Haloperidol (but not ziprasidone) withdrawal potentiates sensitization to the hyperlocomotor effect of cocaine in mice.
One important contributing factor in the high prevalence of drug abuse disorders seen among schizophrenic patients seems to be related to chronic treatment with typical neuroleptics. We have previously demonstrated that withdrawal from long-term treatment with the typical neuroleptic haloperidol, but not the atypical neuroleptic ziprasidone, potentiated the hyperlocomotor effect induced by a single cocaine injection and cocaine-induced conditioned place preference in mice. In the present study we investigated whether withdrawal from long-term treatment with these same neuroleptics would also modify cocaine-induced hyperlocomotion sensitization, which has been proposed as an animal model for the intensification of drug craving in cocaine addiction. Swiss male mice were i.p. treated with haloperidol (1.0 mg/kg) or ziprasidone (4.0 mg/kg) for 15 days. Twenty-four hours after the last injection, animals received an i.p. injection of cocaine (10 mg/kg) for 5 consecutive days, being placed after each injection in the open-field apparatus in order to perform a drug-environment conditioning. Seven days after the last drug-environment conditioning procedure, the animals were challenged with an i.p. injection of cocaine (10 mg/kg), placed in the open-field apparatus and had their locomotor activity quantified. Withdrawal from haloperidol (but not ziprasidone) potentiated cocaine-induced behavioral sensitization. These results are suggested to be a consequence of the development of the dopaminergic supersensitivity phenomenon by long-term treatment with the typical compound. Our findings provide additional support for the use of atypical agents like ziprasidone in the treatment of schizophrenic patients with comorbid substance abuse disorder.


[Multiple stimulation of T lymphocytes in the pathogenesis of multiple sclerosis].
Multiple Sclerosis is a T-cell mediated autoimmune disease leading to demyelination of central nervous system. The precise mechanism by which auto-reactive T cells are activated and tolerance to self-antigens is broken are not fully understood. The dysregulation of costimulatory signals between T cell and antigen presenting cell is taken into consideration in this process. Among many costimulatory molecules presented on activated T cells CD40L and CD28 are of special interest. These molecules are involved in the positive regulation of T cell activation. Contrary to them CTLA-4, the another costimulatory molecule presented on activated T cells down-regulates activation. In this review the role of modification of costimulatory signals on the development and course of EAE, the animal model for multiple sclerosis as well as clinical data documenting the dysregulation of costimulation in Multiple Sclerosis are presented.


Haematopoietic stem cell transplantation: can our genes predict clinical outcome?
Haematopoietic stem cell transplantation (HSCT) is currently the only curative treatment for many patients with malignant and non-malignant haematological diseases. The success of HSCT is greatly reduced by the development of complications, which include graft-versus-host disease (GVHD), relapse and infection. Human leukocyte antigen (HLA) matching of patients and donors is essential, but does not completely prevent these complications; non-HLA genes may also have an impact upon transplant outcome. Polymorphisms within genes that are associated with an individual's capability to mount an immune response to alloantigen and infectious pathogens and/or response to drugs (pharmacogenomics) are all currently being studied for their association with HSCT outcome. This review summarises the potential role of non-HLA polymorphisms in predicting HSCT outcome, from studies on retrospective transplant cohorts of HLA-identical siblings and matched unrelated donors. The clinical relevance and interpretation of non-HLA genetics, and how these could be used alongside clinical risk factors in HSCT, are also discussed.


Comprehensive maternal characteristics associated with birth weight: Bayesian modeling in a prospective cohort study from Iran.
In this study, we aimed to determine comprehensive maternal characteristics associated with birth weight using Bayesian modeling. A total of 526 participants were included in this prospective study. Nutritional status, supplement consumption during the pregnancy, demographic and socioeconomic characteristics, anthropometric measures, physical activity, and pregnancy outcomes were considered as effective variables on the birth weight. Bayesian approach of complex statistical models using Markov chain Monte Carlo approach was used for modeling the data considering the real distribution of the response variable. There was strong positive correlation between infant birth weight and the maternal intake of Vitamin C, folic acid, Vitamin B3, Vitamin A, selenium, calcium, iron, phosphorus, potassium, magnesium as micronutrients, and fiber and protein as macronutrients based on the 95% high posterior density regions for parameters in the Bayesian model. None of the maternal characteristics had statistical association with birth weight. Higher maternal macro- and micro-nutrient intake during pregnancy was associated with a lower risk of delivering low birth weight infants. These findings support recommendations to expand intake of nutrients during pregnancy to high level.


Perifascicular atrophic fibers in childhood dermatomyositis with particular reference to mitochondrial changes.
Ten of 17 muscle biopsy specimens from 15 patients with childhood dermatomyositis (DM) showed distinct perifascicular atrophy. The atrophic fibers showed the following characteristics: (1) decreased cytochrome c oxidase (CCO) activity, (2) rare positive reaction with acridine orange (AO) staining, (3) type 2C reaction in 7.9% (0.4-17.5%) of the fibers, (4) an increased number of activated satellite cells, (5) mitochondria which were increased in number but decreased in size, (6) a significantly decreased CCO activity in isolated mitochondria (51.6 +/- 30.3 nmol/min per mg mitochondrial protein) as compared with that in the controls (103.6 +/- 41.5). The major pathogenetic mechanism in muscles in childhood DM is thought to be ischemia due to involvement of the microvasculature. The presence of type 2C fibers and increased numbers of activated satellite cells reflect a focal repair process taking place concomitantly in the damaged myofibers. Mitochondrial enzyme defect, especially CCO deficiency is present not only in genetic disorders with mitochondrial involvement but in other neuromuscular disorders including inflammatory myopathies.


Optimizing differential identifiability improves connectome predictive modeling of cognitive deficits from functional connectivity in Alzheimer's disease.
Functional connectivity, as estimated using resting state functional MRI, has shown potential in bridging the gap between pathophysiology and cognition. However, clinical use of functional connectivity biomarkers is impeded by unreliable estimates of individual functional connectomes and lack of generalizability of models predicting cognitive outcomes from connectivity. To address these issues, we combine the frameworks of connectome predictive modeling and differential identifiability. Using the combined framework, we show that enhancing the individual fingerprint of resting state functional connectomes leads to robust identification of functional networks associated to cognitive outcomes and also improves prediction of cognitive outcomes from functional connectomes. Using a comprehensive spectrum of cognitive outcomes associated to Alzheimer's disease (AD), we identify and characterize functional networks associated to specific cognitive deficits exhibited in AD. This combined framework is an important step in making individual level predictions of cognition from resting state functional connectomes and in understanding the relationship between cognition and connectivity.


An Artificial Vibrissa-Like Sensor for Detection of Flows.
In nature, there are several examples of sophisticated sensory systems to sense flows, e.g., the vibrissae of mammals. Seals can detect the flow of their prey, and rats are able to perceive the flow of surrounding air. The vibrissae are arranged around muzzle of an animal. A vibrissa consists of two major components: a shaft (infector) and a follicle-sinus complex (receptor), whereby the base of the shaft is supported by the follicle-sinus complex. The vibrissa shaft collects and transmits stimuli, e.g., flows, while the follicle-sinus complex transduces them for further processing. Beside detecting flows, the animals can also recognize the size of an object or determine the surface texture. Here, the combination of these functionalities in a single sensory system serves as paragon for artificial tactile sensors. The detection of flows becomes important regarding the measurement of flow characteristics, e.g., velocity, as well as the influence of the sensor during the scanning of objects. These aspects are closely related to each other, but, how can the characteristics of flow be represented by the signals at the base of a vibrissa shaft or by an artificial vibrissa-like sensor respectively? In this work, the structure of a natural vibrissa shaft is simplified to a slender, cylindrical/tapered elastic beam. The model is analyzed in simulation and experiment in order to identify the necessary observables to evaluate flows based on the quasi-static large deflection of the sensor shaft inside a steady, non-uniform, laminar, in-compressible flow.


Vitamin B12 and Folate Status in Pregnant Females and Their Infants in Norway: Secondary Analysis from the Mommy's Food Study.
Vitamin B12 and folate are essential micronutrients important for normal infant growth and development. The aims were to describe vitamin B12 and folate status in pregnant females and their infants according to commonly used status cutoffs and examine the associations between maternal status, maternal supplement use, and breastfeeding and infant status. Pregnant females were recruited at 18 wk gestation in Bergen, Norway. Maternal vitamin B12 and folate status were measured at gestational weeks 18 (n = 136) and 36 (n = 116), and infant status was measured at ages 3 (n = 73) and 6 (n = 74) mo. At gestational weeks 18 and 36, respectively, 4.4% and 2.6% of the mothers had plasma cobalamin concentrations <148 pmol/L, 0.7% and 6.9% had methylmalonic acid (MMA) concentrations >0.26 μmol/L, and 3.7% and 30% had folate concentrations <10 nmol/L. None of the females had total homocysteine (t-Hcy) concentrations >13 μmol/L or 3 combined indicator of vitamin B12 (cB12) < -0.5. At 3 and 6 mo, respectively, 4.1% and 5.4% of the infants had cobalamin concentrations <148 pmol/L, 63% and 74% had t-Hcy concentrations >6.5 μmol/L, 59% and 66% had MMA concentrations >0.26 μmol/L, and 47% and 60% had cB12 > -0.5. None of the infants had folate concentrations <10 nmol/L. Several of the vitamin B12 biomarkers in infants were associated with maternal vitamin B12 status during pregnancy. Breastfed infants had lower vitamin B12 status (as indicated by plasma cobalamin, t-Hcy, and cB12) than nonbreastfed infants at both 3 and 6 mo. Use of supplements during pregnancy was associated with better vitamin B12 status among infants at 3 and 6 mo, as indicated by infants' cobalamin and t-Hcy concentrations. Subclinical vitamin B12 deficiency among infants was common and associated with maternal vitamin B12 status during pregnancy and breastfeeding. Among the mothers, an increase in biochemical folate deficiency was discovered toward the end of gestation. Further studies are needed to investigate clinical consequences. This trial was registered at clinicaltrials.gov as NCT02610959.


Water Accessibility Refinement of the Extended Structure of KirBac1.1 in the Closed State.
NMR structures of membrane proteins are often hampered by poor chemical shift dispersion and internal dynamics which limit resolved distance restraints. However, the ordering and topology of these systems can be defined with site-specific water or lipid proximity. Membrane protein water accessibility surface area is often investigated as a topological function via solid-state NMR. Here we leverage water-edited solid-state NMR measurements in simulated annealing calculations to refine a membrane protein structure. This is demonstrated on the inward rectifier K+ channel KirBac1.1 found in Burkholderia pseudomallei. KirBac1.1 is homologous to human Kir channels, sharing a nearly identical fold. Like many existing Kir channel crystal structures, the 1p7b crystal structure is incomplete, missing 85 out of 333 residues, including the N-terminus and C-terminus. We measure solid-state NMR water proximity information and use this for refinement of KirBac1.1 using the Xplor-NIH structure determination program. Along with predicted dihedral angles and sparse intra- and inter-subunit distances, we refined the residues 1-300 to atomic resolution. All structural quality metrics indicate these restraints are a powerful way forward to solve high quality structures of membrane proteins using NMR.


Acute Effect of Biomechanical Muscle Stimulation on the Counter-Movement Vertical Jump Power and Velocity in Division I Football Players.
Jacobson, BH, Monaghan, TP, Sellers, JH, Conchola, EC, Pope, ZK, and Glass, RG. Acute effect of biomechanical muscle stimulation on the counter-movement vertical jump power and velocity in division I football players. J Strength Cond Res 31(5): 1259-1264, 2017-Research regarding whole body vibration (WBV) largely supports such training augmentation in attempts to increase muscle strength and power. However, localized biomechanical vibration has not received the same attention. The purpose of this study was to assess peak and average power before and after acute vibration of selected lower-body sites in division I athletes. Twenty-one subjects were randomly assigned to 1 of 2 conditions using a cross-over design. Pretest consisted of a counter-movement vertical jump (VJ) followed by either localized vibration (30 Hz) to 4 selected lower-body areas or 4 minutes of moderately low-resistance stationary cycling (70 rpm). Vibration consisted of 1 minute bouts at each lower-leg site for a total of 4 minutes followed by an immediate post-test VJ. Repeated measures analysis of variance yielded no significant differences (p > 0.05) in either peak power or peak velocity. Similarly, no significant differences were found for average power and velocity between conditions. It should be noted that, while not significant, the vibration condition demonstrated an increase in peak power and velocity while the bike condition registered slight decreases. Comparing each of the post-VJ repetitions (1, 2, and 3) the vibration condition experienced significantly greater peak power and velocity from VJ 1 to VJ 3 compared with the bike condition which demonstrated no significant differences among the post-test VJs. These results yielded similar, although not statistically significant outcomes to previous studies using WBV. However, the novelty of selected site biomechanical vibration merits further investigation with respect to frequency, magnitude, and duration of vibration.


Episode-specific differential gene expression of peripheral blood mononuclear cells in rapid cycling supports novel treatment approaches.
Molecular mechanisms underlying bipolar affective disorders are unknown. Difficulties arise from genetic and phenotypic heterogeneity of patients and the lack of animal models. Thus, we focused on only one patient (n = 1) with an extreme form of rapid cycling. Ribonucleic acid (RNA) from peripheral blood mononuclear cells (PBMC) was analyzed in a three-tiered approach under widely standardized conditions. Firstly, RNA was extracted from PBMC of eight blood samples, obtained on two consecutive days within one particular episode, including two different consecutive depressive and two different consecutive manic episodes, and submitted to (1) screening by microarray hybridizations, followed by (2) detailed bioinformatic analysis, and (3) confirmation of episode-specific regulation of genes by quantitative real-time polymerase chain reaction (qRT-PCR).Secondly, results were validated in additional blood samples obtained one to two years later. Among gene transcripts elevated in depressed episodes were prostaglandin D synthetase (PTGDS) and prostaglandin D2 11-ketoreductase (AKR1C3), both involved in hibernation. We hypothesized them to account for some of the rapid cycling symptoms. A subsequent treatment approach over 5 months applying the cyclooxygenase inhibitor celecoxib (2 x 200 mg daily) resulted in reduced severity rating of both depressed and manic episodes. This case suggests that rapid cycling is a systemic disease, resembling hibernation, with prostaglandins playing a mediator role.


Head and neck lymphoedema-research challenges during the COVID-19 pandemic.
Management of secondary head and neck lymphoedema has undergone little research investigation. Its treatment is time and labour intensive and involves multiple therapeutic modalities without a clear understanding of which is most effective. This study aimed to determine the feasibility of a randomised controlled trial comparing two therapeutic modalities to manage head and neck lymphoedema. The secondary objective was to evaluate the clinical effects of these treatments. Participants were randomised to receive treatment with manual lymphatic drainage or compression over 6 weeks, with the primary outcome-percentage tissue water-measured 12 weeks after treatment. Six participants were recruited until the study was ceased due to restrictions imposed by the COVID-19 pandemic. Some 86% of required attendances were completed. Percentage tissue water increased in all participants at 12 weeks. No consistent trends were identified between internal and external lymphoedema. The small number of people recruited to this study informs its feasibility outcomes but limits any conclusions about clinical implications.


Original research in the classroom: why do zebrafish spawn in the morning?
As part of an upper level undergraduate developmental biology course at the University of Minnesota Duluth, we developed a unit in which students carried out original research as part of a cooperative class project. Students had the opportunity to gain experience in the scientific method from experimental design all of the way through to the preparation of publication on their research that included text, figures, and tables. This kind of inquiry-based learning has been shown to have many benefits for students, including increased long-term learning and a better understanding of the process of scientific discovery. In our project, students designed experiments to explore why zebrafish typically spawn in the first few hours after the lights come on in the morning. The results of our experiments suggest that spawning still occurs when the dark-to-light transition is altered or absent. This is consistent with the work of others that demonstrates that rhythmic spawning behavior is regulated by an endogenous circadian clock. Our successes and failures carrying out original research as part of an undergraduate course should contribute to the growing approaches for using zebrafish to bring the excitement of experimental science to the classroom.


Prior infection with murine cytomegalovirus (MCMV) limits the immunocontraceptive effects of an MCMV vector expressing the mouse zona-pellucida-3 protein.
We have developed a murine cytomegalovirus (MCMV)-vectored vaccine expressing the mouse zona-pellucida-3 gene (rMCMV-ZP3), which successfully induces infertility in experimentally inoculated laboratory or wild-derived mice. However, the future success of this vector as a fully disseminating vaccine in free-living mice may be compromised by pre-existing immunity since there is a high prevalence of naturally acquired MCMV infection in these mice. To evaluate the effect of prior immunity to MCMV on vaccine efficacy, we constructed two new biologically effective recombinant MCMV vectors expressing the mouse ZP3 protein from two MCMV strains (N1 and G4) derived from free-living mice. In wild mice, mixed MCMV infection is common and could be acquired either by simultaneous coinfection or sequential infection with different MCMV strains. Interestingly, while coinfection with both wild-type and rMCMV-ZP3 via the intraperitoneal route reduced the impact of the rMCMV-ZP3, prior infection with the same wild-type strain as that used to construct the rMCMV-ZP3 abrogated the immunocontraceptive effects of either N1-ZP3 or G4-ZP3. However, prior infection with G4 28 days before the introduction of N1-ZP3 had a reduced influence on the efficacy of the rMCMV-ZP3. Thus, the strain of virus and the timing of prior infection are factors that may influence the efficacy of the rMCMV-ZP3. Given that mixed infection of mice with MCMV is common, it is possible that prior immunity acquired by natural mucosal infection may have less a less inhibitory effect on the immunocontraceptive outcome.


Matching of vertebrate cardiac energy demand to energy metabolism.
Concentrations of high-energy phosphates and activities of key enzymes of energy metabolism were assessed in hearts from species with differing levels of cardiac power output. Positive correlations were found between resting power output and the total adenylate pool and between citrate synthase activity and the total adenylate pool. Maximum in vitro activity levels of enzymes from energy metabolism were compared with calculated resting cardiac power output and maximal cardiac power output (as reflected by total oligomycin-insensitive adenosine-triphosphatase activity). Three indexes of carbohydrate metabolism (hexokinase, pyruvate kinase, and L-lactate dehydrogenase) all plateau at relatively low levels of energy demand. In contrast, enzymes required for aerobic fatty acid metabolism, (carnitine palmitoyltransferase and 3-hydroxyacyl-CoA dehydrogenase) and for tricarboxylic acid and electron transport (citrate synthase and cytochrome-c oxidase) show consistent increases as ATP demand is elevated. It appears that as capacity for power development by vertebrate hearts, increases across taxa, the elevated demand for ATP is met by expansion of fatty acid based aerobic metabolism and not carbohydrate metabolism.


Ambulatory electrocardiographic recording in endurance athletes.
Data from ambulatory electrocardiographic recording in 35 highly trained endurance athletes and in 35 non-athletic controls of similar ages are given. The minimal, mean hourly, and maximal heart rates were significantly lower in the athletes. Thirteen athletes (37 . 1%) but only two controls (5 . 7%) had sinus pauses exceeding 2 . 0 seconds. First degree atrioventricular block was observed in 13 athletes (37 . 1%) and five controls (14 . 3%), second degree Wenckebach type block in eight athletes (22 . 9%) and two controls (5 . 7%), and second degree block with Mobitz II-like pattern in three athletes (8 . 6%) and no control. All athletes with Mobitz II-type pattern also had first degree and Wenckebach-type second degree atrioventricular block. The behavior of sinus rate on development of atrioventricular block varied, not only interindividually but also intraindividually, from absence of change to an increase or decrease in most subjects in both study groups. A decrease in sinus rate on appearance of atrioventricular block was found constantly in only two athletes and one control. Atrioventricular dissociation with junctional rhythm occurred in seven athletes (20%) and with ventricular rhythm in one athlete. Neither of these phenomena was seen in the group of controls. The athletes had slightly fewer ventricular extrasystoles than controls, and no athlete had ventricular tachycardia, whereas two controls had ventricular tachycardia.


An electronic enzyme-linked immunosorbent assay platform for protein analysis based on magnetic beads and AlGaN/GaN high electron mobility transistors.
AlGaN/GaN high electron mobility transistor (HEMT) biosensors have attracted attention due to their high sensitivity, stability, and fast response characteristics. Some related studies have been explored but a Debye screening problem exists in physiological solutions hindering the detection of bio-macromolecules. Herein, a novel fast analytical platform for electronic enzyme-linked immunosorbent assay (e-ELISA) is proposed based on AlGaN/GaN HEMT with magnetic beads (MBs); MB-based e-ELISA decouples the modified area from the sensing surface to simplify the assay. Combining the advantages of e-ELISA and MBs, the resulting analytical platform presents a sensing capability beyond the Debye-screening limit and a novel ability to be reused. This platform offers a fast response toward prostate specific antigen (PSA) and the lowest concentration of detection is 1 fg mL-1. Compared with conventional AlGaN/GaN HEMT biosensors, it shows higher sensitivity (3.73 μA dec-1) in a linear range (1 fg mL-1 to 1 pg mL-1), which is within the constraints of emergency care applications. The platform's high sensitivity and fast repeatability endow it with great potential for early and rapid diagnosis.


Efficient assembly of functional cytochrome P450 2C2 requires a spacer sequence between the N-terminal signal anchor and catalytic domains.
Cytochromes P450 (P450) are anchored to the endoplasmic reticulum membrane by an N-terminal transmembrane sequence with the catalytic domain facing the cytoplasmic side. Within the peptide sequence linking these two domains in P450 2C2 is a glycine-rich region from residues 22 to 28. To examine the role of this region, deletion and substitution mutations were constructed, and the activities and spectral properties were determined for the mutant proteins expressed in COS-1 cells, insect cells, and bacteria. Deletion of residues 22 to 28 or substitution of 7 valines for this region inactivated the proteins in COS-1 cells, and no P450 species was detected for these mutations in bacteria or insect cells. Substitution of the three glycine residues with alanine or proline or the entire sequence from 22 to 28 with 7 alanines did not reduce lauric acid hydroxylase activity of the proteins expressed in COS-1 cells. Reducing the number of alanines substituted to 4, 3, and 2 progressively decreased activity in COS-1 cells to undetectable levels when 2 alanines were substituted. The loss of activity in COS-1 cells correlated with decreased expression of hemoprotein with a reduced difference spectrum of 450 nm (P450 species) and a corresponding increase in the inactive P420 species in insect cells and bacteria. The activities expressed per nanomole of P450 in insect microsomes were similar for P450 2C2 and the alanine substitution mutants, including the mutant with 2 alanines which was inactive in COS-1 cells. The rates of conversion of P450 to P420 resulting from incubation at 48 degrees C in vitro were not changed sufficiently to explain the increase in expressed P420 observed for the mutants with 3 or 7 alanines substituted. These data are consistent with a role for the residue 22-28 region as a linker that facilitates the folding of P450; however, once the protein is properly folded into the functional P450 species, this region has little influence on the stability and activity of the enzyme.


Discovery of novel cMET-targeting antibody Fab drug conjugates as potential treatment for solid tumors with highly expressed cMET.
Solid tumors are becoming prevalent affecting both old and young populations. Numerous solid tumors are associated with high cMET expression. The complexity of solid tumors combined with the highly interconnected nature of the cMET/HGF pathway with other cellular pathways make the pursuit of finding an effective treatment extremely challenging. The current standard of care for these malignancies is mostly small molecule-based chemotherapy. Antibody-based therapeutics as well as antibody drug conjugates are promising emerging classes against cMET-overexpressing solid tumors. In this study, we described the design, synthesis, in vitro and in vivo characterization of cMET-targeting Fab drug conjugates (FDCs) as an alternative therapeutic strategy. The format is comprised of a Fab conjugated to a potent cytotoxic drug via a cleavable linker employing lysine-based and cysteine-based conjugation chemistries. We found that the FDCs have potent anti-tumor efficacies in cancer cells with elevated overexpression of cMET. Moreover, they demonstrated a remarkable anti-tumor effect in a human gastric xenograft mouse model. The FDC format has the potential to overcome some of the challenges presented by the other classes of therapeutics. This study highlights the promise of antibody fragment-based drug conjugate formats for the treatment of solid tumors.


Reliability, Validity, and Feasibility of Water Displacement Method, Figure-of-Eight Method, and Circumference Measurements in Determination of Ankle and Foot Edema.
Background: To compare interrater reliability and the time-efficiency for the water displacement method, figure-of-eight method, and circumference measurements of the ankle and foot, and to investigate concurrent validity of the figure-of-eight method and circumference measurements with the water displacement method. Methods and Results: Thirty patients (21 women and 9 men) with primary or secondary lower limb lymphedema were evaluated twice. The volume of the foot and ankle was measured in three different ways: water displacement with a volumeter, figure-of-eight method, and circumference measurements at the level of the malleoli and metatarsals with a tapeline. The intraclass correlation coefficients (ICCs) ranged from good to excellent for all measurements (ICCs = 0.99 for water displacement; 0.94 for figure-of-eight; 0.80 and 0.79 for circumference measurement at malleolus and metatarsals). More time was needed to perform the water displacement method than the figure-of-eight method and the circumference measurements (p < 0.001). There was a statistically significant moderate correlation between the water displacement method and the figure-of-eight method (r = 0.65; p < 0.001), whereas the correlation coefficient between water displacement method and circumference measurements at malleolus and metatarsals was weak (r = 0.51, p < 0.001 and r = 0.34, p = 0.06, respectively). Conclusions: Compared to the water displacement method, reliability of the figure-of-eight method is comparable, time-efficiency is better and concurrent validity is acceptable. So, in clinical practice, the figure-of-eight method is a good alternative for the water displacement method to evaluate the volume of the foot and ankle.


[Expression of MEK/ERK signal pathways in renal cell carcinoma with bone metastasis].
To investigate the expression of MEK/ERK signaling pathways in renal cell carcinoma with bone metastasis, and to analyze the differences of expressions of VEGFR-2, MEK, ERK on the primary and metastasis tissue and its mechanism. The tissue samples were obtained from 7 renal cell carcinoma patients kindly provided by Department of Urology, Peking University People's Hospital from January 1, 2009 to January 1, 2010. The expression of MEK/ERK signaling pathways was detected in the 7 renal cell carcinoma patients` primary and matched metastatic tissues with ICH, The antibody concentrations were 1:200, 1:25, and 1:250, respectively. The mutation of the twentieth exon of the PDGFRA gene, the second exon of the K-ras gene, the fifteenth exon of the Braf gene and the second exon of the MEK1 gene were detected with PCR. The expression intensities of VEGFR-2, MEK, and ERK were measured by H-score [intensity (1, 2, 3, or 4) multiplied by the distribution (%)]. VEGFR-2, MEK, and ERK expressions were divided into 3 groups according to the positive distribution of the tumor cells: 1, 0-5%; 2, 6%-50%; and 3, >50%, To assess intratumor heterogeneity, three distinct microscopic fields (×200) from each specimen were used to evaluate the expressions, Subsequently, the scores were averaged to obtain a single concatenated score for each tissue. VEGFR-2, MEK, and ERK expressions were assessed by 2 independent pathologists who were blinded to the clinicopathological data. The data were expressed as the mean value of the triplicate experiments. The expressions of MEK, and ERK were higher in the metastatic tissues than in the matched RCC tissues (6.10±4.10 vs. 1.33±0.51, P=0.015; 9.10±2.24 vs. 4.43± 2.84, P=0.021) while the expression of VEGFR-2 was not different between the primary and metastatic tissues (P=0.901). No mutation was detected on the twentieth exon of the PDGFRA gene, the second exon of the K-ras gene, the fifteenth exon of the Braf gene and the second exon of the MEK1 gene. MEK/ERK signaling pathways may play an important role in the metastasis and the resistance of sunitinib in RCC patients with bone metastasis.


Mutation of the NADH oxidase gene (nox) reveals an overlap of the oxygen- and acid-mediated stress responses in Streptococcus mutans.
NADH oxidase (Nox) is a flavin-containing enzyme used by Streptococcus mutans to reduce dissolved oxygen encountered during growth in the oral cavity. In this study, we characterized the role of the NADH oxidase in the oxidative and acid stress responses of S. mutans. A nox-defective mutant strain of S. mutans and its parental strain, the genomic type strain UA159, were exposed to various oxygen concentrations at pH values of 5 and 7 to better understand the adaptive mechanisms used by the organism to withstand environmental pressures. With the loss of nox, the activities of oxygen stress response enzymes such as superoxide dismutase and glutathione oxidoreductase were elevated compared to those in controls, resulting in a greater adaptation to oxygen stress. In contrast, the loss of nox led to a decreased ability to grow in a low-pH environment despite an increased resistance to severe acid challenge. Analysis of the membrane fatty acid composition revealed that for both the nox mutant and UA159 parent strain, growth in an oxygen-rich environment resulted in high proportions of unsaturated membrane fatty acids, independent of external pH. The data indicate that S. mutans membrane fatty acid composition is responsive to oxidative stress, as well as changes in environmental pH, as previously reported (E. M. Fozo and R. G. Quivey, Jr., Appl. Environ. Microbiol. 70:929-936, 2004). The heightened ability of the nox strain to survive acidic and oxidative environmental stress suggests a multifaceted response system that is partially dependent on oxygen metabolites.


Down-regulation of nitrergic transmission in the rat striatum after chronic nigrostriatal deafferentation.
Dopamine and NO are physiological stimulators of synthesis of cAMP and cGMP, respectively, and NO synthase-containing interneurons in the striatum are physiologically activated by dopamine-containing neurons in the substantia nigra. This study investigated whether lesioning dopamine neurons has multiple consequences in the striatum consistent with the reported sensitization of cAMP synthesis, including alteration of the NO-cGMP pathway and phosphodiesterase-dependent metabolism of cyclic nucleotides. The substantia nigra of adult Sprague-Dawley rats was unilaterally lesioned with 6-hydroxydopamine. Two months later, we determined expression of NO synthase and evaluated cGMP and cAMP levels of intact and deafferented striatum. Moreover, we evaluated cAMP- and cGMP-phosphodiesterase activities in basal conditions and after Ca2+-calmodulin stimulation and determined the expression of the phosphodiesterase-1B isoform and the levels of phosphodiesterase-1B mRNA. Using immunocytochemistry we characterized the distribution of NO synthase and phosphodiesterase-1B within striatal neurons. In the dopamine-deafferented striatum, NO synthase levels were decreased by 42% while NO synthase-immunopositive intrastriatal fibres but not NO synthase neuronal bodies were reduced in number. In the deafferented striatum basal cGMP levels were reduced, and cAMP levels were increased, but cGMP-phosphodiesterase and cAMP-phosphodiesterase activities were both increased in basal and Ca2+-calmodulin-stimulated conditions. Accordingly, phosphodiesterase-1B expression and phosphodiesterase-1B mRNA were upregulated while a large population of medium-sized striatal neurons showed increased phosphodiesterase-1B immunoreactivity. Dopamine deafferentation led to a complex down-regulation of the NO-cGMP pathway in the striatum and to an up-regulation of phosphodiesterase-1B-dependent cyclic nucleotide metabolism, showing new aspects of neuronal plasticity in experimental hemiparkinsonism.


Two improved methods for preparing ferritin-protein conjugates for electron microscopy.
Two improved procedures were developed for activating ferritin so that the ferritin could be covalently linked to antibodies. One procedure involved use of a water-soluble carbodiimide and N-hydroxysuccinimide to prepare ferritin-containing activated esters. The other involved activation of the ferritin with excess glutaraldehyde. The ferritin-antibody conjugates prepared with the two procedures were shown to have a number of properties which made them suitable for locating antigenic components in cells.


Beyond HIV prevention: a qualitative study of patient-reported outcomes of PrEP among MSM patients in two public STD clinics in Baltimore.
Pre-exposure prophylaxis (PrEP) could have a substantial impact on the HIV epidemic within the US. However, the implementation of PrEP interventions outside of clinical trials has been slow and faces considerable barriers. The aim of the current study was to qualitatively explore PrEP-related patient-reported outcomes (PRO) among MSM patients who enrolled in a PrEP program at two public STD clinics in Baltimore. We conducted in-depth interviews with 18 PrEP patients who self-identified as MSM at two Baltimore City Health Department STD clinics between March and November, 2017. A codebook was developed using an iterative process. During analysis, the study team identified various biomedical and psychosocial PrEP-related PRO among MSM PrEP patients. In addition to HIV prevention, PrEP-related PRO included greater peace of mind, better continuity in care and awareness about health and well-being, relationship solidarity with serodiscordant partners, and access to social services. These findings on PrEP-related PRO can potentially contribute to improving patient-provider communication, leading to increased engagement in care and uptake of PrEP by MSM.


The effect of residual water on antacid properties of sucralfate gel dried by microwaves.
The aim of this work was to study the acid neutralization characteristics of microwave-dried sucralfate gel in relation to the water content and physical structure of the substance. Several dried sucralfate gels were compared with humid sucralfate gel and sucralfate nongel powder in terms of neutralization rate and buffering capacity. Humid sucralfate gel and microwave-dried gel exhibited antacid effectiveness. In particular, the neutralization rate of dried gel powders was inversely related to the water content: as the water content of dried powders decreased, the acid reaction rate linearly increased. The relationship was due to the different morphology of dried sucralfate gels. In fact, the porosity of the dried samples increased with the water reduction. However, the acid neutralization equivalent revealed that the dried sucralfate gel became more resistant to acid attack in the case of water content below 42%. Then, the microwave drying procedure had the opposite effect on the reactivity of the aluminum hydroxide component of dried sucralfate gel powders, since the rate of the reaction increased whereas the buffering capacity decreased as the amount of water was reduced.


The role of steroids in the management of brain metastases: a systematic review and evidence-based clinical practice guideline.
Do steroids improve neurologic symptoms in patients with metastatic brain tumors compared to no treatment? If steroids are given, what dose should be used? Comparisons include: (1) steroid therapy versus none. (2) comparison of different doses of steroid therapy. These recommendations apply to adults diagnosed with brain metastases. Steroid therapy versus no steroid therapy Asymptomatic brain metastases patients without mass effect Insufficient evidence exists to make a treatment recommendation for this clinical scenario. Brain metastases patients with mild symptoms related to mass effect Level 3 Corticosteroids are recommended to provide temporary symptomatic relief of symptoms related to increased intracranial pressure and edema secondary to brain metastases. It is recommended for patients who are symptomatic from metastatic disease to the brain that a starting dose of 4-8 mg/day of dexamethasone be considered. Brain metastases patients with moderate to severe symptoms related to mass effect Level 3 Corticosteroids are recommended to provide temporary symptomatic relief of symptoms related to increased intracranial pressure and edema secondary to brain metastases. If patients exhibit severe symptoms consistent with increased intracranial pressure, it is recommended that higher doses such as 16 mg/day or more be considered. Choice of Steroid Level 3 If corticosteroids are given, dexamethasone is the best drug choice given the available evidence. Duration of Corticosteroid Administration Level 3 Corticosteroids, if given, should be tapered slowly over a 2 week time period, or longer in symptomatic patients, based upon an individualized treatment regimen and a full understanding of the long-term sequelae of corticosteroid therapy. Given the very limited number of studies (two) which met the eligibility criteria for the systematic review, these are the only recommendations that can be offered based on this methodology. Please see "Discussion" and "Summary" section for additional details.


[Comparison of antimicrobial resistance pattern of selected respiratory tract pathogens isolated from different animal species].
The antibiotic resistance pattern of respiratory tract pathogens isolated of different animal species suffering from respiratory tract diseases has been investigated by antibiograms performed by agar diffusion test. The results show that the resistance situation in Switzerland is favourable compared with studies from other countries. However, high resistance rates were found in certain species: 61% of Streptococcus spp. were resistant to erythromycin and 44% to tetracycline, 59% of Bordetella bronchiseptica were resistant to ampicillin and 50% of Mannheimia (Pasteurella) haemolytica were multiresistant to tetracycline, ampicillin and streptomycine. The gram negative isolates were widely resistant to streptomycine.


[The historical experience of medical service in cooperation with state healthcare facilities during the Great Patriotic War 1941-1945].
Cooperation issues between military-medical service and civil healthcare in the field of delivery of medical aid to patients in the rear of country are considered in the artic. The rear is a final stage of the care by echelon and the main medical reserve force for front and army areas. Wide hospital network in the rear consisted mainly of evacuation hospitals of the People's Commissariat of the USSR healthcare. Cooperation between military-medical service and civil healthcare facilities was required. Sometimes necessary cooperation failed and made mutual helming of evacuation hospitals difficult. But despite the problems the main problem - return of maximum wounded soldiers to active duty was solved during the Great Patriotic War.


Cell mediated responses to the mycobacterium tuberculosis specific ESAT-6 antigen in Egyptian tuberculosis patients.
Human T cell responses and gamma interferon (IFN-gamma) secretion by peripheral blood mononuclear cells (PBMCs) in response to M. tuberculosis Early Secretory Antigen Target-6 (ESAT-6) eight synthetic overlapping peptides, were investigated in Egyptian tuberculosis patients as well as a control group. Three cell mediated immunoassays (lymphoproliferative, ELISPOT and intracellular flowcytometry) have been employed in this study to determine the ability of ESAT-6 to induce T-cell responses and IFN-gamma secretion, which play a critical role in protective cell-mediated immunity against tuberculosis (TB). The results revealed that all ESAT-6 peptides (P1-P8) were recognized by 85% of infected TB-patients, indicating that the molecule holds multiple epitopes. However, patients differed in the fine specificity of their peptide responses. Recognition of the N-terminal region of (P2-P7) was predominant. This study demonstrates that ESAT-6 is frequently recognized during infection and holds potential as a component of a future TB-vaccine.


[Effectiveness and toxicity of MOPP, ABVD, BEACOPP chemotherapy in first-diagnosed Hodgkin lymphoma with a poor prognosis].
In a retrospective study during the primary mode MOPP to primary patients LH II/IVAB stages with a poor prognosis rate of CR, 5--and 10-year DFS, OS was 69%, 71% and 68%, 74% and 64%, ABVD--76%, 78%, 83% and 68%, BEACOPP-baseline--73%, 97%, 85% and 82%, respectively. When the program ran BEACOPP-baseline it was revealed higher rates of DFS compared with ABVD and MOPP. Higher OS rates were observed in the primary patients treated with BEACOPP-baseline compared with MOPP (p = 0.04). In terms of DFS and OS MOPP regimen did not differ from ABVD. Program ABVD and BEACOPP-baseline had no differences in terms of OS. The frequency of primary refractory forms of LH did not depend on the conducted regimens of PCT. Significantly less recurrences occurred during the regimen of BEACOPP-baseline compared to MOPP and ABVD. BEACOPP-baseline was accompanied by a more pronounced reversible hematologic toxicity. Against the background of the program BEACOPP-baseline, neutropenia of III-IV degree was detected in 32%, ABVD--16%, MOPP--13%.


A Unified Alternating Direction Method of Multipliers by Majorization Minimization.
Accompanied with the rising popularity of compressed sensing, the Alternating Direction Method of Multipliers (ADMM) has become the most widely used solver for linearly constrained convex problems with separable objectives. In this work, we observe that many existing ADMMs update the primal variable by minimizing different majorant functions with their convergence proofs given case by case. Inspired by the principle of majorization minimization, we respectively present the unified frameworks of Gauss-Seidel ADMMs and Jacobian ADMMs, which use different historical information for the current updating. Our frameworks generalize previous ADMMs to solve the problems with non-separable objectives. We also show that ADMMs converge faster when the used majorant function is tighter. We then propose the Mixed Gauss-Seidel and Jacobian ADMM (M-ADMM) which alleviates the slow convergence issue of Jacobian ADMMs by absorbing merits of the Gauss-Seidel ADMMs. M-ADMM can be further improved by backtracking and wise variable partition. We also propose to solve the multi-blocks problems by Proximal Gauss-Seidel ADMM which is of the Gauss-Seidel type. It convegences for non-strongly convex objective. Experiments on both synthesized and real-world data demonstrate the superiority of our new ADMMs. Finally, we release a toolbox that implements efficient ADMMs for many problems in compressed sensing.


Integrated transcriptomic and metabolomic analysis of the central metabolism of Synechocystis sp. PCC 6803 under different trophic conditions.
Cyanobacteria have received considerable attention as a sustainable energy resource because of their organic material production capacity using light energy and CO2 as a carbon source. Therefore, it is important to understand the cellular metabolism of cyanobacteria for metabolic engineering. In this study, to shed light on the central metabolism of cyanobacteria, we performed transcriptomic and metabolomic analyses of a glucose-tolerant strain of the cyanobacterium Synechocystis sp. PCC 6803, which was cultured under auto- and mixotrophic conditions. Our results indicate that the oxidative pentose phosphate pathway and glycolysis are activated under mixotrophic conditions rather than autotrophic conditions. Moreover, we examined the effect of atrazine, a photosynthesis inhibitor, on the metabolism of PCC 6803 under mixotrophic conditions, which was defined as photoheterotrophic conditions, by transcriptomics and metabolomics. We demonstrated that the activity of the glycolytic pathway decreased due to the indirect effect of atrazine. In addition, the difference in transcriptional and metabolic changes between auto- and photoheterotrophic conditions could also be captured. The omics dataset reported herein provides clues for understanding the metabolism of cyanobacteria.


Donor lymphocyte infusion post-non-myeloablative allogeneic peripheral blood stem cell transplantation for chronic granulomatous disease.
Chronic granulomatous disease (CGD) is a primary immunodeficiency disease symptomized by failure to generate superoxide and recurrent bacterial and fungal infections. Allogeneic bone marrow transplantation (BMT) is one of the therapeutic options available. However, it presents considerable risk to the recipient, especially if the patient is already at an advanced stage of disease, after repeated bacterial and fungal infections and organ damage. We present a case report of a 6-year-old child with long-standing CGD, severe clubbing, and jeopardized pulmonary function after multiple bacterial pulmonary infectious episodes, who had failed treatment with sulphamethazole trimethoprim, multiple antibiotic courses, itraconazole, as well as steroid and interferon-y therapy. He underwent allogeneic peripheral blood stem cell transplantation (alloPBSCT) from his HLA-matched MLC non-reactive sister following non-myeloablative conditioning. His ANC did not fall below 0.2 x 10(9)/l, his lowest WBC was 0.6 x 10(9)/l, and his platelets did not fall below 28 x 10(9)/l. He had normal engraftment, with no mucositis or organ toxicity. Neither parenteral nutrition nor platelet infusions were necessary. Partial donor chimerism following alloPBSCT was converted to full donor chimerism and superoxide production reverted to normal after donor lymphocyte infusions (DLI) from his HLA-matched sister. Twenty four months post transplant the patient is well, with stable and durable engraftment, 100% donor chimerism, normal superoxide production, no GVHD, and stabilization of his pulmonary condition. We suggest that alloPBSCT preceded by non-myeloablative conditioning and followed by DLI may constitute a successful mode of therapy for patients suffering from advanced CGD with recurrent infectious episodes resulting in organ dysfunction, enabling them to achieve full donor chimerism and normal superoxide production with minimal risk of transplant-related toxicity and GVHD.


Steric strain and reactivity: electrophilic bromination of trans-(1-methyl-2-adamantylidene)-1-methyladamantane
trans-(1-Methyl-2-adamantylidene)-1-methyladamantane (DMAD, 1b) reacts with Br(2) in chlorinated hydrocarbon solvents to give either a bromonium polybromide ion pair or a substitution product, depending on bromine concentration. The first intermediate is a 1:1 pi-complex having K(f) = 1.85(0.19) x 10(3) M(-)(1) at 25 degrees C, which rapidly evolves to the bromonium tribromide ion pair. At high bromine concentration, which shifts all equilibria involving the counteranion of the ion pair intermediate toward the pentabromide species, this bromonium ion is stable and unable to further evolve into products. Temperature-dependent NMR spectra indicate chemical exchange of Br(+) between the sides of the plane containing the two carbons of the bromonium ion. At very low bromine concentration, no ionic intermediate is detected and the reaction rapidly yields a rearranged substitution product, identified as 10. Under these conditions the disappearance of the pi-complex follows a first-order rate law, and the observed rate constant increases with increasing olefin concentration, showing that product formation implies Br(-) as counteranion of the ionic intermediate, whose formation is a reversible process. A comparison of the results reported here for the bromination of 1b with those previously found for the parent olefin, adamantylideneadamantane (1a), shows that steric strain markedly affects the reactivity of the double bond.


Microtubule disassembly by ATP-dependent oligomerization of the AAA enzyme katanin.
Katanin, a member of the AAA adenosine triphosphatase (ATPase) superfamily, uses nucleotide hydrolysis energy to sever and disassemble microtubules. Many AAA enzymes disassemble stable protein-protein complexes, but their mechanisms are not well understood. A fluorescence resonance energy transfer assay demonstrated that the p60 subunit of katanin oligomerized in an adenosine triphosphate (ATP)- and microtubule-dependent manner. Oligomerization increased the affinity of katanin for microtubules and stimulated its ATPase activity. After hydrolysis of ATP, microtubule-bound katanin oligomers disassembled microtubules and then dissociated into free katanin monomers. Coupling a nucleotide-dependent oligomerization cycle to the disassembly of a target protein complex may be a general feature of ATP-hydrolyzing AAA domains.


Bicarbonate-activated persulfate oxidation of acetaminophen.
Persulfate (PS) is widely used as an oxidant for in situ chemical remediation of contaminated groundwater. In this study we demonstrated for the first time that PS could be activated by bicarbonate. Acetaminophen was used as the probe compound to examine the reactivity of PS/bicarbonate system. It was found that acetaminophen could be effectively transformed and the reaction rate appeared pseudo-first-order to the concentrations of both acetaminophen and PS. Radical scavenger tests indicated that neither free radicals (SO4- and HO) nor superoxide (O2-) was responsible for acetaminophen transformation. Generation of singlet oxygen (1O2) was verified using furfuryl alcohol (FFA) as a probe. Formation of 1O2 was further quantified in D2O fortified solution based on kinetic solvent isotopic effect (KSIE) but it was found that 1O2 contributed only 51.4% of the total FFA transformation. The other 48.6% was presumed to be ascribed to the reaction with peroxymonocarbonate (HCO4-). However, the transformation of acetaminophen was mostly due to the reaction with HCO4- but not 1O2. Instead of degradation, HCO4- oxidized acetaminophen via a one-electron abstraction mechanism resulting in the generation of acetaminophen radicals which coupled to each other to form dimers and trimers. HCO4- also hydrolyzed rapidly to form hydrogen peroxide (H2O2) which led to the formation of 1O2, during which O2- was a key intermediates. Because bicarbonate is ubiquitously presented in groundwater, the findings of this research provide important insights into the fundamental processes involved in PS oxidation in subsurface.


Nanotechnology in the Olympic Winter Games and Beyond.
Modern Olympics have increasingly benefited from the advancement of materials science. From February 4 to March 13, 2022, the whole world has witnessed spectacular Olympic Winter Games and Winter Paralympic Games held in Beijing, during which nanotechnologies are extensively applied and play indispensable roles in many aspects. Although these nanotechnologies appear in a very small length scale, they have provided tremendous support to all athletes with the pursuit of "faster, higher, stronger". In this Article, we highlight several representative nanotechnologies applied in the Olympic and Paralympic Winter Games including nanoscience in ice skating, wearable monitoring devices, skiing sports equipment, winter protection, and Olympic printing and discuss their physicochemical principles, unique significance for athletes and sports.


Working Memory Has Better Fidelity Than Long-Term Memory: The Fidelity Constraint Is Not a General Property of Memory After All.
How detailed are long-term-memory representations compared with working memory representations? Recent research has found an equal fidelity bound for both memory systems, suggesting a novel general constraint on memory. Here, we assessed the replicability of this discovery. Participants (total N = 72) were presented with colored real-life objects and were asked to recall the colors using a continuous color wheel. Deviations from study colors were modeled to generate two estimates of color memory: the variability of remembered colors-fidelity-and the probability of forgetting the color. Estimating model parameters using both maximum-likelihood estimation and Bayesian hierarchical modeling, we found that working memory had better fidelity than long-term memory (Experiments 1 and 2). Furthermore, within each system, fidelity worsened as a function of time-correlated mechanisms (Experiments 2 and 3). We conclude that fidelity is subject to decline across and within memory systems. Thus, the justification for a general fidelity constraint in memory does not seem to be valid.


Experimental demonstration of continuous quantum error correction.
The storage and processing of quantum information are susceptible to external noise, resulting in computational errors. A powerful method to suppress these effects is quantum error correction. Typically, quantum error correction is executed in discrete rounds, using entangling gates and projective measurement on ancillary qubits to complete each round of error correction. Here we use direct parity measurements to implement a continuous quantum bit-flip correction code in a resource-efficient manner, eliminating entangling gates, ancillary qubits, and their associated errors. An FPGA controller actively corrects errors as they are detected, achieving an average bit-flip detection efficiency of up to 91%. Furthermore, the protocol increases the relaxation time of the protected logical qubit by a factor of 2.7 over the relaxation times of the bare comprising qubits. Our results showcase resource-efficient stabilizer measurements in a multi-qubit architecture and demonstrate how continuous error correction codes can address challenges in realizing a fault-tolerant system.


Effects of dietary energy level and protein source on nutrient digestion and ruminal nitrogen metabolism in steers.
Four Simmental steers with ruminal, duodenal, and ileal cannulas were used to examine effects of dietary forage: concentrate ratio and supply of ruminally degradable true protein on site of nutrient digestion and net ruminal microbial protein synthesis. Steers (345 kg) were fed ammoniated corn cob (high forage; HF)- or corn cob/ground corn/cornstarch (low forage; LF)-based diets supplemented with soybean meal (SBM) or a combination of corn gluten meal and blood meal (CB). Diets were fed at 2-h intervals with average DM intake equal to 2.2% of BW. Feeding LF vs HF increased (P less than .05) OM digestion (percentage of intake) in the stomach, small intestine, and total tract. Efficiency of microbial CP synthesis (EMCP; g of N/kg of OM truly fermented) decreased (P less than .05) for LF vs HF (24.1 vs 26.8), but microbial N and total N flows to the small intestine were similar (P greater than .05) between energy levels (average 112 and 209 g/d, respectively). Total N flows to the small intestine were 13.1% greater (P less than .05) for CB than for SBM because of increased (P less than .05) passage of nonmicrobial N. Feeding SBM vs CB increased (P less than .05) EMCP (27.3 vs 23.3) and microbial N flow to the small intestine (127.5 vs 112.5 g/d), but these increases were not likely due to increased ruminal concentrations of ammonia N (NH3 N). Decreased (P less than .05) incorporation of NH3 N into bacterial N and slower turnover rates of ruminal NH3 N for SBM vs CB suggest that direct incorporation of preformed diet components into cell mass increased when SBM was fed. Results of this study suggest that the inclusion of ruminally degradable protein in the diet may increase the supply of products from proteolysis and that this can increase EMCP and microbial protein flow to the small intestine.


Survival analysis of patients with idiopathic dilated cardiomyopathy in Oman.
To study the 10-year survival of adults with idiopathic dilated cardiomyopathy (IDC) in Oman. Ninety-seven patients aged >13 years with IDC attending the Cardiology Unit, Sultan Qaboos University Hospital, Muscat, Oman from 1992-1995 were prospectively studied, in order to identify the outcome and factors contributing to death. Among 97 patients, 2 died from acute heart failure at presentation. The remaining 95 patients were followed up for periods ranging from 1-10 years (median 7 years). Twenty-four out of 95 patients exhibited clinical deterioration with reduced left ventricular ejection fraction (LVEF), by 5-11%, and 17 of them expired due to resistant heart failure. The remaining 71 patients remained stable and did not show deterioration in LVEF; however, 7 of them died suddenly at home possibly from ventricular arrhythmia. The survival rates were 94% at one year (95% confidence interval [CI] 88% to 99%), 86% at 3 years (95% CI 79% to 93%), and 64% at 10 years (95% CI 51% to 78%). Mean survival was 6.5 years (95% CI 6 to 7 years). Multivariate regression analysis revealed that factors related to death were LVEF <30% (p<0.001) and presence of severe mitral regurgitation (p=0.01). Outcome of dilated cardiomyopathy has improved due to greater understanding of this condition leading to better therapeutic approach. Resistant heart failure and cardiac arrhythmias remain the main causes of mortality. Poor outcome was related to low LVEF and presence of severe mitral regurgitation.


Impaired glycogenic substrate activation of glycogen synthase is associated with depressed synthase phosphatase activity in diabetic rat liver.
Glucose and gluconeogenic substrates promote the activation of hepatic glycogen synthase in vivo and in vitro; activation occurs as inactive glycogen synthase D is dephosphorylated to active glycogen synthase I by glycogen synthase phosphatase. Impairments of glycogen accumulation and glycogen synthase activation in diabetes have been attributed to decreased glycogen synthase phosphatase activity. To determine the role of glycogen synthase phosphatases associated with cytosol and smooth endoplasmic reticulum in the impairment of glycogen synthase activation, livers of normal and streptozotocin-diabetic fed rats were sampled by freeze-clamping before and after perfusion with a mixture of 25 mM glucose, 10 mM glutamine, 4 mM lactate, and 1 mM pyruvate. Perfusion induced activation of glycogen synthase in normal rats, but activation was reduced in the diabetic rats in proportion to the severity of insulin deficiency (r = 0.72, P less than 0.0001). There was also a close correlation between insulin levels and glycogen synthase phosphatase activities of both cytosol (r = 0.76, P less than 0.0001) and SER (r = 0.71, P less than 0.0001) fractions. In contrast, glycogen phosphorylase phosphatase activity and inactivation of glycogen phosphorylase during perfusion were normal in the diabetic livers. This is the first demonstration that glycogen synthase phosphatase activities in both soluble and SER fractions of liver cells are closely related to circulating insulin levels, and that the impairment of glycogen synthesis in diabetes may result from deficient glycogen synthase phosphatase activity in both cell compartments.


[Combination chemotherapy in the adjuvant treatment of disseminated breast cancer].
The results of clinical studies are presented on the combined chemotherapy in spread breast cancer, conducted according to the following three schedules: 1. thiophosphamide, carminomycin, vincristine, ftorafur, prednisolone, 2. diiodobenzoTEPA, carminomycin, vincristine, ftorafur, prednisolone, 3. methotrexate, cyclophosphane, vincristine, ftorafur, prednisolone. The combined chemotherapy in the complex treatment of 91 patients with spread breast cancer was found to be effective in 33-40% of cases. Primary tumors of the mammary gland and metastases in soft tissues, lymph nodes were mostly sensitive to the combined chemotherapy, metastases in bones and visceral organs--less sensitive. Schedules 2 and 3 proved to be the least toxic ones.


Detection of tumor ALK status in neuroblastoma patients using peripheral blood.
New protocols based on ALK-targeted therapy by crizotinib or other ALK-targeting molecules have opened for the treatment of patients with neuroblastoma (NB) if their tumors showed mutation and/or amplification of the ALK gene. However, tumor samples are not always available for analysis of ALK mutational status in particular at relapse. Here, we evaluated the ALK mutational status of NB samples by analysis of circulating DNA, using the droplet digital PCR (ddPCR) system. ddPCR assays was developed for the detection of ALK mutations at F1174 and R1275 hotspots found in NB tumors and was applied for the analysis of circulating DNA obtained from 200 μL of serum or plasma samples collected from 114 patients with NB. The mutations F1174L (exon 23 position 3520, T>C and position 3522, C>A) and the mutation R1275Q (exon 25 position 3824, G>A) were detected in circulating DNA. The sensitivity of our test was 100%, 85%, and 92%, respectively, and the specificity was 100%, 91%, and 98%, respectively. In conclusion, the assay that we have developed offers a reliable, noninvasive blood test to assess ALK mutational status at F1174 and R1275 hotspots and should help clinicians to identify patients showing an ALK mutation in particular when no tumor tissue is available.


Recent progress with functional biosupramolecular systems.
The objective of this account is to summarize our recent progress with functional biosupramolecular systems concisely. The functions covered are artificial photosynthesis, anion transport, and sensing in lipid bilayer membranes. With artificial photosynthesis, the current emphasis is on the construction of ordered and oriented architectures on solid surfaces. Recent examples include the zipper assembly of photosystems with supramolecular n/p-heterojunctions and oriented antiparallel redox gradients. Current transport systems in lipid bilayers reveal new interactions at work. Examples include anion-macrodipole or anion-π interactions. Current attention with membrane-based sensing systems shifts from biosensor approaches with enzymatic signal generation to aptamers (i.e., the DNA version of immunosensing) and differential sensing with dynamic polyion-counterion transporters. The functional diversity accessible with biosupramolecular systems is highlighted, as is the critical importance of cross-fertilization at intertopical convergence zones.


Anuran forelimb muscle tendinous structures and their relationship with locomotor modes and habitat use.
The interaction between organisms and their environment is central in functional morphology. Differences in habitat usage may imply divergent morphology of locomotor systems; thus, detecting which morphological traits are conservative across lineages and which ones vary under environmental pressure is important in evolutionary studies. We studied internal and external morphology in 28 species of Neotropical anurans. Our aim was to determine if internal morphology (muscle and tendons) shows lower phylogenetic signal than external morphology. In addition, we wanted to know if morphology varies in relation to the habitat use and if there are different functional groups. We found differences in the degree of phylogenetic signal on the groups of traits. Interestingly, postaxial regions of the forelimb are evolutionarily more labile than the preaxial regions. Phylomorphospace plots show that arboreal (jumpers and graspers) and swimmer frogs cluster based on length of fingers and the lack of sesamoid, also reflected by the use of habitat. These functional clusters are also related to phylogeny. Sesamoid and flexor plate dimensions together with digit tendons showed to be important to discriminate functional groups as well as use of habitat classification. Our results allow us to identify a "grasping syndrome" in the hand of these frogs, where palmar sesamoid and flexor plate are absent and a third metacarpal with a bony knob are typical. Thus, a lighter skeleton, long fingers and a prensile hand may be key for arboreality.


Bullous pemphigoid and multiple sclerosis: a report of two cases with ELISA test.
We report two patients with longstanding multiple sclerosis (MS) who developed vesicles and bullae consistent with the diagnosis of bullous pemphigoid (BP). Both patients showed linear IgG at the dermal-epidermal junction, located on the epidermal side of patients' skin previously treated with 1M NaCl. In the two cases, the ELISA test was positive for the extracellular fragment of BP 180. However, the indirect immunofluorescence test (IIF) was repeatedly negative. Therapy either with prednisone plus dapsone or prednisone alone was initiated and the disease was controlled after 23 and 15 months of therapy, in patients 1 and 2, respectively. However, the first patient had a flare-up 2 months after treatment was stopped. The association of MS and BP has been described previously in 35 cases. We compare our two cases with the 25 patients previously reported in detail in the literature. We emphasize the role of the ELISA test in establishing the diagnosis of BP.


Acid alpha-glucosidase in Malaysians. Population studies and the occurrence of a new variant.
Acid alpha-glucosidase from the placenta was electrophoretically surveyed in a total of 633 Malaysians, 236 of Malay, 261 of Chinese and 136 of Indian ancestries. A new variant, alpha-glucosidase 3-1 was observed in 1 Malay and 3 Indians. A polymorphism for this enzyme was observed among Indians, but in Chinese and Malays variants are rare. Phenotype 2-1 was observed once in a Chinese and once in a Malay.


A plaque technique for assay and characterization of antibody-dependent cytotoxic effector (K) cells.
Using monolayers of erythrocytes as target cells, a plaque assay was developed by which individual antibody-dependent cytolytic lymphoid cells (K cells) could be identified at the cellular level. The extent of plaque formation depended on the concentration of sensitizing antibody, time of incubation, and number of lymphocytes added. Most of the plaque-forming cells (50% to 70%) were shown to possess receptors for activated complements, a small but variable fraction (5% to 25%) had surface Ig detectable by indirect immunofluorescence, and 5% to 10% bound sheep erythrocytes at 5 degrees C. The plaque-forming cells appeared as villous, small to medium-sized lymphocytes when studied by transmission or scanning electron microscopy. Under standardized experimental conditions the minimal number of active, plaque-forming K cells in purified lymphocyte preparations could be estimated.


ATM and TP53 mutations show mutual exclusivity but distinct clinical impact in mantle cell lymphoma patients.
Mantle cell lymphoma (MCL) is characterized by the hallmark t(11;14)(q13;q32) translocation, leading to cyclin D1 over-expression. Additionally, disrupting the DNA damage response pathway through ATM or TP53 defects plays an important role in MCL pathogenesis. Using deep next-generation sequencing we analyzed the mutual composition of ATM and TP53 mutations in 72 MCL patients, and assessed their impact on progression-free survival (PFS) and overall survival (OS). Mutated ATM and TP53 alleles were found in 51% (37/72) and 22% (16/72) of the cases examined, respectively, with only three patients harboring mutations in both genes. Only a mutated TP53 gene was associated with the significantly reduced PFS and OS and the same output was observed when ATM and TP53 defective groups included also sole deletions 11q and 17p, respectively. Determining the exact ATM/p53 pathway dysfunction may influence the selection of MCL patients for innovative therapies based on the targeted inhibition of selected proteins.


Comparison of Postoperative Respiratory Monitoring by Acoustic and Transthoracic Impedance Technologies in Pediatric Patients at Risk of Respiratory Depression.
In children, postoperative respiratory rate (RR) monitoring by transthoracic impedance (TI), capnography, and manual counting has limitations. The rainbow acoustic monitor (RAM) measures continuous RR noninvasively by a different methodology. Our primary aim was to compare the degree of agreement and accuracy of RR measurements as determined by RAM and TI to that of manual counting. Secondary aims include tolerance and analysis of alarm events. Sixty-two children (2-16 years old) were admitted after tonsillectomy or receiving postoperative patient/parental-controlled analgesia. RR was measured at regular intervals by RAM, TI, and manual count. Each TI or RAM alarm resulted in a clinical evaluation to categorize as a true or false alarm. To assess accuracy and degree of agreement of RR measured by RAM or TI compared with manual counting, a Bland-Altman analysis was utilized showing the average difference and the limits of agreement. Sensitivity and specificity of RR alarms by TI and RAM are presented. Fifty-eight posttonsillectomy children and 4 patient/parental-controlled analgesia users aged 6.5 ± 3.4 years and weighting 35.3 ± 22.7 kg (body mass index percentile 76.6 ± 30.8) were included. The average monitoring time per patient was 15.9 ± 4.8 hours. RAM was tolerated 87% of the total monitoring time. The manual RR count was significantly different from TI (P = .007) with an average difference ± SD of 1.39 ± 10.6 but were not significantly different from RAM (P = .81) with an average difference ± SD of 0.17 ± 6.8. The proportion of time when RR measurements differed by ≥4 breaths was 22% by TI and was 11% by RAM. Overall, 276 alarms were detected (mean alarms/patient = 4.5). The mean number of alarms per patient were 1.58 ± 2.49 and 2.87 ± 4.32 for RAM and TI, respectively. The mean number of false alarms was 0.18 ± 0.71 for RAM and 1.00 ± 2.78 for TI. The RAM was found to have 46.6% sensitivity (95% confidence interval [CI], 0.29-0.64), 95.9% specificity (95% CI, 0.90-1.00), 88.9% positive predictive value (95% CI, 0.73-1.00), and 72.1% negative predictive value (95% CI, 0.61-0.84), whereas the TI monitor had 68.5% sensitivity (95% CI, 0.53-0.84), 72.0% specificity (95% CI, 0.60-0.84), 59.0% positive (95% CI, 0.44-0.74), and 79.5% negative predictive value (95% CI, 0.69-0.90). In children at risk of postoperative respiratory depression, RR assessment by RAM was not different to manual counting. RAM was well tolerated, had a lower incidence of false alarms, and had better specificity and positive predictive value than TI. Rigorous evaluation of the negative predictive value is essential to determine the role of postoperative respiratory monitoring with RAM.


Increased membrane protein phosphorylation and anion transport activity in chorea-acanthocytosis.
Chorea-acanthocytosis is a disorder characterized by neuronal degeneration and the presence of acanthocytic erythrocytes on blood smear. The abnormal function and structure of the membrane protein band 3 are considered to be of pathogenetic relevance in determining the erythrocyte defect. In a clinically evident case of chorea-acanthocytosis, the following parameters were investigated: membrane cholesterol and fatty acid composition, sulphate influx (as a measure of the anion transport activity), membrane protein phosphorylation, membrane casein and tyrosin-kinase activities; moreover, the promoter and all exons of the EPB3 gene were screened for possible mutations by single strand conformational polymorphism (SSCP) study. The sulphate influx, the Ser/Thr phosphorylation level, and the membrane casein-kinase activity were increased in chorea-acanthocytosis compared with normal controls. In the intact vanadate-treated 32P-labelled erythrocytes, Tyr-phosphorylation of the cytoplasmic domain of band 3, as well as the poly(Glu, Tyr) kinase activity in the membranes, were enhanced in the patient's sample. Apparent molecular weight and concentration of band 3 on SDS/PAGE analysis, membrane fatty acid composition and cholesterol/phospholipid molar ratio were normal and the SSCP study of EPB3 exons did not show any abnormal polymorphisms. An abnormal degree of phosphorylation of membrane proteins, in particular of band 2 (beta-subunit) and band 3, may contribute in determining both change of cell shape and increased anion transport in chorea-acanthocytosis.


Comparison of effects of forskolin, cAMP, and vasopressin on Pf/Pd(w) of toad urinary bladder luminal membrane.
We have reported that Pf/Pd(w) (the ratio of osmotic and diffusional water permeabilities) for the luminal membrane of toad urinary bladder is approximately 17 for tissues stimulated with either vasopressin or 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP). In a recent abstract, Kachadorian and co-workers have shown that tissues stimulated with adenosine 3',5'-cyclic monophosphate (cAMP) or forskolin have a lower Pf than would be anticipated from the frequency of aggregates visualized on the flat portion of the luminal membrane using freeze-fracture electron microscopy. We report here measurements of Pf/Pd(w) for the luminal membrane of tissues receiving these agents: Pf/Pd(w) for submaximally stimulated tissues was the same, regardless of whether the stimulant was vasopressin (12.7 +/- 0.3), forskolin (13.7 +/- 0.9), or cAMP (12.0 +/- 1.3). The calculated Pd(w)'s for the series barrier were also identical (6.8 +/- 0.5, 6.5 +/- 0.3, and 8.2 +/- 1.0 X 10(-4) cm/s respectively). Our data, taken together with those of Kachadorian et al. are consistent with a number of possibilities: because our methodology does not permit estimation of Pf for the series barrier, we cannot rule out the possibility of a "post-luminal barrier" that is rate-limiting for Pf, but not for Pd(w) in forskolin- and cAMP-stimulated tissues, Pf and Pd(w) of the luminal surface aggregates could decrease in parallel, so that luminal membrane Pf/Pd(w) remains constant, and there could be a diminished frequency of fused aggregate-rich aggrephores, but not of aggregates that are on the flat portion of the luminal membrane. Only the latter can be unequivocally quantitated using freeze-fracture electron microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)


Mechanical contraction property of the levator veli palatini muscle.
Twenty adult dogs anesthetized with sodium pentobarbital were used as experimental animals in an analysis of the mechanical contraction properties of the levator veli palatini muscle. The following results were obtained: the contraction time and half relaxation time of maximal isometric twitch of the muscle were revealed to be 43 msec and 33 msec, respectively; a summation curve was obtained with stimulation at 15 Hz and maximal tetanic tension was at 70 Hz. The average of the tetanic tension ratio was found to be 5.7. The maximum rate of tension rise was observed at 150 Hz, and the value was twice the rate of the twitch contraction. The duration time of the active state plateau showed values of 3.9 to 4.5 msec. The contraction time of the levator veli palatini muscle closely resembles that of the intrinsic laryngeal and the orbicularis oris muscle.


Plasmid DNA in Treponema pallidum (Nichols): potential for antibiotic resistance by syphilis bacteria.
A plasmid DNA structure (approximate molecular weight = 7.5 X 10(6)) was identified in the human pathogen Treponema pallidum (Nichols). The inability to isolate this plasmid from rabbit host tissue and the total lack of DNA homology of the plasmid with rabbit DNA has confirmed its Treponema pallidum origin. The observation documents a newly recognized and potentially significant genetic capability for Treponema pallidum.


Variation in care for recurrent nonmelanoma skin cancer in a university-based practice and a veterans affairs clinic.
To learn if treatment of recurrent nonmelanoma skin cancer (NMSC) varied in different practice settings. Prospective cohort study of consecutive patients with recurrent NMSC. A university-based dermatology practice and the dermatology clinic at the affiliated Veterans Affairs Medical Center (VAMC). Conventional therapies for NMSC were available at both sites. Patients All 191 patients diagnosed as having recurrent NMSC in 1999 and 2000 were included in the study. Data were collected from medical record review and surveys mailed to patients. Main Outcome Measure Performance of Mohs micrographic surgery (Mohs). Patients at the VAMC were older, less educated, poorer, and had more comorbid illnesses, but their tumors were similar to those of patients at the university-based practice. Treatment choices differed at the 2 sites: the proportions of tumors treated in the VAMC and university sites were 60% and 14%, respectively, for excisional surgery; and 24% and 61%, respectively, for Mohs (P < .001). In multivariate analyses adjusting for patient, tumor, and physician features that may have affected treatment choice, tumors treated at the university-based site remained significantly more likely to be treated with Mohs (odds ratio, 8.68 [95% confidence interval, 3.66-20.55]; P < .001). Substantial variation existed in the treatment of recurrent NMSC in different practice settings. This variation was not explained by measured clinical characteristics of the patients or the tumors.


Identification of a Novel Mutation in the SEC23B Gene Associated With Congenital Dyserythropoietic Anemia Type II Through the Use of Next-generation Sequencing Panel in an Undiagnosed Case of Nonimmune Hereditary Hemolytic Anemia.
Congenital dyserythropoietic anemias (CDAs) are rare hereditary blood disorders characterized by ineffective erythropoiesis, hemolysis, and erythroblast morphologic abnormalities in the bone marrow. The 3 main types of CDA, I to III, and variant types of CDA, IV-VIII, have been described. The causative genes have been identified as CDAN1, C15ORF41, SEC23B, KIF23, KLF1, and GATA1. CDA type II is the most frequent form. Typical symptoms are jaundice, hepatosplenomegaly, mild-to-severe normocytic anemia, and inadequate reticulocyte response. We report an 18-year-old boy who had chronic mild congenital anemia, jaundice, and splenomegaly mimicking nonautoimmune hemolytic anemia since 18 months of age. Compound heterozygous mutations in SEC23B gene were detected by the use of a gene-targeted next-generation sequencing panel: the already reported missense mutation c.40C>T (p.Arg14Trp), and a new frameshift deletion (c.489_489delG, p.Val164Trpfs*3), confirming the diagnosis of CDA type II. The study underlines the molecular heterogeneity of CDA II and the importance of a precise diagnosis in rare congenital diseases such as CDA II. In consequence, it can be difficult to diagnose because of limited resources, financial constraint, and rarity of disease in the developing country. Advanced laboratories and new molecular approaches may help in diagnosing rare anemias.


Cellular physiology of the turtle visual cortex: distinctive properties of pyramidal and stellate neurons.
The electrophysiological properties of neurons in the three-layered dorsal cortex of the turtle, Pseudemys scripta elegans, have been studied in vitro. Intracellular recordings suggested two distinct classes of neuronal behavior. Cell labeling with either Lucifer Yellow or horseradish peroxidase revealed that these behaviors correlated with the two morphological classes of cortical neurons: pyramidal cells and stellate cells. Examination of Golgi-stained neurons of dorsal cortex did not uncover any other obvious classes. Pyramidal cells had their somata in the cell layer, and extended several densely spined apical dendrites through the molecular layer to the pia. They also had spiny basilar dendrites directed through the subcellular layer toward the ependymal border. Physiologically, pyramidal cells had relatively prolonged action potentials that showed marked frequency adaptation during a sustained suprathreshold current pulse. Their most striking characteristic was a tendency to fire two discrete sizes of action potential, one small (mean = 34 mV) and of relatively low threshold, the other large (mean = 76 mV) and of higher threshold. We hypothesize that at least some small spikes arise from distal dendritic sites, whereas large spikes are somatically generated. Both spikes were tetrodotoxin-sensitive, although calcium-dependent electrogenesis occurred when potassium channels were blocked. In contrast to pyramidal cells, the somata of stellate cells were found in the molecular and subcellular zones. Their dendrites tended to be horizontally oriented and spine-free. Stellate cells had relatively brief action potentials, each of which was followed by a large but short-lasting undershoot of membrane potential. Stellate cells showed little or no spike frequency adaptation. Spike amplitudes were always relatively uniform and large (mean = 73 mV). Thus, in the dorsal cortex of turtles, the pyramidal cells, which are projection neurons, and stellate cells, which are local GABAergic inhibitory neurons, have distinctly different membrane characteristics. The physiological properties of the two types of turtle cortical neurons are very similar to their counterparts in cortical structures of the mammalian telencephalon.


Linking fungal communities in roots, rhizosphere, and soil to the health status of Pisum sativum.
Changes in fungal communities associated with healthy and diseased pea roots were investigated using deep amplicon pyrosequencing in three spatial compartments: roots, rhizosphere, and surrounding soil. Thirty root systems were collected from three fields, half of which showing clear symptoms of root rot. In total, 500,461 internal transcribed spacer-1 sequences were obtained that were clustered into 123 (roots), 271 (rhizosphere), and 440 (bulk soil) nonsingleton operational taxonomic units (OTUs). Species richness was highest in bulk soils and lowest in roots; however, no notable differences in richness were observed between samples associated with diseased and healthy roots. Health status and field both had significant effects on fungal community structures in roots, whereas only field had significant effects on communities in rhizosphere and bulk soils. Indicator species analysis across the three fields identified a number of OTUs that were more abundant in healthy roots. Pathogens such as Fusarium oxysporum were abundant in diseased roots in some fields. Patterns of disease and causal agents of root rot were different among the three fields, which were also reflected in fungal communities. In conclusion, health status of roots was only vaguely reflected in rhizosphere and bulk soil fungal communities, whereas health status was more important for shaping root communities.


[Practical proctology (author's transl)].
Hemorrhoidal complaints are the most frequent diseases of the rectal region and form the starting point for practically all other diseases of the rectal area. Acute bleeding often first brings the patient to the doctor, who must establish the source of the hemorrhage with all diagnostic means available. Further stages of the disease are temporary or complete prolapse of the hemorrhoids, under certain circumstances even prolapse of the rectum. Anal cryptitis is the first member of a chain of inflammatory sequelae such as perianal abscess and fistular diseases. It is frequently overlooked, yet in the early stages it could easily be cut short by suitable measures. Proctology has a responsible task to fulfill in the early recognition of diseases of the rectum and malignant tumors of the colon.


Melatonin inhibits the proliferation of human osteosarcoma cell line MG-63.
It seems established that the onset of osteosarcoma and the reduction in melatonin production run in parallel; this suggests that the decline in the cancer-inhibiting agent, melatonin, may contribute to the occurrence of osteosarcoma and that melatonin supplementation may have promise for preventing the development and progression of this condition. There is, however, no direct evidence regarding an antiproliferative effect of melatonin in osteosarcoma cells. In the current study, we examined whether melatonin inhibits the proliferation of human osteosarcoma cell line MG-63. MTT staining showed that at 4 mM-10 mM concentrations, melatonin significantly reduced the MG-63 cell proliferation in a dose-dependent and time-dependent manner. Flow cytometry documented that 4 mM melatonin significantly increased the fraction of cells in the G(0)/G(1) phase of the cell cycle, while simultaneously reducing the proportion in the S and G(2)/M phases. Western blot and real-time PCR analyses further confirmed that melatonin's inhibitory effect was possibly because of downregulation of cyclin D1 and CDK4, related to the G(1) phase, and of cyclin B1 and CDK1, related to the G(2)/M phase. There was no downregulation of cyclin E, CDK2, and cyclin A, which are related to G(1)/S transition and S phase. These findings provide evidence that melatonin may significantly inhibit human osteosarcoma cell proliferation in a dose-dependent and time-dependent manner and this inhibition involves the downregulation of cyclin D1, CDK4, cyclin B1 and CDK1.


[Immunological indicators in children with laryngeal papillomatosis and possible ways of immunocorrection].
Clinicoimmunological characteristics were followed up in the course of laryngeal papillomatosis (LP) in 96 children. It was found that they had suppressed T-cell immunity, phagocytosis which still worsen with the disease aggravation and recurrences. A decrease in nonspecific resistance, properdin, lysozyme, complement and secretory IgA indicated impairment of nasopharyngeal and laryngeal local immunity. A scheme of combined treatment is proposed incorporating basic therapy with reaferon, systemic immunomodulators (thymalin, T-activin) and local immunocorrection. Such approach reduced the time of basic treatment and prolonged remission 2.5-3-fold.


New beta-adrenergic agonists used illicitly as growth promoters in animal breeding: chemical and pharmacodynamic studies.
Clenbuterol and beta-adrenergic receptor agonist drugs are illegally used as growth promoters in animal production. Pharmacologically active residues in edible tissues led to intoxication outbreaks in several countries. Pressure of official controls pulsed synthesis of new compounds to escape analytical procedures. We report two new compounds named 'A' and 'G4', found in feeding stuffs. Chemical structure was studied through nuclear magnetic resonance-imaging and infrared spectroscopy, and beta(1)- and beta(2)-adrenergic activity was evaluated on isolated guinea-pig atrium and trachea in comparison with clenbuterol. Both compounds share with clenbuterol an halogenated aromatic ring with a primary amino group. Main modifications consisted of substitution of secondary amino group with an alkyl chain in compound A and substitution of the ter-butyl group with a benzene ring in compound G4. In guinea-pig trachea these compounds showed myorelaxant potency lower than clenbuterol (EC(50) was 43.8 nM for clenbuterol, 11700 nM for compound A, 2140 nM for G4). On the contrary, in the guinea-pig atrium (heart-beat rate stimulant effect) the compounds were more potent than clenbuterol (EC(50) was 15.2 nM for clenbuterol, 3.4 nM for compound A, 2.8 nM for G4). These pharmacodynamic properties, and stronger lipophilic properties shown by the two compounds may result in increased cardiovascular risk for consumers of illicitly treated animals.


Perceived expectations in daily life among adolescent girls suffering from an eating disorder: a phenomenographic study.
The aim of this study was to describe the variation of perceived expectations in daily life of adolescent girls suffering from eating disorders. Eighteen interviews with girls between the age of 15 and 19 years were analyzed according to the phenomenographic approach. The results could be summarized into three categories: expectations from others, self-imposed expectations, and conflicting expectations. Each of these could be further subdivided into two conceptions. Conflicting and unattainable expectations were severe stressors in the participants' lives. Many participants had problems identifying expectations that were not explicit. The participants internalized these expectations and interpreted them to be self-imposed. Clinical implications are suggested.


Incidence of BK with tacrolimus versus cyclosporine and impact of preemptive immunosuppression reduction.
Our purposes were to determine the incidence of BK viruria, viremia or nephropathy with tacrolimus (FK506) versus cyclosporine (CyA) and whether intensive monitoring and discontinuation of mycophenolate (MMF) or azathioprine (AZA), upon detection of BK viremia, could prevent BK nephropathy. We randomized 200 adult renal transplant recipients to FK506 (n = 134) or CyA (n = 66). Urine and blood were collected weekly for 16 weeks and at months 5, 6, 9 and 12 and analyzed for BK by polymerase chain reaction (PCR). By 1 year, 70 patients (35%) developed viruria and 23 (11.5%) viremia; neither were affected independently by FK506, CyA, MMF or AZA. Viruria was highest with FK506-MMF (46%) and lowest with CyA-MMF (13%), p = 0.005. Viruria >/= 9.5 log(10) copies/mL was associated with a 3-fold increased risk of viremia and a 13-fold increased risk of sustained viremia. After reduction of immunosuppression, viremia resolved in 95%, without increased acute rejection, allograft dysfunction or graft loss. No BK nephropathy was observed. Choice of calcineurin inhibitor or adjuvant immunosuppression, independently, did not affect BK viruria or viremia. Viruria was highest with FK506-MMF and lowest with CyA-MMF. Monitoring and preemptive withdrawal of immunosuppression were associated with resolution of viremia and absence of BK nephropathy without acute rejection or graft loss.


Assessing the Effect of Zinc Supplementation on the Frequency of Migraine Attack, Duration, Severity, Lipid Profile and hs-CRP in Adult Women.
Migraine is a widespread incapacitating neurologic disorder with debilitating headaches which are usually throbbing due to inefficacy and several side effects, complementary therapies recommended as possible alternatives. The current randomized controlled trial was carried out to evaluate the effect of zinc gluconate supplementation on migraine-related symptoms, serum level of high sensitivity C-reactive protein (hs-CRP) and lipid profile in migraineurs. Present study was designed as randomized double-blind, placebo-controlled trial. Sixty women with migraine (mean age of 35.44 ± 7.42 years) were randomly allocated to obtain 15 mg per day of zinc gluconate or placebo for 12 weeks. Frequency, periods of headaches and severity of migraine based on numerical rating scale questionnaire and migraine disability assessment (MIDAS) test were checked. Fasting serum level of lipid profile and hs-CRP were assessed at the beginning and the end of trial. Zinc gluconate supplementation significantly reduced the frequency (p = 0.001), periods of migraine attacks (p < 0.001) and severity of migraine and MIDAS (p < 0.001) compared with control group. The serum level of low-density lipoprotein (p < 0.001), total cholesterol (p < 0.001) and hs-CRP (p < 0.001) decreased following zinc supplementation, but no significant differences in serum level of triglycerides (p = 0.1) and high-density lipoprotein (p = 0.3) was observed. However, after adjustment for baseline values using analysis of covariance test, none of lipid profile components and hs-CRP showed a significant difference. Zinc supplementation has beneficial effect on the migraine related complications like its severity, frequency. Iranian Registry of Clinical Trials Identifier: IRCT20191014045100N1.


Impact of Severe Obesity and Weight Loss on Systolic Left Ventricular Function and Morphology: Assessment by 2-Dimensional Speckle-Tracking Echocardiography.
Obesity is associated with an increased risk of heart failure. Little is known about the impact of dietary changes on the cardiac sequelae in obese patients. Twenty-one obese subjects underwent a 12-week low calorie fasting phase of a formula diet. Transthoracic two-dimensional speckle-tracking echocardiography was performed to obtain systolic left ventricular strain before and after weight loss. Body mass index decreased significantly from 38.6 ± 6.2 to 31.5 ± 5.3 kg/m(2), and the total percentage fat loss was 19%. Weight reduction was associated with a reduction in blood pressure and heart rate. Left ventricular longitudinal global peak systolic strain was in the lower normal range (-18.7 ± 3.2%) before weight loss and was unchanged (-18.8 ± 2.4%) after 12 weeks on diet with substantial weight loss. Also, no significant change in global radial strain after weight loss was noted (41.1 ± 22.0 versus 43.9 ± 23.3, p = 0.09). Left atrial and ventricular dimensions were in normal range before fasting and remained unchanged after weight loss. In our study obesity was associated with normal systolic left ventricular function. A 12-week low calorie diet with successful weight loss can reduce blood pressure and heart rate. Systolic left ventricular function and morphology were not affected by rapid weight reduction.