Age-dependent physiological changes in the histoarchitecture of the articular cartilage of the rabbit mandibular condyle: a morphological and morphometric study.
Mandibular condyle articular cartilage participates in condylar postnatal growth and is responsible for adaptations to anatomical and/or biomechanical alterations throughout life. In a preliminary study in rabbits, differences were observed in the thickness of the layers of articular cartilage in control animals at 5 and 6 months (generally considered adults for this purpose). This study aimed to describe sagittally sectioned condylar cartilages stained with Picrosirius-hematoxylin in rabbits at 40 days and 5, 6, 8, 13, and 18 months to determine when histological maturity is reached. At 40 days, 5 layers were seen: fibrous, proliferative, transition, maturation, and hypertrophic. Older animals (5-18 months) lacked the transition layer. Fibrous, proliferative, and hypertrophic regions were considered for morphometric analysis. The thickness of the fibrous region did not change during the analyzed period (p = 0.1899). When proliferative and hypertrophic regions and the total thickness of the cartilage were compared, a difference was detected (p < 0.001). The thickness of the proliferative region was greatest at 40 days and decreased at 5 months; however, it increased at 6 months, when it was significantly thicker than at 5, 8, 13, and 18 months. Both the hypertrophic region and the total thickness were thickest at 40 days, intermediate at 5, 6, and 8 months, and thinnest at 13 and 18 months. In summary, our data suggest a physiological period of increased cartilage growth at 6 months. Additionally, rabbits at this age should be avoided in experiments involving condylar cartilage. Finally, 13-month-old rabbits have reached histological maturity of the condylar cartilage.


Effects of a three-week vocal exercise program using the Finnish Kuukka exercises on the speaking voice of Norwegian broadcast journalism students.
Nine broadcast journalism students attended 10 hours in Kuukka vocal exercises, which aims at producing a ringing vocal quality. Nine control subjects received no training. A text was read at habitual loudness before and after the course. Five speech specialists evaluated the text samples for perceptual voice quality and analyzed mean fundamental frequency (F0), equivalent sound level (Leq), and long-term average spectrum (LTAS). For the Training Group, voice quality improved and correlated negatively with firmness and timbre (less firm and darker qualities being considered more desirable), and F0 increased slightly. Leq increased significantly in both groups. The results show positive and perceivable differences after the course. However, the aimed ring was not reached, may be due to too short time.


Olmesartan prevents cardiovascular injury and hepatic steatosis in obesity and diabetes, accompanied by apoptosis signal regulating kinase-1 inhibition.
Dietary obesity is associated with type 2 diabetes and cardiovascular diseases, although the underlying mechanism is unknown. This study was undertaken to elucidate the role of angiotensin II and apoptosis signal regulating kinase-1 (ASK1) in obesity/diabetes-associated cardiovascular complications and hepatic steatosis. Mice fed a high-fat diet were treated with olmesartan, an angiotensin II type 1 receptor blocker, to elucidate the role of angiotensin II in diabetic mice. Treatment of mice fed a high-fat diet with olmesartan markedly suppressed cardiac inflammation and fibrosis, as well as vascular endothelial dysfunction and remodeling, induced by obesity/diabetes. Moreover, olmesartan suppressed the disruption of the vascular endothelial NO synthase dimer in diabetic mice. Olmesartan also significantly prevented hepatic steatosis and fibrosis in diabetic mice. These beneficial effects of olmesartan on diabetic mice were associated with the attenuation of ASK1 activation in these mice. ASK1-deficient mice and wild-type mice were compared, regarding the effects of a high-fat diet. A comparison between ASK1-deficient and wild-type mice showed that ASK1 deficiency attenuated cardiac inflammation and fibrosis, as well as vascular endothelial dysfunction and remodeling induced by obesity/diabetes. The amelioration of vascular endothelial impairment by ASK1 deficiency was attributed to the prevention of endothelial NO synthase dimer disruption. ASK1 deficiency also significantly lessened hepatic steatosis in diabetic mice. In conclusion, our work provided the evidence that ASK1 is significantly activated in diet-induced diabetic mice and contributes to cardiovascular diseases and hepatic steatosis in diabetic mice. Moreover, the beneficial effects of angiotensin II inhibition on dietary diabetic mice seem to be mediated by the inhibition of ASK1 activation.


Digital PET vs Analog PET: Clinical Implications?
Positron emission tomography (PET) is a functional imaging technique introduced in 1970s. Over the years, PET was used alone but is in 2000 when the first hybrid PET/CT device was clinically introduced. Since then, PET has continuously been marked by technological developments, being the most recent one the introduction of silicon photomultipliers (SiPMs) as an alternative to standard photomultiplier tubes used in analog PET/CT systems. SiPMs, the basis for the so called digital PET/CT systems, are smaller than standard photomultiplier tubes (enabling higher spatial resolution) and provide up to 100% coverage of the crystal area, as well as high sensitivity, low noise, and fast timing resolution. SiPMs in combination with optimized acquisition and reconstruction parameters improve the localization of the annihilation events, provide high definition PET images, and offer higher sensitivity and higher diagnostic performance. This article summarizes the evidence about the superior performance of the state of the art digital PET and highlights its potential clinical implications. Digital PET opens new perspectives in the quantification and characterization of small lesions, which are mostly undetectable using analog PET systems, potentially changing patient management and improving outcomes in oncological and non-oncological diseases. Moreover, digital PET offers the possibility to reduce radiation dose and scan times which may facilitate the implementation of PET to address unmet clinical needs.


Regulation of carotenoid formation during tomato fruit ripening and development.
Carotenoid biosynthesis and its regulation during tomato fruit development and ripening is a complex process that occurs alongside the differentiation of chloroplasts into chromoplasts and changes to the organoleptic properties of the fruit. Unusually for plants, the ripe tomato fruit accumulates large amounts of lycopene, as the pattern of gene expression found in green fruit changes during fruit ripening. Although the control of gene expression is thought to be the main regulatory mechanism for these alterations in carotenoids, post-transcriptional regulation has also been reported, including feedback inhibition. The use of genetic manipulation of carotenogenesis in tomato has been used primarily for biotechnological reasons, but it has also facilitated investigations into these regulatory mechanisms, as well as into the effects of such perturbations on other isoprenoids such as gibberellins, tocopherols and sterols.


Complications in Placement of a Glaucoma Drainage Device in a Patient With CREST Syndrome.
This case describes difficulty with conjunctival closure in an 80-year-old woman with CREST syndrome, a subset of systemic scleroderma. The patient underwent uneventful glaucoma drainage device implantation, but friability of the conjunctiva was noted during closure. Amniotic membrane was used to ensure secure closure, and the patient had a successful outcome. There is a paucity of existing studies on patients with CREST syndrome and their ocular findings. This report sheds light on ocular findings in this autoimmune disease and the necessity for alternative closure methods in special cases.


Marrow storage techniques: a clinical comparison of refrigeration versus cryopreservation.
Fifty-three patients were evaluated for a comparison of the efficacy, safety, and cost efficiency of bone marrow (BM) transplanted after either refrigeration or cryopreservation. Thirty-eight patients had BM stored at 4 degrees C for an average of 3 days and 15 patients had cryopreserved BM stored for an average of 56 days. The average number of cells harvested was 3.8 x 10(8)/kg. The time to WBC recovery greater than 1 x 10(9)/l was 17 days refrigerated and 23 days for cryopreserved BM. The time to platelet recovery greater than 20 x 10(9)/l was 24 days for refrigeration storage and 51 days for cryopreserved BM. Four of 38 patients with refrigerated vs. 4/15 patients with cryopreserved BM experienced delayed engraftment (p less than 0.05). Refrigeration storage requires no special equipment, is cheaper than and presents a safe and viable alternative to cryopreserved BM in reconstituting hemopoiesis following high-dose chemo-radiotherapy.


Alpha2 nicotine receptors function as a molecular switch to continuously excite a subset of interneurons in rat hippocampal circuits.
Rapid activation of nicotinic acetylcholine receptors (nAChRs) at various anatomical and cellular locations in the hippocampus differentially modulates the operation of hippocampal circuits. However, it is largely unknown how the continued presence of nicotine affects the normal operation of hippocampal circuits. Here, we used single and dual whole-cell recordings to address this question. We found that horizontally oriented interneurons in the stratum oriens/alveus continuously discharged action potentials in the presence of nicotine. In these interneurons, bath application of nicotine produced slow inward currents that were well maintained and inhibited by the non-alpha 7 antagonist dihydro-beta-erythroidine. Single-cell reverse transcription-polymerase chain reaction analysis showed that nicotine-responding interneurons were consistently positive for the alpha2 subunit mRNA. These observations suggest that in the presence of nicotine, a subset of interneurons in the stratum oriens/alveus are continuously excited due to the sustained activation of alpha2* nAChRs. These interneurons were synaptically connected to pyramidal cells, and nicotine increased inhibitory baseline currents at the synapses and suppressed phasic inhibition at the same synapses. Nicotine-induced inhibitory activity increased background noise and masked small phasic inhibition in pyramidal cells, originating from other interneurons in the stratum radiatum. Thus, the continued presence of nicotine alters the normal operation of hippocampal circuits by gating inhibitory circuits through activating a non-desensitizing alpha2 nAChR subtype on a distinct population of interneurons.


Parameterization of the ReaxFF reactive force field for a proline-catalyzed aldol reaction.
A parameterization of the ReaxFF reactive FF is performed using a Monte Carlo Simulated Annealing procedure for the modeling of a proline-catalyzed aldol reaction. Emphasis is put on the accurate reproduction of the relative stabilities of several key intermediates of the reaction, as well as, on the description of the reaction path bridging these intermediates based on quantum mechanical calculations. Our training sets include new criteria based on geometry optimizations and short Molecular Dynamics simulations to ensure that the trained ReaxFF potentials adequately predict the structures of all key intermediates. The transferability of the sets of parameters obtained is assessed for various steps of the considered aldol reaction, as well as for different substrates, catalysts, and reagents. This works indeed highlights the challenge of reaching transferable parameters for several reaction steps. © 2016 Wiley Periodicals, Inc.


Orphan endogenous lipids and orphan GPCRs: a good match.
A large and growing family of over 70 endogenous lipids of the basic structure N-acyl amide has been identified during the last 10 years. Only a few of these lipids have been characterized for biological activity, however, those that have shown a wide-range of activity may act at G-protein coupled receptors (GPCRs). Like orphan GPCRs that are identified as being in the genome and expressed in tissue, the majority of these endogenous lipids many produced throughout the body, some predominately in nervous tissue, remain orphaned. Here, we give a brief history of these orphan lipids and highlight the activity of N-arachidonoyl glycine, and farnesyl pyrophosphate at the orphan receptors GPR18 and GPR92, respectively, as well as summarizing the biological and pharmacological data for the recently identified N-palmitoyl glycine that suggests activity at a novel GPCR. Working to deorphanize both lipids and GPCRs together provides a unique opportunity for a greater understanding of cellular signaling and a challenge to find them all a home.


Crystallographic study of mechanism of ribonuclease T1-catalysed specific RNA hydrolysis.
Ribonuclease T1 (RNase T1) cleaves the phosphodiester bond of RNA specifically at the 3'-end of guanosine. 2'-guanosinemonophosphate (2'-GMP) acts as inhibitor for this reaction and was cocrystallized with RNase T1. X-Ray analysis provided insight in the geometry of the active site and in the parts of the enzyme involved in the recognition of guanosine. RNase T1 is globular in shape and consists of a 4.5 turns alpha-helix lying "below" a four-stranded antiparallel beta-sheet containing recognition center as well as active site. The latter is indicated by the position of phosphate and sugar residues of 2'-GMP and shows that Glu58, His92 and Arg77 are active in phosphodiester hydrolysis. Guanine is recognized by a stretch of protein from Tyr42 to Tyr45. Residues involved in recognition are peptide NH and C = O, guanine O6 and N1H which form hydrogen bonds and a stacking interaction of Tyr45 on guanine. Although, on a theoretical basis, many specific amino acid-guanine interactions are possible, none is employed in the RNase T1.guanine recognition.


Impact Of Hypertension versus Diabetes on Cardiovascular and All-cause Mortality in Iranian Older Adults: Results of 14 Years of Follow-up.
To evaluate the joint effect of hypertension (HTN) and diabetes (DM) on coronary heart disease (CHD), and stroke event, all-cause, and cardiovascular disease (CVD) mortality in Middle Eastern older adults, 2747 people (1436 women) aged ≥ 50 years, free of CVD at baseline, were categorized into four groups (HTN-/DM-, HTN+/DM-, HTN-/DM+, HTN+/DM+). Multivariate Cox proportional hazard models were run for different outcomes. To compare the impact of HTN versus DM, HTN+/DM- was considered as reference. In a median of 13.9 years, incidence rate of CHD, and stroke event, all-cause and CVD mortality in total population were 19.0, 4.7, 13.5, and 6.4 per 1000 person-years, respectively. The multivariate sex-adjusted hazard ratios (HRs) of HTN-/DM+ for CHD, stroke, all-cause mortality and CVD mortality were 1.19 (confidence interval (CI): 0.9-1.57), 1.07 (CI: 0.63-1.82), 1.62 (CI: 1.2-2.18), and 1.28 (CI: 0.83-1.97); the corresponding HRs for HTN+/DM+ were 1.96 (CI: 1.57-2.46), 1.66 (CI: 1.1-2.52), 2.32 (CI: 1.8-2.98), and 2.6 (CI: 1.85-3.65) respectively. The associations between HTN/DM status with stroke incidence and all-cause mortality were stronger among men than in women (P for interaction <0.05). Compared to HTN+/DM-, HTN-/DM+ increases all-cause mortality by 62%, however, they are not considerably different regarding CHD, stroke incidence and CVD mortality.


Spores of two probiotic Bacillus species enhance cellular immunity in BALB/C mice.
Previous studies found that Bacillus subtilis BS02 and B. subtilis subsp. natto BS04 isolated in our laboratory could activate the immune response of murine macrophages in vitro. This study aims to investigate the effects of dietary supplementation with Bacillus species spores on the systemic cellular immune response in BALB/C mice. Results showed that both B. subtilis BS02 and B. subtilis natto BS04 enhanced the phagocytic function of the mononuclear phagocyte system (MPS) and the cytotoxicity of natural killer (NK) cells. In addition, B. subtilis BS02 could increase the respiratory burst activity of blood phagocytes. Furthermore, B. subtilis BS02 and B. subtilis natto BS04 increased the percentage of gamma-interferon-producing CD4+ cells and CD8+ T-cells, but only BS04 increased the percentage of CD3+ cells and CD3+ CD4+ cells in splenocytes. However, there were no effects on other subsets of splenic lymphocytes and mitogen-induced splenic lymphocyte proliferation. All data suggested that oral administration of B. subtilis BS02 or B. subtilis natto BS04 could significantly enhance cellular immunity in BALB/C mice by increasing phagocytic activity of MPS and cytotoxic activity of NK cells in a strain-specific manner.


A fast method based on NESTA to accurately reconstruct CT image from highly undersampled projection measurements.
The CT image reconstruction algorithm based compressed sensing (CS) can be formulated as an optimization problem that minimizes the total-variation (TV) term constrained by the data fidelity and image nonnegativity. There are a lot of solutions to this problem, but the computational efficiency and reconstructed image quality of these methods still need to be improved. To investigate a faster and more accurate mathematical algorithm to settle TV term minimization problem of CT image reconstruction. A Nesterov's algorithm (NESTA) is a fast and accurate algorithm for solving TV minimization problem, which can be ascribed to the use of most notably Nesterov's smoothing technique and a subtle averaging of sequences of iterates, which has been shown to improve the convergence properties of standard gradient-descent algorithms. In order to demonstrate the superior performance of NESTA on computational efficiency and image quality, a comparison with Simultaneous Algebraic Reconstruction Technique-TV (SART-TV) and Split-Bregman (SpBr) algorithm is made using a digital phantom study and two physical phantom studies from highly undersampled projection measurements. With only 25% of conventional full-scan dose and, NESTA method reduces the average CT number error from 51.76HU to 9.98HU on Shepp-Logan phantom and reduces the average CT number error from 50.13HU to 0.32HU on Catphan 600 phantom. On an anthropomorphic head phantom, the average CT number error is reduced from 84.21HU to 1.01HU in the central uniform area. To the best of our knowledge this is the first work that apply the NESTA method into CT reconstruction based CS. Research shows that this method is of great potential, further studies and optimization are necessary.


Pubertal and adult Leydig cell function in Mullerian inhibiting substance-deficient mice.
Mullerian inhibiting substance (MIS) causes Mullerian duct regression during sexual differentiation and regulates postnatal Leydig cell development. MIS knockout (MIS-KO) mice with targeted deletions of MIS develop Leydig cell hyperplasia, but their circulating androgen concentrations are reportedly unaltered. We compared reproductive hormone profiles, androgen biosynthesis, and the expression of key steroidogenic and metabolic enzymes in MIS-KO and wild-type (WT) mice at puberty (36 d) and sexual maturity (60 d). In pubertal animals, basal testosterone and LH concentrations in plasma were lower in MIS-KO than WT mice, whereas human chorionic gonadotropin-stimulated testosterone concentrations were similar. In adults, basal LH, and both basal and human chorionic gonadotropin (hCG)-stimulated testosterone concentrations were similar. Purified Leydig cells from pubertal MIS-KO mice had lower testosterone but higher androstanediol and androstenedione production rates. In contrast, testosterone, androstanediol, and androstenedione production rates were all lower in adult MIS-KO Leydig cells. Steroidogenic acute regulatory protein expression was lower in pubertal MIS-KO mice compared with WT, whereas 17beta-hydroxy-steroid dehydrogenase and 5alpha-reductase were greater, and P450c17 and P450scc were similar. The expression of steroidogenic acute regulatory protein and 17beta-hydroxysteroid dehydrogenase was lower in adult MIS-KO mice, whereas that of 5alpha-reductase, P450c17, and P450scc was similar. Collectively, these results suggest that in the absence of MIS, Leydig cells remain less differentiated, causing an altered intratesticular androgen milieu that may contribute to the infertility of MIS-KO mice. In immature mice, this deficit in steroidogenic capacity appears to be mediated by a direct loss of MIS action in Leydig cells as well as by indirect effects via the hypothalamic-pituitary-gonadal axis.


Detecting dynamical changes in vital signs using switching Kalman filter.
Vital signs contain valuable information about patients' health status during their stay in general wards, when the deterioration process begins. The use of methods to predict and detect regime changes such as switching models can help to understand how vital sign dynamics are altered in disease conditions. However, time series of vital signs are remarkably non-stationary in these scenarios. The objective of this study is to quantify the potential bias of switching models in the presence of non-stationarities, when the inputs are spectral, symbolic and entropy indices. To distinguish stationary from non-stationary periods, a test was used to verify the stability of the mean and variance over short periods. Then, we compared the results from a switching Kalman filter (SKF) model trained using indices obtained over stationary periods with a model trained solely over non-stationary periods. It was observed that indices measured over stationary and non-stationary periods were significantly different. The results of switching models were highly dependent on the indices that were used as inputs. The multi-scale entropy (MSE) approach presented the highest correlation values between non-stationary and stationary switches, an average correlation coefficient of 38%.


Prescribing trends in asthma: a longitudinal observational study.
Inhaled corticosteroids (ICS) are effective treatment for childhood asthma. Cross-sectional studies indicate that some asthmatic children are treated with excessively high doses of ICS and are at risk of serious adverse effects. To describe longitudinal trends in asthma prescribing for children, with particular reference to very-high-dose (unlicensed) ICS prescribing. Retrospective, cross-sectional, observational study of general practitioner prescribing for asthma drugs in children aged under 12 years with a recorded asthma diagnosis between 1992 and 2004 using the General Practice Research Database (GPRD). Data were available for an average of 357,956 children per year. The percentage of children prescribed ICS increased from 2.7 in 1992 to 7.0 in 1997 and 1998 and then fell to 3.3 in 2004. In children under 5 years with asthma, very-high-dose ICS prescriptions (>400 microg/day) fell from 10.6% of all ICS prescriptions in 1992 to 4.5% by 2004. In contrast, very-high-dose ICS prescriptions (>800 microg/day) for asthmatic children aged 5-11 years rose from 1.1% in 1992 to 4.6% in 2004. Oral corticosteroid prescribing in under 5-year-olds who had been prescribed ICS fell from 37.1% in 1992 to 21.7% 1999 and remained constant thereafter; the respective percentages for those aged 5-11 years olds were 20.1 and 12.4. Trends for corticosteroid prescribing in childhood asthma changed dramatically between 1992 and 2004. There are several plausible reasons for this.


Distinct dimer interaction and regulation in nitric-oxide synthase types I, II, and III.
Homodimer formation activates all nitric-oxide synthases (NOSs). It involves the interaction between two oxygenase domains (NOSoxy) that each bind heme and (6R)-tetrahydrobiopterin (H4B) and catalyze NO synthesis from L-Arg. Here we compared three NOSoxy isozymes regarding dimer strength, interface composition, and the ability of L-Arg and H4B to stabilize the dimer, promote its formation, and protect it from proteolysis. Urea dissociation studies indicated that the relative dimer strengths were NOSIIIoxy >> NOSIoxy > NOSIIoxy (endothelial NOSoxy (eNOSoxy) >> neuronal NOSOXY (nNOSoxy) > inducible NOSoxy (iNOSoxy)). Dimer strengths of the full-length NOSs had the same rank order as judged by their urea-induced loss of NO synthesis activity. NOSoxy dimers containing L-Arg plus H4B exhibited the greatest resistance to urea-induced dissociation followed by those containing either molecule and then by those containing neither. Analysis of crystallographic structures of eNOSoxy and iNOSoxy dimers showed more intersubunit contacts and buried surface area in the dimer interface of eNOSoxy than iNOSoxy, thus revealing a potential basis for their different stabilities. L-Arg plus H4B promoted dimerization of urea-generated iNOSoxy and nNOSoxy monomers, which otherwise was minimal in their absence, and also protected both dimers against trypsin proteolysis. In these respects, L-Arg alone was more effective than H4B alone for nNOSoxy, whereas for iNOSoxy the converse was true. The eNOSoxy dimer was insensitive to proteolysis under all conditions. Our results indicate that the three NOS isozymes, despite their general structural similarity, differ markedly in their strengths, interfaces, and in how L-Arg and H4B influence their formation and stability. These distinguishing features may provide a basis for selective control and likely help to regulate each NOS in its particular biologic milieu.


Skeletal Muscle Hypertrophy, Insulin-like Growth Factor 1, Myostatin and Follistatin in Healthy and Sarcopenic Elderly Men: The Effect of Whole-body Resistance Training.
Sarcopenia describes the inevitable deterioration in muscle mass and strength that accompanies biological aging. The purpose of this study was to investigate the effects of resistance training (RT) on quadriceps hypertrophy and related biochemistry in sarcopenic and healthy elderly men. A total of 31 elderly men (55-70 years old) were classified as sarcopenic and nonsarcopenic and were divided into two groups. Both groups participated in a progressive RT program for 8 weeks. Data indicated that the strength in the sarcopenic group increased more than the healthy group (P < 0.05). Quadriceps cross-sectional area also increased more in the healthy group (P < 0.05). Myostatin concentration decreased in both groups after training (P < 0.05). Follistatin and testosterone increased in the healthy group; in contrast, only testosterone increased in the sarcopenic group after training (P < 0.05). The findings from this study suggest that RT improves muscle cross-sectional area and biomarker-related muscle loss in both healthy and sarcopenic elderly men. The findings also demonstrate that growth factor profiles at baseline and changes in testosterone levels play an important role in muscle hypertrophy observed in both groups.


Clinical implication of long non-coding RNA NEAT1 expression in hepatocellular carcinoma patients.
Hepatocellular carcinoma (HCC), a primary malignancy of the liver, is associated with high mortality rate and poor prognosis. Emerging evidence showed that novel biomarkers are required toward a better understanding of the biological mechanisms of HCC. NEAT1 (nuclear paraspeckle assembly transcript 1, also known as MENε/β), a novel long non-coding RNA (lncRNA), serves as a crucial regulator in several cancers. However, the correlation between NEAT1 expression with tumorigenesis and metastasis in HCC tissues remains out of the question so far. In the current study, the aim was to evaluate the potential role of NEAT1 expression in HCC tissues and its relationship with clinicopathological parameters. The expression of NEAT1 was detected by qRT-PCR, in 95 cases of adjacent non-cancerous liver and their paired HCC tissues, respectively. The associations of NEAT1 with clinicopathological features and other biological factors were further analyzed. Our results revealed that NEAT1 appeared to have higher expression in the HCC tissues, compared with the adjacent non-cancerous liver tissues. High levels of NEAT1 promoted the clinical features of HCC, including the number of tumor nodes, metastasis, clinical TNM stage, the status of portal vein tumor embolus, vaso-invasion and the infiltration of tumor cells. Additionally, high NEAT1 expression levels were significantly associated with the expression level of MDTH, NM23 and MALAT1. Our study demonstrates that NEAT1 acts as a pivotal player in tumorigenesis and metastasis of hepatocellular carcinoma.


A permutation-based approach to inference for weighted sum regression with correlated chemical mixtures.
There is a growing demand for methods to determine the effects that chemical mixtures have on human health. One statistical challenge is identifying true "bad actors" from a mixture of highly correlated predictors, a setting in which standard approaches such as linear regression become highly variable. Weighted Quantile Sum regression has been proposed to address this problem, through a two-step process where mixture component weights are estimated using bootstrap aggregation in a training dataset and inference on the overall mixture effect occurs in a held-out test set. Weighted Quantile Sum regression is popular in applied papers, but the reliance on data splitting is suboptimal, and analysts who use the same data for both steps risk inflating the Type I error rate. We therefore propose a modification of Weighted Quantile Sum regression that uses a permutation test for inference, which allows for weight estimation using the entire dataset and preserves Type I error. To minimize computational burden, we propose replacing the bootstrap with L1 or L2 penalization and describe how to choose the appropriate penalty given expert knowledge about a mixture of interest. We apply our method to a national pregnancy cohort study of prenatal phthalate exposure and child health outcomes.


Development of the insect pathogenic alga Helicosporidium.
This study examined the morphogenesis and replication dynamics of the different life stages (cysts, filamentous cells, vegetative cells) of Helicosporidium sp., a non-photosynthetic, entomopathogenic alga. The isolate (SjHe) used originated from an infected black fly larva. Filamentous cell transformation into vegetative cells and autosporulation during vegetative cell replication were observed under controlled in vitro conditions. The transformation process was initiated by a partial swelling of the filamentous cell along with the reorganization of the nuclear material. Two subsequent nuclear and cell divisions resulted in the release of 4 rod-shaped daughter cells, which divided into oval to spherical vegetative cells. These underwent several cycles of autosporogenic cell division. Multiple-passaged vegetative cell cultures formed non-motile, adherent cell clusters (palmelloid colonies). Vegetative replication dynamics were also observed in 2 experimental noctuid hosts, Spodoptera exigua and Helicoverpa zea. The average density of helicosporidial cells produced per microliter hemolymph exceeded cell concentrations obtained in vitro by 15- and 46-fold in S. exigua and H. zea, respectively. Cyst morphogenesis was only observed in the hemolymph, whereas no cysts differentiated at various in vitro conditions.


Visualizing Assembly Dynamics of All-Liquid 3D Architectures.
To better exploit all-liquid 3D architectures, it is essential to understand dynamic processes that occur during printing one liquid in a second immiscible liquid. Here, the interfacial assembly and transition of 5,10,15,20-tetrakis(4-sulfonatophenyl) porphyrin (H6 TPPS) over time provides an opportunity to monitor the interfacial behavior of nanoparticle surfactants (NPSs) during all-liquid printing. The formation of J-aggregates of H4 TPPS2- at the interface and the interfacial conversion of the J-aggregates of H4 TPPS2- to H-aggregates of H2 TPPS4- is demonstrated by interfacial rheology and in situ atomic force microscopy. Equally important are the chromogenic changes that are characteristic of the state of aggregation, where J-aggregates are green in color and H-aggregates are red in color. In all-liquid 3D printed structures, the conversion in the aggregate state with time is reflected in a spatially varying change in the color, providing a simple, direct means of assessing the aggregation state of the molecules and the mechanical properties of the assemblies, linking a macroscopic observable (color) to mechanical properties.


Height and incidence of cardiovascular disease in male physicians.
An inverse association between height and risk of coronary heart disease (CHD) has been reported in several case-control and cohort studies, but the reasons for the association remain uncertain. We evaluated this association among 22,071 male physicians, a population homogeneous for high educational attainment and socioeconomic status in adulthood. The study population was comprised of participants in the Physicians' Health Study, a randomized, double-blind, placebo-controlled trial of low-dose aspirin and beta-carotene in the primary prevention of cardiovascular disease and cancer among US male physicians, aged 40 to 84 years, in 1982. Participants were classified into five height categories at study entry, from shortest to tallest, and were followed an average of 60.2 months to determine the incidence of myocardial infarction (MI), stroke, and death from cardiovascular disease. Men in the tallest (> or = 73 in. or 185.4 cm) compared with the shortest (< or = 67 in. or 170.2 cm) height category had a 35% lower risk of MI (relative risk, 0.65; 95% confidence interval, 0.44 to 0.99; P = .04), after adjusting for known cardiovascular risk factors. Further, a marginally significant inverse trend (P trend = .05) across the height categories was observed. Although the relationship was not strictly linear, for every inch of added height, there was an approximate 2% to 3% decline in risk of MI. In contrast, men in the tallest compared with the shortest height category had only small and nonsignificant decreases in risk of stroke and cardiovascular death. While no significant trend in risks of these end points across the height categories was observed, the numbers of events for these end points were far less than for MI, and thus the confidence intervals were wide. These data indicate that height is inversely associated with subsequent risk of MI. At this time, a few mechanisms are plausible, but none are convincing. Other epidemiological and basic research efforts are needed to explore a variety of physiological correlates of height that may be responsible for mediating the height-MI association. In the meantime, while height is not modifiable, it is easy to measure and may be useful to evaluate CHD disease risk profiles and target lifestyle interventions.


Hypertensive retinopathy in a neonate.
A 23-day-old female child diagnosed as having systemic hypertensive emergency was referred for retinal screening. The fundus examination showed bilateral intraretinal haemorrhages and hard exudates especially at the macula. Venous looping was noted. The ocular features were suggestive of hypertensive retinopathy. Control of systemic hypertension was advised and was managed conservatively with close follow-up. Widefield fundus photography was done at presentation and follow-up to document the change in retinopathy with control of hypertension.The haemorrhages and exudates resolved on follow-up but significant retinal pigment epithelium changes with beaten bronze appearance were noted at the area of previous oedema. Presence of hypertensive retinopathy in a neonate is rare and has long-term effects on visual development. This report describes the course of hypertensive retinopathy in a neonate.


Implementation of an operator intervention system for remote control of the RIKEN 28 GHz superconducting electron cyclotron resonance ion source.
The control system for the RIKEN 28 GHz superconducting electron cyclotron resonance ion source (28 GHz SC-ECRIS) consists of a distributed control system based on the experimental physics and industrial control system. To maintain the beam quality for the long beam-service time at the radioactive isotope beam factory, beam tuning to prevent subtle changes in the 28 GHz SC-ECRIS conditions is required. Once this is achieved, it should then be possible to check conditions and operate the ion source at any time. We have designed a web-based operational interface to remotely control the ion source, but for access and control from several locations, suitable access security, policies, and methods are required. We thus implemented an operator intervention system that makes it possible to safely access the network externally with the permission of on-site accelerator operators in the control room.


Predicting the Survival Time for Bladder Cancer Using an Additive Hazards Model in Microarray Data.
One substantial part of microarray studies is to predict patients' survival based on their gene expression profile. Variable selection techniques are powerful tools to handle high dimensionality in analysis of microarray data. However, these techniques have not been investigated in competing risks setting. This study aimed to investigate the performance of four sparse variable selection methods in estimating the survival time. The data included 1381 gene expression measurements and clinical information from 301 patients with bladder cancer operated in the years 1987 to 2000 in hospitals in Denmark, Sweden, Spain, France, and England. Four methods of the least absolute shrinkage and selection operator, smoothly clipped absolute deviation, the smooth integration of counting and absolute deviation and elastic net were utilized for simultaneous variable selection and estimation under an additive hazards model. The criteria of area under ROC curve, Brier score and c-index were used to compare the methods. The median follow-up time for all patients was 47 months. The elastic net approach was indicated to outperform other methods. The elastic net had the lowest integrated Brier score (0.137±0.07) and the greatest median of the over-time AUC and C-index (0.803±0.06 and 0.779±0.13, respectively). Five out of 19 selected genes by the elastic net were significant (P<0.05) under an additive hazards model. It was indicated that the expression of RTN4, SON, IGF1R and CDC20 decrease the survival time, while the expression of SMARCAD1 increase it. The elastic net had higher capability than the other methods for the prediction of survival time in patients with bladder cancer in the presence of competing risks base on additive hazards model.


Ileal and colonic propulsive behavior: contribution of enteric neural circuits.
Experiments were conducted in vitro to determine the extent to which the intrinsic propulsive behavior of cat terminal ileum and colon is controlled by the enteric nervous system (ENS) and the myogenic control system. Oral and aboral ends of terminal ileal and colonic segments, 17 cm in length, were connected to a propulsion evaluation system that imposed input-output conditions of constant capacitance and negligible resistance. To determine the effect of inhibiting one or more components within the ENS on spontaneously occurring patterns of propulsive behavior, segments were arterially perfused with tetrodotoxin (3.1 X 10(-7) M), atropine sulfate (10(-6) M), tubocurarine chloride (1.5 X 10(-6) M), hexamethonium chloride (2.8 X 10(-5) M), or methysergide (2.8 X 10(-5) M). Results indicated that when cat ileal and colonic segments are evaluated under conditions requiring significant amounts of hydrostatic work to effect fluid ejection 1) the unmodulated myogenic control system is unable to induce significant amounts of fluid expulsion, 2) cholinergic excitatory motor neurons of the ENS are required for the generation of the intrinsic patterns of propulsive behavior normally observed to expel fluid, and 3) the spontaneous occurrence of intrinsic propulsive patterns requires the operation of ENS integrative mechanisms utilizing nicotinic cholinergic and serotonergic synapses.


Immunomodulatory effects of dsRNA and its potential as vaccine adjuvant.
dsRNA can be detected by pattern recognition receptors, for example, TLR3, MDA-5, NLRP3 to induce proinflammatory cytokines responsible for innate/adaptive immunity. Recognized by endosomal TLR3 in myeloid DCs (mDCs), dsRNA can activate mDCs into mature antigen presenting cells (mAPCs) which in turn present antigen epitopes with MHC-I molecules to naïve T cells. Coadministration of protein and synthetic dsRNA analogues can elicit an antigen-specific Th1-polarized immune response which stimulates the CD8+ CTL response and possibly dampen Th17 response. Synthetic dsRNA analogues have been tested as vaccine adjuvant against viral infections in animal models. However, a dsRNA receptor, TLR3 can be expressed in tumor cells while other members of TLR family, for example, TLR4 and TLR2 have been shown to promote tumor progression, metastasis, and chemoresistance. Thus, the promising potential of dsRNA analogues as a tumor therapeutic vaccine adjuvant should be evaluated cautiously.


Delay to mental healthcare in a cohort of Canadian Armed Forces personnel with deployment-related mental disorders, 2002-2011: a retrospective cohort study.
To assess whether the delay to care among Canadian Armed Forces (CAF) personnel who sought care for a mental disorder changed over time and in association with CAF mental health system augmentations. A stratified, random sample (n=2014) was selected for study from an Afghanistan-deployed cohort (N=30 513) and the 415 (weighted N=4108) individuals diagnosed with an Afghanistan service-related mental disorder were further assessed. Diagnosis-related data were abstracted from medical records (22 June 2010 to 30 May 2011). Other data were extracted from administrative databases. Delay to care was assessed across five mental health system eras: 2002/2004, 2005/2006, 2007, 2008 and 2009/2010. Weighted Cox proportional hazards regression assessed the association between era, handled as a time-dependent covariate, and the outcome while controlling for a broad range of potential confounders (ie, sociodemographic, military and clinical characteristics). Taylor series linearisation methods and sample design weights were applied in generating descriptive and regression analysis statistics. The outcome was the delay to mental healthcare, defined as the latency from most recent Afghanistan deployment return date to diagnosis date, among individuals with an Afghanistan service-related mental disorder diagnosis. Mean delay to care was 551 days (95% CI 501 to 602); the median was 400 days. Delay to care decreased in subsequent eras relative to 2002/2004; however, only the most recent era (2009/2010) was statistically significant (adjusted HR (aHR): 3.01 (95% CI 1.91 to 4.73)). Men, operations support occupations, higher ranks, non-musculoskeletal comorbidities and fewer years of military service were also independently associated with longer delays to care. CAF mental health system changes were associated with reduced delays to mental healthcare. Further evaluation research is needed to identify the key system changes that were most impactful.


Notch signaling in the regulation of stem cell self-renewal and differentiation.
Stem cells are rare and unique precursor cells that participate in the building and rebuilding of tissues and organs during embryogenesis, postnatal growth, and injury repair. Stem cells are distinctively endowed with the ability to both self-renew and differentiate, such that they can replenish the stem cell pool while continuing to produce the differentiated daughter cells that are essential for tissue function. Stem cell self-renewal/differentiation decisions must be carefully controlled during organogenesis, tissue homeostasis, and regeneration, as failure in stem cell maintenance or activation can lead to progressive tissue wasting, while unchecked self-renewal is a hallmark of many cancers. Here, we review evidence implicating the Notch signaling pathway, an evolutionarily conserved cell fate determinant with widespread roles in a variety of tissues and organisms, as a crucial regulator of stem cell behavior. As discussed below, this pathway plays varied and critical roles at multiple stages of organismal development, in lineage-specific differentiation of pluripotent embryonic stem cells, and in controlling stem cell numbers and activity in the context of age-related tissue degeneration, injury-induced tissue repair, and malignancy.


Prostatic leiomyosarcoma revealing cytoplasmic virus-like particles and intranuclear paracrystalline structures.
This report concerns an ultramicroscopic study of an autopsy case of primary leiomyosarcoma of the prostate in a patient who died of widespread blood-borne metastases after a rapid clinical course. Ultrastructural observation of tumor cells demonstrated some characteristics of leiomyosarcoma and furthermore revealed frequent intranuclear structures in paracrystalline array together with aggregates of interchromatin granules and cytoplasmic virus-like particles.


[Brain aging, biological and psychosocial aspect].
The world population is aging The frontier between physiological and pathological brain aging is somewhat unclear. Pathological aging can cause both mild cognitive impairments and dementias (vascular and Alzheimer). It is important to understand brain aging in order to identify risk factors and develop protective measures. Proper lifetime brain protection could improve quality of life in old age.


Biological effects of epicuticular flavonoids from Primula denticulata on human leukemia cells.
The biological effects of epicuticular substances in farinose exudates accumulated on inflorescence shafts and calyces of Primula denticulata on human acute myeloid leukemia cells (HL-60) were analyzed. The crude material possessed little antioxidative capacity but strong cytostatic properties. Some of its known components (5-hydroxyflavone, 2'-hydroxyflavone, 5,2'-dihydroxyflavone, and 5,8-dihydroxyflavone) were further tested to identify the biologically active compounds. The effects of these flavones on cell cycle progression, mitochondrial membrane potential, and reactive oxygen species have been investigated by flow cytometry. The flavonol quercetin was included in the study as reference compound because of its known cytostatic properties and its activity as radical scavenger. Compared to quercetin the flavones induced little apoptosis (up to 40 microM), but despite their low toxicity, the Primula flavonoids possessed strong cytostatic properties even at low concentrations. The cell cycle distribution showed a characteristic time-dependent shift, giving evidence of a generally short-lived effect of the test compounds in the exposed cells. The antioxidative properties quantified according to two different methods correlated with the number of hydroxyl groups. Whereas quercetin strongly affected the mitochondrial membrane potential, none of the Primula flavones showed a comparable effect.


[Acquired immunodeficiency syndrome with chronic granulomatous inflammation. Clinically definable special form of the variable immunodeficiency syndrome].
Five cases (3 men, 2 women) of late-onset variable immunodeficiency syndrome (CVID), characterized by similar clinical and immunological findings as well as histological demonstration of chronic granulomatous infection, are reported. All patients had frequent attacks of respiratory infections with recurrent bronchitis and pneumonia. In addition to predominating basally localized streaky-nodular lung changes all patients had hepatosplenomegaly and granulomatous infections of other organs. Immunologically, marked hypogammaglobulinaemia of all Ig classes, lymphopenia, and absence of terminal B-cell maturation were predominant. In-vitro tests under pokeweed-mitogen failed to demonstrate terminal plasma-cell differentiation of B-lymphocytes and thus Ig synthesis. Without pokeweed-mitogen there were largely nonsecretory B-blasts with abnormal granulated cytoplasmic Ig formation. Skin testing with Multitest application revealed almost complete anergy, both in the Arthus (24 h) and the late reactions (48 and 72 h). Nonetheless, T-cell reaction in-vitro was much less affected than B-cell function. "Natural killing" and antibody-dependent cytotoxicity were normal or slightly increased.


Solubilities of trimethoprim and sulfamethoxazole at various pH values and crystallization of trimethoprim from infusion fluids.
Solubilities of sulfamethoxazole (SMX) and trimethoprim (TMP) have been measured in the absence and presence of various concentrations of hydroxypropyl beta-cyclodextrin (HPCD) in buffers over a range of pH values. The solubilities of TMP increased linearly in the presence of HPCD at pH values of 7.0, 7.5, 8.9, and 9.9 indicating the formation of 1 to 1 complexes, whose association values increase as pH does and as the ionization of TMP decreases. For SMX at pH 7.0, a 1:1 complex is formed, but at pH 7.5 HPCD has little effect on the solubility of the highly ionized SMX, presumably since only un-ionized molecules can form inclusion complexes with the HPCD. At pH 7.0 when solubilities of TMP and SMX are measured in the same system containing buffer and HPCD, a massive decrease in the solubility of TMP is found, while that of SMX decreases slightly. A slightly soluble complex of TMP/SMX may be formed. When TMP/SMX injection is diluted 1;10 with phosphate buffer (pH 7.8), an immediate massive precipitation which is mostly TMP is observed. The presence of 10% w/v HPCD in the buffer delays and modifies the precipitation of TMP and SMX but does not prevent it. Mixing the injection (1 to 10) with normal saline (pH 9.5) gives a steadily increasing precipitation of TMP with time, but no discernible change in the concentration of SMX. However, in normal saline containing 10% w/v HPCD there is no crystallization of either drug over a 5 day period.


Chitinolytic and chitosanolytic activities from crude cellulase extract produced by A. niger grown on apple pomace through Koji fermentation.
Enzyme extracts of cellulase [filter paper cellulase (FPase) and carboxymethyl cellulase (CMCase)], chitinase, and chitosanase produced by Aspergillus niger NRRL-567 were evaluated. The interactive effects of initial moisture and different inducers for FP cellulase and CMCase production were optimized using response surface methodology. Higher enzyme activities [FPase 79.24+/- 4.22 IU/gram fermented substrate (gfs) and CMCase 124.04+/-7.78 IU/gfs] were achieved after 48 h fermentation in solid-state medium containing apple pomace supplemented with rice husk [1% (w/w)] under optimized conditions [pH 4.5, moisture 55% (v/w), and inducers veratryl alcohol (2 mM/kg), copper sulfate (1.5 mM/kg), and lactose 2% (w/w)] (p<0.05). Koji fermentation in trays was carried out and higher enzyme activities (FPase 96.67+/-4.18 IU/gfs and CMCase 146.50+/-11.92 IU/gfs) were achieved. The nonspecific chitinase and chitosanase activities of cellulase enzyme extract were analyzed using chitin and chitosan substrates with different physicochemical characteristics, such as degree of deacetylation, molecular weight, and viscosity. Higher chitinase and chitosanase activities of 70.28+/-3.34 IU/gfs and 60.18+/-3.82 to 64.20+/-4.12 IU/gfs, respectively, were achieved. Moreover, the enzyme was stable and retained 92-94% activity even after one month. Cellulase enzyme extract obtained from A. niger with chitinolytic and chitosanolytic activities could be potentially used for making low-molecular-weight chitin and chitosan oligomers, having promising applications in biomedicine, pharmaceuticals, food, and agricultural industries, and in biocontrol formulations.


Endocytosis of Corn Oil-Caseinate Emulsions In Vitro: Impacts of Droplet Sizes.
The relative uptake and mechanisms of lipid-based emulsions of three different particle diameters by Caco-2 cells were studied. The corn oil-sodium caseinate emulsions showed little or no cytotoxicity even at 2 mg/mL protein concentration for any of the three droplet size emulsions. Confocal laser scanning microscopy (CLSM) of Nile red containing emulsions showed that the lipid-based emulsions were absorbed by Caco-2 cells. A negative correlation between the mean droplet size and cellular uptake was observed. There was a time-dependent and energy-dependent uptake as shown by incubation at different times and treatment with sodium azide a general inhibitor of active transport. The endocytosis of lipid-based emulsions was size-dependent. The internalization of nanoemulsion droplets into Caco-2 cells mainly occurred through clathrin- and caveolae/lipid raft-related pathways, while macropinocytosis route played the most important role for 556 nm emulsion endocytosis as shown by the use of specific pathway inhibitors. Permeability of the emulsion through the apical or basal routes also suggested that active transport may be the main route for lipid-based nanoemulsions. The results may assist in the design and application of lipid-based nanoemulsions in nutraceuticals and pharmaceuticals delivery.


Characterization of the steroid-dependence of insulin-like growth factor-binding protein-2 synthesis and mRNA expression in cultured human endometrial stromal cells.
The insulin-like growth factors (IGF-I and -II) are believed to be important in endometrial differentiation and blastocyst nidation, and proteins that regulate IGF action (IGF-binding proteins, IGFBPs) are hormonally regulated in endometrium during the menstrual cycle. To characterize further steroid-dependence of the IGFBPs, we established endometrial stromal cells in culture in the absence and presence of oestradiol (E2) and progesterone (P) and examined the conditioned medium for IGFBPs by Western ligand blot analysis. Stromal cells constitutively synthesized IGFBP-3, IGFBP-2, a 27 kd, and a 24 kd IGFBP. In the presence of E2 and P, a 10- to 15-fold increase in IGFBP-2 was detected in the conditioned medium beginning after about 7 days in culture, when cells decidualized and steroid-mediated prolactin secretion began. Withdrawal of steroids resulted in a marked decrease in IGFBP-2, comparable to control levels, and cells increased their IGFBP-2 production when rechallenged with E2 and P. Total RNA was isolated from stromal cells, and Northern blot analysis using a cDNA probe specific for IGFBP-2 revealed differential expression of a 1.4 kb mRNA transcript in steroid-treated compared to control cells. The effects of progestational agents alone on IGFBP synthesis was also examined. Progesterone, medroxyprogesterone acetate and norethindrone all stimulated IGFBP-2 synthesis 12- to 15-fold compared to controls, and a progesterone receptor antagonist, RU 486, blocked the stimulatory effect of progesterone. IGFBP-2 synthesis was increased two-fold above controls by 17-alpha-hydroxyprogesterone, and RU 486 alone and hydrocortisone were without effect. Identification of IGFBP-2 in conditioned medium was made using IGFBP-specific antiserum. These data show that (a) endometrial stromal cells synthesize and secrete IGFBP-2, (b) IGFBP-2 protein synthesis is controlled by steroid hormones, (c) P, by interacting with its receptor, modulates IGFBP-2 synthesis and (d) expression of IGFBP-2 mRNA is controlled by sex steroids.


Effectiveness of Contour Augmentation with Guided Bone Regeneration: 10-Year Results.
In aesthetic sites, the integrity of the facial bone wall dimension in the anterior maxilla is jeopardized by physiologic and structural changes postextraction. An effective regenerative protocol is key to reestablish and maintain the hard and soft tissue dimensions over time. The present prospective case series study examined the effectiveness of early implant placement with simultaneous contour augmentation through guided bone regeneration with a 2-layer composite graft in postextraction single-tooth sites over an observation period of 10 y among 20 patients. The median peri-implant bone loss was 0.35 mm between the 1- and 10-y examination. A success rate of 95% was obtained, with pleasing aesthetic outcomes and a high median Pink Esthetic Score (8). Implant crowns (ICs) revealed significant median facial recession between IC10y and IC1y (0.17 mm). The facial bone wall dimensions were assessed by preoperative cone beam computed tomography and 2 subsequent scans taken at 6 and 10 y. The median facial bone wall thickness increased significantly from 0 mm at surgery to 1.67 mm at the 10-y examination. The facial vertical bone wall peak (DIC) was located at a median distance of 0.16 mm coronal to the implant shoulder. The facial vertical bone loss of DIC amounted to 0.02 mm between 6 and 10 y. Equivalence testing was performed for the null hypothesis of a difference of >0.2 mm per year between 2 respective time points, showing stable bone conditions. Modulating factors influencing the regenerative outcomes at 10 y were the preoperative proximal crest width and soft tissue thickness. In conclusion, the present study confirmed the long-term effectiveness of early implant placement with simultaneous contour augmentation through guided bone regeneration with a 2-layer composite graft in postextraction single-tooth sites offering stable bone conditions with low risks of mucosal recessions over an observation period of 10 y ( ClinicalTrials.gov NCT03252106).


Substrate specificity and kinetic properties of enzymes belonging to the hormone-sensitive lipase family: comparison with non-lipolytic and lipolytic carboxylesterases.
We have studied the kinetics of hydrolysis of triacylglycerols, vinyl esters and p-nitrophenyl butyrate by four carboxylesterases of the HSL family, namely recombinant human hormone-sensitive lipase (HSL), EST2 from Alicyclobacillus acidocaldarius, AFEST from Archeoglobus fulgidus, and protein RV1399C from Mycobacterium tuberculosis. The kinetic properties of enzymes of the HSL family have been compared to those of a series of lipolytic and non-lipolytic carboxylesterases including human pancreatic lipase, guinea pig pancreatic lipase related protein 2, lipases from Mucor miehei and Thermomyces lanuginosus, cutinase from Fusarium solani, LipA from Bacillus subtilis, porcine liver esterase and Esterase A from Aspergilus niger. Results indicate that human HSL, together with other lipolytic carboxylesterases, are active on short chain esters and hydrolyze water insoluble trioctanoin, vinyl laurate and olive oil, whereas the action of EST2, AFEST, protein RV1399C and non-lipolytic carboxylesterases is restricted to solutions of short chain substrates. Lipolytic and non-lipolytic carboxylesterases can be differentiated by their respective value of K(0.5) (apparent K(m)) for the hydrolysis of short chain esters. Among lipolytic enzymes, those possessing a lid domain display higher activity on tributyrin, trioctanoin and olive oil suggesting, then, that the lid structure contributes to enzyme binding to triacylglycerols. Progress reaction curves of the hydrolysis of p-nitrophenyl butyrate by lipolytic carboxylesterases with lid domain show a latency phase which is not observed with human HSL, non-lipolytic carboxylesterases, and lipolytic enzymes devoid of a lid structure as cutinase.


Fattening Pigs Are a Reservoir of Florfenicol-Resistant Enterococci Harboring Oxazolidinone Resistance Genes.
The use of florfenicol in farm animals may select enterococci that carry resistance genes that confer resistance to linezolid, a critically important oxazolidinone antibiotic used in human medicine. This cross-sectional study aimed to assess the occurrence of oxazolidinone resistance genes in florfenicol-resistant enterococci from fattening pigs in Switzerland and to characterize a subset of the isolates using whole genome sequencing. A total of 31 florfenicol-resistant enterococcal isolates were obtained from 27 (5%) of 565 cecal samples of fattening pigs from seven (11%) of 62 farms. Screening by PCR revealed the presence of cfr-poxtA in 1 of 31, optrA in 15 of 31, and poxtA in 15 of 31 enterococcal isolates. One randomly selected isolate per PCR-positive Enterococcus species and positive farm was selected for further analysis (n = 10). In nine of the 10 isolates, the presence of oxazolidinone resistance genes did not result in phenotypic resistance. Whole genome sequencing analysis showed the presence of E. faecalis (n = 1), E. faecium (n = 1), and E. hirae (n = 1), harboring optrA18, optrA7, and a new optrA allele, respectively. E. durans (n = 1), E. faecium (n = 4), and E. hirae (n = 1) carried the wild-type poxtA, and E. faecalis (n = 1) coharbored cfr(D) and poxtA2. Except for optrA7, all oxazolidinone resistance genes were found on plasmids. Multilocus sequence typing analysis identified E. faecalis ST19 and ST376, E. faecium ST80 belonging to hospital-adapted clade A1, and E. faecium ST21, ST55, ST269, and ST416 belonging to clade A2, which represents human commensals and animal strains. The occurrence of cfr(D), optrA, and poxtA in various porcine Enterococcus spp. demonstrates the spread of oxazolidinone resistance genes among enterococci from fattening pigs in Switzerland. The presence in one sample of poxtA-carrying E. faecium ST80 emphasizes the potential risk to human health through dissemination of strains carrying oxazolidinone resistance genes into the food chain.


RhoA exerts a permissive effect on volume-regulated anion channels in vascular endothelial cells.
Cell swelling triggers in most cell types an outwardly rectifying anion current, I(Cl,swell), via volume-regulated anion channels (VRACs). We have previously demonstrated in calf pulmonary artery endothelial (CPAE) cells that inhibition of the Rho/Rho kinase/myosin light chain phosphorylation pathway reduces the swelling-dependent activation of I(Cl,swell). However, these experiments did not allow us to discriminate between a direct activator role or a permissive effect. We now show that the Rho pathway did not affect VRAC activity if this pathway was activated by transfecting CPAE cells with constitutively active isoforms of Galpha (a Rho activating heterotrimeric G protein subunit), Rho, or Rho kinase. Furthermore, biochemical and morphological analysis failed to demonstrate activation of the Rho pathway during hypotonic cell swelling. Finally, manipulating the Rho pathway with either guanosine 5'-O-(3-thiotriphosphate) or C3 exoenzyme had no effect on VRACs in caveolin-1-expressing Caco-2 cells. We conclude that the Rho pathway exerts a permissive effect on VRACs in CPAE cells, i.e., swelling-induced opening of VRACs requires a functional Rho pathway, but not an activation of the Rho pathway.


[Enzyme immunoassay for secretory immunoglobulin A on skin surface and in sweat].
Recently secretory IgA (S-IgA) was found to be secreted from the eccrine gland. By using a sandwich enzyme immunoassay, we measured the concentration of S-IgA 1) in sweat (sweat S-IgA) and 2) in the extract buffer obtained by pipetting++ on the skin (skin surface S-IgA). Skin surface hydration and skin surface lipid were measured at the sites where the samples of skin surface S-IgA were collected. (These measurements were taken immediately after the buffer was pipetted). The quantity of both sweat S-IgA and skin surface S-IgA differed according to the sites where they were collected. Sites in order of decreasing sweat S-IgA level: face, chest, forearm. Sites in order of decreasing skin surface S-IgA level: face, chest, palm, forearm, sole. (Skin surface S-IgA levels on palm and forearm were approximately equal). Although the amount of skin surface S-IgA was not related to the skin surface hydration, there was a significant correlation between skin surface S-IgA and skin surface lipid.


Concurrent septic arthritis and osteomyelitis in children.
Septic arthritis and osteomyelitis can both independently cause substantial morbidity. With concomitant septic arthritis and osteomyelitis, the septic arthritis may be treated without knowledge of the adjacent osteomyelitis resulting in delayed treatment. This study aims to identify factors that may help to diagnosis concurrent infections (CI) earlier. A retrospective chart review of 200 patients with septic arthritis was performed. Patients with CI were compared with patients with septic arthritis alone using tests determined by the nature of the variable being analyzed (the χ test, the Fisher exact test, the Cochran-Armitage trend test, and the Kruskal-Wallis test.). Two hundred patients were eligible and analyzed, of which 43 (21.5%) had CI. On the basis of age, CI were most common in newborns and adolescents (P<0.0001). On the basis of location, 72% of shoulder infections (P<0.0001) were concurrent, whereas <50% of elbows, hips, knees, and ankle were CI. The most common infective organism was methicillin-sensitive Staphylococcus aureus (P<0.0001). CI were significantly associated with increased median (6) days of clinical symptoms before presentation (P<0.0001), increased duration of median (6) days of hospital stay (P<0.0001), increased number of operative procedures (P=0.005), and increased need for ICU admission (P=0.024). Utilizing advanced imaging (CT scan, bone scan, and/or MRI) in patients with septic arthritis who are younger than 4 months of age, between ages 13 and 20 years, with septic arthritis involving the shoulder, and with symptoms for more than 6 days may shorten hospital stays, decrease the number of operative procedures required, and possibly limit infection-related sequelae by identifying CI earlier. III.


Interactions of dendrimers with biological drug targets: reality or mystery - a gap in drug delivery and development research.
Dendrimers have emerged as novel and efficient materials that can be used as therapeutic agents/drugs or as drug delivery carriers to enhance therapeutic outcomes. Molecular dendrimer interactions are central to their applications and realising their potential. The molecular interactions of dendrimers with drugs or other materials in drug delivery systems or drug conjugates have been extensively reported in the literature. However, despite the growing application of dendrimers as biologically active materials, research focusing on the mechanistic analysis of dendrimer interactions with therapeutic biological targets is currently lacking in the literature. This comprehensive review on dendrimers over the last 15 years therefore attempts to identify the reasons behind the apparent lack of dendrimer-receptor research and proposes approaches to address this issue. The structure, hierarchy and applications of dendrimers are briefly highlighted, followed by a review of their various applications, specifically as biologically active materials, with a focus on their interactions at the target site. It concludes with a technical guide to assist researchers on how to employ various molecular modelling and computational approaches for research on dendrimer interactions with biological targets at a molecular level. This review highlights the impact of a mechanistic analysis of dendrimer interactions on a molecular level, serves to guide and optimise their discovery as medicinal agents, and hopes to stimulate multidisciplinary research between scientific, experimental and molecular modelling research teams.


Athlete's Retention of a Coach's Instruction Before a Judo Competition.
The aim of the present study was to analyze the instruction of the Judo coach immediately before the competition, in the process of preparation for the fights, looking to (1) study the coherency between the information which the coach transmits and that which the athlete retains; (2) identify the correlation between the coherency, the extension and the number of ideas conveyed by the coach; (3) determine if the retention varies in relation to variables such as the form and nature of the information, as well as the gender and practice category of the athletes. The participants were 11 coaches and 58 athletes of 3 categories: under- 15, under-17 and under-20, of both genders. One hundred and sixteen (116) instructional episodes were observed, which corresponds to four hundred and six (406) information units convoyed by the coaches. The coaches' instructions given before the competition were recorded in an audio and video register. After the coaches' instruction, the athletes were approached by the investigator and an interview was accomplished. To determine if the retention varies in relation to form and nature of the information and gender and practice category of the athletes, the non-parametric statistics, U de Mann-Whitney and Kruskal- Wallis, was used. Correlation of Spearman was applied to verify the degree of association between the coherency, the extension and the number of ideas conveyed by the coach. The results showed that a substantial part of the information was not retained by the athletes and the information coherency was inversely related to the number of transmitted ideas. The coaches were, mainly, prescriptive and the form of the information was not important for the retention of the information. Gender was a differentiated variable as the girls showed more coherency in the retained ideas in relation to the ideas transmitted by the coach. Key pointsThe instructions given by the coach are optimized if the athletes retain and understand them well and should be carefully analyzed by researchers and coaches.The ratio between the number of concordant ideas between coach and athlete (coherency) increased when the number of ideas decreased which raises the question of the adequacy of the instructional strategies used by coach.The prescriptive information showed that athletes were able to express a larger number of ideas in fewer words (larger density) while the combined information caused athletes to use more words to reproduce what the coach said.Gender was a differentiated variable as the girls showed more coherency in the retained ideas in relation to the ideas transmitted by the coach. These results indicate a possible tendency for girls to be more attentive when the coach is emitting information. However, to confirm this assumption more research is needed.


Bipolar disorder associated to substance use disorders (dual diagnosis). Systematic review of the scientific evidence and expert consensus.
The present work focuses on the so-called dual diagnosis (DD): bipolar disorder (BD) associated with substance use disorders (SUD). Although the psychiatrists who treat patients with BD and physicians in charge of patients with SUD frequently find this association with DD, unfortunately there are few scientific works that have studied this association. The Spanish Working Group on Bipolar Disorders in Dual Diagnosis reviewed the published material using a Medline search and selected the most relevant articles. Following this, the Work Group developed an expert consensus in DD and finally, a survey was performed among a group of experts in this disorder to cover the areas that were not fully addressed by the scientific evidence or in those areas in which the Work Group was unable to reach a consensus. We conclude that, in view of the above, establishment of a consensus is a valid tool to complement the current scientific evidence.


A diatom light-harvesting pigment-protein complex : purification and characterization.
A light-harvesting pigment-protein complex was isolated from the diatom Phaeodactylum tricornutum using the zwitterionic detergent CHAPS (3-[3-cholamidopropyl)dimethylammonio]-1-propanesulfonate). Detergent-solubilized membranes were fractionated by sucrose density gradient centrifugation into three components. The medium density fraction contained chlorophyll a, chlorophyll c, and fucoxanthin. This fraction was purified by DEAE-ion exchange chromatography, and contained chlorophyll a, chlorophyll c, and fucoxanthin in a molar ratio of 2.4:1.0:4.8. Fluorescence emission and excitation spectra of the isolated complex demonstrated that light energy absorbed by chlorophyll c and fucoxanthin was coupled to chlorophyll a fluorescence. Upon denaturation, the apoprotein yielded a polypeptide doublet at 17.5 to 18.0 kilodaltons which accounted for 30 to 40% of the toal membrane protein. These findings indicate that this pigment-protein complex is a major component of the diatom photosynthetic lammellae. The quantitative amino acid composition of the apoprotein was very similar to those reported for other membrane-bound pigment-protein complexes. Based on the protein to chlorophyll a ratio of 7700 grams protein per mole chlorophyll a for the complex, each apoprotein molecule contains, to the nearest integer, two chlorophyll a, one chlorophyll c, and five fucoxanthin molecules. Polyclonal antibodies raised against the 17.5 to 18.0 kilodaltons apoprotein showed a monospecific reaction with only the 17.5 to 18.0 protein zone from denatured P. tricornutum membranes as well as to the nondenatured pigment-protein complex. It appears that this complex is common to other diatom species.


Comprehensive Identification of Tumor-associated Antigens via Isolation of Human Monoclonal Antibodies that may be Therapeutic.
Although the success of trastuzumab and rituximab for treatment of breast cancer and non-Hodgkins lymphoma, respectively, suggests that monoclonal antibodies (mAbs) will become important therapeutic agents against a wider range of cancers, useful therapeutic Abs are not yet available for the majority of the human cancers because of our lack of knowledge of which antigens (Ags) are likely to become useful targets. We established a procedure for comprehensive identification of such Ags through the extensive isolation of human mAbs that may be therapeutic. Using the phage-display Ab library we isolated a large number of human mAbs that bind to the surface of tumor cells. They were individually screened by immunostaining, and clones that preferentially and strongly stained the malignant cells were chosen. The Ags recognized by those clones were isolated by immunoprecipitation and identified by mass spectrometry (MS). We isolated 2,114 mAbs with unique sequences and identified 25 distinct Ags highly expressed on several carcinomas. Of those 2,114 mAbs 434 bound to specifically to one of the 25 Ags. I am going to discuss how we could select proper target Ags for therapeutic Abs and candidate clones as therapeutic agents.


Effects of Adipose-Derived Stem Cells and Platelet-Rich Plasma for Prevention of Alopecia and Other Skin Complications of Radiotherapy.
Radiotherapy (RT) involves the use of ionizing radiation in treating malignancies and benign disorders. However, RT damages target and healthy surrounding tissues in a dose-dependent manner. This effectively reduces patient compliance and quality of life, thereby warranting the prevention of RT-induced adverse effects on skin. Adipose-derived stem cells (ASCs) are used to treat RT-induced damage and platelet-rich plasma (PRP) provides a scaffold that potentiates the effects of ASCs. Thus, the aim of this study was to determine the mechanism employed by ASCs and PRP in protecting against RT-induced adverse effects. We have established an immunodeficient mouse transplantation model using which human hair follicular units were implanted. When the follicular units were macroscopically and microscopically mature and anagenic, we administered localized RT. Subsequently, the mice were randomly divided into 4 groups based on the subcutaneous injection of the following to the irradiated transplantation site: saline, PRP, ASCs, and a combination of ASCs and PRP. Next, we used macroscopic and microscopic analyses to determine the protective effects of the injected solutions on skin and hair follicles. Adipose-derived stem cells reduced RT-induced adverse effects, such as impaired wound healing, alopecia, skin atrophy, and fibrosis by suppressing inflammation, dystrophy, degeneration, connective tissue synthesis, and apoptosis and increasing cellular proliferation, differentiation, and signaling. Moreover, these effects were augmented by PRP. Thus, co-administering ASCs with PRP in mice prevented RT-induced adverse effects and can be tested for use in clinical practice.


Amphiphilic Janus gold nanoparticles via combining "solid-state grafting-to" and "grafting-from" methods.
Despite the great efforts that have been made toward obtaining Janus architectures, synthesizing sub-10 nanometer Janus nanoparticles (NPs) modified with different types of polymers remains a challenging task. In this Communication, "solid-state grafting-to" and "grafting-from" methods were combined to obtain Janus gold NPs (AuNPs) modified with two types of polymer chains on the opposite sides of the NP. We used functionalized polymer single crystals as the solid substrates to immobilize AuNPs. We then used atom transfer radical polymerization to grow polymer chains on the "free" side of the AuNPs. Amphiphilic polyethylene oxide (PEO)-Au-poly(methyl methacrylate), PEO-Au-poly(tert-butyl acrylate) and hydrophilic PEO-Au-poly(acrylic acid) were synthesized. The Janus nature was demonstrated using a platinum-nanoparticle-decoration method. Using polymer single crystals as the reaction substrates is advantageous because they afford higher throughput compared with self-assembled monolayers. Dissolution of the single crystal also leads to NPs with defined polymer patches. We anticipate that our approach could serve as a generic method for synthesizing polymer-functionalized, sub-10 nm Janus NPs. This unique system holds promises for achieving controlled assembly and tunable optic and electronic properties of NPs.


A Stump of Occluded Posterior Cerebral Artery Mimicking a Ruptured Aneurysm: Case Report.
We report on a 63-year-old female with subarachnoid hemorrhage who had a stump of occluded posterior cerebral artery (PCA) mimicking a ruptured aneurysm of the basilar bifurcation. Intraoperatively, the aneurysmal opacification on preoperative angiograms proved to be the residual lumen of the occluded right P1 segment. Because of the nodular appearance and upward direction of the stump of the right P1 segment, it was misinterpreted as an aneurysm. During operation, a tiny ruptured aneurysm missed on preoperative angiograms was found in the left A1-A2 junction and was clipped safely.


Carbapenemase-producing Enterobacteriaceae in transplant patients.
Carbapenemase-producing Enterobacteriaceae (CPE) are a serious public health concern and represent a major threat to immunocompromised hosts, including solid organ (SOT) and stem cell transplant (HSCT) recipients. Transplant patients are at particular risk of developing CPE colonization and/or infection due to their frequent exposure to prolonged courses of broad-spectrum antibiotics, altered immunocompetence and exposure to invasive procedures and immunosuppressive drugs. Gut colonization with CPE, in particular carbapenem-resistant Klebsiella pneumoniae, may occur before or after SOT in 2%-27% of patients and among 2%-9% of HSCT and has been associated with increased risk of developing CPE infections. In endemic areas, CPE infections occur in up to 18% of SOT, and HSCT patients can account for 5%-18% of all patients with CPE bacteraemia. Mortality rates up to 70% have been associated with CPE infections in both patient populations. The rapid initiation of an active therapy against CPE is advocated in these infections. Therapeutic options, however, are limited by the paucity of novel compounds that are currently available and by potential antibiotic-associated toxicities. Therefore, a multidisciplinary approach involving infection control and antimicrobial stewardship programmes still represents the mainstay for the management of CPE infections among transplant patients. The evidence for the use of prevention strategies such as CPE-targeted perioperative prophylaxis or gut decolonization is still scarce. Large, multicentre trials are required to better define prevention strategies and to guide the management of CPE infections in the transplant setting.


The role of PET in lymphoma.
Malignant lymphomas are a heterogeneous group of diseases whose treatment and prognosis depend on accurate staging and evaluation of histologic features. The conventional imaging procedure is CT; however, nuclear medicine imaging has also had a prominent role. Single-photon imaging with 67Ga-citrate has been widely used for lymphomas. PET with 18F-FDG has gained a role in the staging and follow-up of lymphomas, largely replacing gallium as the nuclear medicine study of choice. 18F-FDG PET has proved useful in the staging and follow-up of Hodgkin's disease and non-Hodgkin's lymphoma (especially more aggressive types), and the widespread use of PET/CT has also increased the sensitivity and specificity. Its usefulness for the staging of slow-growing lymphomas has not been established. After the basics of staging and classification of lymphomas have been outlined, this article will review the role of 18F-FDG PET in the management of patients with lymphoma. PET tracers other than (18)F-FDG, such as positron-emitting isotopes of gallium and the cellular proliferation marker 18F-3'-deoxy-3'-fluorothymidine, will be discussed and future directions for PET in lymphoma proposed.


Note: anti-strong-disturbance signal processing method of vortex flowmeter with two sensors.
Some digital signal processing methods have been used to deal with the output signal of vortex flowmeter for extracting the flow rate frequency from the noisy output of vortex flow rate sensor and achieving the measurement of small flow rate. In applications, however, the power of noise is larger than that of flow rate sometimes. These strong disturbances are caused by pipe vibration mostly. Under this condition the previous digital signal processing methods will be unavailable. Therefore, an anti-strong-disturbance solution is studied for the vortex flowmeter with two sensors in this Note. In this solution, two piezoelectric sensors are installed in the vortex probe. One is called the flow rate sensor for measuring both the flow rate and vibration noise, and the other is called the vibration sensor for detecting the vibration noise and sensing the flow rate signal weakly at the same time. An anti-strong-disturbance signal processing method combining the frequency-domain substation algorithm with the frequency-variance calculation algorithm is proposed to identify the flow rate frequency. When the peak number of amplitude spectrum of the flow rate sensor is different from that of the vibration sensor, the frequency-domain subtraction algorithm will be adopted; when the peak number of amplitude spectrum of the flow rate sensor is the same as that of the vibration sensor, the frequency-variance calculation algorithm will be employed. The whole algorithm is implemented in real time by an ultralow power micro control unit (MCU) to meet requirements of process instrumentation. The experimental results show that this method can obtain the flow rate frequency correctly even if the power of the pipe vibration noise is larger than that of the vortex flow rate signal.


Diaphragm muscle remodeling in a rat model of chronic intermittent hypoxia.
Respiratory muscle remodeling occurs in human sleep apnea--a common respiratory disorder characterized by chronic intermittent hypoxia (CIH) due to recurrent apnea during sleep. We sought to determine if CIH causes remodeling in rat sternohyoid (upper airway dilator) and diaphragm muscles. Adult male Wistar rats were exposed to CIH (n=8), consisting of 90 sec of hypoxia (5% at the nadir; SaO₂ ~80%)/90 sec of normoxia, 8 hr per day, for 7 consecutive days. Sham animals (n=8) were exposed to alternating air/air cycles in parallel. The effect of CIH on myosin heavy-chain (MHC) isoform (1, 2a, 2x, 2b) distribution, sarcoplasmic reticulum calcium ATPase (SERCA) isoform distribution, succinate dehydrogenase activity, glycerol phosphate dehydrogenase activity, and Na⁺/K⁺ ATPase pump content was determined. Sternohyoid muscle structure was unaffected by CIH treatment. CIH did not alter oxidative/glycolytic capacity or the Na⁺/K⁺-ATPase pump content of the diaphragm. CIH significantly increased the areal density of MHC 2b fibers in the rat diaphragm, and this was associated with a shift in SERCA proteins from SERCA2 to SERCA1. We conclude that CIH causes a slow-to-fast fiber transition in the rat diaphragm after just 7 days of treatment. Respiratory muscle functional remodeling may drive aberrant functional plasticity such as decreased muscle endurance, which is a feature of human sleep apnea.


Cutaneous malignant melanoma: association with height, weight and body-surface area. a prospective study in Norway.
To investigate whether anthropometric indices as body height, body-mass index (BMI) and body-surface area (BSA) have any influence on the risk of cutaneous malignant melanoma (CMM), we analyzed height and weight data from a Norwegian screening survey running between 1963 and 1975 and comprising 1.3 million individuals followed for 14 to 26 years. Among females, CMM was diagnosed in 2814 incident cases, while the number among males was 2144 during 22,988,345 person-years of follow-up. With adjustment for age, birth cohort, geographic region and BMI, the risk for CMM increased significantly with increasing quintiles of height in both sexes. The tallest persons were at greater relative risk than the shortest ones: 1.60 and 1.59 in males and females respectively. This positive association was also found for CMM localized on face and trunk in both sexes, and in the lower limbs only in females. Males in the highest quintile of BMI had a relative risk for CMM of 1.26 compared with the lowest quintile, while a non-significant negative association were found in obese females. This negative association increased and was significant for CMM located on the trunk and lower limbs in females. Among males we observed a consistent association between BSA and risk of CMM similar to that for height. These relationships between anthropometric indices and CMM are discussed according to factors which promote growth, among which nutrition, social class and hormones are the most important.


Involvement of protein serine and threonine phosphorylation in human sperm capacitation.
The involvement of serine and threonine phosphorylation in human sperm capacitation was investigated. Anti-phosphoserine monoclonal antibody (mAb) recognized six protein bands in the 43-55-kDa, 94 +/- 2-kDa, 110-kDa, and 190-kDa molecular regions, in addition to a faint band each in the 18-kDa and 35-kDa regions. Anti-phosphothreonine mAb recognized protein bands in six similar regions, except that the 18-kDa, 35-kDa, and 94 +/- 2-kDa protein bands were sharper and thicker, and an additional band was observed in the 110-kDa molecular region. In the 43-55-kDa molecular region, there was a well-characterized glycoprotein, designated fertilization antigen, that showed a further increase in serine/threonine phosphorylation after exposure to solubilized human zona pellucida. In a cell-free in vitro kinase assay carried out on beads or in solution, four to eight proteins belonging to similar molecular regions, namely 20 +/- 2 kDa, 43-55 kDa, 94 +/- 2 kDa, and 110 +/- 10 kDa, as well as in 80 +/- 4 and 210 +/- 10 kDa regions, were phosphorylated at dual residues (serine/tyrosine and threonine/tyrosine). Capacitation increased the intensity of serine/threonine phosphorylation per sperm cell, increased the number of sperm cells that were phosphorylated, and induced a subcellular shift in the serine/threonine-specific fluorescence. These findings indicate that protein serine/threonine phosphorylation is involved and may have a physiological role in sperm capacitation.


Quality of Life in Patients With Diabetes Mellitus.
Diabetes mellitus is a challenging illness that has many lifestyle demands and poses life-threatening and debilitating complications. The treatments are complex and require patient education and constant vigil. Consequently, there has been increasing interest in the effects of diabetes on health-related quality of life (HRQOL). Both Type I and Type II diabetes appear to impact on HRQOL, with Type I diabetes having the greater negative effect. Of interest, the use of intensive treatment methods for Type I diabetes is not associated with decreased HRQOL. This result was reported in the Diabetes Control and Complications Trial, and so may reflect patient selection or the degree of support available in a clinical trial. However, it does suggest that intensive treatment can be undertaken without fear of inevitable decrements in patient well-being. Insulin taking type II patients experience worse HRGOL than those on oral agents or diet alone. Some new measures are now available for assessing HRQOL and related constructs in diabetic patients. In particular, the diabetes quality of life measure has been shown to be a reliable and valid indicator of HRQOL. There is a need to better understand the effects of diabetes and its treatment on HRQOL in the context of clinical practice. Such measures can facilitate this evaluation process.


Ecology of bats and their role in emerging zoonotic diseases: a review.
The order Chiroptera is the second largest order of mammals and shows great physiological and ecological diversity. These animals play significant ecological roles as prey and predator as well as facilitating pollination, seed dispersal, arthropod reduction and nutrient distribution and reutilisation in nature. Bats act as hosts to a range of viral, bacterial, fungal and parasitic zoonoses. Human activities increase the likelihood of exposure to bats, thereby increasing the opportunity for infections to spill over. Continuing ecological processes, emergence and spillover of novel pathogens in naïve hosts, including humans, along with other complex natural phenomena require proper understanding that may help in predicting the next spillover. This review will discuss the ecology of bats and their role in the emergence of different zoonoses, particularly those of viral origin, in an organised manner to increase understanding of the factors that may play significant roles in spillover of these pathogens from bats to other animals, including humans.


Different unfolding pathways for mesophilic and thermophilic homologues of serine hydroxymethyltransferase.
To determine how much information can be transferred from folding and unfolding studies of one protein to another member of the same family or between the mesophilic and thermophilic homologues of a protein, we have characterized the equilibrium unfolding process of the dimeric enzyme serine hydroxymethyltransferase (SHMT) from two sources, Bacillus subtilis (bsSHMT) and Bacillus stearothermophilus (bstSHMT). Although the sequences of the two enzymes are highly identical ( approximately 77%) and homologous (89%), bstSHMT shows a significantly higher stability against both thermal and urea denaturation than bsSHMT. The GdmCl-induced unfolding of bsSHMT was found to be a two-step process with dissociation of the native dimer, resulting in stabilization of a monomeric species, followed by the unfolding of the monomeric species. A similar unfolding pathway has been reported for Escherichia coli aspartate aminotransferase, a member of the type I fold family of PLP binding enzymes such as SHMT, the sequence of which is only slightly identical ( approximately 14%) with that of SHMT. In contrast, for bstSHMT, a highly cooperative unfolding without stabilization of any monomeric intermediate was observed. These studies suggest that mesophilic proteins of the same structural family even sharing a low level of sequence identity may follow a common unfolding mechanism, whereas the mesophilic and thermophilic homologues of the same protein despite having a high degree of sequence identity may follow significantly different unfolding mechanisms.


An Eco-friendly Iron Cathode Electro-Fenton System Coupled With a pH-Regulation Electrolysis Cell for p-nitrophenol Degradation.
The high consumption of salt reagents and strict pH control are still bottlenecks for the full-scale application of the Fenton reaction. In this work, a novel eco-friendly iron cathode electrochemical Fenton (ICEF) system coupled with a pH-regulation divided electrolysis cell was developed. In a pH-regulation divided electrolysis system, the desired pH for an effective Fenton reaction and for a neutral treated media could be obtained by H2O splitting into H+ and OH- at the anode and cathode, respectively. In an ICEF system, an iron plate was used as the cathode to inhibit the release of iron ions and promote the reduction of Fe3+ to Fe2+. It was found that when a potential of 1.2 V/SCE was applied on the iron cathode, 98% of p-nitrophenol was removed in the combined system after 30 min with continuously adding 200 mg/L of H2O2. Meanwhile, a COD and TOC removal efficiency of 79 and 60% was obtained, respectively. In this case, the conductivity just slightly increased from 4.35 to 4.37 mS/cm, minimizing the increase of water salinity, as compared with the conventional Fenton process. Generally, this combined system was eco-friendly, energy-efficient, and has the potential of being a promising technology for the removal of bio-refractory organic pollutants from wastewaters.


The contribution of an avoidance response to contralateral neglect.
Displacement of line bisection to the right of midline in a patient with a right hemisphere lesion has been attributed to either contralesional attentional or intentional hemispatial neglect. However, it has been suggested that patients with hemispheric lesions might also have an avoidance response, and the ipsilesional eye and hand deviation attributed to neglect might, at least in part, be related to this response. The authors report a patient with a net contralateral neglect produced by the combination of intention and avoidance factors.


Photoactivated drug delivery and bioimaging.
Among the various types of diseases, cancer remains one of the most leading causes of mortality that people are always suffering from and fighting with. So far, the effective cancer treatment demands accurate medical diagnosis, precise surgery, expensive medicine administration, which leads to a significant burden on patients, their families, and the whole national healthcare system around the world. In order to increase the therapeutic efficiency and minimize side effects in cancer treatment, various kinds of stimuli-responsive drug delivery systems and bioimaging platforms have been extensively developed within the past decades. Among them, the strategy of photoactivated approach has attracted considerable research interest because light enables the precise control, in a highly spatial and temporal manner, the release of drug molecules as well as the activation of bioimaging agents. In general, several appropriate photoresponsive systems, which are normally sensitive to ultraviolet (UV) or visible light irradiation to undergo the multiple reaction pathways such as photocleavage and photoisomerization strategy etc. have been mainly involved in the light activated cancer therapies. Considering the potential issues of poor tissue penetration and high photoctotoxicity of short wavelength light, the recently emerged therapies based on long-wavelength irradiation, e.g., near-infrared (NIR) light (700-1000 nm), have displayed distinct advantages in biomedical applications. The light irradiation at NIR window indicates minimized photodamage, deep penetration, and low autofluorescence in living cells and tissues, which are of clinical importance in the desired diagnosis and therapy. In this review article, we introduce the recent advances in light-activated drug release and biological imaging mainly for anticancer treatment. Various types of strategies such as photocage, photo-induced isomerization, optical upconversion, and photothermal release by which different wavelength ranges of light can play the important roles in the controlled therapeutic or imaging agents delivery, and activation will be systemically discussed. In addition, the challenges and future perspectives for photo-based cancer theranostics will be also summarized. WIREs Nanomed Nanobiotechnol 2017, 9:e1408. doi: 10.1002/wnan.1408 For further resources related to this article, please visit the WIREs website.


Heterologous Expression of the Melatonin-Related Gene HIOMT Improves Salt Tolerance in Malus domestica.
Melatonin, a widely known indoleamine molecule that mediates various animal and plant physiological processes, is formed from N-acetyl serotonin via N-acetylserotonin methyltransferase (ASMT). ASMT is an enzyme that catalyzes melatonin synthesis in plants in the rate-determining step and is homologous to hydroxyindole-O-methyltransferase (HIOMT) melatonin synthase in animals. To date, little is known about the effect of HIOMT on salinity in apple plants. Here, we explored the melatonin physiological function in the salinity condition response by heterologous expressing the homologous human HIOMT gene in apple plants. We discovered that the expression of melatonin-related gene (MdASMT) in apple plants was induced by salinity. Most notably, compared with the wild type, three transgenic lines indicated higher melatonin levels, and the heterologous expression of HIOMT enhanced the expression of melatonin synthesis genes. The transgenic lines showed reduced salt damage symptoms, lower relative electrolyte leakage, and less total chlorophyll loss from leaves under salt stress. Meanwhile, through enhanced activity of antioxidant enzymes, transgenic lines decreased the reactive oxygen species accumulation, downregulated the expression of the abscisic acid synthesis gene (MdNCED3), accordingly reducing the accumulation of abscisic acid under salt stress. Both mechanisms regulated morphological changes in the stomata synergistically, thereby mitigating damage to the plants' photosynthetic ability. In addition, transgenic plants also effectively stabilized their ion balance, raised the expression of salt stress-related genes, as well as alleviated osmotic stress through changes in amino acid metabolism. In summary, heterologous expression of HIOMT improved the adaptation of apple leaves to salt stress, primarily by increasing melatonin concentration, maintaining a high photosynthetic capacity, reducing reactive oxygen species accumulation, and maintaining normal ion homeostasis.