Genes affecting the productivity of alpha-amylase in Bacillus subtilis Marburg.
Genetic control of alpha-amylase (alpha-1,4-glucan glucanohydrolase, EC 126.96.36.199.) production by Bacillus subtilis 168 was studied from the standpoint that alpha-amylase production by bacteria is dependent on a long-lived messenger ribonucleic acid and obeys the following equation: E = kappa integral of X-DT where x = cell mass at time t, E = alpha amylase produced, t = culture time, and kappa = productivity constant. So a productivity constand (kappa) is obtained from the slope of the straight line plot of alpha-amylase formed versus the total mass of cells accumulated over that time during the culture process. The following results were obtained. (i) Two sequential mutants, derived from the 168(kappa = 20) strain and having improved alpha-amylase productivity (168 leads to 196), were analyzed for their serine and metal protease production. Strain 128 (kappa = 40) produced half the amount of both proteases, but strain 196 (kappa = 60 similar to 80) produced 20 times that in the original strain. (ii) Amy+ transformants, using the 196 strain as the other three had higher productivity (kappa = 37 similar to 46). These transformants (J71, J47, groups. Seventy-one of 74 Amy+ transformants had a kappa value of 21.0 plus or minus 2.1 and the other three had higher productivity (kappa = 37 similar to 46). These transformants (J71,J47, and J10) produced levels of serine and metal proteases 20 times higher than the other transformants. (iii) Strains 196, J71, J47, and J10 were found to be nonmotile and resistant to phage PBS1, whereas other strains, including strains 168, 128, 3 revertants of strain J71 and 2 revertants of strain 196, were all motile and sensitive to the phage. (iv) Strains 196 and J71 were nonflagellated under electron microscopic observation but strain 168, 128 and a revertant of J71 were flagellated. From the above experimental results, the existence of a quality controlling gene (amyB) was deduced, which is loosely linked to the structural gene and controls productivities of alpha-amylase and proteases, and flagellation. The probable existence of another regulatory gene, amyC, is also discussed.


'A worse nightmare than expected' - a Swedish qualitative study of women's experiences two months after obstetric anal sphincter muscle injury.
this study explores women's experiences of the first two months after obstetric anal sphincter injury (OASIS) during childbirth with a focus on problematic recovery. this qualitative study used inductive qualitative content analysis to investigate open-ended responses from 1248 women. The data consists of short and comprehensive written responses to open-ended questions focusing on recovery in the national quality register, the Perineal Laceration Register, two months after OASIS at childbirth. the theme 'A worse nightmare than expected' illustrated women's experiences of their life situation. Pain was a constant reminder of the trauma, and the women had to face physical and psychological limitations as well as crushed expectations of family life. Furthermore, navigating healthcare services for help added further stress to an already stressful situation. we found that women with problematic recovery two months after OASIS experienced their situation as a worse nightmare than expected. Extensive pain resulted in physical and psychological limitations, and crushed expectations of family life. Improved patient information for women with OASIS regarding pain, psychological and personal aspects, sexual function, and subsequent pregnancy delivery is needed. Also, there is a need for clear organizational structures and information to guide help-seeking women to needed care.


Regulation of dendritic spine morphology by the rho family of small GTPases: antagonistic roles of Rac and Rho.
Dendritic spines mediate most excitatory transmission in the mammalian CNS and have been traditionally considered stable structures. Following the suggestion that spines may 'twitch', it has been recently shown that spines are capable of rapid morphological rearrangements. Because of the role of the small GTPases from the Rho family in controlling neuronal morphogenesis, we investigated the effects of several members of this biochemical signaling pathway in the maintenance of the morphology of extant dendritic spines by combining biolistic transfection of pyramidal neurons in cultured cortical and hippocampal slices with two-photon microscopy. We find a variety of effects on the density and morphology of dendritic spines by expressing either constitutively active or dominant negative forms of several small GTPases of the Rho family, by blocking the entire pathway with Clostridium difficile toxin B or by blocking Rho with C3 transferase. We propose a model where Rac promotes spine formation, while Rho prevents it. We conclude that the small GTPases provide antagonistic control mechanisms of spine maintenance in pyramidal neurons.


Inflammatory myofibroblastic tumor in children.
The authors presented the cases of two children with inflammatory myofibroblastic (IMF) tumor and reviewed the literature to facilitate the preoperative recognition, delineate the clinical features, and describe the natural history of this entity. The first child had IMF tumor arising from the mesentery of the small intestine. He presented with an abdominal mass associated with severe inflammatory response manifested by fever, impaired growth, thrombocytosis, and microcytic, hypochromic anemia. After surgical resection, his fever resolved and his growth rate and the laboratory abnormalities normalized. Five months after initial diagnosis, the fever, anemia, and thrombocytosis recurred along with two tumors arising from the omentum and the abdominal soft tissue. After the second surgery, he remains free of recurrent disease for 30 months. The second child presented with a lung mass that was radiologically indistinguishable from pulmonary sequestration. After surgical resection, she remains free of recurrent disease for 18 months. IMF tumor should be considered in any solid tumor that occurs in association with a chronic inflammatory response. IMF tumor should also be considered in the differential diagnosis of pulmonary sequestration.


All-fiber Q-switched single-frequency Tm-doped laser near 2 mum.
We present an all-fiber Q-switched single-frequency laser oscillator operating in the eye-safe region at 1950 nm. It is based on the stress-induced polarization modulation in a Tm-doped distributed Bragg reflector fiber laser. The laser emits Q-switched single-frequency laser pulses with a pulse repetition rate ranging from tens of hertz to hundreds of kilohertz and an average power of several milliwatts. Pulse duration and laser spectral linewidth have been characterized. This is, to our best knowledge, the first demonstration of a Q-switched single-frequency fiber laser near 2 microm.


Differential gene-expression of metallothionein 1M and 1G in response to zinc in sertoli TM4 cells.
Zinc (Zn) as an important trace element is essential for testicular development and spermatogenesis. Molecular mechanism of Zn action in the reproductive system may be related to metal binding low-molecular weight proteins, metallothioneins (MT). Our objective was to determine the effect of Zn on two important isoforms of MT, MT1M and MT1G genes expression on testicular sertoli cells. Cultured sertoli TM4 cells were exposed to different concentrations of Zn at different time points. Cellular uptake of Zn was tested using flame atomic absorption spectrometry. The cellular viability and gene expression were assessed by MTT and real-time PCR methods, respectively. The treated cells resulted in higher Zn concentration and cellular viability. The expression of MT1M and MT1G genes in the treated cells were greater than those of the untreated cells (P less than 0.05). In the high dosage treated group (100 and 500 muM), Zn concentration and expression of MT1M and MT1G genes increased three h after treatment; MT1G gene expression increased more at sixth h. At 18th h of treatment, the expression of both genes especially MT1G, increased dramatically while Zn concentration decreased. Since the increase of MT1G mRNA was coincident with cellular Zn level, it seems that MT1G has a more prominent role than MT1M in the homeostasis of Zn. In addition, Zn at dosage of 50 muM (pharmacologic concentration) may protect cells by increasing the expression of MT genes at longer periods.


Roll-to-roll cohesive, coated, flexible, high-efficiency polymer light-emitting diodes utilizing ITO-free polymer anodes.
This paper reports solution-processed, high-efficiency polymer light-emitting diodes fabricated by a new type of roll-to-roll coating method under ambient air conditions. A noble roll-to-roll cohesive coating system utilizes only natural gravity and the surface tension of the solution to flow out from the capillary to the surface of the substrate. Because this mechanism uses a minimally cohesive solution, the roll-to-roll cohesive coating can effectively realize an ultra-thin film thickness for the electron injection layer. In addition, the roll-to-roll cohesive coating enables the fabrication of a thicker polymer anode film more than 250 nm at one time by modification of the surface energy and without wasting the solution. It is observed that the standard sheet resistance deviation of the polymer anode is only 2.32 Ω/□ over 50 000 bending cycles. The standard sheet resistance deviation of the polymer anode in the different bending angles (0 to 180°) is 0.313 Ω/□, but the case of the ITO-PET is 104.93 Ω/□. The average surface roughness of the polymer anode measured by atomic force microscopy is only 1.06 nm. Because the surface of the polymer anode has a better quality, the leakage current of the polymer light-emitting diodes (PLEDs) using the polymer anode is much lower than that using the ITO-PET substrate. The luminous power efficiency of the two devices is 4.13 lm/W for the polymer anode and 3.21 lm/W for the ITO-PET. Consequently, the PLEDs made by using the polymer anode exhibited 28% enhanced performance because the polymer anode represents not only a higher transparency than the ITO-PET in the wavelength of 560 nm but also greatly reduced roughness. The optimized the maximum current efficiency and power efficiency of the device show around 6.1 cd/A and 5.1 lm/W, respectively, which is comparable to the case of using the ITO-glass.


Physician awareness regarding contraceptive counselling in women with chronic disease.
Contraceptive counselling is essential in women with chronic disease, though it is frequently overlooked. We aimed to assess practices and awareness in contraceptive counselling in hospital physicians caring for reproductive age women with chronic disease. A questionnaire was distributed in April 2017 to physicians from 16 medical specialties in a tertiary care hospital in Portugal. Statistical analysis performed using Microsoft Office Excel® and IBM SPSS-Statistics®. Two-hundred physicians participated in the survey: 59.5% were female, 55% under 35 years-old and 48.5% were residents. Only 26.5% practiced in surgical specialties. Two-thirds consistently inquired about the use of contraception. Most referred patients to Family Planning services or performed contraceptive counselling when pregnancy posed significant health risks (89.0%), when prescribing teratogenic medication (79.5%) or drugs with potential interaction with contraceptives (75.0%). Lack of training in contraception was the main reason for not referring patients to Family Planning services. While 83% considered counselling very important, only 5% had recently undergone training and 12.5% felt knowledgeable enough to perform it. Only 12.5% were aware of the institution's Family Planning resources. Female gender, non-surgical specialty, age over 35 years-old and over 10 years since specialisation were positively associated with awareness. Most physicians addressed contraceptive management in women with comorbidities, but adequate training is lacking. Our results highlight the importance of communication between health care providers to achieve an adequate and multidisciplinary approach to reproductive and general health goals.


A Comparison of Solar Ultraviolet Radiation Exposure in Urban Canyons in Venice, Italy and Johannesburg, South Africa.
Urban environments can have high-risk spaces that can provide excess personal sun exposure, such as urban or street canyons, and the spaces between buildings, among others. In these urban spaces, sun exposure can be high or low depending on several factors. Polysulphone film (PSF) was used to assess possible daily solar ultraviolet radiation (UVR) exposure in urban canyons in Venice, Italy and, for the first time in Africa, in Johannesburg, South Africa. The photodegradation of PSF upon solar exposure was monitored at a wavelength of 330 nm by ultraviolet-visible spectrophotometry, and the resultant change was converted to standard erythemal dose (SED) units (1 SED = 100 J m-2 ). Mean daily ambient solar UVR exposure measured for Venice and Johannesburg ranged between 20-28 SED and 33-43 SED, respectively. Canyon-located PSF exposures were lower in Venice (1-9 SED) than those in Johannesburg (9-39 SED), depending mainly on the sky view factor and orientation to the sun. There was large variation in solar UVR exposure levels in different urban canyons. These preliminary results should be bolstered with additional studies for a better understanding of excess personal exposure risk in urban areas, especially in Africa.


Molecular Response to Neoadjuvant Chemotherapy in High-Grade Serous Ovarian Carcinoma.
While high-grade serous ovarian carcinoma (HGSOC) is the most common histologic subtype of ovarian cancer, significant tumor heterogeneity exists. In addition, chemotherapy induces changes in gene expression and alters the mutational profile. To evaluate the notion that patients with HGSOC could be better classified for optimal treatment based on gene expression, we compared genetic variants [by DNA next-generation sequencing (NGS) using a 50 gene Ion Torrent panel] and gene expression (using the NanoString PanCancer 770 gene Panel) in the tumor from 20 patients with HGSOC before and after neoadjuvant chemotherapy (NACT). NGS was performed on plasma cell free DNA (cfDNA) on a select group of patients (n = 14) to assess the utility of using cfDNA to monitor these changes. A total of 86 genes had significant changes in RNA expression after NACT. Thirty-eight genetic variants (including SNPs) from 6 genes were identified in tumors pre-NACT, while 59 variants from 19 genes were detected in the cfDNA. The number of DNA variants were similar after NACT. Of the 59 variants in the plasma pre-NACT, only 6 persisted, whereas 33 of 38 specific variants in the tumor DNA remained unchanged. Pathway analysis showed the most significant alterations in the cell cycle and DNA damage pathways.Implications: Gene expression profiles at the time of interval debulking provide additional genetic information that could help impact treatment decisions after NACT; although, continued collection and analysis of matched tumor and cfDNA from multiple time points are needed to determine the role of cfDNA in the management of HGSOC. Mol Cancer Res; 16(5); 813-24. ©2018 AACR.


Atypical presentations of multiple evanescent white dot syndrome without white dots: a case series.
To describe three unusual clinical cases of patients who were diagnosed with multiple evanescent white dot syndrome at the onset of their disease but lacked the characteristic white dots on clinical examination, autofluorescence, and angiographic testing at any time during their follow-up. Three eyes of three patients with clinical symptoms of multiple evanescent white dot syndrome seen at an academic clinical practice were included. Retinal imaging, including fluorescein angiography, indocyanine green imaging, fundus autofluorescence, and optical coherence tomography, was used to further characterize and describe the clinical findings. All three patients presented with classic multiple evanescent white dot syndrome complaints. All patients demonstrated classic foveal granularity and mild disk swelling and/or peripapillary whitening. No characteristic hyperfluorescent lesions were seen on fluorescein angiography, and indocyanine green imaging did not demonstrate typical patterns of hypofluorescence and/or leakage. Optical coherence tomography documented focal disruption or loss of the inner segment/outer segment photoreceptor line in all cases. Our case series demonstrates another atypical presentation of multiple evanescent white dot syndrome that has not been described previously: foveal involvement at the onset of the disease without white spots.


Malignant transformation of cultured rat hepatocytes by (+)- and (-)-trans-7,8-dihydro-7,8-dihydroxybenzo(a)pyrene.
Rat hepatocyte cultures (RL-PR-C) were tested with the (+)- or (-)-trans-7,8-dihydrodiol isomers of benzo(a)pyrene [B(a)P] and were assessed for growth inhibition, chromosomal damage, growth in soft agar, and tumor formation. Because early-passage cells have a noninducible low specific activity, aryl hydrocarbon hydroxylase parallel inhibition studies were performed on aryl hydrocarbon hydroxylase-inducible late-passage cultures. Early-passage cells exhibited little inhibition in the presence of B(a)P or either isomer. Comparable later-passage cells demonstrated inhibitory effects with B(a)P and the (+)- and (-)-trans-7,8-dihydrodiol metabolites. No treated cultures grew in soft agar, and the modal number of chromosomes was unaffected by carcinogen treatment. However, both (+) and (-) isomer-treated early-passage cells formed tumors in isogeneic animals, the (+) isomer being more efficient in this regard. These results indicate that noninhibitory doses of either the (+)- or (-)-trans-7,8-dihydrodiol isomer of B(a)P are nonetheless capable of malignantly transforming hepatocyte cells in vitro.


Incidence of Postoperative Deep Venous Thrombosis Is Higher among Cardiac and Vascular Surgery Patients as Compared with General Surgery Patients.
Unlike general surgery patients, most of vascular and cardiac surgery patients receive therapeutic anticoagulation during operations. The purpose of this study was to report the incidence of deep venous thrombosis (DVT) among cardiac and vascular surgery patients, compared with general surgery. The American College of Surgeons National Surgical Quality Improvement Program database was queried for all patients who underwent surgical procedures from 2005 to 2010. Patients who developed DVT within 30 days of an operation were identified. The incidence of DVT was compared among vascular, general, and cardiac surgery patients. Risk factors for developing postoperative DVT were identified and compared among these patients. Of total 2,669,772 patients underwent surgical operations in the period between 2005 and 2010. Of all the patients, 18,670 patients (0.69%) developed DVT. The incidence of DVT among different surgical specialties was cardiac surgery (2%), vascular surgery (0.99%), and general surgery (0.66%). The odds ratio for developing DVT was 1.5 for vascular surgery patients and 3 for cardiac surgery patients, when compared with general surgery patients (P < 0.001). The odds ratio for developing DVT after cardiac surgery was 2, when compared with vascular surgery (P < 0.001). The incidence of DVT is higher among vascular and cardiac surgery patients as compared with that of general surgery patients. Intraoperative anticoagulation does not prevent the occurrence of DVT in the postoperative period. These patients should receive DVT prophylaxis in the perioperative period, similar to other surgical patients according to evidence-based guidelines.


3D Co-culture of hiPSC-Derived Cardiomyocytes With Cardiac Fibroblasts Improves Tissue-Like Features of Cardiac Spheroids.
Purpose: Both cardiomyocytes and cardiac fibroblasts (CF) play essential roles in cardiac development, function, and remodeling. Properties of 3D co-cultures are incompletely understood. Hence, 3D co-culture of cardiomyocytes and CF was characterized, and selected features compared with single-type and 2D culture conditions. Methods: Human cardiomyocytes derived from induced-pluripotent stem cells (hiPSC-CMs) were obtained from Cellular Dynamics or Ncardia, and primary human cardiac fibroblasts from ScienCell. Cardiac spheroids were investigated using cryosections and whole-mount confocal microscopy, video motion analysis, scanning-, and transmission-electron microscopy (SEM, TEM), action potential recording, and quantitative PCR (qPCR). Results: Spheroids formed in hanging drops or in non-adhesive wells showed spontaneous contractions for at least 1 month with frequent media changes. SEM of mechanically opened spheroids revealed a dense inner structure and no signs of blebbing. TEM of co-culture spheroids at 1 month showed myofibrils, intercalated disc-like structures and mitochondria. Ultrastructural features were comparable to fetal human myocardium. We then assessed immunostained 2D cultures, cryosections of spheroids, and whole-mount preparations by confocal microscopy. CF in co-culture spheroids assumed a small size and shape similar to the situation in ventricular tissue. Spheroids made only of CF and cultured for 3 weeks showed no stress fibers and strongly reduced amounts of alpha smooth muscle actin compared to early spheroids and 2D cultures as shown by confocal microscopy, western blotting, and qPCR. The addition of CF to cardiac spheroids did not lead to arrhythmogenic effects as measured by sharp-electrode electrophysiology. Video motion analysis showed a faster spontaneous contraction rate in co-culture spheroids compared to pure hiPSC-CMs, but similar contraction amplitudes and kinetics. Spontaneous contraction rates were not dependent on spheroid size. Applying increasing pacing frequencies resulted in decreasing contraction amplitudes without positive staircase effect. Gene expression analysis of selected cytoskeleton and myofibrillar proteins showed more tissue-like expression patterns in co-culture spheroids than with cardiomyocytes alone or in 2D culture. Conclusion: We demonstrate that the use of 3D co-culture of hiPSC-CMs and CF is superior over 2D culture conditions for co-culture models and more closely mimicking the native state of the myocardium with relevance to drug development as well as for personalized medicine.


The effects of prolonged epinephrine infusion on the physiology of the rainbow trout, Salmo gairdneri. I. Blood respiratory, acid-base and ionic states.
Rainbow trout were infused continuously for 24 h with epinephrine in order to elevate circulating levels of this hormone to those measured during periods of acute extracellular acidosis (approximately 5 X 10(-8) mol l-1). Concomitant effects on selected blood respiratory acid-base and ionic variables were evaluated. Infusion of epinephrine caused a transient respiratory acidosis as a result of hypoventilation and/or inhibition of red blood cell (RBC) bicarbonate dehydration. The acidosis was regulated by gradual accumulation of plasma bicarbonate. Even though whole blood pH (pHe) was depressed by 0.16 units, RBC pH (pHi) remained constant, thereby causing the transmembrane pH gradient (pHe-pHi) to decrease. A similar effect of epinephrine on RBC pH was observed in vitro, although the response required a higher concentration of epinephrine (2.0 X 10(-7) mol l-1). We speculate that the release of epinephrine during periods of depressed blood pH is important for preventing excessive shifts in RBC pH and for initiating a series of responses leading to plasma HCO3- accumulation and eventual restoration of blood acid-base status.


Enrichment of in vivo transcription data from dietary intervention studies with in vitro data provides improved insight into gene regulation mechanisms in the intestinal mucosa.
Gene expression profiles of intestinal mucosa of chickens and pigs fed over long-term periods (days/weeks) with a diet rich in rye and a diet supplemented with zinc, respectively, or of chickens after a one-day amoxicillin treatment of chickens, were recorded recently. Such dietary interventions are frequently used to modulate animal performance or therapeutically for monogastric livestock. In this study, changes in gene expression induced by these three interventions in cultured "Intestinal Porcine Epithelial Cells" (IPEC-J2) recorded after a short-term period of 2 and 6 hours, were compared to the in vivo gene expression profiles in order to evaluate the capability of this in vitro bioassay in predicting in vivo responses. Lists of response genes were analysed with bioinformatics programs to identify common biological pathways induced in vivo as well as in vitro. Furthermore, overlapping genes and pathways were evaluated for possible involvement in the biological processes induced in vivo by datamining and consulting literature. For all three interventions, only a limited number of identical genes and a few common biological processes/pathways were found to be affected by the respective interventions. However, several enterocyte-specific regulatory and secreted effector proteins that responded in vitro could be related to processes regulated in vivo, i.e. processes related to mineral absorption, (epithelial) cell adherence and tight junction formation for zinc, microtubule and cytoskeleton integrity for amoxicillin, and cell-cycle progression and mucus production for rye. Short-term gene expression responses to dietary interventions as measured in the in vitro bioassay have a low predictability for long-term responses as measured in the intestinal mucosa in vivo. The short-term responses of a set regulatory and effector genes, as measured in this bioassay, however, provided additional insight into how specific processes in piglets and broilers may be modulated by "early" signalling molecules produced by enterocytes. The relevance of this set of regulatory/effector genes and cognate biological processes for zinc deficiency and supplementation, gluten allergy (rye), and amoxicillin administration in humans is discussed.


High-dose chemoradiotherapy and anti-B-cell monoclonal antibody-purged autologous bone marrow transplantation in mantle-cell lymphoma: no evidence for long-term remission.
The role for high-dose therapy and autologous stem-cell transplantation in mantle-cell lymphoma (MCL) is unknown. We retrospectively analyzed patients with chemosensitive disease who underwent high-dose chemoradiotherapy and anti-B-cell monoclonal antibody-purged autologous bone marrow transplantation (ABMT) for MCL in first remission, as well as following relapse from conventional therapy. Between August 1985 and April 1996, 28 patients underwent ABMT using a uniform ablative regimen with cyclophosphamide and total-body irradiation (TBI) and a bone marrow-purging regimen. Re-review of original tissue demonstrated that all patients had morphologic, phenotypic, and genotypic characteristics of MCL. MCL was the original diagnosis in 21 patients, whereas seven patients had a prior diagnosis of diffuse small cleaved-cell lymphoma. Twenty patients received multiple regimens before ABMT, while eight underwent ABMT in first complete remission (CR)/partial remission (PR) following CHOP induction. At bone marrow harvest, only 18% of patients were in CR and overt BM infiltration was present in 57%. Following cyclophosphamide/TBI, no treatment-related deaths were seen. Nineteen of 28 patients have relapsed at a median time of 21 months (range, 3 to 70). Of eight patients transplanted in first CR/PR, five have relapsed. Nine patients are in continuous CR with a median follow-up time of 24 months (range, 10 to 135). Disease-free survival (DFS) and overall survival (OS) are estimated to be 31% and 62% at 4 years, respectively. ABMT using cyclophosphamide/TBI conditioning may at best be effective in only a small fraction of patients with relapsed MCL. The lack of plateau with a median follow-up time of 24 months suggests cure may not be achievable. The role of this therapy in patients in first remission requires more study using better induction therapy to enhance the CR rate before ABMT.


[Chemical modification of lysine residues of beta-1,3-glucanase from Spisula sachalinensis].
The effects of chemical modification of the amino groups of lysine residues on the activity of beta-1.3-glucanase from Spisula sachalinensis were studied. Modification of two lysine residues per molecule did not affect either the enzyme activity with respect to laminarine, nor the Km value. The modified beta-1.3-glucanase retains the ability to catalyze the transglycosylation and cleaves the high molecular weight CM-pachyman at the same rate as does the native enzyme. No significant changes in the enzyme thermal stability were observed. Thus, the modified enzyme groups cannot be involved in the enzyme active center and are exposed on the surface of the protein globule. The chemical modification was shown to have no effect on the enzyme kinetics, which is essential for its immobilization.


A new method for studying population genetics of cyst nematodes based on Pool-Seq and genomewide allele frequency analysis.
Cyst nematodes are important agricultural pests responsible for billions of dollars of losses each year. Plant resistance is the most effective management tool, but it requires a close monitoring of population genetics. Current technologies for pathotyping and genotyping cyst nematodes are time-consuming, expensive and imprecise. In this study, we capitalized on the reproduction mode of cyst nematodes to develop a simple population genetic analysis pipeline based on genotyping-by-sequencing and Pool-Seq. This method yielded thousands of SNPs and allowed us to study the relationships between populations of different origins or pathotypes. Validation of the method on well-characterized populations also demonstrated that it was a powerful and accurate tool for population genetics. The genomewide allele frequencies of 23 populations of golden nematode, from nine countries and representing the five known pathotypes, were compared. A clear separation of the pathotypes and fine genetic relationships between and among global populations were obtained using this method. In addition to being powerful, this tool has proven to be very time- and cost-efficient and could be applied to other cyst nematode species.


Purification and aminopyrine monooxygenase activity of liver microsomal cytochrome P-450 from alloxan-induced diabetic rats.
We purified two diabetes-inducible and insulin-sensitive forms of cytochrome P-450, named P-450AL-1 and AL-2, from the liver microsomes of alloxan-diabetic male rats, using sodium cholate solubilization, octylamino-Sepharose 4B chromatography, and HPLC with diethylaminoethyl-5PW and hydroxyapatite columns. The purified forms gave a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular weight of 50,000 for P-450AL-1 or 48,500 for P-450AL-2. The CO-reduced spectral maximum of these forms was at 452 nm for P-450AL-1 and 451 nm for P-450AL-2. The two purified forms had the low-spin state of heme in the oxidized form. Both P-450AL-1 and AL-2 were active in the metabolism of aniline, benzphetamine, and 7-ethoxycoumarin. However, the catalytic activity of P-450AL-2 for these substrates was obviously higher than that of AL-1. The NH2-terminal sequences of P-450AL-1 and AL-2 differed from each other, and did not agree with those of the other P-450 forms purified from diabetic rats previously. Furthermore, we examined the metabolism of aminopyrine in a reconstituted system with the purified cytochromes P-450. The diabetes-inducible forms of P-450 had high aminopyrine 3-hydroxylation and low N-demethylation activities. These findings provide clear evidence supporting our previous results, which have shown an increase in 3-hydroxymethyl-2-methyl-4-dimethylamino-1-phenyl-3-pyrazolin-5-on e and a decrease in 4-monomethylaminoantipyrine in intact diabetic rats.


[Gitelman syndrome: a rare cause of hypokalemia].
We report a case of hypokaliemia whose origin is a urinary potassium loss associated with syndrome of Gitelman. The syndrome of Gitelman is a rare familiar autosomal recessive nephropathy. We will discuss the pathophysiology of this syndrome and differential diagnoses.


Sleep-disordered breathing and cardiovascular disease: cross-sectional results of the Sleep Heart Health Study.
Disordered breathing during sleep is associated with acute, unfavorable effects on cardiovascular physiology, but few studies have examined its postulated association with cardiovascular disease (CVD). We examined the cross-sectional association between sleep- disordered breathing and self-reported CVD in 6,424 free-living individuals who underwent overnight, unattended polysomnography at home. Sleep-disordered breathing was quantified by the apnea-hypopnea index (AHI)-the average number of apneas and hypopneas per hour of sleep. Mild to moderate disordered breathing during sleep was highly prevalent in the sample (median AHI: 4.4; interquartile range: 1.3 to 11.0). A total of 1,023 participants (16%) reported at least one manifestation of CVD (myocardial infarction, angina, coronary revascularization procedure, heart failure, or stroke). The multivariable-adjusted relative odds (95% CI) of prevalent CVD for the second, third, and fourth quartiles of the AHI (versus the first) were 0.98 (0.77-1.24), 1.28 (1.02-1.61), and 1.42 (1.13-1.78), respectively. Sleep-disordered breathing was associated more strongly with self-reported heart failure and stroke than with self-reported coronary heart disease: the relative odds (95% CI) of heart failure, stroke, and coronary heart disease (upper versus lower AHI quartile) were 2.38 (1.22-4.62), 1.58 (1.02- 2.46), and 1.27 (0.99-1.62), respectively. These findings are compatible with modest to moderate effects of sleep-disordered breathing on heterogeneous manifestations of CVD within a range of AHI values that are considered normal or only mildly elevated.


[Anaemia in hospital patients: transfusion or not?]
Increasingly restrictive red blood cell transfusion strategies have led to growing numbers of patients being discharged from hospital with moderate anaemia, and doubts about the effects of this trend on the outcomes of patients with anaemia are growing. One advantage of doubts is that they lead to variation in practice, which, in theory, allows the study of routinely collected data on the effects of an intervention (transfusion) on patient-relevant outcomes. To do this in a reliable way, we need solid data on outcomes and on all the potentially relevant clinical characteristics that are associated with both the decision to transfuse and the outcomes. Currently, such data are not available. As part of routine practice, clinicians should be encouraged to document the arguments for their treatment decisions and the intended outcomes. This will allow learning from daily practice in the field of transfusion medicine and in other health care disciplines.


Angiotension receptor blockers reduce the risk of dementia.
Recent studies implied that angiotension receptor blockers (ARBs) not only have an antihypertensive effect but also have beneficial effects on dementia. The purpose of this study was to investigate the effects of ARBs on dementia and the subtypes. We conducted a population-based cohort study with data from the Taiwan National Health Insurance Research Database. A total of 24 531 matching pairs (1 : 1) of ARB-exposed and non-ARB-exposed patients were included. Each patient was individually tracked from 1997 to 2009 to identify incident cases of dementia (onset in 1999 or later). Cox proportional hazard regressions were employed to calculate the hazard ratios and 95% confidence intervals (CIs) for the association between ARBs and dementia, Alzheimer's disease and vascular dementia, conditional for matching pairs. There were 1322 cases (5.4%) of dementia in the ARB cohort and 2181 cases (8.9%) in the non-ARB cohort identified during the 11-year follow-up period. The multivariate-adjusted hazard ratios for dementia, Alzheimer's disease and vascular dementia were 0.54 (95% CI 0.51-0.59), 0.53 (95% CI 0.43-0.64) and 0.63 (95% CI 0.54-0.73) for patients with ARB treatments, respectively. In terms of cumulative dosage, patients with more than 1460 defined daily dose of ARBs had less risk than those patients with less than 1460 defined daily dose (hazard ratio 0.37 vs. 0.61; P < 0.05). These results suggest that ARB may be associated with a reduced risk of dementia in high vascular-risk individuals. Patients exposed to ARBs for higher cumulative doses experienced more protection from dementia and the subtypes.


Epidemiology of Posterior Glenohumeral Instability in a Young Athletic Population.
While several studies have observed the incidence of posterior glenohumeral instability in selected populations, there are no data from large-scale population-based studies with corresponding athletic exposure data to calculate incidence rates (IRs) and associated risk factors. To determine risk factors for posterior glenohumeral instability within the physically active population at the United States Military Academy. Descriptive epidemiology study. A longitudinal cohort study was performed over a 6-year period from 2006 to 2012 at the United States Military Academy utilizing the Cadet Illness and Injury Tracking System. Exposure data were collected from daily attendance data documented for every practice and game at the intramural, club, or varsity sport level. The primary outcomes calculated were the IRs of posterior glenohumeral joint instability per 1000 person-years at risk and per 1000 athlete-exposures. IR ratios and confidence intervals were calculated between male and female cadets and between intercollegiate and intramural athletes. Between 2006 and 2012, there were 1348 shoulder injuries in total, with 633 instability events. During the study period, 113 posterior shoulder instability injuries (17.9% of instability events) and 26,408 person-years at risk were documented, for an overall IR of 4.28 per 1000 person-years. The overall IR of posterior instability was 0.08 per 1000 athlete exposures. In male athletes, 105 sustained a posterior shoulder instability event, for an IR of 4.67 per 1000 person-years. Female athletes were less likely to sustain a posterior instability event, accounting for only 8 events, for an IR of 2.04 per 1000 person-years. A total of 6670 athletes were at risk for posterior shoulder instability injury during the study period. A total of 55 injuries were documented, for an IR of 8.25 per 1000 person-years. Intercollegiate wrestlers had the highest IR: 34.15 per 1000 person-years. The sport with the highest injury rate among intramural sports was football, at 2.79 per 1000 person-years. Three athletes had a history of a posterior shoulder dislocation, while no significant difference was found for subluxation versus pain as the presenting symptom. Of the 113 posterior shoulder instability injuries, 77.0% required surgical stabilization. All injuries associated with weight lifting required surgical stabilization. Among athletes, intercollegiate athletes are at an increased risk of posterior glenohumeral instability when compared with intramural athletes. Intercollegiate athletes and weight lifters demonstrate a high likelihood of requiring surgical treatment as compared with intramural athletes.


In vitro cultivation of the hymenoptera genetic model, Nasonia.
The wasp genus Nasonia is a genetic model with unique advantages for the study of interspecific differences, including haplodiploidy and interfertile species. However, as a parasitoid, Nasonia is confined within a fly host, thus restricting direct observations and manipulation of development over time. Here, we present the first in vitro cultivation method for this system that decouples Nasonia from its host, allowing continuous observations from embryo to adulthood. Using transwell plates and a simple Nasonia rearing medium, we demonstrate a technique that will significantly expand the utility of the Nasonia model.


A suite of Lotus japonicus starch mutants reveals both conserved and novel features of starch metabolism.
The metabolism of starch is of central importance for many aspects of plant growth and development. Information on leaf starch metabolism other than in Arabidopsis (Arabidopsis thaliana) is scarce. Furthermore, its importance in several agronomically important traits exemplified by legumes remains to be investigated. To address this issue, we have provided detailed information on the genes involved in starch metabolism in Lotus japonicus and have characterized a comprehensive collection of forward and TILLING (for Targeting Induced Local Lesions IN Genomes) reverse genetics mutants affecting five enzymes of starch synthesis and two enzymes of starch degradation. The mutants provide new insights into the structure-function relationships of ADP-glucose pyrophosphorylase and glucan, water dikinase1 in particular. Analyses of the mutant phenotypes indicate that the pathways of leaf starch metabolism in L. japonicus and Arabidopsis are largely conserved. However, the importance of these pathways for plant growth and development differs substantially between the two species. Whereas essentially starchless Arabidopsis plants lacking plastidial phosphoglucomutase grow slowly relative to wild-type plants, the equivalent mutant of L. japonicus grows normally even in a 12-h photoperiod. In contrast, the loss of GLUCAN, WATER DIKINASE1, required for starch degradation, has a far greater effect on plant growth and fertility in L. japonicus than in Arabidopsis. Moreover, we have also identified several mutants likely to be affected in new components or regulators of the pathways of starch metabolism. This suite of mutants provides a substantial new resource for further investigations of the partitioning of carbon and its importance for symbiotic nitrogen fixation, legume seed development, and perenniality and vegetative regrowth.


Analysis of Unplanned Postoperative Admissions to the Intensive Care Unit.
To investigate factors associated with unplanned postoperative admissions to the intensive care unit (ICU). Data from the National Anesthesia Clinical Outcomes Registry (NACOR) were analyzed. We performed univariate and multivariate logistic regression to identify patient- and surgery-specific characteristics associated with unplanned postoperative ICU admission. We also recorded the prevalence of Current Procedural Terminology (CPT) and International Classification of Diseases, ninth revision ( ICD-9) billing codes and outcomes for unplanned postoperative ICU admissions. Of 23 341 130 records in the database, 2 910 738 records met our inclusion criteria. A higher American Society of Anesthesiologists physical status (ASA PS) class, case duration greater than 4 hours, and advanced age were associated with a greater likelihood of unplanned ICU admission. Vascular and thoracic surgery patients were more likely to have an unplanned ICU admission. The most common CPT and ICD-9 codes involved repair of femur/hip fracture, bowel resection, and acute postoperative pain. Large community hospitals were more likely than university hospitals to have unplanned postoperative ICU admissions. Patients in the unplanned postoperative ICU admission group were more likely to have experienced intraoperative cardiac arrest, hemodynamic instability, or respiratory failure and were more likely to die in the immediate perioperative period. Our study is the first diverse analysis of unplanned postoperative ICU admissions in the literature across multiple specialties and practice models. We found an association of advanced age, higher ASA PS class, and duration of procedure with unplanned ICU admission after surgery. Surgical specialties and procedures with the most unplanned ICU admissions could be areas for quality improvement and clinical pathways in the future.


The NHERF1 PDZ2 domain regulates PKA-RhoA-p38-mediated NHE1 activation and invasion in breast tumor cells.
Understanding the signal transduction systems governing invasion is fundamental for the design of therapeutic strategies against metastasis. Na(+)/H(+) exchanger regulatory factor (NHERF1) is a postsynaptic density 95/disc-large/zona occludens (PDZ) domain-containing protein that recruits membrane receptors/transporters and cytoplasmic signaling proteins into functional complexes. NHERF1 expression is altered in breast cancer, but its effective role in mammary carcinogenesis remains undefined. We report here that NHERF1 overexpression in human breast tumor biopsies is associated with metastatic progression, poor prognosis, and hypoxia-inducible factor-1alpha expression. In cultured tumor cells, hypoxia and serum deprivation increase NHERF1 expression, promote the formation of leading-edge pseudopodia, and redistribute NHERF1 to these pseudopodia. This pseudopodial localization of NHERF1 was verified in breast biopsies and in three-dimensional Matrigel culture. Furthermore, serum deprivation and hypoxia stimulate the Na(+)/H(+) exchanger, invasion, and activate a protein kinase A (PKA)-gated RhoA/p38 invasion signal module. Significantly, NHERF1 overexpression was sufficient to induce these morphological and functional changes, and it potentiated their induction by serum deprivation. Functional experiments with truncated and binding groove-mutated PDZ domain constructs demonstrated that NHERF1 regulates these processes through its PDZ2 domain. We conclude that NHERF1 overexpression enhances the invasive phenotype in breast cancer cells, both alone and in synergy with exposure to the tumor microenvironment, via the coordination of PKA-gated RhoA/p38 signaling.


Antibody-independent capture of circulating tumor cells of non-epithelial origin with the ApoStream® system.
Circulating tumor cells (CTCs) are increasingly employed for research and clinical monitoring of cancer, though most current methods do not permit the isolation of non-epithelial tumor cells. Furthermore, CTCs isolated with antibody-dependent methods are not suitable for downstream experimental uses, including in vitro culturing and implantation in vivo. In the present study, we describe the development, validation, and transfer across laboratories of a new antibody-independent device for the enrichment of CTCs from blood samples of patients with various cancer diagnoses. The ApoStream® device uses dielectrophoresis (DEP) field-flow assist to separate non-hematopoietic cells from the peripheral blood mononuclear fraction by exposing cells in a laminar flow stream to a critical alternating current frequency. The ApoStream® device was calibrated and validated in a formal cross-laboratory protocol using 3 different cancer cell lines spanning a range of distinct phenotypes (A549, MDA-MB-231, and ASPS-1). In spike-recovery experiments, cancer cell recovery efficiencies appeared independent of spiking level and averaged between 68% and 55%, depending on the cell line. No inter-run carryover was detected in control samples. Moreover, the clinical-readiness of the device in the context of non-epithelial cancers was evaluated with blood specimens from fifteen patients with metastatic sarcoma. The ApoStream® device successfully isolated CTCs from all patients with sarcomas examined, and the phenotypic heterogeneity of the enriched cells was demonstrated by fluorescence in situ hybridization or with multiplex immunophenotyping panels. Therefore, the ApoStream® technology expands the clinical utility of CTC evaluation to mesenchymal cancers.


Quercetin and Related Analogs as Therapeutics to Promote Tissue Repair.
Quercetin is a polyphenol of the flavonoid class of secondary metabolites that is widely distributed in the plant kingdom. Quercetin has been found to exhibit potent bioactivity in the areas of wound healing, neuroprotection, and anti-aging research. Naturally found in highly glycosylated forms, aglycone quercetin has low solubility in aqueous environments, which has heavily limited its clinical applications. To improve the stability and bioavailability of quercetin, efforts have been made to chemically modify quercetin and related flavonoids so as to improve aqueous solubility while retaining bioactivity. In this review, we provide an updated overview of the biological properties of quercetin and proposed mechanisms of actions in the context of wound healing and aging. We also provide a description of recent developments in synthetic approaches to improve the solubility and stability of quercetin and related analogs for therapeutic applications. Further research in these areas is expected to enable translational applications to improve ocular wound healing and tissue repair.


ACAULIS5 controls Arabidopsis xylem specification through the prevention of premature cell death.
Cell size and secondary cell wall patterning are crucial for the proper functioning of xylem vessel elements in the vascular tissues of plants. Through detailed anatomical characterization of Arabidopsis thaliana hypocotyls, we observed that mutations in the putative spermine biosynthetic gene ACL5 severely affected xylem specification: the xylem vessel elements of the acl5 mutant were small and mainly of the spiral type, and the normally predominant pitted vessels as well as the xylem fibers were completely missing. The cell-specific expression of ACL5 in the early developing vessel elements, as detected by in situ hybridization and reporter gene analyses, suggested that the observed xylem vessel defects were caused directly by the acl5 mutation. Exogenous spermine prolonged xylem element differentiation and stimulated cell expansion and cell wall elaboration in xylogenic cell cultures of Zinnia elegans, suggesting that ACL5 prevents premature death of the developing vessel elements to allow complete expansion and secondary cell wall patterning. This was further supported by our observations that the vessel elements of acl5 seemed to initiate the cell death program too early and that the xylem defects associated with acl5 could be largely phenocopied by induction of premature, diphtheria toxin-mediated cell death in the ACL5-expressing vessel elements. We therefore provide, for the first time, mechanistic evidence for the function of ACL5 in xylem specification through its action on the duration of xylem element differentiation.


A rapamycin-sensitive signaling pathway is essential for the full expression of persistent pain states.
Translational control through the mammalian target of rapamycin (mTOR) is critical for synaptic plasticity, cell growth, and axon guidance. Recently, it was also shown that mTOR signaling was essential for the maintenance of the sensitivity of subsets of adult sensory neurons. Here, we show that persistent pain states, but not acute pain behavior, are substantially alleviated by centrally administered rapamycin, an inhibitor of the mTOR pathway. We demonstrate that rapamycin modulates nociception by acting on subsets of primary afferents and superficial dorsal horn neurons to reduce both primary afferent sensitivity and central plasticity. We found that the active form of mTOR is present in a subpopulation of myelinated dorsal root axons, but rarely in unmyelinated C-fibers, and heavily expressed in the dorsal horn by lamina I/III projection neurons that are known to mediate the induction and maintenance of pain states. Intrathecal injections of rapamycin inhibited the activation of downstream targets of mTOR in dorsal horn and dorsal roots and reduced the thermal sensitivity of A-fibers. Moreover, in vitro studies showed that rapamycin increased the electrical activation threshold of Adelta-fibers in dorsal roots. Together, our results imply that central rapamycin reduces neuropathic pain by acting both on an mTOR-positive subset of A-nociceptors and lamina I projection neurons and suggest a new pharmacological route for therapeutic intervention in persistent pain states.


Multitarget Approach for the Treatment of Alzheimer's Disease: Inhibition of Phosphodiesterase 9 (PDE9) and Histone Deacetylases (HDACs) Covering Diverse Selectivity Profiles.
Here, we present a series of dual-target phosphodiesterase 9 (PDE9) and histone deacetylase (HDAC) inhibitors devised as pharmacological tool compounds for assessing the implications of these two targets in Alzheimer's disease (AD). These novel inhibitors were designed taking into account the key pharmacophoric features of known selective PDE9 inhibitors as well as privileged chemical structures, bearing zinc binding groups (hydroxamic acids and ortho-amino anilides) that hit HDAC targets. These substituents were selected according to rational criteria and previous knowledge from our group to explore diverse HDAC selectivity profiles (pan-HDAC, HDAC6 selective, and class I selective) that were confirmed in biochemical screens. Their functional response in inducing acetylation of histone and tubulin and phosphorylation of cAMP response element binding (CREB) was measured as a requisite for further progression into complete in vitro absorption, distribution, metabolism and excretion (ADME) and in vivo brain penetration profiling. Compound 31b, a selective HDAC6 inhibitor with acceptable brain permeability, was chosen for assessing in vivo efficacy of these first-in-class inhibitors, as well as studying their mode of action (MoA).


Retinal ganglion cell responses to voltage and current stimulation in wild-type and rd1 mouse retinas.
Retinal prostheses are being developed to restore vision for those with retinal diseases such as retinitis pigmentosa or age-related macular degeneration. Since neural prostheses depend upon electrical stimulation to control neural activity, optimal stimulation parameters for successful encoding of visual information are one of the most important requirements to enable visual perception. In this paper, we focused on retinal ganglion cell (RGC) responses to different stimulation parameters and compared threshold charge densities in wild-type and rd1 mice. For this purpose, we used in vitro retinal preparations of wild-type and rd1 mice. When the neural network was stimulated with voltage- and current-controlled pulses, RGCs from both wild-type and rd1 mice responded; however the temporal pattern of RGC response is very different. In wild-type RGCs, a single peak within 100 ms appears, while multiple peaks (approximately four peaks) with ∼ 10 Hz rhythm within 400 ms appear in RGCs in the degenerated retina of rd1 mice. We find that an anodic phase-first biphasic voltage-controlled pulse is more efficient for stimulation than a biphasic current-controlled pulse based on lower threshold charge density. The threshold charge densities for activation of RGCs both with voltage- and current-controlled pulses are overall more elevated for the rd1 mouse than the wild-type mouse. Here, we propose the stimulus range for wild-type and rd1 retinas when the optimal modulation of a RGC response is possible.


Unmethylated E-cadherin gene expression is significantly associated with metastatic human prostate cancer cells in bone.
The concurrent determination of methylation status of E-cadherin gene and E-cadherin protein expression remains scant in metastatic prostate cancer cells in bone, the most prevalent site for metastatic growth. Therefore, the study was undertaken to ascertain the methylation status of E-cadherin gene, a most frequent and known epigenetic mechanism of its regulation, and the protein expression in prostate tissue biopsy specimen. The methylation of E-cadherin gene was determined by methylation specific-PCR and the protein expression by immunohistochemical method in the consecutive sections of each prostate tissue biopsy specimen. The unmethylated E-cadherin gene and homogeneous E-cadherin protein expression was significantly associated with BPH as compared to the primary prostate carcinoma (Fisher's Exact P < 0.001). A significant association was observed between the concurrent methylated gene and markedly reduced expression of the protein in the primary prostate cancer cells as compared to the BPH cells, suggesting methylation-dependent regulation of the gene expression in these cases. In contrast to the primary cancer, a highly significant increase in the frequency of metastatic prostate cancer cells in bone exhibited the concurrent expression of unmethylated gene and homogeneous protein (Fisher's Exact P < 0.001). The study clearly demonstrated a significant association of the concurrent expression of unmethylated E-cadherin gene and E-cadherin protein with metastatic prostate cancer cells in bone, and that its expression may have a role in the intercellular adhesion in the formation of metastatic lesions in bone.


Privacy versus open science.
Pervasive internet and sensor technologies promise to revolutionize psychological science. However, the data collected using these technologies are often very personal-indeed, the value of the data is often directly related to how personal they are. At the same time, driven by the replication crisis, there is a sustained push to publish data to open repositories. These movements are in fundamental conflict. In this article, we propose a way to navigate this issue. We argue that there are significant advantages to be gained by ceding the ownership of data to the participants who generate the data. We then provide desiderata for a privacy-preserving platform. In particular, we suggest that researchers should use an interface to perform experiments and run analyses, rather than observing the stimuli themselves. We argue that this method not only improves privacy but will also encourage greater compliance with good research practices than is possible through open repositories.


Pooled rates and demographics of POTS following SARS-CoV-2 infection versus COVID-19 vaccination: Systematic review and meta-analysis.
To address recent concerns of postural orthostatic tachycardia syndrome (POTS) occurring after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and coronavirus disease 2019 (COVID-19) vaccination. We searched PubMed, Web of Science, and Scopus as of 1st June 2023. We performed a systematic review and meta-analysis of pooled POTS rate in SARS-CoV-2-infected and COVID-19-vaccinated groups from epidemiological studies, followed by subgroup analyses by characteristic. Meta-analysis of risk ratio was conducted to compare POTS rate in infected versus uninfected groups. Meta-analysis of demographics was also performed to compare cases of post-infection and post-vaccination POTS from case reports and series. We estimated the pooled POTS rate of 107.75 (95 % CI: 9.73 to 273.52) and 3.94 (95 % CI: 0 to 16.39) cases per 10,000 (i.e., 1.08 % and 0.039 %) in infected and vaccinated individuals based on 5 and 2 studies, respectively. Meta-regression revealed age as a significant variable influencing 86.2 % variance of the pooled POTS rate in infected population (P < 0.05). Moreover, POTS was 2.12-fold more likely to occur in infected than uninfected individuals (RR = 2.12, 95 % CI: 1.71 to 2.62, P < 0.001). Meta-analyzed demographics for cases of post-infection (n = 43) and post-vaccination (n = 17) POTS found no significant differences in several variables between groups, except that the time from exposure to symptom onset was shorter for cases of post-vaccination POTS (P < 0.05). Although evidence is limited for post-vaccination POTS, our study showed that POTS occur more frequently following SARS-CoV-2 infection than COVID-19 vaccination.


Gut feelings: the relations between depression, anxiety, psychotropic drugs and the gut microbiome.
The gut microbiome is involved in the bi-directional relationship of the gut - brain axis. As most studies of this relationship are small and do not account for use of psychotropic drugs (PTDs), we explored the relations of the gut microbiome with several internalizing disorders, while adjusting for PTDs and other relevant medications, in 7,656 Lifelines participants from the Northern Netherlands (5,522 controls and 491 participants with at least one internalizing disorder). Disorders included dysthymia, major depressive disorder (MDD), any depressive disorder (AnyDep: dysthymia or MDD), generalized anxiety disorder (GAD) and any anxiety disorder (AnyAnx: GAD, social phobia and panic disorder). Compared to controls, 17 species were associated with depressive disorders and 3 were associated with anxiety disorders. Around 90% of these associations remained significant (FDR <0.05) after adjustment for PTD use, suggesting that the disorders, not PTD use, drove these associations. Negative associations were observed for the butyrate-producing bacteria Ruminococcus bromii in participants with AnyDep and for Bifidobacterium bifidum in AnyAnx participants, along with many others. Tryptophan and glutamate synthesis modules and the 3,4-Dihydroxyphenylacetic acid synthesis module (related to dopamine metabolism) were negatively associated with MDD and/or dysthymia. After additional adjustment for functional gastrointestinal disorders and irritable bowel syndrome, these relations remained either statistically (FDR <0.05) or nominally (P < 0.05) significant. Overall, multiple bacterial species and functional modules were associated with internalizing disorders, including gut - brain relevant components, while associations to PTD use were moderate. These findings suggest that internalizing disorders rather than PTDs are associated with gut microbiome differences relative to controls.


Comparison of different polymerase chain reaction-based approaches for clonality assessment of immunoglobulin heavy-chain gene rearrangements in B-cell neoplasia.
Several frequently applied polymerase chain reaction strategies for analysis of immunoglobulin heavy-chain gene rearrangements were compared by analyzing 70 B-cell lymphoproliferative disorders and 24 reactive lymphoid lesions. Southern blot analysis was used as the "gold standard" for clonality assessment. For polymerase chain reaction analysis, primers directed against framework (FR) 3 (FR3-A and FR3-B), FR2, and FR1 of the variable gene segments and against joining gene segments of the immunoglobulin heavy-chain gene were used. Polymerase chain reaction products were analyzed by high-resolution fingerprinting analysis using radiolabeled nucleotides, gene scanning using fluorochrome-labeled primers, and heteroduplex analysis. All of the assays generated polyclonal patterns in the reactive tissues. The sensitivity in detecting monoclonality as defined by Southern blotting varied between 60% (heteroduplex analysis with FR3 primers) and 77% (high-resolution fingerprinting analysis with FR2 primers). Comparison of the three FR3 assays revealed that gene scanning had the highest sensitivity (69%), probably because it could detect small aberrant monoclonal amplicons. False-negative results were especially frequent in follicular center lymphoma (n = 20), but also in diffuse large B-cell lymphoma (n = 18), both renowned for having mutated variable gene segments, potentially leading to primer mismatching. For example, in follicular center lymphoma, the FR3, FR2, and FR1 region primer sets detected clonality in only 35 to 40, 65, and 50%, respectively. Combining these techniques, 17 (85%) of 20 follicular center lymphoma samples showed monoclonality. Therefore, especially in follicular center lymphoma, diffuse large B-cell lymphoma, and, to a lesser extent, in other lymphomas, multiple variable gene segment primer sets must be used for a reliable assessment of clonality. Our results also suggest that gene scanning is somewhat more sensitive than other read-out systems. Heteroduplex analysis, however, is a reliable alternative within a diagnostic setting, avoiding the use of radioactivity or expensive automated sequencing equipment and fluorochrome-labeled (oligo)nucleotides.


Recent advances for treatment of upper gastrointestinal malignancy.
Recent prospective trials for esophageal cancer, gastric cancer, and gastrointestinal stromal tumor (GIST) are encouraging. This manuscript reviews selected recently published studies. Not surprisingly, immunotherapy dominates the current clinical trial landscape. However, targeted biologic therapies and standard chemotherapy remain critical to the treatment of gastric and esophageal cancer while imatinib remains the backbone for advanced or metastatic GISTs. For all three cancers, surgical resection remains important when intent of treatment is potential cure.


Mussel disturbance dynamics: signatures of oceanographic forcing from local interactions.
Local interactions, biotic and abiotic, can have a strong influence on the large-scale properties of ecosystems. However, ecological models often explore the influence of local biotic interactions where physical disturbance is included as a large-scale and imposed source of variability but is not allowed to interact with biotic processes at the local scale. In marine intertidal communities dominated by mussels, wave disturbances create gaps in the mussel bed that recover through a successional sequence. We present a lattice model of mussel disturbance dynamics that allows local interactions between wave disturbance and mussel recolonization, in which each cell of the lattice can be empty, occupied by a mussel bed element, or disturbed (which corresponds to a newly disturbed cell that has unstable edges). As in natural ecosystems, wave disturbance can also spread from disturbed to adjacent occupied cells, and recolonization can also spread from occupied to adjacent empty cells. We first validate the local rules from artificial gap experiments and from natural gap monitoring along the Oregon coast. We analyze the properties of the model system as a function of different oceanographic forcings of productivity and disturbance. We show that the mussel bed can go through phase transitions characterized by a large sensitivity of mussel cover and patterns to oceanographic forcings but also that criticality (scale invariance) is observed over wide ranges of parameters, which suggests self-organization. We also show that spatial patterns in the intertidal can provide a robust signature of local processes and can inform about oceanographic regimes. We do so by comparing the large-scale patterns of the simulation (scaling exponents) with field data, which suggest that some experimental sites are close to criticality. Our results suggest that regional patterns in disturbed populations can be explained by local biotic and abiotic processes submitted to oceanographic forcing.


Neural mapping of anhedonia across psychiatric diagnoses: A transdiagnostic neuroimaging analysis.
Anhedonia has been associated with abnormal reward-related striatal dopamine functioning in patients with different psychiatric disorders. Here, we tested whether anhedonia expression mapped onto striatal volume across several psychiatric diagnoses. T1-weighted images from 313 participants including 89 healthy controls (HC), 22 patients with opioid use disorder (OUD), 50 patients with major depressive disorder (MDD), 45 patients with borderline personality disorder (BPD), 49 patients with first-episode psychosis (FEP), 43 patients with cocaine use disorder (CUD) and 15 patients with schizophrenia (SZ) were included. Anhedonia was assessed with subscores of the Beck Depression Inventory (BDI) and/or the Scale for the Assessment of Negative Symptoms (SANS). Voxel-based morphometry (VBM) was conducted for identifying dimensional symptom-structure associations using region of interest (ROI, dorsal and ventral striatum) and whole-brain analyses, as well as for group comparisons of striatal volume. ROI analyses revealed significant negative relationships between putamen volume and BDI and SANS anhedonia scores across OUD, MDD, BPD, CUD and SZ patients (n = 175) and MDD, FEP and SZ patients (n = 114), respectively. Whole-brain VBM analyses confirmed these associations and further showed negative relationships between anhedonia severity and volume of the bilateral cerebellum. There were group differences in right accumbens volume, which however were not related to anhedonia expression across the different diagnoses. Our findings indicate volumetric abnormalities in the putamen and cerebellum as a common neural substrate of anhedonia severity that cut across psychiatric entities.


Quantitative analysis of neonatal skeletal muscle functional improvement in the mouse.
Postnatal skeletal muscle growth is classically attributed to fiber hypertrophy and myogenic differentiation, but these processes do not account for the size-independent increase of muscle mechanical performance that occurs during postnatal growth. There is also little knowledge about the precise time-course of contractile function or the underlying factors that affect it. The present study investigated morphological factors (muscle fiber size and myofibrillar packing), biochemical factors (myosin heavy chain isoform and desmin intermediate filament protein expression), and muscle architecture during postnatal development in mice. Physiological testing of the mouse tibialis anterior revealed that maximum isometric stress increased from 27+/-3 kPa at postnatal day 1 to 169+/-10 kPa by postnatal day 28, roughly a sixfold increase. Morphological measurements revealed a robust increase in the size-independent packing of myofibrillar matrix material occurring with the functional improvement, with just 48.1+/-5.5% of the cross-sectional area filled with myofibrils at postnatal day 1 whereas 92.5+/-0.9% was filled by day 28. Expression of four myosin heavy chain isoforms (embryonic, neonatal, IIX and IIB), as well as desmin, correlated significantly with muscle mechanical function. Stepwise multiple regression showed that, of the variables measured, percentage content of neonatal myosin heavy chain was the best predictor of mechanical function during the postnatal time-course. These data provide the first specific structural basis for increases in muscle tension development during growth. Therefore, models of muscle growth must be modified to include an intrinsic quality enhancement component.


Generation of 1/f noise in locked systems working in nonlinear mode.
Properties of the phase fluctuations of a nonlinear oscillator were investigated. The multiscale aspect of synchronization is experimentally confirmed with the observation of lockings at any rational ratio. This global property of synchronization is associated with a local one by the study of the phase locked loop (PLL) under nonlinear conditions. We show that the discontinuity between the locking and the free-run state is characterized by the presence of slow temporal variations that induce a low frequency spectrum. An experimental verification of the increase of fluctuations confirms the classical results, which associate 1/f noise to nonlinearity, autosimilarity and regularity, represented here by the synchronization.


Monitoring Fatty Liver Disease with MRI Following Bariatric Surgery: A Prospective, Dual-Center Study.
Purpose To longitudinally monitor liver fat before and after bariatric surgery by using quantitative chemical shift-encoded (CSE) MRI and to compare with changes in body mass index (BMI), weight, and waist circumference (WC). Materials and Methods For this prospective study, which was approved by the internal review board, a total of 126 participants with obesity who were undergoing evaluation for bariatric surgery with preoperative very low calorie diet (VLCD) were recruited from June 27, 2010, through May 5, 2015. Written informed consent was obtained from all participants. Participants underwent CSE MRI measuring liver proton density fat fraction (PDFF) before VLCD (2-3 weeks before surgery), after VLCD (1-3 days before surgery), and 1, 3, and 6-10 months following surgery. Linear regression was used to estimate rates of change of PDFF (ΔPDFF) and body anthropometrics. Initial PDFF (PDFF0), initial anthropometrics, and anthropometric rates of change were evaluated as predictors of ΔPDFF. Mixed-effects regression was used to estimate time to normalization of PDFF. Results Fifty participants (mean age, 51.0 years; age range, 27-70 years), including 43 women (mean age, 50.8 years; age range, 27-70 years) and seven men (mean age, 51.7 years; age range, 36-62 years), with mean PDFF0 ± standard deviation of 18.1% ± 8.6 and mean BMI0 of 44.9 kg/m2 ± 6.5 completed the study. By 6-10 months following surgery, mean PDFF decreased to 4.9% ± 3.4 and mean BMI decreased to 34.5 kg/m2 ± 5.4. Mean estimated time to PDFF normalization was 22.5 weeks ± 11.5. PDFF0 was the only strong predictor for both ΔPDFF and time to PDFF normalization. No body anthropometric correlated with either outcome. Conclusion Average liver proton density fat fraction (PDFF) decreased to normal (< 5%) by 6-10 months following surgery, with mean time to normalization of approximately 5 months. Initial PDFF was a strong predictor of both rate of change of PDFF and time to normalization. Body anthropometrics did not predict either outcome. Online supplemental material is available for this article. © RSNA, 2018.


Initial state training procedure improves dynamic recurrent networks with time-dependent weights.
The problem of learning multiple continuous trajectories by means of recurrent neural networks with (in general) time-varying weights is addressed. The learning process is transformed into an optimal control framework where both the weights and the initial network state to be found are treated as controls. For such a task, a learning algorithm is proposed which is based on a variational formulation of Pontryagin's maximum principle. The convergence of this algorithm, under reasonable assumptions, is also investigated. Numerical examples of learning nontrivial two-class problems are presented which demonstrate the efficiency of the approach proposed.


Modulation of actA gene expression in Listeria monocytogenes by iron.
This study analysed the invasiveness of Listeria monocytogenes into enterocyte-like Caco-2 cells in which iron depletion was achieved by picolinic acid treatment. Both entry and intracellular multiplication varied depending on the endogenous iron content of bacterial and eukaryotic cells. The behaviour within enterocytes was correlated with a 10-fold increased transcription of the actA gene observed in bacterial cells grown under conditions of iron stress.


Meta-analysis: the effect of age on immunological response to hepatitis B vaccine in end-stage renal disease.
Patients on maintenance dialysis typically show a suboptimal immune response to hepatitis B virus vaccine compared with the non-uraemic population. A variety of inherited or acquired factors have been implicated in this diminished response. Age-associated changes in immune status may contribute to decreased vaccine efficacy in older individuals although contradictory results have been reported in individuals with normal kidney function. To evaluate the relationship between age and immune response to hepatitis B vaccine in patients with end-stage renal disease by performing a systematic review of the literature with a meta-analysis of clinical trials. We used the random effects model of DerSimonian and Laird; sources of heterogeneity in effect estimates were explored by performing sensitivity analyses. We identified 17 clinical trials (1800 unique patients); six (35%) were controlled studies. Pooling of study results demonstrated a significantly decreased risk of response to hepatitis B vaccine among older dialysis patients (overall risk ratio: 0.74; 95% confidence intervals: 0.70-0.79). The P-value was 0.0139 for our test of study heterogeneity. A lowered risk of response to hepatitis B vaccine persisted after exclusion of trials based on plasma-derived vaccines; it was present even when 'older' individuals were defined as being as 50 years (RR: 0.85, 95% CI: 0.75-0.96) or more (cut-off 60 years RR: 0.75; 95% CI: 0.66-0.85). An effect of age on seroprotection rate was present in all clinical reports, irrespective of the geographic origin of the study group: Europe (RR: 0.76; 95% CI: 0.70-0.83) North America (RR: 0.67; 95% CI: 0.60-0.74) or other countries (RR: 0.83; 95% CI: 0.71-0.97). Additional doses of vaccine did not appear to have an impact on RR of response by age. Our meta-analysis showed a clear association between older age and impaired response to hepatitis B virus vaccine in end-stage renal disease patients. Such a relationship is biologically plausible. Vaccination schedules with adapted vaccine doses and frequent serum testing for loss of immunity against hepatitis B virus are recommended in elderly patients on maintenance dialysis.


Silencing of UCHL1 by CpG promoter hyper-methylation is associated with metastatic gastroenteropancreatic well-differentiated neuroendocrine (carcinoid) tumors.
Well-differentiated gastroenteropancreatic neuroendocrine tumors (GEP-NETs) are rare tumors with varying metastatic potential. The underlying molecular basis for metastasis by GEP-NETs remains undefined. Quantitative PCR and immunohistochemistry (IHC) staining for ubiquitin carboxyl-terminal esterase L1 (UCHL1) gene and protein expression was performed on a group of localized and metastatic well-differentiated GEP-NET samples acquired from a prospectively maintained tissue bank. The ability of extent of UCHL1 IHC staining to differentiate localized and metastatic tumors was compared with Ki-67 index. Among 46 total samples, UCHL1 expression at both the gene and protein level was significantly greater among localized GEP-NETs compared with metastatic tumors and metastases (p < 0.001). Hypermethylation of the UCHL1 promoter was commonly observed among metastatic primary tumors and metastases (those with the lowest UCHL1 expression) but not among localized tumors (p < 0.001). Poor staining (<50 %) for UCHL1 was observed in 27 % of localized tumors compared with 87 % of metastatic tumors (p = 0.001). The presence of <50 % staining for UCHL1 was 88 % sensitive and 73 % specific for identifying metastatic disease. In contrast, there was no association between Ki-67 index and metastatic disease. In multivariable analysis, only UCHL1 staining <50 % [odds ratio (OR) 24.5, p = 0.035] and vascular invasion (OR 38.4, p = 0.03) were independent risk factors for metastatic disease at the time of initial surgery. Loss of UCHL1 expression by CpG promoter hypermethylation is associated with metastatic GEP-NETs. Extent of UCHL1 staining should be explored as a potentially clinically useful adjunct to Ki-67 index in evaluating GEP-NETs for aggressive features.


Characterization and assembly of the Pseudomonas aeruginosa aspartate transcarbamoylase-pseudo dihydroorotase complex.
Pseudomonas aeruginosa is a virulent pathogen that has become more threatening with the emergence of multidrug resistance. The aspartate transcarbamoylase (ATCase) of this organism is a dodecamer comprised of six 37 kDa catalytic chains and six 45 kDa chains homologous to dihydroorotase (pDHO). The pDHO chain is inactive but is necessary for ATCase activity. A stoichiometric mixture of the subunits associates into a dodecamer with full ATCase activity. Unlike other known ATCases, the P. aeruginosa catalytic chain does not spontaneously assemble into a trimer. Chemical-crosslinking and size-exclusion chromatography showed that P. aeruginosa ATCase is monomeric which accounts for its lack of catalytic activity since the active site is a composite comprised of residues from adjacent monomers in the trimer. Circular dichroism spectroscopy indicated that the ATCase chain adopts a structure that contains secondary structure elements although neither the ATCase nor the pDHO subunits are very stable as determined by a thermal shift assay. Formation of the complex increases the melting temperature by about 30°C. The ATCase is strongly inhibited by all nucleotide di- and triphosphates and exhibits extreme cooperativity. Previous studies suggested that the regulatory site is located in an 11-residue extension of the amino end of the catalytic chain. However, deletion of the extensions did not affect catalytic activity, nucleotide inhibition or the assembly of the dodecamer. Nucleotides destabilized the dodecamer which probably accounts for the inhibition and apparent cooperativity of the substrate saturation curves. Contrary to previous interpretations, these results suggest that P. aeruginosa ATCase is not allosterically regulated by nucleotides.


The pathology of diverticulosis: classical concepts and mucosal changes in diverticula.
Left-sided diverticular disease affects upwards of 50% of the population over the age of 60 years in western countries and is becoming more common as the population ages. Studies from the 1960s to 1980s related its occurrence to the use of low fiber diets, and to the prolonged colonic transit time and increased intraluminal pressure associated with low-volume stools. Pulsion diverticula (pseudodiverticula) emerge through the muscularis propria of the left colon at points of penetration of the vasa recta that supply the submucosa and mucosa. Cardinal features of the sigmoid colon in diverticular disease are abnormalities of the muscularis propria, including thickening and elastosis of the teniae, shortening of the bowel, and thickening and folding of the circular muscle with the development of transverse semilunar ridges between the mesenteric and antimesenteric teniae. Complications of diverticular disease such as hemorrhage, diverticulitis, peridiverticular abscess, fistula, and perforation are well recognized.


Genetic variation in competition traits at different ages and time periods and correlations with traits at field tests of 4-year-old Swedish Warmblood horses.
For many years, the breeding value estimation for Swedish riding horses has been based on results from Riding Horse Quality Tests (RHQTs) of 4-year-olds only. Traits tested are conformation, gaits and jumping ability. An integrated index including competition results is under development to both get as reliable proofs as possible and increases the credibility of the indexes among breeders, trainers and riders. The objectives of this study were to investigate the suitability of competition data for use in genetic evaluations of horses and to examine how well young horse performance agrees with performance later in life. Competition results in dressage and show jumping for almost 40 000 horses from the beginning of the 1960s until 2006 were available. For RHQT data of 14 000 horses judged between 1988 and 2007 were used. Genetic parameters were estimated for accumulated competition results defined for different age groups (4 to 6 years of age, 4 to 9 years of age and lifetime), and for different birth year groups. Genetic correlations were estimated between results at RHQT and competitions with a multi-trait animal model. Heritabilities were higher for show jumping than dressage and increased with increasing age of the horse and amount of information. For dressage, heritabilities increased from 0.11 for the youngest group to 0.16 for lifetime results. For show jumping corresponding values increased from 0.24 to 0.28. Genetic correlations between competition results for the different age groups were highly positive (0.84 to 1.00), as were those between jumping traits at RHQT and competition results in show jumping (0.87 to 0.89). For dressage-related traits as 4-year-old and dressage competition results the estimated genetic correlations were between 0.47 and 0.77. We suggest that lifetime results from competitions should be integrated into the genetic evaluation system. However, genetic parameters showed that traits had changed during the over 35-year period covered due to the development of the sport, which needs to be considered in future genetic evaluations.


Salicylic acid and jasmonic acid are essential for systemic resistance against tobacco mosaic virus in Nicotiana benthamiana.
Systemic resistance is induced by pathogens and confers protection against a broad range of pathogens. Recent studies have indicated that salicylic acid (SA) derivative methyl salicylate (MeSA) serves as a long-distance phloem-mobile systemic resistance signal in tobacco, Arabidopsis, and potato. However, other experiments indicate that jasmonic acid (JA) is a critical mobile signal. Here, we present evidence suggesting both MeSA and methyl jasmonate (MeJA) are essential for systemic resistance against Tobacco mosaic virus (TMV), possibly acting as the initiating signals for systemic resistance. Foliar application of JA followed by SA triggered the strongest systemic resistance against TMV. Furthermore, we use a virus-induced gene-silencing-based genetics approach to investigate the function of JA and SA biosynthesis or signaling genes in systemic response against TMV infection. Silencing of SA or JA biosynthetic and signaling genes in Nicotiana benthamiana plants increased susceptibility to TMV. Genetic experiments also proved the irreplaceable roles of MeSA and MeJA in systemic resistance response. Systemic resistance was compromised when SA methyl transferase or JA carboxyl methyltransferase, which are required for MeSA and MeJA formation, respectively, were silenced. Moreover, high-performance liquid chromatography-mass spectrometry analysis indicated that JA and MeJA accumulated in phloem exudates of leaves at early stages and SA and MeSA accumulated at later stages, after TMV infection. Our data also indicated that JA and MeJA could regulate MeSA and SA production. Taken together, our results demonstrate that (Me)JA and (Me)SA are required for systemic resistance response against TMV.


Intracerebellar vermis histamine facilitates memory consolidation in the elevated T maze model.
Experimental evidence suggests that the cerebellum plays a more complex role in learning than simply regulating the motor response. Rather, it is thought to play a significant role in the consolidation of emotional memory in mice. Due to the difficulty of interpreting fear and anxiety behaviors-the standard methodology for the study of the histaminergic system and emotional memory-in mice, we propose a behavioral assessment of mice subjected to the Elevated T-maze after histamine microinjection of the cerebellar vermis. Young male Swiss albino mice weighing 25-35g were used. In addition, locomotor activity was tested in an open field test. Our data suggest that histamine did not affect memory consolidation during escape or open field behavior at the doses used in this study. However, we observed a significant increase in inhibitory avoidance on the second day in animals receiving a dose of 6.8nmol/0.5μl, suggesting that histamine facilitates the consolidation of inhibitory avoidance in mice.


Effects of the T-type calcium channel blockade with oral mibefradil on the electrophysiologic properties of the human heart.
As the calcium T-channel blockade is a new pharmacological category with presumably unique electrophysiological effects, the influence of its only representative yet mibefradil was tested after the single oral dose. 10 patients underwent the electrophysiologic examination. Normal baseline values in sinus node cycle length (SNCL), sinus node recovery time (SNRT), corrected sinus node recovery time (CSNRT), PA interval, atrial effective refractory period, AH interval, Wenckebach point (WP), atrioventricular nodal refractory period, and HV interval were measured using standard techniques. After that a single dose of 100 mg mibefradil was given and the testing repeated in 90 minutes. Though non-significantly in a study-group limited in size due to global withdrawal of mibefradil, sinus node automaticity was suppressed (prolongation of SNRT by 5.1% and CSNRT by 11.5%) and heart rate lowered (SNCL prolonged by 2.8%) comparatively more than was the negative dromotropic effect on the atrioventricular node (negligible prolongation of AH interval by 1.1% and WP cycle by 0.4%). Demonstrated electrophysiological effects of oral mibefradil with more pronounced influence on the automaticity of the sinus node seem to be in agreement with the preclinical data on the predominant role of T-channels in the pacemaker activity of the sinus node. According to the Framingham data on the risk of heart rate for the cardiovascular as well as all-cause mortality, calcium T-channel blockade offers a desirable profile for antihypertensive treatment. From this point of view development of new representatives of calcium T-channel blockers could be a useful contribution to clinical practice.


Wearable Nail Deformation Sensing for Behavioral and Biomechanical Monitoring and Human-Computer Interaction.
The dynamics of the human fingertip enable haptic sensing and the ability to manipulate objects in the environment. Here we describe a wearable strain sensor, associated electronics, and software to detect and interpret the kinematics of deformation in human fingernails. Differential forces exerted by fingertip pulp, rugged connections to the musculoskeletal system and physical contact with the free edge of the nail plate itself cause fingernail deformation. We quantify nail warpage on the order of microns in the longitudinal and lateral axes with a set of strain gauges attached to the nail. The wearable device transmits raw deformation data to an off-finger device for interpretation. Simple motions, gestures, finger-writing, grip strength, and activation time, as well as more complex idioms consisting of multiple grips, are identified and quantified. We demonstrate the use of this technology as a human-computer interface, clinical feature generator, and means to characterize workplace tasks.


The Mice Drawer System (MDS) experiment and the space endurance record-breaking mice.
The Italian Space Agency, in line with its scientific strategies and the National Utilization Plan for the International Space Station (ISS), contracted Thales Alenia Space Italia to design and build a spaceflight payload for rodent research on ISS: the Mice Drawer System (MDS). The payload, to be integrated inside the Space Shuttle middeck during transportation and inside the Express Rack in the ISS during experiment execution, was designed to function autonomously for more than 3 months and to involve crew only for maintenance activities. In its first mission, three wild type (Wt) and three transgenic male mice over-expressing pleiotrophin under the control of a bone-specific promoter (PTN-Tg) were housed in the MDS. At the time of launch, animals were 2-months old. MDS reached the ISS on board of Shuttle Discovery Flight 17A/STS-128 on August 28(th), 2009. MDS returned to Earth on November 27(th), 2009 with Shuttle Atlantis Flight ULF3/STS-129 after 91 days, performing the longest permanence of mice in space. Unfortunately, during the MDS mission, one PTN-Tg and two Wt mice died due to health status or payload-related reasons. The remaining mice showed a normal behavior throughout the experiment and appeared in excellent health conditions at landing. During the experiment, the mice health conditions and their water and food consumption were daily checked. Upon landing mice were sacrificed, blood parameters measured and tissues dissected for subsequent analysis. To obtain as much information as possible on microgravity-induced tissue modifications, we organized a Tissue Sharing Program: 20 research groups from 6 countries participated. In order to distinguish between possible effects of the MDS housing conditions and effects due to the near-zero gravity environment, a ground replica of the flight experiment was performed at the University of Genova. Control tissues were collected also from mice maintained on Earth in standard vivarium cages.


Ethnic and sex differences in the longitudinal association between heart rate variability and blood pressure.
Elevated blood pressure is a risk factor for increased cardiovascular morbidity and mortality. Decreased vagally-mediated heart rate variability has previously been prospectively linked with increased blood pressure; however, to date, no such prospective data exist regarding this relationship among Blacks. We examined this association in 387 normotensive young adults (mean age, 23 years, 52% female, 54% Black) who participated in two laboratory evaluations spanning approximately six years. Blood pressure was measured at both timepoints with a non-invasive oscillometric device and heart rate variability was assessed via bio-impedance. In the total sample, heart rate variability significantly predicted systolic (p = .022) and diastolic (p < .001) blood pressure increases six years into the future. However, this pattern varied as a function of ethnicity and sex with the effect of heart rate variability on Time 2 systolic blood pressure only significant among White males (p = .007). Heart rate variability was also predictive of Time 2 diastolic blood pressure in White males (p = .038) as well as among both White (p = .032) and Black (p = .015) females, but was not related to blood pressure among Black males. We report for the first time significant ethnic and sex differences in the prospective relationship between heart rate variability and blood pressure change. These findings may give clues as to the underlying mechanisms that are involved in the well-known health disparities in blood pressure and hypertension-related cardiovascular diseases.


Influence of pharmorubicin on the left ventricular ejection fraction of patients with breast cancer: A mechanism study.
Breast cancer is a great public health problem. It remains unclear how pharmorubicin induces cardiac dysfunction in patients with breast cancer. Our study was aimed to explore the influence of pharmorubicin on the left ventricular ejection fraction (EF) of patients with breast cancer and its potential mechanism. Patients with breast cancer were enrolled at the same hospital. Group I consisted of 135 samples, who were under treatment of pharmorubicin (200 mg/m2). Group II consisted of 144 samples, who were under treatment the of pharmorubicin (360 mg/m2). Group III was used as control group, which consists of 120 samples without treatment of pharmorubicin. Color Doppler ultrasonic inspection and measurement were performed to examine EF. Flow cytometry was performed to assess oxygen free radical level in hemocytes. Further combination therapy (N-acetylcysteine [NAC] + pharmorubicin) was provided for patients, and the same examinations were performed for the assessment of cardiac function and oxygen free radical level. The ultrasound results showed that pharmorubicin treatment significantly jeopardized cardiac function, verified by decrease of both EF and fractional shortening (FS) (P < 0. 05). Moreover, such effect was dose-dependent. Oxygen free radical level was remarkably increased after pharmorubicin treatment (P < 0. 05), verified by flow cytometry. Adjunctive therapy of NAC decreased oxygen free radical level and improved cardiac function of patients with breast cancer, suggesting NAC ameliorated side effect of pharmorubicin treatment. Pharmorubicin treatment decreased EF and FS of patients with breast cancer through increasing oxygen free radical level in hemocytes. Adjunctive therapy of NAC could be a potential treatment to ameliorated side effect pharmorubicin treatment.


Review. Transcriptional mechanisms of addiction: role of DeltaFosB.
Regulation of gene expression is considered a plausible mechanism of drug addiction, given the stability of behavioural abnormalities that define an addicted state. Among many transcription factors known to influence the addiction process, one of the best characterized is DeltaFosB, which is induced in the brain's reward regions by chronic exposure to virtually all drugs of abuse and mediates sensitized responses to drug exposure. Since DeltaFosB is a highly stable protein, it represents a mechanism by which drugs produce lasting changes in gene expression long after the cessation of drug use. Studies are underway to explore the detailed molecular mechanisms by which DeltaFosB regulates target genes and produces its behavioural effects. We are approaching this question using DNA expression arrays coupled with the analysis of chromatin remodelling--changes in the posttranslational modifications of histones at drug-regulated gene promoters--to identify genes that are regulated by drugs of abuse via the induction of DeltaFosB and to gain insight into the detailed molecular mechanisms involved. Our findings establish chromatin remodelling as an important regulatory mechanism underlying drug-induced behavioural plasticity, and promise to reveal fundamentally new insight into how DeltaFosB contributes to addiction by regulating the expression of specific target genes in brain reward pathways.


K(+)-neutral amino acid symport of Bombyx mori larval midgut: a system operative in extreme conditions.
The K(+)-dependent symporter for leucine and other neutral amino acids expressed along the midgut of the silkworm Bombyx mori operates with best efficiency in the presence of a steep pH gradient across the brush-border membrane, with external alkaline pH values up to 11, and an electrical potential difference (delta psi) of approximately 200 mV. Careful determinations of leucine kinetics as a function of external amino acid concentrations between 50 and 1,000 microM, performed with brush-border membrane vesicles (BBMV) obtained from the middle and posterior midgut regions, revealed that the kinetic parameter affected by the presence of a delta pH was the maximal rate of transport. The addition of delta psi caused a further marked increase of the translocation rate. At nonsaturating leucine concentrations in the solution bathing the external side of the brush-border membrane, leucine accumulation within BBMV and midgut cells was not only driven by the gradient of the driver cation K+ and delta psi but occurred also in the absence of K+. The ability of the symporter to translocate the substrate in its binary form allows the intracellular accumulation of leucine in the absence of K+, provided that a pH gradient, with alkaline outside, is present. The mechanisms involved in this accumulation are discussed.