Chondroitin for osteoarthritis.
Osteoarthritis, a common joint disorder, is one of the leading causes of disability. Chondroitin has emerged as a new treatment. Previous meta-analyses have shown contradictory results on the efficacy of chondroitin. This, in addition to the publication of more trials, necessitates a systematic review. To evaluate the benefit and harm of oral chondroitin for treating osteoarthritis compared with placebo or a comparator oral medication including, but not limited to, nonsteroidal anti-inflammatory drugs (NSAIDs), analgesics, opioids, and glucosamine or other "herbal" medications. We searched seven databases up to November 2013, including the Cochrane Central Register of Controlled Trials (CENTRAL), Ovid MEDLINE, CINAHL, EMBASE, Science Citation Index (Web of Science) and Current Controlled Trials. We searched the US Food and Drug Administration (FDA) and European Medicines Agency (EMEA) websites for adverse effects. Trial registers were not searched. All randomized or quasi-randomized clinical trials lasting longer than two weeks, studying adults with osteoarthritis in any joint, and comparing chondroitin with placebo, an active control such as NSAIDs, or other "herbal" supplements such as glucosamine. Two review authors independently performed all title assessments, data extractions, and risk of bias assessments. Forty-three randomized controlled trials including 4,962 participants treated with chondroitin and 4,148 participants given placebo or another control were included. The majority of trials were in knee OA, with few in hip and hand OA. Trial duration varied from 1 month to 3 years. Participants treated with chondroitin achieved statistically significantly and clinically meaningful better pain scores (0-100) in studies less than 6 months than those given placebo with an absolute risk difference of 10% lower (95% confidence interval (CI), 15% to 6% lower; number needed to treat (NNT) = 5 (95% CI, 3 to 8; n = 8 trials) (level of evidence, low; risk of bias, high); but there was high heterogeneity between the trials (T(2) = 0.07; I(2) = 70%, which was not easily explained by differences in risk of bias or study sample size). In studies longer than 6 months, the absolute risk difference for pain was 9% lower (95% CI 18% lower to 0%); n = 6 trials; T(2) = 0.18; I(2) = 83% ), again with low level of evidence.For the Western Ontario and McMaster Universities Osteoarthritis Index Minimal Clinically Important Improvement (WOMAC MCII Pain subscale) outcome, a reduction in knee pain by 20% was achieved by 53/100 in the chondroitin group versus 47/100 in the placebo group, an absolute risk difference of 6% (95% CI 1% to 11%), (RR 1.12, 95% CI 1.01 to 1.24; T(2) = 0.00; I(2) = 0%) (n = 2 trials, 1253 participants; level of evidence, high; risk of bias, low).Differences in Lequesne's index (composite of pain,function and disability) statistically significantly favoured chondroitin as compared with placebo in studies under six months, with an absolute risk difference of 8% lower (95% CI 12% to 5% lower; T(2)= 0.78; n = 7 trials) (level of evidence, moderate; risk of bias, unclear), also clinically meaningful. Loss of minimum joint space width in the chondroitin group was statistically significantly less than in the placebo group, with a relative risk difference of 4.7% less (95% CI 1.6% to 7.8% less; n = 2 trials) (level of evidence, high; risk of bias, low). Chondroitin was associated with statistically significantly lower odds of serious adverse events compared with placebo with Peto odds ratio of 0.40 (95% CI 0.19 to 0.82; n = 6 trials) (level of evidence, moderate). Chondroitin did not result in statistically significant numbers of adverse events or withdrawals due to adverse events compared with placebo or another drug. Adverse events were reported in a limited fashion, with some studies providing data and others not.Comparisons of chondroitin taken alone or in combination with glucosamine or another supplement showed a statistically significant reduction in pain (0-100) when compared with placebo or an active control, with an absolute risk difference of 10% lower (95% CI 14% to 5% lower); NNT = 4 (95% CI 3 to 6); T(2) = 0.33; I(2) = 91%; n = 17 trials) (level of evidence, low). For physical function, chondroitin in combination with glucosamine or another supplement showed no statistically significant difference from placebo or an active control, with an absolute risk difference of 1% lower (95% CI 6% lower to 3% higher with T(2) = 0.04; n = 5 trials) (level of evidence, moderate). Differences in Lequesne's index statistically significantly favoured chondroitin as compared with placebo, with an absolute risk difference of 8% lower (95% CI, 12% to 4% lower; T(2) = 0.12; n = 10 trials) (level of evidence, moderate). Chondroitin in combination with glucosamine did not result in statistically significant differences in the numbers of adverse events, withdrawals due to adverse events, or in the numbers of serious adverse events compared with placebo or with an active control.The beneficial effects of chondroitin in pain and Lequesne's index persisted when evidence was limited to studies with adequate blinding or studies that used appropriate intention to treat (ITT) analyses. These beneficial effects were uncertain when we limited data to studies with appropriate allocation concealment or a large study sample (> 200) or to studies without pharmaceutical funding. A review of randomized trials of mostly low quality reveals that chondroitin (alone or in combination with glucosamine) was better than placebo in improving pain in participants with osteoarthritis in short-term studies. The benefit was small to moderate with an 8 point greater improvement in pain (range 0 to 100) and a 2 point greater improvement in Lequesne's index (range 0 to 24), both seeming clinically meaningful. These differences persisted in some sensitivity analyses and not others. Chondroitin had a lower risk of serious adverse events compared with control. More high-quality studies are needed to explore the role of chondroitin in the treatment of osteoarthritis. The combination of some efficacy and low risk associated with chondroitin may explain its popularity among patients as an over-the-counter supplement.


The key role of R-NHC coupling (R = C, H, heteroatom) and M-NHC bond cleavage in the evolution of M/NHC complexes and formation of catalytically active species.
Complexes of metals with N-heterocyclic carbene ligands (M/NHC) are typically considered the systems of choice in homogeneous catalysis due to their stable metal-ligand framework. However, it becomes obvious that even metal species with a strong M-NHC bond can undergo evolution in catalytic systems, and processes of M-NHC bond cleavage are common for different metals and NHC ligands. This review is focused on the main types of the M-NHC bond cleavage reactions and their impact on activity and stability of M/NHC catalytic systems. For the first time, we consider these processes in terms of NHC-connected and NHC-disconnected active species derived from M/NHC precatalysts and classify them as fundamentally different types of catalysts. Problems of rational catalyst design and sustainability issues are discussed in the context of the two different types of M/NHC catalysis mechanisms.


Corticosteroid use in COVID-19 patients: a systematic review and meta-analysis on clinical outcomes.
In the current SARS-CoV-2 pandemic, there has been worldwide debate on the use of corticosteroids in COVID-19. In the recent RECOVERY trial, evaluating the effect of dexamethasone, a reduced 28-day mortality in patients requiring oxygen therapy or mechanical ventilation was shown. Their results have led to considering amendments in guidelines or actually already recommending corticosteroids in COVID-19. However, the effectiveness and safety of corticosteroids still remain uncertain, and reliable data to further shed light on the benefit and harm are needed. The aim of this systematic review and meta-analysis was to evaluate the effectiveness and safety of corticosteroids in COVID-19. A systematic literature search of RCTS and observational studies on adult patients was performed across Medline/PubMed, Embase and Web of Science from December 1, 2019, until October 1, 2020, according to the PRISMA guidelines. Primary outcomes were short-term mortality and viral clearance (based on RT-PCR in respiratory specimens). Secondary outcomes were: need for mechanical ventilation, need for other oxygen therapy, length of hospital stay and secondary infections. Forty-four studies were included, covering 20.197 patients. In twenty-two studies, the effect of corticosteroid use on mortality was quantified. The overall pooled estimate (observational studies and RCTs) showed a significant reduced mortality in the corticosteroid group (OR 0.72 (95%CI 0.57-0.87). Furthermore, viral clearance time ranged from 10 to 29 days in the corticosteroid group and from 8 to 24 days in the standard of care group. Fourteen studies reported a positive effect of corticosteroids on need for and duration of mechanical ventilation. A trend toward more infections and antibiotic use was present. Our findings from both observational studies and RCTs confirm a beneficial effect of corticosteroids on short-term mortality and a reduction in need for mechanical ventilation. And although data in the studies were too sparse to draw any firm conclusions, there might be a signal of delayed viral clearance and an increase in secondary infections.


[Behavior of caliptrate Diptera in relation to oviposition substrates under laboratory conditions in Rio de Janeiro, RJ, Brazil].
How adult females of calyptrate Diptera recognize the appropriate breeding substrate is a matter of controversy. Among holometabolic insects, the feeding opportunities of immature stages are generally determined by the adult female choice of an oviposition site. The ovipositional and larvipositional substrate preference for the synanthropic flies (Chrysomya megacephala, C. putoria, Phaenicia cuprina: Calliphoridae; Atherigona orientalis, Synthesiomyia nudiseta: Muscidae; Ravinia belforti, Parasacophaga ruficornis, Peckia chrysostoma: Sarcophagidae) is presented in this work. The substrate used for testing were the following: bovine minced meat, fish (sardine), bovine liver, shrimp, squid, human faeces and banana. Bovine minced meat was the ovipositional and larvipositional substrate preferred by seven species. Human faces were preferred by R. belforti.


FOVEAL EXUDATE AND CHOROIDAL NEOVASCULARIZATION IN ATYPICAL CASES OF MULTIPLE EVANESCENT WHITE DOT SYNDROME.
To describe atypical cases of multiple evanescent white dot syndrome (MEWDS) associated with foveal exudation, increased choroidal thickness, and secondary Type 2 (subretinal) neovascularization. Four cases of atypical MEWDS were studied at a retina referral center. Patients underwent evaluation with multimodal retinal imaging, including fluorescein angiography, indocyanine green angiography, spectral-domain and enhanced depth imaging optical coherence tomography (OCT). Two patients were imaged with OCT angiography. Four patients (3 female, 1 male) with a median age of 23.5 years presented with acute onset, painless, decreased central vision. All cases demonstrated fundus findings consistent with MEWDS on color photography, indocyanine green angiography, fluorescein angiography, fundus autofluorescence, and structural OCT imaging. On structural OCT, all 4 patients were noted to have hyperreflective subretinal material and increased subfoveal choroidal thickness ranging from 307 μm to 515 μm. Type 2 neovascularization was diagnosed in all four patients using fluorescein angiography, indocyanine green angiography, and/or OCT angiography. Two patients had poor visual acuity at the last follow-up despite resolution of characteristic clinical findings of MEWDS. A subset of patients with atypical MEWDS may develop persistent poor vision due to subfoveal exudation and secondary Type 2 neovascularization. Patients showing increased choroidal thickness at presentation may be more susceptible to this unusual presentation.


Enhanced cyclophosphamide and ifosfamide activation in primary human hepatocyte cultures: response to cytochrome P-450 inducers and autoinduction by oxazaphosphorines.
The anticancer oxazaphosphorine prodrugs cyclophosphamide and ifosfamide are activated in human liver by a 4-hydroxylation reaction catalyzed by multiple cytochrome P450 (CYP) enzymes. In the present study, we used a cultured human hepatocyte model to identify possible inducers of the CYP-catalyzed activation of these two anticancer prodrugs. Treatment of primary cultures of human hepatocytes with phenobarbital, dexamethasone, or rifampin elevated hepatocyte microsomal oxazaphosphorine 4-hydroxylation by up to 200-400% of control for both drug substrates. These inductions were associated with corresponding increases in immunoreactive CYP2B6, CYP2C8, CYP2C9, and CYP3A4, all previously shown to catalyze oxazaphosphorine activation. Rifampin (1 microM, 96-h exposure) was a particularly potent inducer of ifosfamide and cyclophosphamide 4-hydroxylation, as well as of CYP3A protein levels and CYP3A-dependent testosterone 6beta-hydroxylation. CYP3A4, CYP2C8, and CYP2C9 protein levels were also increased by exposure of the hepatocytes to cyclophosphamide or ifosfamide (50 microM), which thereby enhanced their own rates of 4-hydroxylation in the cultured hepatocytes. In one human hepatocyte culture that contained the polymorphically expressed CYP3A5 in addition to the more widely expressed CYP3A4, only CYP3A4 was induced by cyclophosphamide, ifosfamide, and rifampin. These studies: (a) demonstrate an underlying metabolic basis for the clinically important oxazaphosphorine autoinduction pharmacokinetics seen with these drugs in cancer patients; and (b) identify rifampin and other CYP inducers as potentially useful for increasing the rates of cyclophosphamide 4-hydroxylation and ifosfamide 4-hydroxylation in human liver in a manner that could favorably impact the clinical pharmacokinetics of these anticancer prodrugs.


The use of agricultural substrates to improve methane yield in anaerobic co-digestion with pig slurry: effect of substrate type and inclusion level.
Anaerobic co-digestion of pig slurry with four agricultural substrates (tomato, pepper, persimmon and peach) was investigated. Each agricultural substrate was tested in co-digestion with pig slurry at four inclusion levels: 0%, 15%, 30% and 50%. Inclusion levels consisted in the replacement of the volatile solids (VS) from the pig slurry with the VS from the agricultural substrate. The effect of substrate type and inclusion level on the biochemical methane potential (BMP) was evaluated in a batch assay performed at 35 °C for 100 days. Agricultural substrate's chemical composition was also analyzed and related with BMP. Additionally, Bacteria and Archaea domains together with the four main methanogenic archaeal orders were quantified using quantitative real-time TaqMan polymerase chain reaction (qPCR) at the end of the experiment to determine the influence of agricultural substrate on sludge's microbial composition. Results showed that vegetable substrates (pepper and tomato) had higher lipid and protein content and lower carbohydrates than fruit substrates (persimmon and peach). Among substrates, vegetable substrates showed higher BMP than fruit substrates. Higher BMP values were obtained with increasing addition of agricultural substrate. The replacement of 50% of VS from pig slurry by tomato and pepper increased BMP in 41% and 44%, respectively compared with pig slurry only. Lower increments in BMP were achieved with lower inclusion levels. Results from qPCR showed that total bacteria and total archaea gene concentrations were similar in all combinations tested. Methanomicrobiales gene concentrations dominated over the rest of individual archaeal orders.


Myoinositol metabolism in diabetes mellitus. Effect of insulin treatment.
The metabolism of myoinositol has been studied in 10 nondiabetic subjects and in six patients with diabetes mellitus before and after insulin therapy. While dietary myoinositol intake and fecal myoinositol excretion were similar in both groups, urinary myoinositol excretion was increased 10-fold in the untreated diabetic and accounted for a significant fraction of his dietary myoinositol intake. Insulin treatment restored the urinary myoinositol excretion toward normal. Despite increased myoinositol excretion, plasma myoinositol concentrations were significantly higher in the diabetics following the ingestion of a standard diet or of a 3.0-gm. myoinositol load. This abnormality in oral myoinositol tolerance was also corrected by insulin treatment. The size of the rapidly equilibrating myoinositol pool was significantly decreased in the untreated diabetic and returned to normal following a brief period of insulin treatment. The elevated plasma myoinositol concentrations observed following myoinositol ingestion in the uncontrolled diabetic presumably represents a combination of enhanced gastrointestinal absorption and impaired intracellular transport of myoinositol. The decreased space of distribution of myoinositol also suggests an impairment of intracellular myoinositol transport in the untreated diabetic. These observations are consistent with the speculation that hyperglycemia may condition a widespread relative intracellular myoinositol deficiency in man and suggest that restoration of normal intracellular myoinositol concentrations might prove to be of benefit in the prevention and treatment of certain of the complications associated with human diabetes mellitus.


Interrelationships among Grain Size, Surface Composition, Air Stability, and Interfacial Resistance of Al-Substituted Li7La3Zr2O12 Solid Electrolytes.
The interfacial resistances of symmetrical lithium cells containing Al-substituted Li7La3Zr2O12 (LLZO) solid electrolytes are sensitive to their microstructures and histories of exposure to air. Air exposure of LLZO samples with large grain sizes (∼150 μm) results in dramatically increased interfacial impedances in cells containing them, compared to those with pristine large-grained samples. In contrast, a much smaller difference is seen between cells with small-grained (∼20 μm) pristine and air-exposed LLZO samples. A combination of soft X-ray absorption (sXAS) and Raman spectroscopy, with probing depths ranging from nanometer to micrometer scales, revealed that the small-grained LLZO pellets are more air-stable than large-grained ones, forming far less surface Li2CO3 under both short- and long-term exposure conditions. Surface sensitive X-ray photoelectron spectroscopy (XPS) indicates that the better chemical stability of the small-grained LLZO is related to differences in the distribution of Al and Li at sample surfaces. Density functional theory calculations show that LLZO can react via two different pathways to form Li2CO3. The first, more rapid, pathway involves a reaction with moisture in air to form LiOH, which subsequently absorbs CO2 to form Li2CO3. The second, slower, pathway involves direct reaction with CO2 and is favored when surface lithium contents are lower, as with the small-grained samples. These observations have important implications for the operation of solid-state lithium batteries containing LLZO because the results suggest that the interfacial impedances of these devices is critically dependent upon specific characteristics of the solid electrolyte and how it is prepared.


Renal graft survival and calcineurin inhibitor.
Tacrolimus is associated with fewer acute rejections than cyclosporine, but a greater risk of new onset diabetes mellitus. When compared to no tacrolimus among nondiabetics in a large patient registry, it is associated with improved graft survival. The current study used the same patient registry to compare more correctly graft survival between nondiabetic renal transplant recipients initially immunosuppressed with either of the two most frequently used calcineurin inhibitors, tacrolimus or modified cyclosporine (CsA). We examined data provided by the United States Renal Data System (USRDS) on all first, single-organ, renal transplants to nondiabetic recipients that occurred during the years 1996 to 2000. Importantly, we then limited the study to patients on CsA (n = 7,867) or tacrolimus (n = 3,082) as the initial agent. Patients with both or neither were excluded. We used Cox proportional hazards regressions to estimate the tacrolimus-related relative risk of graft failure, controlling for other significant donor, recipient, and transplant characteristics We found that tacrolimus patients had graft failure rates equivalent to those of CsA patients (hazard ratio= 1.031, P = 0.631) Although tacrolimus is being used with increasing frequency, analyses of the USRDS data show no net advantage in the ultimate transplantation outcome, graft survival. Given the higher acquisition price of tacrolimus compared to CsA and the similar risk of graft failure, further studies should be conducted to define those patient groups for which tacrolimus might be cost-effective.


Neuronal regulation of the development of the alpha-adrenergic chronotropic response in the rat heart.
During development, there are changes in the response of automatic cardiac fibers to alpha-adrenergic agonists. In neonatal rat ventricle, in vitro phenylephrine (1 X 10(-8) M) induces an increase in automatic rate from 115 +/- 12 (mean +/- SEM) to 168 +/- 10 beats/min, P less than 0.05. In contrast, in adult rat ventricle, the rate decreases from 36 +/- 8 to 12 +/- 12 beats/min, P less than 0.05. At both ages, the response is attenuated by the alpha 1-antagonist, prazosin (1 X 10(-6) M). We used cultures of neonatal rat myocytes to determine whether maturation of innervation contributes to the ontogeny of this response. All non-innervated cultures showed a positive chronotropic response to alpha-stimulation; phenylephrine (1 X 10(-8) M) increased the rate from 40 +/- 2 to 52 +/- 2 beats/min, P less than 0.05. In contrast, 60% of the myocytes innervated with sympathetic neurons showed a decrease in rate in response to phenylephrine, from 78 +/- 6 to 67 +/- 6 beats/min, P less than 0.05. The negative chronotropic effect of phenylephrine did not result from the release of acetylcholine or adenosine, or the inhibition of presynaptic norepinephrine release by phenylephrine. Furthermore, exposure to neuronal norepinephrine is not responsible for the alteration in muscle cell responsiveness. In conclusion, we have demonstrated the modulation of the myocardial response to alpha-adrenergic stimulation by the occurrence of innervation in tissue culture. This provides an explanation for the previously identified ontogenetic change in alpha-adrenergic effects on intact cardiac fibers from excitation to inhibition.


Mycobacterium celeriflavum sp. nov., a rapidly growing scotochromogenic bacterium isolated from clinical specimens.
Six strains of a rapidly growing scotochromogenic mycobacterium were isolated from pulmonary specimens of independent patients. Biochemical and cultural tests were not suitable for their identification. The mycolic acid pattern analysed by HPLC was different from that of any other mycobacterium. Genotypic characterization, targeting seven housekeeping genes, revealed the presence of microheterogeneity in all of them. Different species were more closely related to the test strains in various regions: the type strain of Mycobacterium moriokaense showed 99.0 % 16S rRNA gene sequence similarity, and 91.5-96.5 % similarity for the remaining six regions. The whole genome sequences of the proposed type strain and that of M. moriokaense presented an average nucleotide identity (ANI) of 82.9 %. Phylogenetic analysis produced poorly robust trees in most genes with the exception of rpoB and sodA where Mycobacterium flavescens and Mycobacterium novocastrense were the closest species. This phylogenetic relatedness was confirmed by the tree inferred from five concatenated genes, which was very robust. The polyphasic characterization of the test strains, supported by the ANI value, demonstrates that they belong to a previously unreported species, for which the name Mycobacterium celeriflavum sp. nov. is proposed. The type strain is AFPC-000207(T) ( = DSM 46765(T) = JCM 18439(T)).


Linezolid for treatment of chronic extensively drug-resistant tuberculosis.
Linezolid has antimycobacterial activity in vitro and is increasingly used for patients with highly drug-resistant tuberculosis. We enrolled 41 patients who had sputum-culture-positive extensively drug-resistant (XDR) tuberculosis and who had not had a response to any available chemotherapeutic option during the previous 6 months. Patients were randomly assigned to linezolid therapy that started immediately or after 2 months, at a dose of 600 mg per day, without a change in their background regimen. The primary end point was the time to sputum-culture conversion on solid medium, with data censored 4 months after study entry. After confirmed sputum-smear conversion or 4 months (whichever came first), patients underwent a second randomization to continued linezolid therapy at a dose of 600 mg per day or 300 mg per day for at least an additional 18 months, with careful toxicity monitoring. By 4 months, 15 of the 19 patients (79%) in the immediate-start group and 7 of the 20 (35%) in the delayed-start group had culture conversion (P=0.001). Most patients (34 of 39 [87%]) had a negative sputum culture within 6 months after linezolid had been added to their drug regimen. Of the 38 patients with exposure to linezolid, 31 (82%) had clinically significant adverse events that were possibly or probably related to linezolid, including 3 patients who discontinued therapy. Patients who received 300 mg per day after the second randomization had fewer adverse events than those who continued taking 600 mg per day. Thirteen patients completed therapy and have not had a relapse. Four cases of acquired resistance to linezolid have been observed. Linezolid is effective at achieving culture conversion among patients with treatment-refractory XDR pulmonary tuberculosis, but patients must be monitored carefully for adverse events. (Funded by the National Institute of Allergy and Infectious Diseases and the Ministry of Health and Welfare, South Korea; ClinicalTrials.gov number, NCT00727844.).


Molecular pathways: IL11 as a tumor-promoting cytokine-translational implications for cancers.
Emerging evidence suggests that cytokines produced by inflammatory cells act as rheostats to link the degree of wounding and local inflammation to epithelial cell survival, proliferation, and metabolism that collectively underpin the repair response. Among these cytokines, the GP130 family, which encompasses, among others, IL6 and IL11, plays a major role in orchestrating these complex processes through the activation of the latent signal transducer and activator of transcription 3 (STAT3) in the epithelium. However, many of the molecular mechanisms that govern and ensure effective epithelial wound healing and regeneration renewal also promote tumorigenesis and the progression of established cancers. Accordingly, GP130 cytokines endow the inflammatory tumor microenvironment with a capacity to promote "cancer hallmark capabilities" of the malignant epithelium, while simultaneously suppressing the antitumor response of innate and adaptive immune cells. Here, we review some recent insights derived from genetic and therapeutic inhibition of the IL6/IL11-GP130-STAT3 signaling cascade in the context of preclinical mouse models of cancer, which are likely to have implications to other solid malignancies.


A-type voltage-gated K+ currents influence firing properties of isolectin B4-positive but not isolectin B4-negative primary sensory neurons.
Voltage-gated K+ channels (Kv) in primary sensory neurons are important for regulation of neuronal excitability. The dorsal root ganglion (DRG) neurons are heterogeneous, and the types of native Kv currents in different groups of nociceptive DRG neurons are not fully known. In this study, we determined the difference in the A-type Kv current and its influence on the firing properties between isolectin B4 (IB4)-positive and -negative DRG neurons. Whole cell voltage- and current-clamp recordings were performed on acutely dissociated small DRG neurons of rats. The total Kv current density was significantly higher in IB+-positive than that in IB(4)-negative neurons. Also, 4-aminopyridine (4-AP) produced a significantly greater reduction in Kv currents in IB4-positive than in IB4-negative neurons. In contrast, IB4-negative neurons exhibited a larger proportion of tetraethylammonium-sensitive Kv currents. Furthermore, IB4-positive neurons showed a longer latency of firing and required a significantly larger amount of current injection to evoke action potentials. 4-AP significantly decreased the latency of firing and increased the firing frequency in IB4-positive but not in IB4-negative neurons. Additionally, IB4-positive neurons are immunoreactive to Kv1.4 but not to Kv1.1 and Kv1.2 subunits. Collectively, this study provides new information that 4-AP-sensitive A-type Kv currents are mainly present in IB4-positive DRG neurons and preferentially dampen the initiation of action potentials of this subpopulation of nociceptors. The difference in the density of A-type Kv currents contributes to the distinct electrophysiological properties of IB4-positive and -negative DRG neurons.


Condition-dependent trade-offs maintain honest signalling.
How and why animals and humans signal reliably is a key issue in biology and social sciences that needs to be understood to explain the evolution of communication. In situations in which the receiver needs to differentiate between low- and high-quality signallers, once a ruling paradigm, the Handicap Principle has claimed that honest signals have to be costly to produce. Subsequent game theoretical models, however, highlighted that honest signals are not necessarily costly. Honesty is maintained by the potential cost of cheating: by the difference in the marginal benefit to marginal cost for low versus high-quality signallers; i.e. by differential trade-offs. Owing to the difficulties of manipulating signal costs and benefits, there is lack of empirical tests of these predictions. We present the results of a laboratory decision-making experiment with human participants to test the role of equilibrium signal cost and signalling trade-offs for the development of honest communication. We found that the trade-off manipulation had a much higher influence on the reliability of communication than the manipulation of the equilibrium cost of signal. Contrary to the predictions of the Handicap Principle, negative production cost promoted honesty at a very high level in the differential trade-off condition.


Immunobiological factors aggravating the fatty infiltration on tendons and muscles in rotator cuff lesions.
Rotator cuff lesions (RCLs) are a common cause of shoulder pain and dysfunction. The rotator cuff tendons can degenerate and/or tear from the greater tuberosity of the humerus, which is associated with several anatomical, physiological, biochemical, and molecular changes in tendon and muscle. In this article, these pathways are critically reviewed and discussed with various management strategies of RCLs. The article also highlights the immunobiological responses following the RCL and the inherent repair mechanisms elicited by the body. The greatest difficulty in treating this pathology is that the muscle can undergo irreversible fatty infiltration in the setting of chronic tears that is associated with poor surgical outcomes. The article also investigates the key molecular pathways of the muscle homeostasis (mTOR, Rho kinase, AMPK, and Ca(2+)) with the energy metabolism to propose a possible mechanism for fatty infiltration. Future research is warranted to target the key players of these pathways in the management of fatty infiltration and thus RCL.


Assaying autophagic activity in transgenic GFP-Lc3 and GFP-Gabarap zebrafish embryos.
Autophagy mediates the bulk turnover of cytoplasmic constituents in lysosomes. During embryonic development in animals, a dramatic degradation of yolk proteins and synthesis of zygotic proteins takes place, leading to intracellular remodeling and cellular differentiation. Zebrafish represents a unique system to study autophagy due in part to its rapid embryonic development relative to other vertebrates. The technical advantages of this organism make it uniquely suited to various studies including high-throughput drug screens. To study autophagy in zebrafish, we identified two zebrafish Atg8 homologs, lc3 and gabarap, and generated two transgenic zebrafish lines expressing GFP-tagged versions of the corresponding proteins. Similar to yeast Atg8 and mammalian LC3, zebrafish Lc3 undergoes post-translational modification starting at the pharyngula stage during embryonic development. We observed a high level of autophagy activity in zebrafish embryos, which can be further upregulated by the TOR inhibitor rapamycin or the calpain inhibitor calpeptin. In addition, zebrafish Gabarap accumulates within lysosomes upon autophagy induction. Thus, we established a convenient zebrafish tool to assay autophagic activity during embryogenesis in vivo.


Incorporation of molecular data into the Revised International Prognostic Scoring System in treated patients with myelodysplastic syndromes.
The Revised International Prognostic Scoring System (IPSS-R) was developed for untreated myelodysplastic syndrome (MDS) patients based on clinical data. We created and validated a new model that incorporates mutational data to improve the predictive capacity of the IPSS-R in treated MDS patients. Clinical and mutational data from treated MDS patients diagnosed between January 2000 and January 2012 were used to develop the new prognostic system. A total of 508 patients were divided into training (n=333) and validation (n=175) cohorts. Independent significant prognostic factors for survival included age, IPSS-R, EZH2, SF3B1 and TP53. Weighted coefficients for each factor were used to build the new linear predictive model, which produced four prognostic groups: low, intermediate-1, intermediate-2 and high with a median overall survival of 37.4, 23.2, 19.9 and 12.2 months, respectively, P<0.001. Significant improvement in the C-index of the new model (0.73) was observed compared with the IPSS-R (0.69). The new model predicted outcome both in a separate validation cohort and in another cohort of patients with paired samples at different time points during their disease course. The addition of mutational data to the IPSS-R makes it dynamic and enhances its predictive ability in treated MDS patients regardless of their initial or subsequent therapies.


Associations of candidate genes to age-related macular degeneration among racial/ethnic groups in the multi-ethnic study of atherosclerosis.
To describe the relationships of selected candidate genes to the prevalence of early age-related macular degeneration (AMD) in a cohort of whites, blacks, Hispanics, and Chinese Americans. Cross-sectional study. setting: Multicenter study. study population: A total of 2456 persons aged 45-84 years with genotype information and fundus photographs. procedures: Twelve of 2862 single nucleotide polymorphisms (SNPs) from 11 of 233 candidate genes for cardiovascular disease were selected for analysis based on screening with marginal unadjusted P value <.001 within 1 or more racial/ethnic groups. Logistic regression models tested for association in case-control samples. main outcome measure: Prevalence of early AMD. Early AMD was present in 4.0% of the cohort and varied from 2.4% in blacks to 6.0% in whites. The odds ratio increased from 2.3 for 1 to 10.0 for 4 risk alleles in a joint effect analysis of Age-Related Maculopathy Susceptibility 2 rs10490924 and Complement Factor H Y402H (P for trend = 4.2×10(-7)). Frequencies of each SNP varied among the racial/ethnic groups. Adjusting for age and other factors, few statistically significant associations of the 12 SNPs with AMD were consistent across all groups. In a multivariate model, most candidate genes did not attenuate the comparatively higher odds of AMD in whites. The higher frequency of risk alleles for several SNPs in Chinese Americans may partially explain their AMD frequency's approaching that of whites. The relationships of 11 candidate genes to early AMD varied among 4 racial/ethnic groups, and partially explained the observed variations in early AMD prevalence among them.


Ceramic-dentin bonding: effect of adhesive systems and light-curing units.
To investigate the effect of different light-curing units and adhesive systems on the bonding of leucite ceramic to dentin. Flat dentin surfaces were ground occlusally on human molars (n = 60). Hot-pressed ceramic blocks of IPS Empress (Ivoclar-Vivadent) were fabricated, air-particle abraded, and cleaned in distilled water. Following acid etching and silane treatment of ceramic, bonding procedures were performed. The teeth were divided into two groups according to the type of the adhesive system: (1) total-etch: etchant (Etch 37) and adhesive (One Step Plus); (2) self-etching: self-priming etchant (Tyrian SPE) and adhesiveOne Step Plus) (n = 30). Ceramic blocks were bonded to the dentin surfaces with dual-polymerizing resin luting cement (Duolink). For polymerization, the QTH (Blue Swan Digital, Dentanet) was used in soft-up and high-power mode, the LED (Elipar Freelight 2, 3M Espe) in exponential and standard mode, and the PAC (PlasmaStar, SP-2000, Monitex) in normal and ramp-curing mode (n = 5). Following storage in distilled water for 24 h, the samples were thermocycled for 6000 cycles. The bonded specimens were serially sectioned and trimmed to hourglass shapes with approximately 1.6 +/- 0.16 mm2 cross-sectional areas, then tested with the microtensile tester at a rate of 1 mm/min. Fracture surfaces (were analyzed with SEM. The data were analyzed with three-way analysis of variance (ANOVA). ANOVA revealed that adhesive systems (p < 0.001) and light-curing units (p = 0.015) had a significant effect on bond strength values. Bond strength means (+/- SD) in MPa were as follows: total-etch system: QTH/soft-up mode = 16.2 (5.4); QTH/high power mode = 15 (5.4); LED/standard mode = 12.1 (3.3); LED/exponential mode = 15 (5); PAC/normal mode = 19.3 (7); PAC/ramp-curing mode = 19.6 (7.4). Self-etching system: QTH/soft-up mode = 9.9 (2.1); QTH/high-power mode = 12.5 (4.3); LED/standard mode = 8.6 (2); LED/exponential mode = 13 (3.8); PAC/normal mode = 13.1 (3.9); PAC/ramp-curing mode = 8.9 (2.2). Results indicated that the conventional total-etch system provided more reliable bonding compared to the self-etching system. The PAC light-curing unit together with the total-etch adhesive system showed the highest mean micro-TBS values with either normal mode or ramp-curing mode.


Dietary fibers influence the intestinal SCFAs and plasma metabolites profiling in growing pigs.
Dietary fibers (DFs), widely used as food additives to replace fat, can benefit metabolic disorders. Soluble and highly fermentable inulin (INU) and insoluble and less fermentable microcrystalline cellulose (MCC) have hypolipidemic or cholesterol-lowering effects, respectively. The current study was aimed at identifying plasma metabolite signatures and metabolic pathways associated with inclusion of DFs with different physiochemical properties, which will lead to a better understanding of functional pathways for DFs to improve metabolic health. Pigs were fed with diets containing 5% INU or MCC or cornstarch (CON) for 60 days. Lipid parameters, fasting hormones and gut SCFAs were measured, and metabolites were profiled with GC-MS. The results showed that the inclusion of 5% MCC decreased the total and LDL-cholesterol in the plasma, while INU only displayed a tendency. Fasting glucagon was decreased by the inclusion of INU and MCC when insulin and leptin remained unchanged. In the cecum and colon, INU increased propionate while MCC increased the colonic branched chain fatty acids. For the first time, specific patterns of metabolite profiles were identified for the INU and MCC inclusion. Four signatures (isoleucine, proline, ornithine and erythrose) were associated with the inclusion of INU, while six metabolites (isoleucine, proline, ornithine, erythrose, hydroxyproline and 5-MIAA) were associated with MCC when compared with the CON group. The direct comparison between the INU and MCC groups revealed significant differences in 5-oxoproline, hydroxyproline, azelaic acid and dodecanoic acid. Identified signatures were enriched in metabolic pathways related to amino acid (arginine and proline; phenylalanine), carbohydrate (pyruvate; galactose) and linoleic acid metabolism. Some amino acid metabolites were also found to correlate with plasma lipid parameters. Our data provided new insights for mechanisms in amino acid and lipid metabolism, by which different DFs could benefit metabolic disorders.


A Glimpse into the World of Integrative and Mobilizable Elements in Streptococci Reveals an Unexpected Diversity and Novel Families of Mobilization Proteins.
Recent analyses of bacterial genomes have shown that integrated elements that transfer by conjugation play an essential role in horizontal gene transfer. Among these elements, the integrative and mobilizable elements (IMEs) are known to encode their own excision and integration machinery, and to carry all the sequences or genes necessary to hijack the mating pore of a conjugative element for their own transfer. However, knowledge of their prevalence and diversity is still severely lacking. In this work, an extensive analysis of 124 genomes from 27 species of Streptococcus reveals 144 IMEs. These IMEs encode either tyrosine or serine integrases. The identification of IME boundaries shows that 141 are specifically integrated in 17 target sites. The IME-encoded relaxases belong to nine superfamilies, among which four are previously unknown in any mobilizable or conjugative element. A total of 118 IMEs are found to encode a non-canonical relaxase related to rolling circle replication initiators (belonging to the four novel families or to MobT). Surprisingly, among these, 83 encode a TcpA protein (i.e., a non-canonical coupling protein (CP) that is more closely related to FtsK than VirD4) that was not previously known to be encoded by mobilizable elements. Phylogenetic analyses reveal not only many integration/excision module replacements but also losses, acquisitions or replacements of TcpA genes between IMEs. This glimpse into the still poorly known world of IMEs reveals that mobilizable elements have a very high prevalence. Their diversity is even greater than expected, with most encoding a CP and/or a non-canonical relaxase.


Pattern of expression of highly polysialylated neural cell adhesion molecule in the developing and adult rat striatum.
In rats, morphological and synaptic maturation of the striatum, a brain area involved in the control of movement and in cognitive behaviour, proceeds for several weeks postnatally. Little is known, however, about the molecular events associated with the final maturation of the striatum. In particular, there is little information on molecules playing a role in cell adhesion, a phenomenon of particular importance for neuronal development. We have examined the time course and topography of expression of the highly polysialylated form of the neural cell adhesion molecule in the rat striatum during postnatal development and in the adult, and compared it to growth-associated protein-43, a marker of axonal growth. As earlier during development [Aaron L. I. and Chesselet M.-F. (1989) Neuroscience 28, 701-710], immunolabelling for polysialylated neural cell adhesion molecule was very intense in the entire striatum at postnatal days 17-19. At postnatal days 21 and 22, loss of polysialylated neural cell adhesion molecule immunoreactivity in the caudal part of the striatum contrasted with the persistence of immunoreactivity at more rostral levels. Most of the striatum was devoid of polysialylated neural cell adhesion molecule immunoreactivity by postnatal day 25. At this age, as well as in the striatum of adult rats, immunolabelling was only observed along the ventricular edge of the striatum. In contrast to polysialylated neural cell adhesion molecule immunoreactivity, immunolabelling for growth-associated protein-43 had reached its adult pattern by postnatal day 17, indicating that polysialylated neural cell adhesion molecule persists beyond the period of major axonal growth. In the adult, an area of stronger growth associated protein-43 immunoreactivity overlapped with the region which retained immunoreactivity to polysialylated neural cell adhesion molecule. The results indicate that, in the developing rat striatum, the neural cell adhesion molecule remains highly sialylated not only during the ingrowth of cortical and nigral inputs but also during the formation of dendritic spine and synaptogenesis. Loss of polysialyated neural cell adhesion molecule occurs at the time of emerging spontaneous activity in cerebral cortex, and precedes the development of mature responses to cortical stimulation and adult membrane properties in a majority of striatal neurons.


Comparison of health parameters in normal cats fed a limited iodine prescription food vs a conventional diet.
Objectives The objective was to compare the effect on thyroid function in healthy, adult cats fed a limited-iodine food or conventional diet for 24 months. Methods Cats in the limited-iodine group (n = 14) were fed a commercial, dry food containing 0.2 ppm iodine on a dry-matter basis (DMB). Cats in the conventional diet group (n = 12) were fed an identical diet except that the iodine content was 3.2 ppm on a DMB. Both groups were maintained on their respective diets for 24 months. The median age of the cats at baseline was 3.2 years (interquartile range 2.4-4.3). Diagnostic samples were obtained for measurement of serum biochemistry parameters, thyroid hormone concentrations, complete blood count and urinalysis, and thyroid ultrasound examination was performed at baseline, 6, 12, 18 and 24 months. Results Median serum concentrations of free and total thyroxine and thyroid-stimulating hormone were within respective reference intervals for both test groups at all sampling intervals. Median urine iodine concentrations in the limited-iodine group declined significantly ( P = 0.0001) from baseline and were significantly different than conventional diet ( P ⩽0.0007). Ultrasound examination revealed no significant change in median thyroid height in the limited-iodine group at any time point. Conclusions and relevance Normal serum thyroid hormone concentrations and results of urinalysis and routine biochemical testing confirmed that the limited-iodine food was safely fed for 2 years to healthy, adult cats. Ultrasound examination showing that cats on limited-iodine did not develop a change in thyroid gland height was a further indication of the absence of iatrogenic hypothyroidism in the limited iodine diet group. These results support the lack of obvious side effects associated with feeding a limited-iodine diet for 2 years to healthy, adult cats, a situation that may occur in multi-cat households where healthy cats have access to a limited-iodine diet being used to manage cats with feline hyperthyroidism.


Langerhans cell deficiency impairs Ixodes scapularis suppression of Th1 responses in mice.
Ixodes scapularis ticks transmit a number of human pathogens, including the Lyme disease spirochete Borrelia burgdorferi. I. scapularis suppresses host immunity in the skin to promote feeding and systemically skew T-helper (Th)-cell differentiation toward Th2 cells in secondary lymphoid organs. Although components of tick saliva are known to influence Th-cell polarization, the mechanism whereby tick feeding in the skin modulates regional and systemic Th-cell responses is unknown. In this study, the role of the epidermal Langerhans cell (LC) subset of skin dendritic cells in tick-mediated Th1/Th2-cell immunomodulation was assessed. Mice deficient in LCs (Langerin-DTA mice) exhibited enhanced lymph node (LN) concanavalin A (ConA)-induced Th1 responses after tick infestation in comparison to results for uninfested Langerin-DTA or wild-type (WT) mice, whereas effects on Th2-cell production of interleukin 4 were more variable. Nonetheless, the altered T-cell response did not impact tick feeding or refeeding. Gamma interferon production by ConA-stimulated LN cells of both WT and LC-deficient mice was enhanced by as much as fourfold after B. burgdorferi-infected-tick feeding, indicating that immunomodulatory effects of tick saliva were not able to attenuate the Th1 immune responses induced by this pathogen. Taken together, these findings show a requirement for LCs in the tick-mediated attenuation of Th1 responses in regional lymph nodes but not in the spleens of mice and show that the presence of a pathogen can overcome the Th1-inhibitory effects of tick feeding on the host.


Optimization of extraction conditions for polyphenols from the stem bark of Funtumia elastica (Funtum) utilizing response surface methodology.
Background: The recovery of phenolic compounds is seen as an arduous task because phenolic compounds are available as free aglycones, as sugar or ester conjugates, or as polymers with several monomeric components. Furthermore, phenolic compounds do not disperse evenly and may be connected to cell walls, carbohydrates, or proteins. This study looks at the optimization of factors that affect the efficiency for the extraction of phenolic compounds from the stem-bark of Funtumia elastica. Methods: Five independent variables (solvent concentration, time, the temperature, solid-liquid ratio, and pH) of the extraction process were selected. Single factor analysis as well as the response surface method was used to evaluate the impact of the selected factors on the total phenolic content. The effect of the extraction factors on the phenolic content was statistically significant (p <0.05). For the response surface method, a five/factor, five/level central composite design used, and a fitted second-order polynomial regression model equation was used to show how the extraction parameters affected the total phenolic recovery. Results: The predicted value (R² of 0.5917) agreed with the adjusted value (R² of 0.7707). The residuals for response predictions were less than 5%. The optimal factors for the extraction were ethanol concentration of 75.99% v/v, extraction time of 193.86 minutes, temperature of 63.66°C, pH of 5.62, and solid-liquid ratio of 1:21.12 g/mL. Actual overall content of the phenolic compounds was validated at 82.83 ± 3.335 mg gallic acid equivalent (GAE) /g weight of extract, which agreed with the predicted response of 89.467 mg GAE/g of the dried extract under the optimal factors. Conclusions: The rich phenolic content of stem-bark of Funtumia elastica points to its potential as a functional medicinal product to alleviate diseases caused by oxidative stress such as asthma, breathing disorders, inflammation, and cardiovascular diseases.


Generalized dephasing relation for fidelity and application as an efficient propagator.
The dephasing relation (DR), a linearization of semiclassical fidelity, is generalized to include the overlap of "off-diagonal" elements. The accuracy of the formulation is tested in integrable and chaotic systems and its scaling with dimensionality is studied in a Caldeira-Leggett model with many degrees of freedom. It is shown that the DR is often in very good agreement with numerically analytic quantum results and frequently outperforms an alternative semiclassical treatment. Most importantly, since there is no computationally expensive prefactor, and Monte Carlo Metropolis sampling is used to facilitate the calculation, the DR is found to scale remarkably well with increasing dimension. We further demonstrate that a propagator based on the DR can include more quantum coherence and outperform other popular linearized semiclassical methods, such as forward-backward semiclassical dynamics (FBSD) and the linearized semiclassical initial value representation (LSC-IVR).


Further neuroimaging evidence for the deficit subtype of schizophrenia: a cortical connectomics analysis.
The clinical heterogeneity of schizophrenia has hindered neurobiological investigations aimed at identifying neural correlates of the disorder. To identify network-based biomarkers across the spectrum of impairment present in schizophrenia by separately evaluating individuals with deficit and nondeficit subtypes of this disorder. A university hospital network-based neuroimaging study was conducted between February 1, 2007, and February 28, 2012. Participants included patients with schizophrenia (n = 128) and matched healthy controls (n = 130) from two academic centers and patients with bipolar I disorder (n = 39) and matched healthy controls (n = 43) from a third site. Patients with schizophrenia at each site in the top quartile on the proxy scale for the deficit syndrome were classified as having deficit schizophrenia and those in the bottom quartile were classified as having nondeficit schizophrenia. All participants underwent magnetic resonance brain imaging. Network-level properties of cortical thickness were assessed in each group. Interregional cortexwide coupling was compared among the groups, and graph theoretical approaches were used to assess network density and node degree, betweenness, closeness, and eigenvector centrality. Stronger frontoparietal and frontotemporal coupling was found in patients with deficit schizophrenia compared with those with nondeficit schizophrenia (17 of 1326 pairwise relationships were significantly different, P < .05; 5% false discovery rate) and in patients with deficit schizophrenia compared with healthy controls (49 of 1326 pairwise relationships were significantly different, P < .05; 5% false discovery rate). Participants with nondeficit schizophrenia and bipolar I disorder did not show significant differences in coupling relative to those in the control groups (for both comparisons, 0 of 1326 pairwise relationships were significantly different, P > .05; 5% false discovery rate). The networks formed from patients with deficit schizophrenia demonstrated increased density of connections relative to controls and nondeficit patients (range, 0.07-0.45 in controls, 0.09-0.43 in the nondeficit group, and 0.18-0.67 in the deficit group). High centrality nodes were identified in the supramarginal, middle, and superior temporal and inferior frontal regions in deficit schizophrenia networks based on ranking of 4 centrality metrics. High centrality regions were identified as those that ranked in the top 10 in 50% or more of the thresholded networks in 3 or more of the centrality measures. Network properties were similar in patients with deficit schizophrenia from both study sites. Patients with schizophrenia at one end of a spectrum show characteristic signatures of altered intracortical relationships compared with those at the other end of that spectrum, patients with bipolar I disorder, and healthy individuals. Cortical connectomic approaches can be used to reliably identify neural signatures in clinically heterogeneous groups of patients.


Rational performance metrics for operating theatres, principles of efficiency, and how to achieve it.
Several performance metrics are commonly used by National Health Service (NHS) organizations to measure the efficiency and productivity of operating lists. These include: start time, utilization, cancellations, number of operations and gap time between operations. The authors describe reasons why these metrics are flawed, and use clinical evidence and mathematics to define a rational, balanced efficiency metric. A narrative review of literature on the efficiency and productivity of elective NHS operating lists was undertaken. The aim was to rationalize how best to define and measure the efficiency of an operating list, and describe strategies to achieve it. There is now a wealth of literature on how optimally to measure the performance of elective surgical lists. Efficiency may be defined as the completion of all scheduled operations within the allocated time with no over- or under-runs. Achieving efficiency requires appropriate scheduling using specific procedure mean (or median) times and their associated variance (standard deviation or interquartile range) to calculate the probability they can be completed on time. The case mix may be adjusted to yield better time management. This review outlines common misconceptions applied to managing scheduled operating theatre lists and the challenges of measuring unscheduled operations in emergency settings.


A stearoyl-acyl carrier protein desaturase, NbSACPD-C, is critical for ovule development in Nicotiana benthamiana.
Stearoyl-acyl carrier protein desaturase (SACPD) activity is essential for production of the major unsaturated fatty acids (UFAs) in plant lipids. We report here the characterization of three SACPD genes from Nicotiana benthamiana, NbSACPD-A, -B, and -C. All three genes share high similarity to AtSSI2/FAB2 (Suppressor of Salicylic acid-Insensitivity2/Fatty Acid Biosynthesis2), the primary SACPD isoform in Arabidopsis. Knocking down the expression of individual or combinations of NbSACPDs by an artificial microRNA approach resulted in significantly reduced accumulation of 18C UFAs and elevated levels of 18:0-FA (Fatty acids) in leaves, indicating that all three genes participated in fatty acid desaturation. The triple knockdown (KD) plants displayed severe growth phenotypes, including spontaneous cell death and dwarfing. While no vegetative morphologic abnormality was observed in NbSACPD-A, -B, or -C KD plants, strikingly, NbSACPD-C KD plants produced small fruits with aborted ovules. Reciprocal crosses with wild-type and NbSACPD-C KD plants revealed that knocking down NbSACPD-C expression caused female, but not male, sterility. Furthermore, arrested ovule development and significantly altered lipid composition in ovaries were observed in NbSACPD-C KD plants, consistent with the predominant NbSACPD-C expression in ovules. The ovule development defect was fully complemented by coexpressing an amiRNA-resistant NbSACPD-C variant in the NbSACPD-C KD background, further supporting a specific requirement for NbSACPD-C in female fertility. Our results thus indicated that NbSACPD-C plays a critical role maintaining membrane lipid composition in ovule development for female fertility in N. benthamiana, complementing and extending prior understanding on the well-demonstrated roles of SACPDs in biotic and abiotic stresses.


A centrosome-autonomous signal that involves centriole disengagement permits centrosome duplication in G2 phase after DNA damage.
DNA damage can induce centrosome overduplication in a manner that requires G2-to-M checkpoint function, suggesting that genotoxic stress can decouple the centrosome and chromosome cycles. How this happens is unclear. Using live-cell imaging of cells that express fluorescently tagged NEDD1/GCP-WD and proliferating cell nuclear antigen, we found that ionizing radiation (IR)-induced centrosome amplification can occur outside S phase. Analysis of synchronized populations showed that significantly more centrosome amplification occurred after irradiation of G2-enriched populations compared with G1-enriched or asynchronous cells, consistent with G2 phase centrosome amplification. Irradiated and control populations of G2 cells were then fused to test whether centrosome overduplication is allowed through a diffusible stimulatory signal, or the loss of a duplication-inhibiting signal. Irradiated G2/irradiated G2 cell fusions showed significantly higher centrosome amplification levels than irradiated G2/unirradiated G2 fusions. Chicken-human cell fusions demonstrated that centrosome amplification was limited to the irradiated partner. Our finding that only the irradiated centrosome can duplicate supports a model where a centrosome-autonomous inhibitory signal is lost upon irradiation of G2 cells. We observed centriole disengagement after irradiation. Although overexpression of dominant-negative securin did not affect IR-induced centrosome amplification, Plk1 inhibition reduced radiation-induced amplification. Together, our data support centriole disengagement as a licensing signal for DNA damage-induced centrosome amplification.


Mesenchymal stem cells accelerate bone allograft incorporation in the presence of diabetes mellitus.
Allograft (Allo) incorporation in the presence of a systemic disease like diabetes mellitus (DM) is becoming a major issue in the orthopedic community. Mesenchymal stem cells (MSC) are multipotent stem cells that may be derived from adult, whole bone marrow and have been shown to induce bone formation in segmental defects when combined with the appropriate carrier/scaffold. The objectives of this study were to analyze the effect of DM upon Allo incorporation in a segmental rat femoral defect and to also investigate MSC augmentation of Allo incorporation. Segmental (5 mm) femoral defects were created in non-DM and DM rats and treated with Allo containing demineralized bone matrix (DBM) or DBM with MSC augmentation. Histological scoring at 4 weeks demonstrated less mature bone in the DM/DBM group compared to its non-DM counterpart (p < 0.001). However, there was significantly more mature bone in the DM/MSC group when compared to the DM/DBM group at both 4 and 8 weeks (p < 0.001 and p = 0.004). Furthermore, significantly more bone formation was observed in the DM/MSC group compared to the DM/DBM group at the 4-week time point (p < 0.001). The results of this study suggest that MSC are a potential adjunct for bone regeneration when implanted in an orthotopic site in the presence of DM.


An alternative splice variant of human αA-crystallin modulates the oligomer ensemble and the chaperone activity of α-crystallins.
In humans, ten genes encode small heat shock proteins with lens αA-crystallin and αB-crystallin representing two of the most prominent members. The canonical isoforms of αA-crystallin and αB-crystallin collaborate in the eye lens to prevent irreversible protein aggregation and preserve visual acuity. α-Crystallins form large polydisperse homo-oligomers and hetero-oligomers and as part of the proteostasis system bind substrate proteins in non-native conformations, thereby stabilizing them. Here, we analyzed a previously uncharacterized, alternative splice variant (isoform 2) of human αA-crystallin with an exchanged N-terminal sequence. This variant shows the characteristic α-crystallin secondary structure, exists on its own predominantly in a monomer-dimer equilibrium, and displays only low chaperone activity. However, the variant is able to integrate into higher order oligomers of canonical αA-crystallin and αB-crystallin as well as their hetero-oligomer. The presence of the variant leads to the formation of new types of higher order hetero-oligomers with an overall decreased number of subunits and enhanced chaperone activity. Thus, alternative mRNA splicing of human αA-crystallin leads to an additional, formerly not characterized αA-crystallin species which is able to modulate the properties of the canonical ensemble of α-crystallin oligomers.


The effect of age on the expression of apoptosis biomarkers in human spermatozoa.
To evaluate the impact of age on the expression of apoptotic biomarkers in human spermatozoa. Cross sectional, prospective study. Academic centers. Healthy volunteers with proven fertility, stratified by age (n = 25, range: 20-68 years). Examination of serum hormone levels and basic semen parameters, and assessment of early (plasma membrane translocation of phosphatidylserine) and late (DNA fragmentation) sperm apoptotic markers by flow cytometry (using Annexin-V binding and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling). Apoptosis markers. Advancing male age was significantly and positively correlated with Annexin-V binding results. Although not significant, there was a clear trend for increased DNA fragmentation in the older groups. The age threshold for these observations appears to be 40 years. Advancing male age was positively correlated with FSH and sex hormone-binding globulin (SHBG) levels, and negatively correlated with sperm concentration. Advancing male age is associated with the expression of early apoptotic markers as evidenced by significantly increased plasma membrane translocation of phosphatidylserine, as well as with a more subtle proportion of sperm carrying DNA fragmentation. This study confirmed that male age is also associated with a decline in sperm concentration.


Further evaluation of the mechanism by which amphetamine reduces striatal dopamine metabolism: a brain dialysis study.
An intracerebral dialysis method was used in the halothane-anaesthetized rat to further clarify the site which mediates the amphetamine-induced decrease of the striatal dopamine (DA) metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA). Following subcutaneous injection of amphetamine (0.1-5.0 mg/kg), DOPAC and HVA in striatal perfusates decreased over the 2 h time course, with 0.5 mg/kg of the drug having maximal effect. In comparison, amphetamine (0.1-5.0 mg/kg) caused a strictly dose-dependent increase of DA in striatal perfusates. Following low (0.1-5.0 mg/kg) but not high (2.0-5.0 mg/kg) doses of amphetamine there was a negative correlation between the increase of DA and decrease of DOPAC in the striatum. Amphetamine (0.5 mg/kg) caused a reduction in DA metabolism in the ibotenic acid-lesioned striatum. Intranigral injection of 10 micrograms, but not of 1 microgram amphetamine, decreased DOPAC (-27%) in the striatal perfusates. However, injection of 1 microgram amphetamine into the striatum caused a strong decrease in striatal DOPAC (-46%) and HVA (-22%). The possible mechanisms of action of amphetamine are discussed in the light of these data.


Idiopathic eosinophilic esophagitis in adults: the ringed esophagus.
To retrospectively assess the findings of idiopathic eosinophilic esophagitis (IEE) at barium studies and determine the frequency of the ringed esophagus in patients with this condition. The institutional review board approved all aspects of this retrospective study and did not require informed consent from patients whose records were included in the study. The study was compliant with the Health Insurance Portability and Accountability Act. A review of the pathology and radiology databases at the authors' institution revealed 14 patients (11 men and three women; mean age, 41.3 years) with IEE (defined as more than 20 eosinophils per high-power field in biopsy specimens) who had undergone barium studies and endoscopy. The radiographs were reviewed for strictures, esophagitis, or other abnormalities. The endoscopic reports, clinical records, and laboratory data were also reviewed and compared with the radiographic findings. Seven of the 14 patients (50%) had a history of allergies, and two of nine patients with complete blood cell counts (22%) had peripheral eosinophilia. Thirteen patients (93%) had dysphagia, six (43%) had food impactions, and six (43%) had reflux symptoms. Ten patients (71%) had a total of 11 strictures at barium studies (two in the upper part of the esophagus, two in the middle part, three in the distal part, one in the middle and distal parts, and three at the gastroesophageal junction). The strictures had a mean length of 5.1 cm. In seven patients (50%), the strictures contained multiple fixed ringlike indentations that produced a ringed esophagus. The ringlike indentations appeared as multiple, fixed, closely spaced, concentric rings traversing the stricture. Four patients (28%) had esophagitis. Of 13 patients who underwent recumbent imaging, 10 (77%) had hiatal hernias and nine (69%) had reflux. Eight of the 10 patients (80%) with strictures underwent endoscopic dilation procedures, which resulted in only temporary relief of dysphagia. The findings suggest that most patients with IEE have esophageal strictures, often with distinctive ringlike indentations that produce a ringed esophagus.


Malaria: targeting parasite and host cell kinomes.
Malaria still remains one of the deadliest infectious diseases, and has a tremendous morbidity and mortality impact in the developing world. The propensity of the parasites to develop drug resistance, and the relative reluctance of the pharmaceutical industry to invest massively in the developments of drugs that would offer only limited marketing prospects, are major issues in antimalarial drug discovery. Protein kinases (PKs) have become a major family of targets for drug discovery research in a number of disease contexts, which has generated considerable resources such as kinase-directed libraries and high throughput kinase inhibition assays. The phylogenetic distance between malaria parasites and their human host translates into important divergences in their respective kinomes, and most Plasmodium kinases display atypical properties (as compared to mammalian PKs) that can be exploited towards selective inhibition. Here, we discuss the taxon-specific kinases possessed by malaria parasites, and give an overview of target PKs that have been validated by reverse genetics, either in the human malaria parasite Plasmodium falciparum or in the rodent model Plasmodium berghei. We also briefly allude to the possibility of attacking Plasmodium through the inhibition of human PKs that are required for survival of this obligatory intracellular parasite, and which are targets for other human diseases.


Absenteeism among hospital nurses: an idiographic-longitudinal analysis.
For several months, nurses completed ratings of the degree to which certain events relevant to absence were present during each of their scheduled workdays. The event ratings for days when the nurses decided to be absent were then compared with those for days when the nurses attended. As expected, certain events, such as ill health and tiredness, tended to covary and proved to be consistently related to absenteeism across nurses. Also as expected, some events that were not especially relevant for the nurses as a whole, like having a sick family member or friend and concerns about previous poor attendance, nonetheless emerged as being relevant to the absence behavior of certain individuals. Finally, some events were consistently related to the nurses' expressed desire to be absent but not to actual absences. We discuss these differences from two perspectives, one emphasizing the role of attribution bias and the other, a two-stage process in which such bias has no major role.


Hematopoiesis from human embryonic stem cells: overcoming the immune barrier in stem cell therapies.
The multipotency and proliferative capacity of human embryonic stem cells (hESCs) make them a promising source of stem cells for transplant therapies and of vital importance given the shortage in organ donation. Recent studies suggest some immune privilege associated with hESC-derived tissues. However, the adaptability of the immune system makes it unlikely that fully differentiated tissues will permanently evade immune rejection. One promising solution is to induce a state of immune tolerance to a hESC line using tolerogenic hematopoietic cells derived from it. This could provide acceptance of other differentiated tissues from the same line. However, this approach will require efficient multilineage hematopoiesis from hESCs. This review proposes that more efficient differentiation of hESCs to the tolerogenic cell types required is most likely to occur through applying knowledge gained of the ontogeny of complex regulatory signals used by the embryo for definitive hematopoietic development in vivo. Stepwise formation of mesoderm, induction of definitive hematopoietic stem cells, and the application of factors key to their self-renewal may improve in vitro production both quantitatively and qualitatively.


Model-based evaluation of fouling mechanisms in powdered activated carbon/membrane bioreactor system.
Identifying the fouling degree of a membrane bioreactor (MBR) provides guidance on the determination of suitable membrane cleaning methods. There is still a lack of knowledge on the effects of powdered activated carbon (PAC) refresh ratio on the MBR fouling mechanism. Major fouling mechanisms of an MBR with constant flow rate at different PAC replenishment ratios were investigated by individual and combined mechanistic fouling models. The root mean square errors were employed to assess the prediction accuracy of the used fouling models. The combined models showed better prediction. The cake-complete model provided far better fits of the transmembrane pressure data, and provided good fits of other individual model predictions regardless of the PAC refreshment ratio. Fourier transform infrared spectroscopy, scanning electron microscopy and energy dispersive X-ray spectroscopy confirmed that the cake layer clogging was the main fouling mechanism followed by complete blockage and standard plugging. The cake-complete model may be used to predict the fouling mechanisms in PAC/MBR systems.


Experimental fire increases soil carbon dioxide efflux in a grassland long-term multifactor global change experiment.
Numerous studies have demonstrated that soil respiration rates increase under experimental warming, although the long-term, multiyear dynamics of this feedback are not well constrained. Less is known about the effects of single, punctuated events in combination with other longer-duration anthropogenic influences on the dynamics of soil carbon (C) loss. In 2012 and 2013, we assessed the effects of decadal-scale anthropogenic global change - warming, increased nitrogen (N) deposition, elevated carbon dioxide (CO2 ), and increased precipitation - on soil respiration rates in an annual-dominated Mediterranean grassland. We also investigated how controlled fire and an artificial wet-up event, in combination with exposure to the longer-duration anthropogenic global change factors, influenced the dynamics of C cycling in this system. Decade-duration surface soil warming (1-2 °C) had no effect on soil respiration rates, while +N addition and elevated CO2 concentrations increased growing-season soil CO2 efflux rates by increasing annual aboveground net primary production (NPP) and belowground fine root production, respectively. Low-intensity experimental fire significantly elevated soil CO2 efflux rates in the next growing season. Based on mixed-effects modeling and structural equation modeling, low-intensity fire increased growing-season soil respiration rates through a combination of three mechanisms: large increases in soil temperature (3-5 °C), significant increases in fine root production, and elevated aboveground NPP. Our study shows that in ecosystems where soil respiration has acclimated to moderate warming, further increases in soil temperature can stimulate greater soil CO2 efflux. We also demonstrate that punctuated short-duration events such as fire can influence soil C dynamics with implications for both the parameterization of earth system models (ESMs) and the implementation of climate change mitigation policies that involve land-sector C accounting.


Adalimumab: A Review in Hidradenitis Suppurativa.
Subcutaneous adalimumab (Humira®) is a tumour necrosis factor-α blocker that is the only approved agent for the treatment of moderate to severe hidradenitis suppurativa (HS) in several countries worldwide. This article reviews the clinical efficacy and safety of subcutaneous adalimumab in patients with moderate to severe HS. In clinical trials (PIONEER I and II), a greater proportion of adalimumab than placebo recipients reached HS clinical response (HiSCR) at week 12. The main secondary endpoints, such as the proportion of patients with an abscess and inflammatory nodule count of ≤2 at week 12, were significantly greater with adalimumab than with placebo in PIONEER II, but not in PIONEER I. In addition, adalimumab showed the potential to reduce the high health-related quality of life burden of HS and increase patient satisfaction. HiSCR rates were generally maintained in the longer term, and the safety profile of adalimumab in patients with moderate to severe HS was consistent with the known safety profile of the drug for other indications, with no new emerging safety signals. Adalimumab is an effective and generally well tolerated treatment for patients with moderate to severe HS, and is the first agent approved for this difficult-to-treat disease.


Myths and Misconceptions: Terminating Treatment.
This ongoing column is dedicated to providing information to our readers on managing legal risks associated with medical practice. We invite questions from our readers. The answers are provided by PRMS, Inc. (www.prms.com), a manager of medical professional liability insurance programs with services that include risk management consultation, education and on-site risk management audits, and other resources offered to health care providers to help improve patient outcomes and reduce professional liability risk. The answers published in this column represent those of only one risk management consulting company. Other risk management consulting companies or insurance carriers might provide different advice, and readers should take this into consideration. The information in this column does not constitute legal advice. For legal advice, contact your personal attorney. Note: The information and recommendations in this article are applicable to physicians and other health care professionals so "clinician" is used to indicate all treatment team members.


Comparative analysis of boar seminal plasma proteome from different freezability ejaculates and identification of Fibronectin 1 as sperm freezability marker.
Variation in boar sperm freezability (i.e. capacity to withstand cryopreservation) between ejaculates is a limitation largely reported in the literature. Prediction of sperm freezability and classification of boar ejaculates into good (GFEs) and poor freezability ejaculates (PFEs) before cryopreservation takes place may increase the use of frozen-thawed spermatozoa. While markers of boar sperm freezability have been found from sperm cell extracts, little attention has been paid to seminal plasma. On this basis, the present study compared the fresh seminal plasma proteome of 9 GFEs and 9 PFEs through two-dimensional difference gel electrophoresis (2D-DIGE) and liquid chromatography mass spectrometry (LC-MS/MS). The ejaculates were previously classified as GFE or PFE upon their sperm viability and progressive motility assessments at 30 and 240 min post thawing. From a total of 51 spots, four were found to significantly (p < 0.05) differ between GFEs and PFEs, and two were identified as fibronectin-1 (FN1) and glutathione peroxidase 5 (GPX5). These two potential markers were further studied by western blot and correlation analysis between protein relative abundances in fresh seminal plasma and regression factors from principal component analyses (PCA) run using post-thawing sperm quality parameters. Results confirmed that FN1 is a reliable marker of boar sperm freezability, because GFEs presented significantly (p < 0.05) higher FN1-amounts than PFEs and FN1 was found to be correlated with the first PCA component at 240 min post thawing. In contrast, GPX5 was not validated as a boar sperm freezability marker. We can thus conclude that levels of FN1 in fresh seminal plasma from boar semen may be used as a sperm freezability marker, thereby facilitating the use of frozen-thawed boar spermatozoa.


Race, socioeconomic status, and return migration to New Orleans after Hurricane Katrina.
Hurricane Katrina struck New Orleans on the 29th of August 2005 and displaced virtually the entire population of the city. Soon after, observers predicted the city would become whiter and wealthier as a result of selective return migration, although challenges related to sampling and data collection in a post-disaster environment have hampered evaluation of these hypotheses. In this article, we investigate return to the city by displaced residents over a period of approximately 14 months following the storm, describing overall return rates and examining differences in return rates by race and socioeconomic status. We use unique data from a representative sample of pre-Katrina New Orleans residents collected in the Displaced New Orleans Residents Pilot Survey. We find that black residents returned to the city at a much slower pace than white residents even after controlling for socioeconomic status and demographic characteristics. However, the racial disparity disappears after controlling for housing damage. We conclude that blacks tended to live in areas that experienced greater flooding and hence suffered more severe housing damage which, in turn, led to their delayed return to the city. The full-scale survey of displaced residents being fielded in 2009-2010 will show whether the repopulation of the city was selective over a longer period.


Serum creatine kinase levels in spinobulbar muscular atrophy and amyotrophic lateral sclerosis.
We compared serum creatine kinase (CK) levels between spinobulbar muscular atrophy (SBMA) and amyotrophic lateral sclerosis (ALS) and reviewed available histochemical studies of frozen sections of muscle biopsies. CK levels and the frequency of patients with elevated CK levels were significantly higher in the SBMA group when compared with the ALS group. CK levels occasionally approached values up to 8 times the upper limit of normal in the SBMA group. In addition to the chronic neurogenic changes in the muscle biopsy, all SBMA patients showed one or more myopathic changes. Increased numbers of markedly hypertrophic fibers were consistently seen in all patients. It is not clear whether the elevated CK level is directly related to the increased number of hypertrophic fibers or to other myopathic features. Based on these findings, we recommend genetic testing for SBMA in cases of male patients with motor neuron disease who present with a significantly elevated serum creatine kinase level, even when other characteristic clinical features of SBMA are absent.


Phosphorylation of αB-crystallin in the myocardium: Analysis of relations with aging and cardiomyopathy.
Phosphorylation is a major post-translational modification of αB-crystallin (CryaB) and determines this protein's chaperone activity, intracellular distribution, translocation, and cytoprotective functions. Phosphorylation of CryaB manifests itself as either beneficial or deleterious consequences depending on the extent of phosphorylation and interaction with the cytoskeleton. Herein, for the first time, we compared the age-related alterations of the expression and phosphorylation (on Ser59: pS59) of CryaB in the myocardium of Wistar and senescence-accelerated OXYS rats. The latters, as we demonstrated here, develop cardiomyopathy by the age of 12 months against the background of hypertension. Rats at the age of 20 days, 3, 12, and 24 months were used. The expression of CryaB mRNA (studied by RT-PCR) and of the CryaB protein (analyzed by western blotting) increased with age in the myocardium of both Wistar and OXYS rats, but only at the age of 24 months did their levels become lower in OXYS rats. Phosphorylation of CryaB increased with age in all rats. There was no association of cardiomyopathy with the pS59-CryaB amount in the detergent-soluble fraction either. Moreover, immunostaining of the myocardium revealed that the amount of pS59-CryaB was greater in OXYS rats than in the control animals. This phenomenon was the result of translocation of pS59-CryaB from the detergent-soluble protein fraction to the detergent-insoluble one. The amount of pS59-CryaB in striated sarcomeres (detergent-insoluble) of the myocardium increased with age in both strains but faster in the myocardium of OXYS rats, and its accumulation preceded the development of cardiomyopathy. Translocation of phosphorylated CryaB to sarcomeres affects functional and structural properties (of cardiomyocytes) that are crucial for contractile function and myofibrillar organization and may be an important component of an endogenous mechanism of aging of the myocardium.


Exercise and Conjugated Linoleic Acid Supplementation Induce Changes in the Composition of Liver Fatty Acids.
Exercise and supplementation with conjugated linoleic acid (CLA) are used to reduce body weight and to improve health. Applied together, they may exert a synergistic effect. However, the effects of exercise and CLA supplementation on liver metabolism are poorly understood. The aim of this study was to examine the influence of exercise and CLA supplementation on fatty acids (FA) composition in mouse liver. We analyzed 44 of known FAs of this organ by gas chromatography-mass spectrometry. Our results demonstrated that exercise contributed to a decrease in odd-chain FA and an increase in n-6 polyunsaturated FA contents. In turn, CLA stimulated an increase in branched-chain FAs and n-3 polyunsaturated FAs. Exercise combined with CLA supplementation caused a substantial increase in branched-chain FA content and a slight increase in n-6 polyunsaturated FAs. The elevated liver content of branched-chain FAs after the exercise combined with CLA supplementation, as well as the increase in n-3 polyunsaturated FAs after CLA may be favorable since both these FA groups were previously shown to produce health benefits. However, the expression pattern of enzymes involved in fatty acid synthesis did not match the changes in FA composition. Thus, the mechanism of exercise- and CLA-induced changes in liver FA profile is yet to be established. Also, the consequences of CLA- and/or exercise-induced changes in the composition of liver FAs need to be elucidated.


The time course of emotional responses to music.
Two empirical studies investigate the time course of emotional responses to music. In the first one, musically trained and untrained listeners were required to listen to 27 musical excerpts and to group those that conveyed a similar emotional meaning. In one condition, the excerpts were 25 seconds long on average. In the other condition, excerpts were as short as 1 second. The groupings were then transformed into a matrix of emotional dissimilarity that was analyzed with multidimensional scaling methods (MDS). We compared the outcome of these analyses for the 25-s and 1-s duration conditions. In the second study, we presented musical excerpts of increasing duration, varying from 250 to 20 seconds. Participants were requested to evaluate on a subjective scale how "moving" each excerpt was. On the basis of the responses given for the longer duration, excerpts were then sorted into two groups: highly moving and weakly (or less) moving. The main purpose of the analysis was to identify the point in time where these two categories of excerpts started to be differentiated by participants. Both studies provide consistent findings that less than 1 s of music is enough to instill elaborated emotional responses in listeners.


DENEDDYLASE1 Protein Counters Automodification of Neddylating Enzymes to Maintain NEDD8 Protein Homeostasis in Arabidopsis.
In eukaryotes, the conjugation of the ubiquitin-like protein NEDD8 onto protein targets is an important post-translational modification. The best understood neddylation targets are the cullins, scaffold subunits of E3 ubiquitin ligases, where neddylation as well as deneddylation, facilitated by the protease activity of the CSN (COP9 signalosome), are required to control ubiquitin ligase assembly, function, and ultimately substrate degradation. Little is known about the role of other deneddylating enzymes besides CSN and the role of neddylation and deneddylation of their substrates. We previously characterized Arabidopsis thaliana mutants with defects in the conserved NEDD8-specific protease DEN1 (DENEDDYLASE1). These mutants display only subtle growth phenotypes despite the strong accumulation of a broad range of neddylated proteins. Specifically, we identified AXR1 (AUXIN-RESISTANT1), a subunit of the heterodimeric NAE (E1 NEDD8-ACTIVATING ENZYME), as highly neddylated in den1 mutants. Here, we examined the mechanism and consequences of AXR1 neddylation in more detail. We find that AXR1 as well as other neddylation enzymes are autoneddylated at multiple lysines. NAE autoneddylation can be linked to reduced NCE (E2 NEDD8-CONJUGATING ENZYME) NEDD8 thioester levels, either by critically reducing the pool of free NEDD8 or by reducing NAE activity. In planta, increasing NEDD8 gene dosage is sufficient to suppress den1 mutant phenotypes. We therefore suggest that DEN1 serves to recover diverted NEDD8 moieties from autoneddylated NAE subunits, and possibly also other neddylated proteins, to maintain NEDD8 pathway activity toward other NEDD8-dependent processes such as cullin E3 ligase regulation.


Reversal of endothelial dysfunction reduces white matter vulnerability in cerebral small vessel disease in rats.
Dementia is a major social and economic problem for our aging population. One of the most common of dementia in the elderly is cerebral small vessel disease (SVD). Magnetic resonance scans of SVD patients typically show white matter abnormalities, but we do not understand the mechanistic pathological link between blood vessels and white matter myelin damage. Hypertension is suggested as the cause of sporadic SVD, but a recent alternative hypothesis invokes dysfunction of the blood-brain barrier as the primary cause. In a rat model of SVD, we show that endothelial cell (EC) dysfunction is the first change in development of the disease. Dysfunctional ECs secrete heat shock protein 90α, which blocks oligodendroglial differentiation, contributing to impaired myelination. Treatment with EC-stabilizing drugs reversed these EC and oligodendroglial pathologies in the rat model. EC and oligodendroglial dysfunction were also observed in humans with early, asymptomatic SVD pathology. We identified a loss-of-function mutation in ATPase11B, which caused the EC dysfunction in the rat SVD model, and a single-nucleotide polymorphism in ATPase11B that was associated with white matter abnormalities in humans with SVD. We show that EC dysfunction is a cause of SVD white matter vulnerability and provide a therapeutic strategy to treat and reverse SVD in the rat model, which may also be of relevance to human SVD.


Design and fabrication of diffractive optical elements by use of gray-scale photolithography.
Diffractive optical elements (DOEs) are key components in the miniaturization of optical systems because of their planarity and extreme thinness. We demonstrate the fabrication of DOEs by use of gray-scale photolithography with a high-energy-beam sensitive glass photomask. We obtained DOE lenses with continuous phase profiles as small as 800 microm in diameter and 5.9 microm in the outermost grating pitch by selecting a suitable optical density for each height level and optimizing the process variables. Microlenses patterned with eight levels and replicated by UV embossing with the polymer master mold showed a diffraction efficiency of 81.5%, which was sufficiently high for the devices to be used as optical pickups. The effects of deviations in diffraction efficiency between the DOE height and profile design were analyzed.


Modulation of serotonergic transmission by eltoprazine in L-DOPA-induced dyskinesia: Behavioral, molecular, and synaptic mechanisms.
L-3,4-dihydroxyphenylalanine (L-DOPA)-induced dyskinesias (LIDs) represent the main side effect of Parkinson's Disease (PD) therapy. Among the various pharmacological targets for novel therapeutic approaches, the serotonergic system represents a promising one. In experimental models of PD and in PD patients the development of abnormal involuntary movements (AIMs) and LIDs, respectively, is accompanied by the impairment of bidirectional synaptic plasticity in key structures such as striatum. Recently, it has been shown that the 5-HT1A/1B receptor agonist, eltoprazine, significantly decreased LIDs in experimental PD and human patients. Despite the fact that several papers have tested this and other serotonergic drugs, nothing is known about the electrophysiological consequences on this combined serotonin receptors modulation at striatal neurons. The present study demonstrates that activation of 5-HT1A/1B receptors reduces AIMs via the restoration of Long-Term Potentiation (LTP) and synaptic depotentiation in a sub-set of striatal spiny projection neurons (SPNs). This recovery is associated with the normalization of D1 receptor-dependent cAMP/PKA and ERK/mTORC signaling pathways, and the recovery of NMDA receptor subunits balance, indicating these events as key elements in AIMs induction. Moreover, we analyzed whether the manipulation of the serotonergic system might affect motor behavior and cognitive performances. We found that a defect in locomotor activity in parkinsonian and L-DOPA-treated rats was reversed by eltoprazine treatment. Conversely, the impairment in the striatal-dependent learning was found exacerbated in L-DOPA-treated rats and eltoprazine failed to recover it.


Genetic disruption of 2-arachidonoylglycerol synthesis reveals a key role for endocannabinoid signaling in anxiety modulation.
Endocannabinoid (eCB) signaling has been heavily implicated in the modulation of anxiety and depressive behaviors and emotional learning. However, the role of the most-abundant endocannabinoid 2-arachidonoylglycerol (2-AG) in the physiological regulation of affective behaviors is not well understood. Here, we show that genetic deletion of the 2-AG synthetic enzyme diacylglycerol lipase α (DAGLα) in mice reduces brain, but not circulating, 2-AG levels. DAGLα deletion also results in anxiety-like and sex-specific anhedonic phenotypes associated with impaired activity-dependent eCB retrograde signaling at amygdala glutamatergic synapses. Importantly, acute pharmacological normalization of 2-AG levels reverses both phenotypes of DAGLα-deficient mice. These data suggest 2-AG deficiency could contribute to the pathogenesis of affective disorders and that pharmacological normalization of 2-AG signaling could represent an approach for the treatment of mood and anxiety disorders.


Immunostimulant adjuvant patch enhances humoral and cellular immune responses to DNA immunization.
The focus of this report is on the development of an improved DNA immunization protocol, which takes advantage of the strengths of DNA immunization, as well as those associated with adjuvant delivered by transcutaneous immunostimulatory (IS) patches. Because transcutaneous delivery of adjuvants to the skin at the vaccination site has been shown to amplify the immune response to protein antigens, we hypothesized that the same IS patch when placed on the skin at the site of DNA injection could further enhance the immune response to a DNA influenza vaccine. We have combined an influenza DNA vaccine, hemagglutinin fused with three copies of complement C3d, to enhance uptake and antigen presentation, with an IS patch containing heat-labile enterotoxin from Escherichia coli. Coadministration of a potent adjuvant in IS patches placed on the skin at the site of DNA vaccination dramatically amplifies anti-influenza antibody immune response. Supplementing DNA vaccines with IS patches may be a particularly valuable strategy because DNA vaccines can be rapidly modified in response to mutations in pathogens, and individuals with compromised immune systems such as transplant patients and the elderly will benefit from the enhanced antibody response induced by the IS patches.


Early detection of schizophrenia.
This article will evaluate the rationale and feasibility of detecting psychosis and schizophrenia earlier than is currently the case. Schizophrenia incidence may vary more than has been believed previously. Early detection studies fall into two groups. Firstly, operational criteria now exist for prodromal or at risk mental states which predict transition to psychosis of 20-40% over 1 year. The first randomized trials of antipsychotic drug and psychological interventions aimed at reducing this transition rate have shown promising results. Secondly, duration of untreated psychosis in the first episode seems genuinely to be associated with clinical outcome but how much of the association is truly causal remains disputed. Shortening duration of untreated psychosis by early detection of full psychosis appears feasible but its benefits are not yet unequivocally demonstrated.


Validity of using tri-axial accelerometers to measure human movement - Part I: Posture and movement detection.
A robust method for identifying movement in the free-living environment is needed to objectively measure physical activity. The purpose of this study was to validate the identification of postural orientation and movement from acceleration data against visual inspection from video recordings. Using tri-axial accelerometers placed on the waist and thigh, static orientations of standing, sitting, and lying down, as well as dynamic movements of walking, jogging and transitions between postures were identified. Additionally, subjects walked and jogged at self-selected slow, comfortable, and fast speeds. Identification of tasks was performed using a combination of the signal magnitude area, continuous wavelet transforms and accelerometer orientations. Twelve healthy adults were studied in the laboratory, with two investigators identifying tasks during each second of video observation. The intraclass correlation coefficients for inter-rater reliability were greater than 0.95 for all activities except for transitions. Results demonstrated high validity, with sensitivity and positive predictive values of greater than 85% for sitting and lying, with walking and jogging identified at greater than 90%. The greatest disagreement in identification accuracy between the algorithm and video occurred when subjects were asked to fidget while standing or sitting. During variable speed tasks, gait was correctly identified for speeds between 0.1m/s and 4.8m/s. This study included a range of walking speeds and natural movements such as fidgeting during static postures, demonstrating that accelerometer data can be used to identify orientation and movement among the general population.