated regions. The distribution of these two gene products as well as that of the beta/slow MHC gene was analyzed in human skeletal muscles by in situ hybridization. The distribution of beta/slow, IIa, and IIx MHC transcripts defines three major muscle fiber types expressing a single MHC mRNA, i.e., either beta/slow, IIa, or IIx MHC mRNA, and two populations of hybrid fibers coexpressing beta/slow with IIa or IIa with IIx MHC mRNA. Fiber typing by ATPase histochemistry shows that IIa MHC transcripts are more abundant in histochemical type IIa fibers, whereas IIx MHC transcripts are more abundant in histochemical type IIb fibers.


Insulin-stimulated glucose uptake in skeletal muscle, adipose tissue and liver: a positron emission tomography study.
Insulin resistance is reflected by the rates of reduced glucose uptake (GU) into the key insulin-sensitive tissues, skeletal muscle, liver and adipose tissue. It is unclear whether insulin resistance occurs simultaneously in all these tissues or whether insulin resistance is tissue specific. We measured GU in skeletal muscle, adipose tissue and liver and endogenous glucose production (EGP), in a single session using 18F-fluorodeoxyglucose with positron emission tomography (PET) and euglycemic-hyperinsulinemic clamp. The study population consisted of 326 subjects without diabetes from the CMgene study cohort. Skeletal muscle GU less than 33 µmol/kg tissue/min and subcutaneous adipose tissue GU less than 11.5 µmol/kg tissue/min characterized insulin-resistant individuals. Men had considerably worse insulin suppression of EGP compared to women. By using principal component analysis (PCA), BMI inversely and skeletal muscle, adipose tissue and liver GU positively loaded on same principal component explaining one-third of the variation in these measures. The results were largely similar when liver GU was replaced by EGP in PCA. Liver GU and EGP were positively associated with aging. We have provided threshold values, which can be used to identify tissue-specific insulin resistance. In addition, we found that insulin resistance measured by GU was only partially similar across all insulin-sensitive tissues studied, skeletal muscle, adipose tissue and liver and was affected by obesity, aging and gender.


Psychopathology, social adjustment and personality correlates of schizotypy clusters in a large nonclinical sample.
Correlational methods, unlike cluster analyses, cannot take into account the possibility that individuals score highly on more than one symptom dimension simultaneously. This may account for some of the inconsistency found in the literature of correlates of schizotypy dimensions. This study explored the clustering of positive and negative schizotypy dimensions in nonclinical subjects and whether schizotypy clusters have meaningful patterns of adjustment in terms of psychopathology, social functioning, and personality. Positive and negative schizotypy dimensional scores were derived from the Chapman Psychosis-Proneness Scales for 6137 college students and submitted to cluster analysis. Of these, 780 completed the NEO-PI-R and Social Adjustment Scale-self report version, and a further 430 were interviewed for schizophrenia-spectrum, mood, and substance use psychopathology. Four clusters were obtained: low (nonschizotypic), high positive, high negative, and mixed (high positive and negative) schizotypy. The positive schizotypy cluster presented high rates of psychotic-like experiences, schizotypal and paranoid symptoms, had affective and substance abuse pathology, and was open to experience and extraverted. The negative schizotypy cluster had high rates of negative and schizoid symptoms, impaired social adjustment, high conscientiousness and low agreeableness. The mixed cluster was the most deviant on almost all aspects. Our cluster solution is consistent with the limited cluster analytic studies reported in schizotypy and schizophrenia, indicating that meaningful profiles of schizotypy features can be detected in nonclinical populations. The clusters identified displayed a distinct and meaningful pattern of correlates in different domains, thus providing construct validity to the schizotypy types defined.


Grandiose delusions: an experimental investigation of the delusion as defense.
Two distinct roles for emotion in the development of delusions have been outlined. Some authors argue that delusions defend against low self-esteem and negative emotion (the delusion-as-defense account). Other authors hypothesize that delusions are not a defense but are a direct reflection of emotion and associated processes (the emotion-consistent account). An empirical investigation was conducted of the delusion-as-defense account with reference to grandiose delusions. Twenty individuals with grandiose delusions and 21 individuals without mental illness were compared on overt and covert measures of self-esteem. No evidence for a discrepancy between overt and covert self-esteem in individuals with grandiose delusions was found. One potential interpretation of the results is that the tasks were not able to penetrate defensive processes. However, we argue that in this group, the grandiose delusions do not currently defend against low self-esteem. Instead, grandiose delusions may in part be direct exaggerations of the emotional state of individuals.


Endotoxin modulation of hepatocyte secretory and cellular protein synthesis is mediated by Kupffer cells.
We hypothesize that alterations of hepatocyte function in sepsis are modulated by endotoxin (lipopolysaccharide)-triggered Kupffer cells. In the present experiments the effect of lipopolysaccharide on secreted and cellular proteins synthesized by hepatocytes cultured alone or cocultured with Kupffer cells was investigated using polyacrylamide gel electrophoresis. Lipopolysaccharide had no direct effect on the types or amounts of secreted proteins synthesized by hepatocytes alone. Kupffer cells coculturing resulted in increased synthesis of some hepatocyte proteins (68k and 23k) whose production was not altered by lipopolysaccharide. In contrast, addition of lipopolysaccharide to the hepatocyte-Kupffer cell coculture substantially decreased synthesis of several proteins (73k, 66k, and 35k). Despite an overall decrease in protein synthesis of hepatocytes cocultured with Kupffer cells after the addition of lipopolysaccharide, there was increased synthesis of several individual proteins (58k and 44k). Similar effects were seen in synthesis of cellular protein. The addition of recombinant interleukin 1 to hepatocytes alone or in coculture had no effect on the amount or type of protein synthesized. We conclude that Kupffer cells regulate the types and amounts of individual proteins synthesized by hepatocytes via mediators other than interleukin 1.


Epidemiological evidence on health risks of cellular telephones.
It is too soon for a verdict on the health risks from cellular telephones, especially in view of changing technology. From the Interphone project and some other large studies in progress, better information may emerge. Based on the epidemiological evidence available now, the main public-health concern is clearly motor vehicle collisions, a behavioural effect rather than an effect of radiofrequency exposure as such. Neither the several studies of occupational exposure to radiofrequencies nor the few of cellular telephone users offer any clear evidence of an association with brain tumours or other malignancies. Even if the studies in progress were to find large relative effects for brain cancer, the absolute increase in risk would probably be much smaller than the risk stemming from motor vehicle collisions. Cellular telephones affect the quality of our lives in myriad ways, for good and ill; the health risk is just one part of a picture that is slowly coming into focus.


Intraoperative transfusion practices in Europe.
Transfusion of allogeneic blood influences outcome after surgery. Despite widespread availability of transfusion guidelines, transfusion practices might vary among physicians, departments, hospitals and countries. Our aim was to determine the amount of packed red blood cells (pRBC) and blood products transfused intraoperatively, and to describe factors determining transfusion throughout Europe. We did a prospective observational cohort study enrolling 5803 patients in 126 European centres that received at least one pRBC unit intraoperatively, during a continuous three month period in 2013. The overall intraoperative transfusion rate was 1.8%; 59% of transfusions were at least partially initiated as a result of a physiological transfusion trigger- mostly because of hypotension (55.4%) and/or tachycardia (30.7%). Haemoglobin (Hb)- based transfusion trigger alone initiated only 8.5% of transfusions. The Hb concentration [mean (sd)] just before transfusion was 8.1 (1.7) g dl(-1) and increased to 9.8 (1.8) g dl(-1) after transfusion. The mean number of intraoperatively transfused pRBC units was 2.5 (2.7) units (median 2). Although European Society of Anaesthesiology transfusion guidelines are moderately implemented in Europe with respect to Hb threshold for transfusion (7-9 g dl(-1)), there is still an urgent need for further educational efforts that focus on the number of pRBC units to be transfused at this threshold. NCT 01604083.


The functional role of the inferior parietal lobe in the dorsal and ventral stream dichotomy.
Current models of the visual pathways have difficulty incorporating the human inferior parietal lobe (IPL) into dorsal or ventral streams. Some recent proposals have attempted to integrate aspects of IPL function that were not hitherto dealt with well, such as differences between the left and right hemisphere and the role of the right IPL in responding to salient environmental events. However, we argue that these models also fail to capture adequately some important findings regarding the functions of the IPL. Here we critically appraise existing proposals regarding the functional architecture of the visual system, with special emphasis on the role of this region, particularly in the right hemisphere. We review evidence that shows the right IPL plays an important role in two different, but broadly complementary, aspects of attention: maintaining attentive control on current task goals as well as responding to salient new information or alerting stimuli in the environment. In our view, findings from functional imaging, electrophysiological and lesion studies are all consistent with the view that this region is part of a system that allows flexible reconfiguration of behaviour between these two alternative modes of operation. Damage to the right IPL leads to deficits in both maintaining attention and also responding to salient events, impairments that contribute to hemineglect, the classical syndrome that follows lesions of this region.


Elevated Steady State WBC and Platelet Counts Are Associated with Frequent Emergency Room Use in Adults with Sickle Cell Anemia.
Sickle cell anemia has many sequelae that result in emergency department (ED) use, but a minority of patients with sickle cell disease are frequent utilizers and make up the majority of ED visits. If patients who are likely to be frequent ED can be identified in steady state, they can be treated with disease modifying agents in an attempt to reduce ED use frequency. We sought to identify steady state markers for frequent ED use. We identified all patients with SS/Sβ0 seen at our facilities in 2012. Health care utilization over the entire year was calculated and ED visit numbers categorized as either 0-1, 2-5, or 6 or more visits a year. Steady state and acutely active laboratory parameters were collected and analyzed using analysis of variance models and odds ratios. 432 adult sickle cell patients were identified, ages 18-87, 54% female, and 38% had been prescribed hydroxyurea. Of the 432 patients,192 had 0-1 visits in the year, 144 had 2-5 visits in the year, and 96 had >6 visits for a total of 2259 visits. Those who had >6 visits accounted for 1750 (77%) of the total visits for the year. When steady state laboratory markers were examined, each additional 50x10(9)/L platelets was associated with 22% greater risk (p < .001); each 1x10(9)/L of WBC was associated with 11% greater risk (p = .003), and each 1g/dL Hb was associated with 23% lower risk (p = .007) of >6 ED visits/year. We did not observe a relationship between baseline HbF, LDH or reticulocyte count with >6 ED visits. Patients with elevated white blood cell counts, elevated platelet counts, and low hemoglobin levels exhibited higher risk for frequent ED utilization and could be candidates for early and aggressive therapy with disease modifying agents.


[Detectability and diagnostic criteria of p53 gene mutations in human oral squamous cell carcinoma using yeast functional assay].
In yeast functional assay (YF assay), a newly developed screening system for p53 mutation, wild-type p53 gives white yeast colonies and transcriptionally inactive mutant p53 gives red colonies. In the present study, the author applied YF assay to the detection of p53 mutations in human oral squamous cell carcinoma (SCC). Total RNA was extracted from samples and YF assay was performed. Four SCC cell lines (SAS, HSC-2, HSC-3 and Ca9-22) known to have p53 mutations all gave 100% red colonies, whereas nine oral non-tumor tissues gave 2.9-10% (average 5.2 +/- 2.7%) red colonies. Furthermore, a rat hepatoma cell line, WHp53, which had been transfected with human wild-type p53 expression vector, presented 7.8% red colonies. Thus the functional assays of tissues or cells containing only wild-type p53 give 3-10% red colonies as a background. To assess the detectability of p53 mutations, YF assay was performed on mixtures of wild-type and mutant p53 PCR products at serial ratios. The result showed that the mutation was detectable if 6% population of transcriptionally inactive mutant p53 mRNA were present in the total p53 mRNA. Twenty-two clinical samples of human oral SCC were then tested by YF assay. Fourteen out of 22 cases gave more than 20% red colonies. In these 14 cases, clonal p53 mutations with deletion, nonsense mutation or missense mutation were identified. In a case which gave 17% red colonies, identical p53 mutation was found in 2 out of 6 independent red colonies. However, no identical mutations were found in the cases giving 13, 9 and 8% red colonies. Based on these results, the author proposes that 20% of red colonies is the minimal value for the diagnosis of p53 mutation in YF assay under PCR conditions using Pfu polymerase and hot start method.


Evolutionary Analysis of the B56 Gene Family of PP2A Regulatory Subunits.
Protein phosphatase 2A (PP2A) is an abundant serine/threonine phosphatase that functions as a tumor suppressor in numerous cell-cell signaling pathways, including Wnt, myc, and ras. The B56 subunit of PP2A regulates its activity, and is encoded by five genes in humans. B56 proteins share a central core domain, but have divergent amino- and carboxy-termini, which are thought to provide isoform specificity. We performed phylogenetic analyses to better understand the evolution of the B56 gene family. We found that B56 was present as a single gene in eukaryotes prior to the divergence of animals, fungi, protists, and plants, and that B56 gene duplication prior to the divergence of protostomes and deuterostomes led to the origin of two B56 subfamilies, B56αβε and B56γδ. Further duplications led to three B56αβε genes and two B56γδ in vertebrates. Several nonvertebrate B56 gene names are based on distinct vertebrate isoform names, and would best be renamed. B56 subfamily genes lack significant divergence within primitive chordates, but each became distinct in complex vertebrates. Two vertebrate lineages have undergone B56 gene loss, Xenopus and Aves. In Xenopus, B56δ function may be compensated for by an alternatively spliced transcript, B56δ/γ, encoding a B56δ-like amino-terminal region and a B56γ core.


Integrating transcriptomic and metabolomic analysis to understand natural leaf senescence in sunflower.
Leaf senescence is a complex process, which has dramatic consequences on crop yield. In sunflower, gap between potential and actual yields reveals the economic impact of senescence. Indeed, sunflower plants are incapable of maintaining their green leaf area over sustained periods. This study characterizes the leaf senescence process in sunflower through a systems biology approach integrating transcriptomic and metabolomic analyses: plants being grown under both glasshouse and field conditions. Our results revealed a correspondence between profile changes detected at the molecular, biochemical and physiological level throughout the progression of leaf senescence measured at different plant developmental stages. Early metabolic changes were detected prior to anthesis and before the onset of the first senescence symptoms, with more pronounced changes observed when physiological and molecular variables were assessed under field conditions. During leaf development, photosynthetic activity and cell growth processes decreased, whereas sucrose, fatty acid, nucleotide and amino acid metabolisms increased. Pathways related to nutrient recycling processes were also up-regulated. Members of the NAC, AP2-EREBP, HB, bZIP and MYB transcription factor families showed high expression levels, and their expression level was highly correlated, suggesting their involvement in sunflower senescence. The results of this study thus contribute to the elucidation of the molecular mechanisms involved in the onset and progression of leaf senescence in sunflower leaves as well as to the identification of candidate genes involved in this process.


Homologous-pairing activity of the human DNA-repair proteins Xrcc3.Rad51C.
The human Xrcc3 protein is involved in the repair of damaged DNA through homologous recombination, in which homologous pairing is a key step. The Rad51 protein is believed to be the only protein factor that promotes homologous pairing in recombinational DNA repair in mitotic cells. In the brain, however, Rad51 expression is extremely low, whereas XRCC3, a human homologue of Saccharomyces cerevisiae RAD57 that activates the Rad51-dependent homologous pairing with the yeast Rad55 protein, is expressed. In this study, a two-hybrid analysis conducted with the use of a human brain cDNA library revealed that the major Xrcc3-interacting protein is a Rad51 paralog, Rad51C/Rad51L2. The purified Xrcc3.Rad51C complex, which shows apparent 1:1 stoichiometry, was found to catalyze the homologous pairing. Although the activity is reduced, the Rad51C protein alone also catalyzed homologous pairing, suggesting that Rad51C is a catalytic subunit for homologous pairing. The DNA-binding activity of Xrcc3.Rad51C was drastically decreased in the absence of Xrcc3, indicating that Xrcc3 is important for the DNA binding of Xrcc3.Rad51C. Electron microscopic observations revealed that Xrcc3.Rad51C and Rad51C formed similar filamentous structures with circular single-stranded DNA.


Clinical and genetic features of eight Chinese autosomal-dominant optic atrophy pedigrees with six novel OPA1 pathogenic variants.
Autosomal-dominant optic atrophy (ADOA) is one of the most common types of inherited optic atrophy. We identify OPA1 pathogenic variants and assess the clinical features of a cohort of Chinese ADOA patients Materials and Methods: Detailed clinical evaluations were performed and genomic DNA was extracted from peripheral blood for all the participants. Sanger sequencing was used to analyze all exons and exon/intron junctions of OPA1 for eight pedigrees. Target exome capture plus next-generation sequencing (NGS) were applied for one atypical family with photophobia. Reverse transcription polymerase chain reaction was carried out to further characterize the mRNA change of selected splicing alteration. All 17 patients had impaired vision and optic-disk pallor; however, the clinical severity varied markedly. Two patients complicated with hearing loss. Six novel and two reported pathogenic variants in OPA1 (GenBank Accession No. NM_130837.2) were identified including four nonsynonymous variants (c.2400T > G, c.1468T > C, c.1567A > G and c.1466T > C), two splicing variants (c.2984-1_2986delGAGA and c.2983 + 5G > A), one small deletion (c.2960_2968delGCGTTCAAC), and one small insertion (c.3009_3010insA). RNA analysis revealed the splicing variant c.2984-1_2986delGAGA caused small deletion of mRNA (r.2983_2988del). ADOA patients presented variable clinical manifestations. Novel OPA1 pathogenic variants are the main genetic defect for Chinese ADOA cases. NGS may be a useful molecular testing tool for atypical ADOA.


A novel method for the synthesis of the PEG-crosslinked chitosan with a pH-independent swelling behavior.
In this study, a simple method was developed to crosslink chitosan using poly(ethylene glycol) (PEG) with different molecular weights. Crosslinking of chitosan was confirmed by various spectral analyses. The differential scanning calorimetric (DSC) study indicated that the rigid crystalline structure of chitosan was decreased after crosslinking with PEG. The PEG-crosslinked chitosan (PEG-Ch) showed a pH-independent swelling behavior: swelled in both the simulated stomach (pH 1.1) and intestinal (pH 7.4) solutions. The swelling ratio of PEG-Ch increased significantly with a higher molecular weight of PEG used. In contrast, chitosan dissolved completely in a simulated stomach solution and showed a comparatively less swelling in a simulated intestinal solution. Thus, the prepared PEG-Ch could be a better biomaterial than chitosan in the development of orally sustained drug-delivery devices.


Multimer technologies for detection and adoptive transfer of antigen-specific T cells.
Identification and purification of antigen-specific T cells without altering their functional status are of high scientific and clinical interest. Staining with major histocompatibility complex (MHC)-peptide multimers constitutes a very powerful method to study antigen-specific T-cell subpopulations, allowing their direct visualization and quantification. MHC-peptide multimers, such as dimers, tetramers, pentamers, streptamers, dextramers and octamers have been used to evaluate the frequency of CD8(+) T cells, specific for tumor/leukemia-associated antigens as well as for viral antigens, e.g., CMVpp65 and EBV-EBNA. Moreover, MHC-peptide multimers have been used for rapid and efficient ex vivo isolation and expansion of T cells. A recent development in the field of MHC-peptide multimers led to the purification of CD8(+) T cells specific for leukemia antigens. This might help to select leukemia-specific donor lymphocyte infusions (DLIs), thus allowing dissection of the noxious graft-versus-host disease (GvHD) from beneficial anti-viral and even anti-leukemic effects. This review covers different types of MHC-peptide multimers and their applications, as well as the impact that multimers might have on further development of DLIs.


Recommendations for Institutions Transitioning to High-Sensitivity Troponin Testing: JACC Scientific Expert Panel.
High-sensitivity cardiac troponin (hs-cTn) I or T methods have been in use in certain regions for years but are now increasingly globally adopted, including in the United States. Accordingly, inevitable challenges are created for institutions transitioning from conventional cardiac troponin (cTn) assays. hs-cTn assays have higher analytic precision at lower concentrations, yielding greater clinical sensitivity for myocardial injury and allowing accurate recognition of small changes in troponin concentration (rise or fall) within a short time frame. Although much of the knowledge regarding troponin biology that was applicable with older troponin assays still holds true, considerable education regarding the differences between conventional cTn and hs-cTn is needed before medical systems convert to the newer methods. This includes a basic understanding of how hs-cTn testing differs from conventional cTn testing and how it is best deployed in different settings, such as the emergency department and inpatient services. This Expert Panel will review important concepts for institutional transition to hs-cTn methodology, providing recommendations useful for education before implementation.


Practical management of patients with myelofibrosis receiving ruxolitinib.
Myelofibrosis (MF) is characterized by bone marrow fibrosis, progressive anemia and extramedullary hematopoiesis, primarily manifested as splenomegaly. Patients also experience debilitating constitutional symptoms, including sequelae of splenomegaly, night sweats and fatigue. Ruxolitinib (INC424, INCB18424, Jakafi, Jakavi), a JAK1 and JAK2 inhibitor, was approved in November 2011 by the US FDA for the treatment of intermediate- or high-risk MF, and more recently in Europe and Canada for the treatment of MF-related splenomegaly or symptoms. These approvals were based on data from two randomized Phase III studies: COMFORT-I randomized against placebo, and COMFORT-II randomized against best available therapy. In these studies, ruxolitinib rapidly improved multiple disease manifestations of MF, reducing splenomegaly and improving quality of life of patients and potentially prolonging survival. However, as with other chemotherapies, ruxolitinib therapy is associated with some adverse events, such as anemia and thrombocytopenia. The aims of this article are to provide a brief overview of ruxolitinib therapy, to discuss some common adverse events associated with ruxolitinib therapy and to provide clinical management recommendations to maximize patients' benefit from ruxolitinib.


MHCWeb: converting a WWW database into a knowledge-based collaborative environment.
The World Wide Web (WWW) is useful for distributing scientific data. Most existing web data resources organize their information either in structured flat files or relational databases with basic retrieval capabilities. For databases with one or a few simple relations, these approaches are successful, but they can be cumbersome when there is a data model involving multiple relations between complex data. We believe that knowledge-based resources offer a solution in these cases. Knowledge bases have explicit declarations of the concepts in the domain, along with the relations between them. They are usually organized hierarchically, and provide a global data model with a controlled vocabulary. We have created the OWEB architecture for building online scientific data resources using knowledge bases. OWEB provides a shell for structuring data, providing secure and shared access, and creating computational modules for processing and displaying data. In this paper, we describe the translation of the online immunological database MHCPEP into an OWEB system called MHCWeb. This effort involved building a conceptual model for the data, creating a controlled terminology for the legal values for different types of data, and then translating the original data into the new structure. The OWEB environment allows for flexible access to the data by both users and computer programs.


Genetic mapping of centromeres in the nine Citrus clementina chromosomes using half-tetrad analysis and recombination patterns in unreduced and haploid gametes.
Mapping centromere locations in plant species provides essential information for the analysis of genetic structures and population dynamics. The centromere's position affects the distribution of crossovers along a chromosome and the parental heterozygosity restitution by 2n gametes is a direct function of the genetic distance to the centromere. Sexual polyploidisation is relatively frequent in Citrus species and is widely used to develop new seedless triploid cultivars. The study's objectives were to (i) map the positions of the centromeres of the nine Citrus clementina chromosomes; (ii) analyse the crossover interference in unreduced gametes; and (iii) establish the pattern of genetic recombination in haploid clementine gametes along each chromosome and its relationship with the centromere location and distribution of genic sequences. Triploid progenies were derived from unreduced megagametophytes produced by second-division restitution. Centromere positions were mapped genetically for all linkage groups using half-tetrad analysis. Inference of the physical locations of centromeres revealed one acrocentric, four metacentric and four submetacentric chromosomes. Crossover interference was observed in unreduced gametes, with variation seen between chromosome arms. For haploid gametes, a strong decrease in the recombination rate occurred in centromeric and pericentromeric regions, which contained a low density of genic sequences. In chromosomes VIII and IX, these low recombination rates extended beyond the pericentromeric regions. The genomic region corresponding to a genetic distance < 5cM from a centromere represented 47% of the genome and 23% of the genic sequences. The centromere positions of the nine citrus chromosomes were genetically mapped. Their physical locations, inferred from the genetic ones, were consistent with the sequence constitution and recombination pattern along each chromosome. However, regions with low recombination rates extended beyond the pericentromeric regions of some chromosomes into areas richer in genic sequences. The persistence of strong linkage disequilibrium between large numbers of genes promotes the stability of epistatic interactions and multilocus-controlled traits over successive generations but also maintains multi-trait associations. Identification of the centromere positions will allow the development of simple methods to analyse unreduced gamete formation mechanisms in a large range of genotypes and further modelling of genetic inheritance in sexual polyploidisation breeding schemes.


Plasma versus whole blood perfusion in a bioartificial liver assist device.
The ramifications of using whole blood or plasma for perfusion off an hepatocyte containing bioartificial liver bioreactor in which the hepatocytes are separated by a membrane or other physical barrier from the perfusate stream on the rate of change of patient blood concentrations are explored through dynamic modeling of whole blood perfusion as a two compartment system (patient tissue and blood compartments), and plasma perfusion as a three compartment system (patient tissue and blood compartments, and a plasma reservoir). The whole blood perfusion model is described by three dimensionless parameters: the Damkohler number, Da, which represents the ratio of the rate of conversion by the bioreactor to the rate of perfusion; kappa, which represents the ratio of the rate of internal reequilibration between the tissue and blood compartments and the rate of perfusion; and Vtb, the tissue/blood volume ratio. The plasma perfusion model has three additional dimensionless parameters: f, the fraction of plasma withdrawn from the blood in a plasma separator; alpha, the ratio of the plasma perfusion rate in the bioreactor to the blood draw rate; and Vbr, the blood/plasma reservoir volume ratio. Within the physiologic range of parameters, the rate of reduction in blood concentration in both the whole blood-perfused and plasma-perfused systems are sensitive to Damkohler number up to Da approximately 2. Neither system is sensitive to variations in kappa, and the plasma perfusion system has little sensitivity to alpha. Given bioreactors of equivalent activity, a greater rate of blood concentration reduction and lower endpoint blood concentration at equivalent perfusion times will be achieved with whole blood perfusion. There are two physical reasons for this. The first is that the plasma perfused system is only processing a fraction, f, of the blood compared with the whole blood perfusion system. The second reason is that, although the blood-perfused system is limited by overall bioreactor performance, the plasma-perfused system is mass transfer limited to the rate of blood concentration dilution into the plasma reservoir rather than limited by the overall bioreactor performance.


Subdividing Y-chromosome haplogroup R1a1 reveals Norse Viking dispersal lineages in Britain.
The influence of Viking-Age migrants to the British Isles is obvious in archaeological and place-names evidence, but their demographic impact has been unclear. Autosomal genetic analyses support Norse Viking contributions to parts of Britain, but show no signal corresponding to the Danelaw, the region under Scandinavian administrative control from the ninth to eleventh centuries. Y-chromosome haplogroup R1a1 has been considered as a possible marker for Viking migrations because of its high frequency in peninsular Scandinavia (Norway and Sweden). Here we select ten Y-SNPs to discriminate informatively among hg R1a1 sub-haplogroups in Europe, analyse these in 619 hg R1a1 Y chromosomes including 163 from the British Isles, and also type 23 short-tandem repeats (Y-STRs) to assess internal diversity. We find three specifically Western-European sub-haplogroups, two of which predominate in Norway and Sweden, and are also found in Britain; star-like features in the STR networks of these lineages indicate histories of expansion. We ask whether geographical distributions of hg R1a1 overall, and of the two sub-lineages in particular, correlate with regions of Scandinavian influence within Britain. Neither shows any frequency difference between regions that have higher (≥10%) or lower autosomal contributions from Norway and Sweden, but both are significantly overrepresented in the region corresponding to the Danelaw. These differences between autosomal and Y-chromosomal histories suggest either male-specific contribution, or the influence of patrilocality. Comparison of modern DNA with recently available ancient DNA data supports the interpretation that two sub-lineages of hg R1a1 spread with the Vikings from peninsular Scandinavia.


Meditation and auditory attention: An ERP study of meditators and non-meditators.
The findings of a study by Cahn and Polich (2009) suggests that there is an effect of a meditative state on three event-related potential (ERP) brain markers of "low-level" auditory attention (i.e., acoustic representations in sensory memory) in expert meditators: the N1, the P2, and the P3a. The current study built on these findings by examining trait and state effects of meditation on the passive auditory mismatch negativity (MMN), N1, and P2 ERPs. We found that the MMN was significantly larger in meditators than non-meditators regardless of whether they were meditating or not (a trait effect), and that N1 amplitude was significantly attenuated during meditation in non-meditators but not expert meditators (an interaction between trait and state). These outcomes suggest that low-level attention is superior in long-term meditators in general. In contrast, low-level attention is reduced in non-meditators when they are asked to meditate for the first time, possibly due to auditory fatigue or cognitive overload.


Suppression of antitumor immunity by IL-10 and TGF-beta-producing T cells infiltrating the growing tumor: influence of tumor environment on the induction of CD4+ and CD8+ regulatory T cells.
We examined the hypothesis that a failure of the immune system to eradicate tumors is due to the immunosuppressive environment created by the growing tumor, which is influenced by the site of tumor growth. We demonstrated that T cell responses to a bystander Ag in mice were suppressed by a growing CT26 tumor. T cells purified from the growing tumor expressed mRNA for IL-10, TGF-beta, and Foxp3. Intracellular cytokine staining revealed a high frequency of IL-10-secreting macrophages, dendritic cells, and CD4+ and CD8+ T cells infiltrating the tumor. In contrast, T cell IFN-gamma production was weak and CD8+ CTL responses were undetectable in mice with CT26 lung metastases and weak and transient following s.c. injection of CT26 cells, but were enhanced in the presence of anti-IL-10 and anti-TGF-beta. Consistent with this, removal of CD8+ T cells abrogated CTL responses and promoted progression of the s.c. tumor. However, in the lung model, depletion of CD8+ T cells significantly reduced the tumor burden. Furthermore, depletion of CD4+ or CD25+ T cells in vivo reduced tumor burden in s.c. and lung models, and this was associated with significantly enhanced IFN-gamma production by CD8+ T cells. These findings suggest that tumor growth facilitates the induction or recruitment of CD4+ regulatory T cells that secrete IL-10 and TGF-beta and suppress effector CD8+ T cell responses. However, CD8+ T regulatory cells expressing IL-10 and TGF-beta are also recruited or activated by the immunosuppressive environment of the lung, where they may suppress the induction of antitumor immunity.


Chronic kidney disease and premature ageing of the adaptive immune response.
Los of renal function is associated with uremia-associated immune deficiency, which contributes significantly to the mortality and morbidity of end-stage renal disease (ESRD) patients. In this review, the effect of ESRD on the adaptive cellular immune system is discussed. Progressive loss of renal function causes a preferential loss of number and function of lymphoid cells. More in depth analysis of these changes reveals a loss of thymic function, attrition of telomeres, and expanded memory T cell population, which is compatible with the concept of premature immunological ageing. Latency for cytomegalovirus is associated with more profound changes and the expansion of a unique pro-inflammatory, cytotoxic subset of CD4-positive CD28null T cells. Epigenetically, modifications in hematopoietic stem cells may underlie uremia-associated immunological ageing, which is not reversed by kidney transplantation. Possible therapeutic options to reverse or halt uremia-associated immunological ageing are discussed.


Membrane attack complex-associated molecules from redlip mullet (Liza haematocheila): Molecular characterization and transcriptional evidence of C6, C7, C8β, and C9 in innate immunity.
The redlip mullet (Liza haematocheila) is one of the most economically important fish in Korea and other East Asian countries; it is susceptible to infections by pathogens such as Lactococcus garvieae, Argulus spp., Trichodina spp., and Vibrio spp. Learning about the mechanisms of the complement system of the innate immunity of redlip mullet is important for efforts towards eradicating pathogens. Here, we report a comprehensive study of the terminal complement complex (TCC) components that form the membrane attack complex (MAC) through in-silico characterization and comparative spatial and temporal expression profiling. Five conserved domains (TSP1, LDLa, MACPF, CCP, and FIMAC) were detected in the TCC components, but the CCP and FIMAC domains were absent in MuC8β and MuC9. Expression analysis of four TCC genes from healthy redlip mullets showed the highest expression levels in the liver, whereas limited expression was observed in other tissues; immune-induced expression in the head kidney and spleen revealed significant responses against Lactococcus garvieae and poly I:C injection, suggesting their involvement in MAC formation in response to harmful pathogenic infections. Furthermore, the response to poly I:C may suggest the role of TCC components in the breakdown of the membrane of enveloped viruses. These findings may help to elucidate the mechanisms behind the complement system of the teleosts innate immunity.


Postantibiotic effect of beta-lactam antibiotics on Escherichia coli evaluated by bioluminescence assay of bacterial ATP.
The in vitro postantibiotic effects (PAE) of aztreonam, ceftazidime, cefuroxime, imipenem, and piperacillin on Escherichia coli ATCC 25922 were studied by a bioluminescence assay of bacterial ATP. In parallel with the PAE investigation, viability and morphology studies were performed. The strain was exposed for 2 h to different concentrations of beta-lactam antibiotics. The antibiotic activity was eliminated by 10(-4) dilutions, and regrowth of bacteria was monitored hourly by the bioluminescence assay of bacterial ATP. The length of PAE was dose dependent for ceftazidime (0.5 to 2.6 h), cefuroxime (0.4 to 2.6 h), and imipenem (0.3 to 4.5 h). The long PAE for these antibiotics at higher concentrations was associated with a potent initial killing and the presence of spheroplasts. Aztreonam and piperacillin produced a short, non-dose-dependent PAE (0.4 to 0.95 h). Short PAEs (below 1 h) were seen concomitantly with production of filaments, except in the case of imipenem, which only produced spheroplasts. The bioluminescence method was not jeopardized by filament formation, in contrast to the viable count assay which is normally used for PAE investigations. This makes it possible to study PAE for beta-lactam antibiotics on gram-negative bacteria with bioluminescence.


Distribution and chemical form of selenium in Neptunia amplexicaulis from Central Queensland, Australia.
Selenium (Se), a trace element essential for human and animal biological processes, is deficient in many agricultural soils. Some extremely rare plants can naturally accumulate extraordinarily high concentrations of Se. The native legume Neptunia amplexicaulis, endemic to a small area near Richmond and Hughenden in Central Queensland, Australia, is one of the strongest Se hyperaccumulators known on Earth, with foliar concentrations in excess of 4000 μg Se g-1 previously recorded. Here, we report on the Se distribution at a whole plant level using laboratory micro X-ray Fluorescence Microscopy (μXRF) and scanning electron microscopy (SEM-EDS), as well as on chemical forms of Se in various tissues using liquid chromatography-mass spectrometry (LC-MS) and synchrotron X-ray absorption spectroscopy (XAS). The results show that Se occurs in the forms of methyl-selenocysteine and seleno-methionine in the foliar tissues, with up to 13 600 μg Se g-1 total in young leaves. Selenium was found to accumulate primarily in the young leaves, flowers, pods and taproot, with lower concentrations present in the fine-roots and stem and the lowest present in the oldest leaves. Trichomes were not found to accumulate Se. We postulate that Se is (re)distributed in this plant via the phloem from older leaves to newer leaves, using the taproot as the main storage organ. High concentrations of Se in the nodes (pulvini) indicate this structure may play an important a role in Se (re)distribution. The overall pattern of Se distribution was similar in a non-Se tolerant closely related species (Neptunia gracilis), although the prevailing Se concentrations were substantially lower than in N. amplexicaulis.


Two families of exocomets in the β Pictoris system.
The young planetary system surrounding the star β Pictoris harbours active minor bodies. These asteroids and comets produce a large amount of dust and gas through collisions and evaporation, as happened early in the history of our Solar System. Spectroscopic observations of β Pictoris reveal a high rate of transits of small evaporating bodies, that is, exocomets. Here we report an analysis of more than 1,000 archival spectra gathered between 2003 and 2011, which provides a sample of about 6,000 variable absorption signatures arising from exocomets transiting the disk of the parent star. Statistical analysis of the observed properties of these exocomets allows us to identify two populations with different physical properties. One family consists of exocomets producing shallow absorption lines, which can be attributed to old exhausted (that is, strongly depleted in volatiles) comets trapped in a mean motion resonance with a massive planet. Another family consists of exocomets producing deep absorption lines, which may be related to the recent fragmentation of one or a few parent bodies. Our results show that the evaporating bodies observed for decades in the β Pictoris system are analogous to the comets in our own Solar System.


Towards automated discrimination of lipids versus peptides from full scan mass spectra.
Although physicochemical fractionation techniques play a crucial role in the analysis of complex mixtures, they are not necessarily the best solution to separate specific molecular classes, such as lipids and peptides. Any physical fractionation step such as, for example, those based on liquid chromatography, will introduce its own variation and noise. In this paper we investigate to what extent the high sensitivity and resolution of contemporary mass spectrometers offers viable opportunities for computational separation of signals in full scan spectra. We introduce an automatic method that can discriminate peptide from lipid peaks in full scan mass spectra, based on their isotopic properties. We systematically evaluate which features maximally contribute to a peptide versus lipid classification. The selected features are subsequently used to build a random forest classifier that enables almost perfect separation between lipid and peptide signals without requiring ion fragmentation and classical tandem MS-based identification approaches. The classifier is trained on in silico data, but is also capable of discriminating signals in real world experiments. We evaluate the influence of typical data inaccuracies of common classes of mass spectrometry instruments on the optimal set of discriminant features. Finally, the method is successfully extended towards the classification of individual lipid classes from full scan mass spectral features, based on input data defined by the Lipid Maps Consortium.


Effects of Cytokine Activation and Oxidative Stress on the Function of the Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells.
In several retinal complications, such as age-dependent macular degeneration (AMD), oxidative stress is increased and cytokine level is elevated. These are shown to alter the activation and expression of matrix metalloproteinase (MMP) both in human primary and immortalized retinal pigment epithelial (RPE) cells. However, the effects on human embryonic stem cell (hESC)-derived RPE cells remain to be elucidated. The mature hESC-RPE cells were exposed to inflammatory cytokines (IFN-γ or TNF-α) for 24 hours or oxidative stress (H2O2) for 1 hour. Effects on barrier properties were analyzed with transepithelial electrical resistance (TEER), the expression of MMP-1, MMP-2, MMP-3, MMP-9, collagen I, and collagen IV genes with quantitative RT-PCR, and the expression of MMP-1 and MMP-3 proteins with Western blot or ELISA, respectively. Also, activation and secretion of MMP-2 and -9 proteins were analyzed with zymography. In normal state, mature hESC-RPE cells expressed MMP-1, -2, -3, and -9 genes in low levels, respectively. Tumor necrosis factor-α increased MMP-1 and -2 gene expression, and H2O2 increased MMP-3 and -9 gene expression. Zymography revealed IFN-γ- and TNF-α-induced secretion of MMP-2 and high-molecular-weight species of MMP (HMW MMP), but H2O2 decreased their secretion. Furthermore, TNF-α and H2O2 significantly decreased barrier properties. Here, cytokines induced the MMP-1 and -2 gene and protein expression. Also, H2O2 induced MMP-3 and -9 gene expression, but not their protein secretion. These data propose that under oxidative stress and cytokine stimuli, mature hESC-RPE cells resemble their native counterpart in the human eye in regard to MMP secretion and expression and could be used to model retinal disorders involving alterations in MMP activity such as AMD, diabetic retinopathy, or proliferative vitreoretinopathy in vitro.


Dosage-dependent phenotypes in models of 16p11.2 lesions found in autism.
Recurrent copy number variations (CNVs) of human 16p11.2 have been associated with a variety of developmental/neurocognitive syndromes. In particular, deletion of 16p11.2 is found in patients with autism, developmental delay, and obesity. Patients with deletions or duplications have a wide range of clinical features, and siblings carrying the same deletion often have diverse symptoms. To study the consequence of 16p11.2 CNVs in a systematic manner, we used chromosome engineering to generate mice harboring deletion of the chromosomal region corresponding to 16p11.2, as well as mice harboring the reciprocal duplication. These 16p11.2 CNV models have dosage-dependent changes in gene expression, viability, brain architecture, and behavior. For each phenotype, the consequence of the deletion is more severe than that of the duplication. Of particular note is that half of the 16p11.2 deletion mice die postnatally; those that survive to adulthood are healthy and fertile, but have alterations in the hypothalamus and exhibit a "behavior trap" phenotype-a specific behavior characteristic of rodents with lateral hypothalamic and nigrostriatal lesions. These findings indicate that 16p11.2 CNVs cause brain and behavioral anomalies, providing insight into human neurodevelopmental disorders.


Adverse events after smallpox immunizations are associated with alterations in systemic cytokine levels.
The immunization of healthy adults with vaccinia virus (VV) induces a protective response against smallpox in most individuals but is also reactogenic in a significant number of vaccinees. The immunological mechanisms underlying the protective response or adverse events in humans are not well defined. Although cytokines contribute to antiviral immunity and, in some cases, cause systemic adverse effects, their role in the human response to VV is unknown. We investigated the effect of smallpox immunization on systemic cytokine concentrations in a cohort of VV-naive individuals. We found that smallpox immunization induces an interferon (IFN)- gamma -dominant response in the systemic compartment 1 week after immunization, with concentrations returning to baseline during convalescence. The level of IFN- gamma induced was not affected by the dilution of vaccine used. We also found that particular adverse events correlated with systemic cytokine patterns, which suggests a role for these molecules in the pathogenesis of adverse events.


Changes in myosin heavy chain mRNA and protein expression in human skeletal muscle with age and endurance exercise training.
Aging is associated with reduced muscle strength and atrophy of type II muscle fibers. Muscle fiber type and contractile function are primarily determined by myosin heavy chain (MHC) isoforms. There are few data available on the effects of aging on MHC isoform expression in humans. In the present study, we tested the hypothesis that MHC isoform protein composition and mRNA abundance would favor a fast-to-slow isoform shift with aging and in response to endurance exercise training. Muscle biopsies were obtained from previously sedentary, healthy men and women, aged 21-87 yr before (n = 77) and after (n = 65) 16 wk of bicycle training (up to 45 min at 80% peak heart rate, 3-4 days/wk). At baseline, MHC I mRNA was unchanged with age, whereas IIa and IIx declined by 14 and 10% per decade, respectively (P < 0.001). MHC IIa and IIx protein declined by 3 and 1% per decade with a reciprocal increase in MHC I (P < 0.05). After training, MHC I and IIa mRNA increased by 61 and 99%, respectively, and IIx decreased by 50% (all P < 0.001). The increase in MHC I mRNA was positively associated with age, whereas the changes in MHC IIa and IIx mRNA were similar across age. MHC I protein increased by 6% and was positively related to age, whereas IIx decreased by 5% and was inversely related to age. These results suggest that the altered expression of MHC isoforms with aging is transcriptionally regulated. In response to endurance exercise, regulation of MHC isoform transcripts remains robust in older muscle, but this did not result in corresponding changes in MHC protein expression.


M-Cells Contribute to the Entry of an Oral Vaccine but Are Not Essential for the Subsequent Induction of Protective Immunity against Francisella tularensis.
M-cells (microfold cells) are thought to be a primary conduit of intestinal antigen trafficking. Using an established neutralizing anti-RANKL (Receptor Activator of NF-κB Ligand) antibody treatment to transiently deplete M-cells in vivo, we sought to determine whether intestinal M-cells were required for the effective induction of protective immunity following oral vaccination with ΔiglB (a defined live attenuated Francisella novicida mutant). M-cell depleted, ΔiglB-vaccinated mice exhibited increased (but not significant) morbidity and mortality following a subsequent homotypic or heterotypic pulmonary F. tularensis challenge. No significant differences in splenic IFN-γ, IL-2, or IL-17 or serum antibody (IgG1, IgG2a, IgA) production were observed compared to non-depleted, ΔiglB-vaccinated animals suggesting complementary mechanisms for ΔiglB entry. Thus, we examined other possible routes of gastrointestinal antigen sampling following oral vaccination and found that ΔiglB co-localized to villus goblet cells and enterocytes. These results provide insight into the role of M-cells and complementary pathways in intestinal antigen trafficking that may be involved in the generation of optimal immunity following oral vaccination.


Brain and skeletal muscle bioenergetic failure in familial hypobetalipoproteinaemia.
To determine whether a multisystemic bioenergetic deficit is an underlying feature of familial hypobetalipoproteinaemia. Brain and skeletal muscle bioenergetics were studied by in vivo phosphorus MR spectroscopy (31P-MRS) in two neurologically affected members (mother and son) and in one asymptomatic member (daughter) of a kindred with familial hypobetalipoproteinaemia. Plasma concentrations of vitamin E and coenzyme Q10 (CoQ10) were also assessed. Brain 31P-MRS disclosed in all patients a reduced phosphocreatine (PCr) concentration whereas the calculated ADP concentration was increased. Brain phosphorylation potential was reduced in the members by about 40%. Skeletal muscle was studied at rest in the three members and during aerobic exercise and recovery in the son and daughter. Only the mother showed an impaired mitochondrial function at rest. Both son and daughter showed an increased end exercise ADP concentration whereas the rates of postexercise recovery of PCr and ADP were slow in the daughter. The rate of inorganic phosphate recovery was reduced in both cases. Plasma concentration of vitamin E and CoQ10 was below the normal range in all members. Structural changes in mitochondrial membranes and deficit of vitamin E together with reduced availability of CoQ10 can be responsible for the multisystemic bioenergetic deficit. Present findings suggest that CoQ10 supplementation may be important in familial hypobetalipoproteinaemia.


Performance characteristics of the MultiCode-RTx hepatitis C virus load test targeting the 3' untranslated region.
Viral load testing for hepatitis C virus (HCV) RNA has become a key parameter in the diagnosis of infection and treatment monitoring. This study evaluated the performance characteristics of the MultiCode-RTx HCV assay (MultiCode; EraGen Biosciences, Inc., Madison, WI), a real-time PCR test targeting the 3' untranslated region (UTR) of the HCV genome, compared to the TaqMan HCV load ASR assay (TaqMan; Roche Diagnostics, Indianapolis, IN) that targets the 5' UTR. For plasma specimens, the MultiCode assay had a limit of detection of 2.3 log10 IU/ml and a linear range of at least 6.7 log10 IU/ml. Comparison of plasma viral loads obtained by the MultiCode and TaqMan tests showed that they were in very close agreement (mean difference, -0.1 log10 IU/ml). No genotype bias was observed for genotypes 1, 2, and 3. When the MultiCode assay was evaluated with the MagNA Pure and easyMAG extraction methods, the viral loads for the easyMAG extraction were consistently higher for all specimens tested (mean difference, 0.45 log10 IU/ml). Aside from the limit of detection, the performance characteristics of the MultiCode assay were similar to the TaqMan assay for the clinical application of HCV load testing.