Oxygen toxicity in neonatal rats: the effect of endotoxin treatment on survival during and post-O2 exposure.
Neonatal rats were treated with low doses of bacterial lipopolysaccharide (endotoxin) to test for a protective effect of endotoxin against O2 toxicity and the severe inhibition of normal lung development which occurs during prolonged exposure to hyperoxia. The rationale for the prophylactic use of endotoxin included its marked protective effect against pulmonary O2 toxicity in adult rats and its lung growth-promoting effect in experimental pulmonary stress models. Neonatal rats (4-5 days old) survived a 14-day exposure to greater than 95% O2 equally well whether treated with saline (39/51 = 76%) or with endotoxin (41/51 = 80%). However, during the following 24 h of gradual weaning to room air breathing, there was a marked difference in survival between the endotoxin group (32/41 = 78%) and the saline pups (14/39 = 36%) (p less than 0.001). Both groups showed inhibition of lung development (alveolarization) during O2 exposure, but endotoxin treatment compared to saline was associated with increased specific lung volume (5.33 versus 4.50 ml/100 g) (air control = 4.08), smaller mean airspace diameter (mean linear intercept = 49.0 versus 55.8 microns) (air control = 43.3), increased specific internal surface area (4393 versus 3232 cm2/100 g) (air control = 3753), and greater preservation of alveolar wall capillary patency (24.83 versus 18.52% "capillary density") (air control = 27.70%). We conclude that endotoxin treatment resulted in significant protection against O2 toxicity in neonatal rats which was manifested during readaptation to room air breathing. The protective effect was likely due to a combination of reduced inhibition of lung growth and development and reduced hyperoxic damage to the respiratory membrane of the lung.


Proteomics of the neurotoxic fraction from the sea anemone Bunodosoma cangicum venom: Novel peptides belonging to new classes of toxins.
In contrast to the many studies on the venoms of scorpions, spiders, snakes and cone snails, up to now there has been no report of the proteomic analysis of sea anemones venoms. In this work we report for the first time the peptide mass fingerprint and some novel peptides in the neurotoxic fraction (Fr III) of the sea anemone Bunodosoma cangicum venom. Fr III is neurotoxic to crabs and was purified by rp-HPLC in a C-18 column, yielding 41 fractions. By checking their molecular masses by ESI-Q-Tof and MALDI-Tof MS we found 81 components ranging from near 250 amu to approximately 6000 amu. Some of the peptidic molecules were partially sequenced through the automated Edman technique. Three of them are peptides with near 4500 amu belonging to the class of the BcIV, BDS-I, BDS-II, APETx1, APETx2 and Am-II toxins. Another three peptides represent a novel group of toxins (~3200 amu). A further three molecules (~ approximately 4900 amu) belong to the group of type 1 sodium channel neurotoxins. When assayed over the crab leg nerve compound action potentials, one of the BcIV- and APETx-like peptides exhibits an action similar to the type 1 sodium channel toxins in this preparation, suggesting the same target in this assay. On the other hand one of the novel peptides, with 3176 amu, displayed an action similar to potassium channel blockage in this experiment. In summary, the proteomic analysis and mass fingerprint of fractions from sea anemone venoms through MS are valuable tools, allowing us to rapidly predict the occurrence of different groups of toxins and facilitating the search and characterization of novel molecules without the need of full characterization of individual components by broader assays and bioassay-guided purifications. It also shows that sea anemones employ dozens of components for prey capture and defense.


Side effects and compliance with low- and conventional-dose oral contraceptives among adolescents.
Oral contraceptives are the most popular birth control method for teenagers, yet many teens discontinue use of these contraceptives prematurely. The need to minimize any potential long-term medical complications from the use of contraceptive hormones must be balanced with the desirability of increasing acceptance of contraceptives by adolescents. There has been concern that the use of so-called "low-dose" estrogen preparations, although decreasing the likelihood of complications, may lead to side effects that make compliance less certain. The present study comparing two commonly used oral contraceptive preparations, one low dose, one conventional dose, tests the hypothesis that among adolescents an association exists between oral contraceptive side effects and compliance. Using a double-blind crossover method, 55 sexually active adolescent females received two months each of a preparation containing 35 micrograms ethinyl estradiol and 0.5 mg norethindrone and another containing 50 micrograms mestranol and 1.0 mg norethindrone. The 50-microgram preparation was associated with fewer side effects when administered during the first two months. No differences in side effects were noted in the latter two months, but there was a slight increase in weight gain when compared with the 35-microgram preparation. The most common side effect was inter-menstrual bleeding with the 35-microgram pill. There was no documented relationship between the occurrence of side effects and compliance.


Correlates of cognitive impairment in first episode schizophrenia: the EUFEST study.
Profile and correlates of cognitive deficits in first episode (FE) schizophrenia patients are still debated. The present study is aimed to clarify in a large sample of FE patients the extent of impairment in key cognitive domains and its relationships with demographic and clinical variables. The European First Episode Schizophrenia Trial collected demographic, clinical and neurocognitive baseline data in 498 FE patients with minimal or no prior exposure to antipsychotics. Two-hundred-twenty healthy subjects (HS) were also evaluated. Neurocognitive assessment included the Rey Auditory Verbal Learning Test; Trail Making A and B, Purdue Pegboard and Digit-Symbol Coding. Patients performed worse than HS on all tests (effect sizes from -0.88 to -1.73). Correlations with psychopathological dimensions were weak and involved reality distortion and disorganization. The duration of untreated psychosis (DUP) was not associated with cognitive impairment. Subjects living alone had a better neurocognitive performance, while the occupation status did not reveal any association with cognition. A moderate/severe impairment of processing speed, motor dexterity, verbal memory and cognitive flexibility was found in the largest sample of FE patients analyzed so far. The impairment was largely independent from psychopathology and not associated with DUP.


Pseudodystonia: A new perspective on an old phenomenon.
Pseudodystonia represents a wide range of conditions that mimic dystonia, including disorders of the peripheral nervous system, spinal cord, brainstem, thalamus, cortex and non-neurological conditions such as musculoskeletal diseases. Here, we propose a definition of pseudodystonia and suggest a classification based on underlying pathophysiological mechanisms. We describe phenomenology of different forms of pseudodystonia and point to distinctions between dystonia and pseudodystonia as well as challenging issues that may arise in clinical practice. The term pseudodystonia can be used to describe abnormal postures, repetitive movements or both, in which results of clinical, imaging, laboratory or electrophysiological investigations provide definite explanation of symptoms which is not compatible with dystonia. Pseudodystonia can be classified into non-neurological disorders of the musculoskeletal system, disorders of sensory pathways, disorders of motor pathways and compensatory postures in other neurological diseases. Presence of associated neurological findings in the affected body part is the key towards diagnosis of pseudodystonia. Additional supporting features are the presence of fixed postures, the absence of sensory trick, acute mode of onset and severe pain. Worsening on eye closure, traditionally considered typical for pseudodystonia, is not always present and can also appear in dystonia. It is challenging to separate dystonia and pseudodystonia in patients with thalamic lesions or corticobasal syndrome, where abnormal postures coexist with sensory loss. Many cases of pseudodystonia are treatable. Therefore, it is essential to consider pseudodystonia in a differential diagnosis of abnormal postures until a detailed neurological examination rules it out.


Purification and identification of dTDP-oleandrose, the precursor of the oleandrose units of the avermectins.
The nucleotide sugar precursor of the oleandrose units of the avermectins has been purified from a mutant of Streptomyces avermitilis, which does not synthesize any avermectins but which converts avermectin aglycones to their respective disaccharides. This precursor has been identified as dTDP-oleandrose. The purification was achieved by anion exchange and reverse phase high performance liquid chromatography. The purified nucleotide sugar had an absorption spectra characteristic of thymidine, released dTMP when treated with phosphodiesterase, and possessed an NMR spectrum in which three resonances characteristic of oleandrose were seen in addition to the thymidine signals. The enzyme, avermectin aglycone dTDP-oleandrose glycosyltransferase, which catalyzes the stepwise addition of oleandrose to the avermectin aglycones, has been demonstrated in cell-free extracts and (NH4)2SO4 fractions of cell-free extracts of S. avermitilis. The enzyme is specific for dTDP-oleandrose as the glycosyl donor but utilizes all avermectin aglycones as glycosyl acceptors. The stoichiometry between dTDP-oleandrose consumed in the reaction and oleandrose units transferred to the avermectin mono- and disaccharide was found to be 1:1.


Effects of sulfur-containing analogues of stearic acid on growth and fatty acid biosynthesis in the protozoan Crithidia fasciculata.
A variety of analogues of stearic acid in which one of the methylene groups was replaced by a sulfur atom were examined as inhibitors of growth and fatty acid biosynthesis in the trypanosomatid protozoan Crithidia fasciculata. The 8-, 9-, 10-, and 11-thiastearic acids were found to suppress the synthesis of the cyclopropane-containing fatty acid dihydrosterculic acid (9,10-methyleneoctadecanoic acid) at micromolar concentrations in the growth medium, and all but the 9-thiastearate were found to inhibit the growth of the protozoa at concentrations. The most potent inhibitor, 8-thiastearic acid (I50 for growth = 0.8 microM; I50 dihydrosterculate synthesis = 0.4 microM), was also observed to inhibit the synthesis of gamma-linolenic acid at a similar concentration. The sulfoxide derivatives of the 9- and 10-thiastearates were found to have little effect on growth or fatty acid synthesis, and several long-chain amides of 3-amino-1,2-propanediol were found to have effects similar to those of the fatty acids from which they were derived.


Respiratory viral infections in immunocompetent and immunocompromised persons.
The acute respiratory illnesses are the most common type of acute illness in the United States today. The respiratory viruses--which include influenza viruses, parainfluenza viruses, respiratory syncytial virus (RSV), rhinoviruses, coronaviruses, and adenoviruses--cause the majority of these illnesses. Some of these viruses cause illness throughout the year, whereas others are most common in winter. All population groups experience these infections and illnesses. As the number of elderly persons and those with underlying disease increases, awareness is growing that these common infections can have serious consequences. This has recently been emphasized for immunocompromised persons. At the M.D. Anderson Cancer Center (MDACC), infection surveillance of mostly hospitalized adults with leukemia or a recent bone marrow transplant yielded a respiratory virus from 181 of 668 (27.1%) respiratory illness episodes. In descending order of frequency, infections with RSV, rhinoviruses, influenza viruses, parainfluenza viruses, and adenoviruses were detected in each of three surveillance years. High frequencies of nosocomial acquisition occurred, as has been noted in prior reports. Similarly, persistence of infection and high frequencies of pneumonia and death among infected patients occurred, which have also been noted earlier. At MDACC, pneumonia occurred in 58-78% of infected patients, and 22-44% died. The role of the virus infection in many cases of pneumonia is uncertain, but death from pure viral pneumonia is well documented. A number of immune deficiencies in this patient population and options for control of these infections have been described that can, respectively, account for the medical problem and provide ways to approach prevention and treatment.


Parathyroid hormone-related protein (PTHrP) signal cascade modulates myocardial dysfunction in the pressure overloaded heart.
Pressure overload induces the cardiac expression of parathyroid hormone-related protein (PTHrP). Plasma levels are elevated in patients with heart disease. It is unknown whether this represents an epiphenomenon or suggests involvement in hypertrophy. To identify a potential role of PTHrP in pressure induced hypertrophy and heart failure. Pressure load was produced via thoracic aortic constriction (TAC) and application of a PTHrP antagonist (PTHrP(7-34)) via osmotic minipumps in mice. Main findings were confirmed in vitro by exposing isolated adult ventricular mice cardiomyocytes to PTHrP(1-34) (100 nmol/l). TAC treated animals developed myocardial hypertrophy within 2 weeks. The heart weight to body weight ratio increased from 5.02+/-0.14 mg/g (sham/vehicle) and 5.16+/-0.19 mg/g (sham/antagonist) to 6.59+/-0.85 mg/g (TAC/vehicle) and 7.07+/-0.80 mg/g (TAC/antagonist) (each n=6-8; p<0.05 for TAC vs. sham; not significantly different between TAC groups). In parallel, the expression of atrial natriuretic factor increased. Cardiac dysfunction (+dP/dt, -dP/dt), however, was significantly lower in TAC mice receiving the antagonist, and SERCA2 expression was higher. Isolated cardiomyocytes exposed to PTHrP(1-34) developed reduced cell shortening. This reduction in cell function was abolished in the co-presence of the antagonist. PTHrP contributes to the progression of cardiac dysfunction in the pressure overloaded heart.


Some statistical issues in project prioritization in the pharmaceutical industry.
Various aspects of portfolio management and project prioritization within the pharmaceutical industry are examined. It is shown that the cost and probability architecture of a project is a crucial aspect of its value. An appropriate simple tool for ranking projects is the Pearson index. Various difficulties are considered.


Cystic fibrosis: a disease of altered protein folding.
Cystic fibrosis (CF) is caused by mutations in the gene that encodes the cystic fibrosis transmembrane conductance regulator, CFTR. Previously we demonstrated that the common delta F508 mutation in the first nucleotide binding domain (NBD1) alters the ability of the domain to fold into a functional three-dimensional structure, providing a molecular explanation for the observation that the mutant CFTR is retained in the endoplasmic reticulum and does not traffic to the apical membrane of affected epithelial cells. Notably, when conditions are altered to promote folding of the mutant protein, it can assume a functional conformation. Correcting the folding defect may have therapeutic benefit for the treatment of cystic fibrosis. Here we summarize these results and discuss the implications in vitro folding studies have for understanding the pathobiology of CF.


A qubit coupled with confined phonons: the interplay between true and fake decoherence.
The decoherence of a qubit coupled with the phonons of a finite-size lattice is investigated. The confined phonons no longer behave as a reservoir. They remain sensitive to the qubit so that the origin of the decoherence is twofold. First, a qubit-phonon entanglement yields an incomplete true decoherence. Second, the qubit renormalizes the phonon frequency resulting in fake decoherence when a thermal average is performed. To account for the initial thermalization of the lattice, the qua- ntum Langevin theory is applied so that the phonons are viewed as an open system coupled with a thermal bath of harmonic oscillators. Consequently, it is shown that the finite lifetime of the phonons does not modify fake decoherence but strongly affects true decoherence. Depending on the values of the model parameters, the interplay between fake and true decoherence yields a very rich dynamics with various regimes.


Assessment of CFTR function in homozygous R117H-7T subjects.
R117H is a frequent missense mutation included in most CFTR mutation panels. However knowledge about the residual function of R117H-CFTR channels in cystic fibrosis-affected organs, e.g. airways, intestines and sweat glands is presently lacking. We evaluated clinical CF symptoms and assessed CFTR function by sweat tests, nasal potential difference and intestinal current measurements in 2 homozygous R117H individuals (7T variant). The CFTR activity in airways and intestine was within the normal range. However both individuals presented with a borderline sweat test and the male patient was infertile. The lack of impact of the R117H mutation on chloride secretion in intestine and nose contrasts with the ~80% loss of CFTR activity reported in patch clamp studies. Apparently CFTR activity is not rate-limiting for chloride secretion in both tissues at levels >20% of normal, or compensatory factors may operate that are absent in heterologous host cells in vitro.


Potential for adhesive wear in friction couples of UHMWPE running against oxidized zirconium, titanium nitride coatings, and cobalt-chromium alloys.
The classical wear mechanisms abrasion, fatigue, and adhesion are the most frequent causes of surface changes of ultra high molecular weight polyethylene (UHMWPE) in artificial joints. The counterpart material has a strong influence on the wear and friction behavior of artificial joints due to its abrasive properties and adhesive interaction with UHMWPE. The formation of a transfer layer on the counterpart in UHMWPE bearing systems is often described as being a clear indication of strong adhesive forces. The influence of using a cobalt-chromium-molybdenum (CoCrMo) alloy, a titanium nitride plasma coating or an oxidized zirconium alloy on adhesive wear was studied. The surface free energy and the bonding forces of these counterpart materials to UHMWPE were investigated. Catalytic effects on the degradation behavior of polyethylene, the micro friction behavior, and the build-up and constitution of a transfer layer deposited under loads, and relative velocities that are relevant in knee joints were analyzed.


Surgical management and evaluation of prognostic factors influencing postoperative visual outcome of suprasellar meningiomas.
To study the microsurgical technique and the prognostic factors influencing postoperative visual outcome in patients with suprasellar meningiomas (SMs). The clinical materials of 45 patients with SMs treated microsurgically between January 2002 and October 2008 were analyzed retrospectively. Patients received ophthalmologic and radiologic assessment before and after operation. Follow-up ranged from 12 to 93 months (median, 39 months). As far as monocular vision was concerned, univariate and multivariate statistical analysis was performed among factors that might influence postoperative visual outcome. In this group, the mean age of the patients was 51 years. Median tumor size was 3.5 cm. The duration of symptoms ranged from 10 days to 35 years (median, 18 months). Total tumor resection was achieved in 40 cases (88.9%) and subtotal in 5 cases (11.1%). With respect to visual outcome of the 90 eyes, 54 eyes (60%) improved, 24 (26.7%) remained unchanged, and 12 (13.3%) had worsened. It has been illustrated that the postoperative visual improvement was determined by various factors, including age, recurrence, duration and severity of visual disturbance, preoperative condition of optic disc, tumor size, location, peritumoral edema, involvement with optic canal, arachnoid membrane interface, and extent of tumor removal. However, the multivariate analysis showed that recurrence, preoperative condition of optic disc, duration of impaired vision, and peritumoral edema are the most influential factors. Skillful microsurgical techniques are key to resect SMs while getting the desired visual outcome. The recurrence, preoperative condition of optic disc, duration of impaired vision, and peritumoral edema might be the most important factors influencing postoperative visual outcome.


Altered tyrosine phosphorylation via the very late antigen (VLA)/beta1 integrin stimulation is associated with impaired T-cell signaling through VLA-4 after allogeneic bone marrow transplantation.
Our previous study showed that the cross-linking of very late antigen (VLA)/beta1 with anti-CD29 monoclonal antibody (MoAb), or interactions with extracellular matrix (ECM) proteins through VLA/beta1, failed to induce T-cell costimulation via the CD3/T cell receptor (TCR) pathway for over 1 year after allogeneic bone marrow transplantation (allo-BMT), although normal CD29 and CD3 expression was observed after 3 months following allo-BMT. Molecular analysis revealed altered tyrosine phosphorylation of cellular proteins by the solid-phase cross-linking of VLA/beta1 molecules in T cells from patients after allo-BMT. In T cells from early allo-BMT patients (<4 months), various sizes of highly tyrosine phosphorylated proteins were observed as high background even without the stimulation through VLA/beta1 integrin. The high tyrosine phosphorylation pattern gradually disappeared and it was finally returned to normal tyrosine phosphorylation patterns by 2 years after BMT. Interestingly, poor expression of focal adhesion kinase (pp125FAK), a VLA/beta1-mediated signaling molecule, was observed within 1 year after BMT. These results suggest that these molecular defects appear to be implicated in the impaired VLA/beta1-mediated signaling in T cells from patients after allo-BMT, and it could explain, in part, the persistent immunoincompetent state after allo-BMT at least 1 year.


Potential role of MG53 in the regulation of transforming-growth-factor-β1-induced atrial fibrosis and vulnerability to atrial fibrillation.
Atrial fibrosis plays a critical role in atrial fibrillation (AF) by the transforming growth factor (TGF)-β1/Smad pathway. The disordered differentiation, proliferation, migration and collagen deposition of atrial fibroblasts play significant roles in atrial fibrosis. Mitsugumin (MG)53 is predominantly expressed in myocardium of rodents and has multiple biological functions. However, the role of MG53 in cardiac fibrosis remains unclear. This study provided clinical and experimental evidence for the involvement of MG53 in atrial fibrosis in humans and atrial fibrosis phenotype in cultured rat atrial fibroblasts. In atrial tissue from patients we demonstrated that MG53 was expressed in human atrium. Expression of MG53 increased with the extent of atrial fibrosis, which could induce AF. In cultured atrial fibroblasts, depletion of MG53 by siRNA caused down-regulation of the TGF-β1/Smad pathway, while overexpression of MG53 by adenovirus up-regulated the pathway. MG53 regulated the proliferation and migration of atrial fibroblasts. Besides, exogenous TGF-β1 suppressed expression of MG53. In conclusion, we demonstrated that MG53 was expressed in human atrium, and may be a potential upstream of the TGF-β1/Smad pathway in human atrium and rat atrial fibroblasts. This suggests that MG53 is a potential regulator of atrial fibrosis induced by the TGF-β1/Smad pathway in patients with AF.


Predicting early signs of dyslexia at a preliterate age by combining behavioral assessment with structural MRI.
Recent studies suggest that neurobiological anomalies are already detectable in pre-school children with a family history of developmental dyslexia (DD). However, there is a lack of longitudinal studies showing a direct link between those differences at a preliterate age and the subsequent literacy difficulties seen in school. It is also not clear whether the prediction of DD in pre-school children can be significantly improved when considering neurobiological predictors, compared to models based on behavioral literacy precursors only. We recruited 53 pre-reading children either with (N=25) or without a family risk of DD (N=28). Quantitative T1 MNI data and literacy precursor abilities were assessed at kindergarten age. A subsample of 35 children was tested for literacy skills either one or two years later, that is, either in first or second grade. The group comparison of quantitative T1 measures revealed significantly higher T1 intensities in the left anterior arcuate fascicle (AF), suggesting reduced myelin concentration in preliterate children at risk of DD. A logistic regression showed that DD can be predicted significantly better (p=.024) when neuroanatomical differences between groups are used as predictors (80%) compared to a model based on behavioral predictors only (63%). The Wald statistic confirmed that the T1 intensity of the left AF is a statistically significant predictor of DD (p<.05). Our longitudinal results provide evidence for the hypothesis that neuroanatomical anomalies in children with a family risk of DD are related to subsequent problems in acquiring literacy. Particularly, solid white matter organization in the left anterior arcuate fascicle seems to play a pivotal role.


[Duration of the cell cycles and mitotic phases during the cleavage period of the salmon Salmo salar L].
The temporal, parameters of the cell cycle in the Atlantic salmon embryos were studied at the temperature 4.8 +/- 0.1 degrees from 10.5 hrs after fertilization (end of interphase of the 1st cleavage division) till 93 hrs (beginning of blastulation). The period of cleavage comprises 11 cell cycles. The II--V cell cycles equal each other by their duration, then the divisions accelerate and the VII--X cell cycles are shorter than the former by 10%, on the average. The duration of some phases (pro-, ana- and telophase) in different cycles proved to be variable and all mitotic phases varied by their relative duration from cycle to cycle; however, in all cell cycles of the cleavage period the mitosis occupied 2/3 of each cycle. The synchroneous character of the passage of cell cycle phases by all the cells is preserved only up to 8-blastomere stage. Beginning from the 16-blastomere stage when two cell layers appear along the animal--vegetative axis, the gradient is initiated: the animal layer cells in the cycle phases leave behind the vegetative layer cells which border the yolk.


Advances in upper airway diseases and allergen immunotherapy.
Evidence of an increased prevalence of rhinitis in patients with asthma, and asthma in patients with rhinitis, supports the 1 airway concept. However, there are basic differences between the upper and lower airways, such as the virtual absence of remodeling in the nose compared with the bronchi, despite the presence of similar inflammation. Etiologic factors in chronic rhinosinusitis (CRS) attract increasing interest. Peripheral blood monocytes from patients with CRS release IL-4, IL-13, and IFN-gamma on stimulation with fungal antigens, especially those from Alternaria. This is not seen with cells from normal controls. However, a double-blind trial of intranasal amphotericin in CRS was negative. Evidence continues to accumulate of the pivotal role of regulatory T-lymphocytes secretion of IL-10 in the response to allergen immunotherapy. In patients with asthma and house dust mite allergy who are receiving appropriate pharmacotherapy and have instituted environmental controls, house dust mite immunotherapy provides marginal additional benefits in asthma control. Immunotherapy with cat dander extract at a maintenance dose containing 15 microg Fel d 1 produces a more consistent immunologic response than with maintenance doses containing 3.0 microg, whereas doses containing only 0.6 microg are no more effective than placebo. Sublingual immunotherapy for seasonal grass allergy can be safely administered by general practitioners, but symptom relief begins only in the second season of therapy. Sublingual immunotherapy for seasonal grass allergy in children reduced symptoms and onset of new asthma symptoms but, again, beginning only in the second year of treatment. A course of 6 weekly injections of ragweed Amb a 1 bound to cytosine phosphorothionate guanosine containing DNA produced a shift from T H 2 to T H 1 cytokine release both in peripheral blood cells and in the nose after allergen challenge. No symptom improvement was seen the first year, but symptoms were reduced the second year without further treatment.


EVALUATION OF VITRECTOMY AND REIMPLANTATION FOLLOWING LATE DISLOCATION OF THE INTRAOCULAR LENS-CAPSULAR BAG COMPLEX: A 3-Year Follow-up Study.
To study the predisposition factors for dislocation of the intraocular lens (IOL)-capsular bag complex and analyze the results of subsequent reimplantation surgery. The exclusion criteria were complicated cataract surgery and minor dislocations that appeared in the first year after cataract surgery. Thirty-six months of monitoring of the reimplantation surgery was required. The primary measurements of results were factors concerning dislocation, the interval between cataract surgery and dislocation, and postoperative complications. Thirty-six patients with dislocation of the capsular bag and lens were identified. Pseudoexfoliation was mentioned in 17 cases (47.2%) and was the main predisposition factor. The average interval between cataract surgery and dislocation was 11.5 years. The dislocated IOL was replaced by a posterior chamber implant in 38% of cases and an anterior chamber implant in 62%. The average best-corrected visual acuity improved significantly after the reposition surgery (P < 0.01). The average monitoring period after secondary reimplantation was 5.95 years (min. 3.03-max. 8.46). Three patients developed bullous keratopathy, all of them with an anterior chamber IOL implantation. According to our knowledge, this article is the one with the longest monitoring time to date. Pseudoexfoliation was the main risk factor for the dislocation of the IOL. Surgery significantly improved best-corrected visual acuity, and the reimplantation of the IOL in the posterior chamber was associated with less serious complications.


The role of stem cell transplantation in the management of relapsed follicular lymphoma in the era of targeted therapies.
Patients with relapsed follicular lymphoma (FL) following first-line therapy have an increasing number of management strategies ranging from observation through to stem cell transplantation. There has been an exciting expansion in novel agents to treat FL, but at present there is not a universally accepted standard of care in the relapse setting. Decision-making can be difficult for the clinician as there is a paucity of data to compare the various relapse therapies available. This review will discuss conventional therapy for relapsed FL, consider the use of novel agents and explore the role and timing of autologous and allogeneic stem cell transplantation.


Influence of abiraterone acetate on circulating neuromediators in chemotherapy-naïve castration-resistant prostate cancer.
Abiraterone Acetate (AA) represents a highly effective androgen-receptor (AR) axis targeted agent. Treatment with AA in castration-resistant prostate cancer (CRPC) may partly mediate neuroendocrine differentiation (NED) as an escape mechanism, which may have implications for the choice of sequential therapy in CRPC. We evaluated how treatment with AA influences circulating neuromediators chromogranin A (CGA), neuron-specific enolase (NSE), and pro-gastrin-releasing peptide (Pro-GRP) in chemotherapy-naïve CRPC patients. We conducted an analysis in chemotherapy-naïve CRPC patients with clinical or radiographic progression of disease. A total of 35 patients were included at five institutions between February 2013 and December 2014. Sixteen of them had received AA. Serum samples were obtained before a docetaxel-based chemotherapy and analyzed in a reference laboratory. Univariable and multivariable analyses were performed to test the influence of AA treatment, its duration of treatment, and other clinicopathological variables on circulating neuromediators. CGA and NSE levels were above the upper limit of normal (ULN) in n = 20 (57.1%) and n = 13 (37.1%), respectively. Treatment with AA and duration of treatment were not associated with levels above the ULN (CGA and NSE) or higher levels (Pro-GRP) of neuromediators. CGA levels were associated with age (P = 0.092), lymph node metastasis (P = 0.014), duration of androgen deprivation therapy (ADT; P = 0.083), and intake of proton pump inhibitors (P = 0.069). Pro-GRP levels were significantly associated with PSA levels (P = 0.002). On multivariate analysis, CGA levels above the ULN were significantly correlated with ADT (P = 0.01) and intake of proton pump inhibitors (P = 0.03). Circulating neuromediators in chemotherapy-naïve CRPC patients were elevated in a high percentage of patients. ADT was found to be a relevant NED driver in this cohort. Our results may imply that patients with CRPC after first-line treatment with AA in CRPC are not at a higher risk for developing NED. The major limitation of the study represents the one-time analysis of neuromediators. Larger studies with serial blood measurements or biopsy analysis before and after treatment are needed to confirm our results.


Activation of the masseter muscle during normal smile production and the implications for dynamic reanimation surgery for facial paralysis.
In cases of unilateral facial paralysis, free muscle transfer with coaptation to the motor nerve of the Masseter is gaining popularity as a primary alternative to cross-facial nerve grafting. Despite initial expectations, a majority of these subjects can achieve a spontaneous smile. The mechanism behind this spontaneity is unclear. Plasticity of the cerebral cortex as well as the relative proximity of the motor centres of the mimetic and Masseter muscles has been used in explanation. This study demonstrates the involvement of the Masseter muscle during normal smile production, suggesting a more direct explanation for the spontaneous smile seen following reanimation procedures innervated by the Masseter nerve. Twenty healthy volunteers were subjected to electromyography of the Masseter muscle bilaterally to demonstrate whether contraction of the Masseter muscle occurred during voluntary and involuntary smile production. Patient age ranged from 20 to 61 years (mean 41.6 years) with an equal male to female ratio. Activation of the Masseter occurred in 40 percent of individual muscles during smile production, occurring bilaterally in six participants, and unilaterally in four. There was no correlation between muscle activation and patient age or gender. Natural contraction of the Masseter muscle during normal smile production helps to explain the high rate of spontaneous smile development in subjects with facial paralysis who have undergone a free muscle reanimation procedure powered by the nerve to the Masseter muscle.


Variation of zeta-potential with temperature in fused-silica capillaries used for capillary electrophoresis.
The temperature variation of electroosmotic mobility corrected for the effects of Joule heating (muEOF0) was employed to investigate the variation of the zeta-potential (zeta) with temperature in fused-silica capillaries. Experimentally determined values for zeta increased at 0.39% per degrees C, a rate that is about four to five times smaller than reported previously. Experimentally determined values of zeta were directly proportional to the absolute temperature although values were also influenced slightly by changes to the dielectric constant. It was found that the effective charge density at the inner surface of the capillary was independent of temperature.


Facilitating an understanding of integrative physiology: emphasis on the composition of body fluid compartments.
As a teaching exercise, we used deductive reasoning and a quantitative analysis to convert a number of facts into a series of concepts to facilitate an understanding of integrative physiology and shed light on the composition of the different body fluid compartments. The starting point was the central need to regenerate ATP to perform biologic work. Because a large quantity of O2 must be delivered to cells at a sufficiently high concentration to aid its diffusion into mitochondria, approximately one third of the O2 in inspired air was extracted; this led to a P(CO2) in arterial blood of 40 mmHg (1 mmHg = 133.322 Pa). Blood flow to individual organs must be adjusted precisely to avoid having too low or too high a P(O2) in mitochondria--the latter augments the formation of reactive O2 species. The extracellular fluid (ECF) bicarbonate concentration (E(HCO3)) must be high to minimize H+ buffering by proteins. This high E(HCO3) sets the ECF concentrations of ionized calcium (Ca2+) and inorganic phosphate (HPO4(2-)) because of solubility issues. Three features defined the intracellular fluid (ICF) volume and composition. First, expelling monovalent anions minimized its mass (volume). Second, controlling the tissue P(CO2) ensured a relatively constant net valence on intracellular proteins. Third, the range of ICF Ca2+ concentrations must both induce regulatory signals and avoid Ca3(PO4)2 formation. All the above were incorporated into the integrated response that optimized the capacity for vigorous exercise.


Non-markovian dynamics of spectral narrowing for excitons in the layered semiconductor GaSe observed using optical four-wave mixing spectroscopy.
We have investigated non-markovian dephasing by using time-resolved and spectrally resolved four-wave mixing measurements in a layered semiconductor GaSe. When the time interval between the first and second excitation pulses is increased, photon echo spectra exhibit narrowing only in a regime of a few picoseconds. In the initial dephasing of these signals, fast damping is observed. The narrowing of the spectrally resolved signal is consistent with the Fourier transformation of the time-resolved signals. Spectral narrowing is crucial evidence of the transition from non-markovian to markovian dynamics. By comparing experimental data with calculation results based on the non-markovian theory, we have found that the correlation time of the exciton-phonon interaction is 1.1 ps.


Blood pressure control and knowledge of target blood pressure in coronary patients across Europe: results from the EUROASPIRE III survey.
Blood pressure management is a key issue among patients with coronary heart disease (CHD). The present study aimed to identify particular patient groups that may need to be specifically targeted in secondary prevention of CHD. EUROASPIRE III is a cross-sectional study conducted in 2006-2007 among patients up to 80 years of age hospitalized for CHD. Patients from 76 centres in 22 European regions were examined on average 15 months after hospitalization. Logistic regression analysis was applied to investigate factors associated with blood pressure control and knowledge of target blood pressure using the cut-point of less than 140/90 mmHg. Among 7649 patients using antihypertensive medication 50.4% achieved blood pressure control and 49.4% provided accurate knowledge of target blood pressure. Obese patients were less likely to show controlled blood pressure [odds ratio (OR) 0.72, 95% confidence interval (CI) 0.65-0.80] and accurate knowledge of blood pressure target values (OR 0.80, 95% CI 0.72-0.90). Dyslipidaemia was negatively associated with blood pressure control and accurate target knowledge. Patients with diabetes mellitus less frequently achieved blood pressure control (OR 0.89, 95% CI 0.79-0.99). Accurate knowledge of target blood pressure was positively related to blood pressure control (OR 1.12, 95% CI 1.00-1.24). Patients who received advice by a health professional to reduce salt intake, to reduce weight, and to increase physical activity more frequently showed accurate knowledge of blood pressure target values. Blood pressure control and knowledge of target blood pressure are inappropriate in the European high-risk population of coronary patients. Particularly CHD patients with obesity, diabetes, and dyslipidaemia need better management and control of elevated blood pressure.


Argyria secondary to ingestion of homemade silver solution.
Argyria is a rare skin disease caused by cutaneous deposition of silver granules in the skin as a result of exposure to silver substrate or ingestion of silver salt. This report describes a patient with generalized argyria caused by ingestion of homemade colloidal silver solution. The patient learned about the uses of the silver solution and its preparation at a convention for "natural medicine."


Circulating desmosine levels do not predict emphysema progression but are associated with cardiovascular risk and mortality in COPD.
Elastin degradation is a key feature of emphysema and may have a role in the pathogenesis of atherosclerosis associated with chronic obstructive pulmonary disease (COPD). Circulating desmosine is a specific biomarker of elastin degradation. We investigated the association between plasma desmosine (pDES) and emphysema severity/progression, coronary artery calcium score (CACS) and mortality.pDES was measured in 1177 COPD patients and 110 healthy control subjects from two independent cohorts. Emphysema was assessed on chest computed tomography scans. Aortic arterial stiffness was measured as the aortic-femoral pulse wave velocity.pDES was elevated in patients with cardiovascular disease (p<0.005) and correlated with age (rho=0.39, p<0.0005), CACS (rho=0.19, p<0.0005) modified Medical Research Council dyspnoea score (rho=0.15, p<0.0005), 6-min walking distance (rho=-0.17, p<0.0005) and body mass index, airflow obstruction, dyspnoea, exercise capacity index (rho=0.10, p<0.01), but not with emphysema, emphysema progression or forced expiratory volume in 1 s decline. pDES predicted all-cause mortality independently of several confounding factors (p<0.005). In an independent cohort of 186 patients with COPD and 110 control subjects, pDES levels were higher in COPD patients with cardiovascular disease and correlated with arterial stiffness (p<0.05).In COPD, excess elastin degradation relates to cardiovascular comorbidities, atherosclerosis, arterial stiffness, systemic inflammation and mortality, but not to emphysema or emphysema progression. pDES is a good biomarker of cardiovascular risk and mortality in COPD.


The role of ultrastructural cytochemistry and monoclonal antibodies in clarifying the nature of undifferentiated cells in acute leukaemia.
The nature of the cells in 21 cases of acute leukaemia with blasts which were undifferentiated by light microscopy criteria was investigated by immunophenotyping, ultrastructural cytochemistry and DNA analysis. Two groups of cases were recognized. Fourteen cases were negative with B and T lymphoid markers and expressed one or two myeloid antigens detected by the monoclonal antibodies (McAb) MCS2 (CD13) and MY9 (CD33). Peroxidase activity was demonstrated at ultrastructural level by the method of Roels on unfixed cells in eight out of 10 cases; rearrangement of the immunoglobulin (Ig) genes was demonstrated in one of the three cases investigated. These cases are proliferations of early, MO, myeloblasts which can only be recognized by immunological and ultrastructural cytochemical methods. The remaining seven cases revealed a complex phenotype with expression of myeloid and lymphoid antigens. Peroxidase activity was detected in blasts from two cases with rearrangement of the Ig-heavy chain gene; in one of them the T cell receptor beta and gamma chain genes were also found in rearranged configuration. This group comprises cases of biphenotypic and mixed acute leukaemia which probably involve multipotent stem cells. This study demonstrates that the expression of myeloid antigens on blast cells parallels closely the presence of peroxidase activity and that lymphoid markers correlate with gene rearrangements at DNA level. Our findings are reassuring with respect to the specificity of the antimyeloid McAb for the diagnosis of cases which are unclassifiable by conventional methods.


Ethics and human values committee survey: (AMI Denver Hospitals: Saint Luke's, Presbyterian Denver, Presbyterian Aurora: Summer 1989). A study of physician attitudes and perceptions of a hospital ethics committee.
By studying physician perceptions of the Hospital Ethics Committee (HEC), the committee can determine (1) what areas the committee should concentrate on, and (2) how it is perceived by the physician community. The topics covered by the study included whether or not a HEC is needed in any hospital system, as well as the specific need within AMI Denver hospitals. In addition, the physician participants discussed what departments should be represented on the HEC and what topics it should study and discuss. By discussing the perceived effectiveness and the ideal function of HECs the physicians were able to provide the committee with many concrete suggestions to improve its effectiveness and visibility within the hospital community. Most of the advice is captured in one physician's words: "Be Visible and Credible."


Structure/function relationships in nickel metallobiochemistry.
Among the many highlights of nickel metallobiochemistry in 1998 were the discoveries that Escherichia coli glyoxalase I is the first example of a nickel isomerase, and that the superoxide dismutase isolated from Streptomyces seoulensis is a new structural class of superoxide dismutase that features thiolate ligation.


Evidence for gene-nutrient interaction at the PPARgamma locus.
The importance of the nuclear receptor peroxisome proliferator-activated receptor-gamma (PPARgamma) in regulating insulin resistance and blood pressure has been demonstrated in families with loss of function mutations. Gain of function mutations has been associated with severe obesity. However, previous population studies of the common variant Pro12Ala have produced conflicting results. As it is likely that the natural ligands for this receptor may include fatty acids, we hypothesized that the effect of this common variant may be altered by the character of the diet, particularly the ratio of dietary polyunsaturated fat to saturated fat (P:S ratio). We studied 592 nondiabetic participants in an ongoing population-based cohort study who were genotyped for the Pro12Ala polymorphism in the PPAR gamma2 isoform. As the Ala homozygotes were uncommon (2.0%), all analyses were conducted comparing Pro homozygotes (79.1%) to Ala allele carriers. There was no difference in fasting insulin concentration or BMI between Ala allele carriers and Pro homozygotes. The fasting insulin concentration was negatively associated with the P:S ratio (P = 0.0119) after adjustment for age and sex, and a strong interaction was evident between the P:S ratio and the Pro12Ala polymorphism for both BMI (P = 0.0038) and fasting insulin (P = 0.0097). The data suggest that when the dietary P:S ratio is low, the BMI in Ala carriers is greater than that in Pro homozygotes, but when the dietary ratio is high, the opposite is seen. This gene-nutrient interaction emphasizes the difficulty of examining the effect of common polymorphisms in the absence of data on nongenetic exposures, and may explain the heterogeneity of findings in previous studies.


Molecular spectra of HPRT deletion mutations in circulating T-lymphocytes in Fanconi anemia patients.
The principal cellular feature of Fanconi anemia (FA), an inherited cancer prone disorder, is a high level of chromosomal breakage, amplified after treatment with crosslinking agents. Three of the eight genes involved in FA have been cloned: FANCA, FANCC and FANCG. However, their biological functions remain unknown. We previously observed an excessive production of deletions at the HPRT locus in FA lymphoblasts belonging to the relatively rare complementation group D(1) and an increased frequency of glycophorin A (GPA) variants in erythrocytes derived from FA patients (2). In thi study, we examined the molecular nature of 31 HPRT mutations formed in vivo in circulating T-lymphocytes isolated from 9 FA male patients. The results show that in all FA patients investigated the deletions are by far the most prevalent mutational event in contrast to age matched healthy donors, in which point mutations predominate. The complementation group in the FA patients examined in the present study has not yet been defined. However, knowing that mutations in the FANCA and FANCC gene are found to be involved in at least 70% of the FA patients, it can be expected that the excessive production of deletions is a general feature of the FA phenotype. In addition, the spectrum of HPRT deletions observed in FA patients differs from that of healthy children: there is a high frequency of 3'-terminal deletions and a strikingly low proportion of V(D)J mediated events. Based on previous findings, a decreased fidelity of coding V(D)J joint formation (3) and an inaccurate repair of specific DNA double strand breaks via Non-Homologous End Joining (4), we propose that FA genes play a role in the control of the fidelity of rejoining of specific DNA ends. Such a defect may explain several basic features of FA, such as chromosomal instability and deletion pronenness.


Suppressor T cells control the HLA-linked low responsiveness to streptococcal antigen in man.
We have previously reported that low immune responsiveness to the streptococcal cell wall (SCW) antigen is controlled by an HLA-linked dominant gene which we designated as an immune suppression gene to the SCW antigen (Is-SCW) without knowing its function. We have extended the study of the genetic control of the immune response to the SCW antigen and confirmed both the bimodal distribution of immune responsiveness and HLA-linked dominant inheritance of low responsiveness. Here we report an analysis of the function and expression of Is-SCW at the cellular level and demonstrate that the Is-SCW controls the generation of antigen-specific suppressor T cell in low responders.


A study of patients receiving no billed nursing care.
Although home health care traditionally is conceptualized as nursing care, in today's environment care at home may be delivered by a myriad of professional and nonprofessional practitioners. In fact, many patients who receive home care do not receive billed nursing visits. We studied a group of patients (n = 200) who received no billed nursing care, but rather received billed care from therapists, social workers and home health aides. This cohort of patients differed from patients who received billed nursing care in several ways: they were more frequently referred to home care from the community, their prognoses on admission to home care were significantly better, and their outcomes from home care services were considerably more favorable. Clearly patients not receiving billed nursing care have service and resource needs different from those of the population receiving billed nursing care. Therefore, new perspectives must be used when considering how to assess, deliver and perhaps pay for the requirements of these patients.


LncRNA ST3Gal6-AS1/ST3Gal6 axis mediates colorectal cancer progression by regulating α-2,3 sialylation via PI3K/Akt signaling.
Sialylation is associated with cancer progression. Long noncoding RNAs (lncRNAs) have important roles in diverse diseases including cancer. The lncRNA ST3Gal6 antisense 1 (ST3Gal6-AS1) derives from the promoter region of sialyltransferase ST3Gal6. However, the mechanisms by which ST3Gal6-AS1 modulates colorectal cancer (CRC) development through sialylation remain largely unknown. Here, we found that ST3Gal6-AS1 and ST3Gal6 levels were lower in tumor tissues than adjacent normal tissues of CRC patients. The correlation between ST3Gal6-AS1 and ST3Gal6 was further validated in several types of CRC cell lines. In addition, ST3Gal6 was dysregulated and positively correlated to ST3Gal6-AS1. ST3Gal6-AS1 recruited histone methyltransferase MLL1 to the promoter region of ST3Gal6, induced H3K4me3 modification and activated ST3Gal6 transcription. Furthermore, ST3Gal6-AS1/ST3Gal6 axis mediated α-2, 3 sialylation and inhibited the activation of PI3K/Akt signaling, thereby resulting in Foxo1 nuclear translocation in CRC cells. ST3Gal6-AS1 was a target of transcription factor Foxo1 and regulated by Foxo1. ST3Gal6-AS1 also inhibited CRC cell proliferation, metastasis, and promoted cell apoptosis in vitro. Overexpression of ST3Gal6-AS1 significantly decreased the tumorigenesis, lung and liver metastasis of SW620 cells in vivo. ST3Gal6-AS1 expression was negatively correlated with tumor size, lymphatic metastasis, distant metastasis and tumor stage in CRC patients. Collectively, these data indicated that ST3Gal6-AS1, ST3Gal6, PI3K/Akt, and Foxo1 formed a positive feedback loop, which might play a key role in CRC progression.


[Dominance analysis on the association between reasons for smoking and results related to successful smoking cessation].
To study the association between reasons for smoking and results related to successful smoking cessation. A community-based case-control study was conducted with 642 male adults recognized as successful spontaneous smoking quitters being the subject cases while another 700 male adults who had failed to quit smoking were served as the controls. Russell Reason for Smoking Questionnaire (RRSQ) was used to investigate the reasons for smoking. Dominance logistic regression was performed to determine the relative importance of reasons for starting smoking and the endpoint of successful quit smoking. After adjusting the potential confounders of age, age of smoking initiation, marital status, profession, and education, the adjusted means of sedative and stimulation of RRSQ in the successful quitters were significant lower than that in the failed quitters. Results from the dominance logistic regression showed that the first two important factors for smoking cessation were sedative and stimulation of RRSQ, followed by psychological image, automatic, hand-mouth activity, indulgence, and addiction. This study demonstrated that factors as dependence-addiction showed more important impacts than the social-psychological ones on the endpoint of successful smoking cessation.


Original multivalent copper(II)-conjugated phosphorus dendrimers and corresponding mononuclear copper(II) complexes with antitumoral activities.
Novel multivalent copper(II)-conjugated phosphorus dendrimers and their corresponding mononuclear copper(II) complexes were synthesized, characterized, and screened for antiproliferative activity against human cancer cell lines. Selected copper ligands were grafted on the surface of phosphorus dendrimers of generation G(n) (n = 1 to 3): N-(pyridin-2-ylmethylene)ethanamine for dendrimers 1G(n), N-(di(pyridin-2-yl)methylene)ethanamine for dendrimers 2G(n), and 2-(2-methylenehydrazinyl)pyridine for dendrimers 3G(n). The results indicated that the most potent derivatives are 1G(n) and 1G(n)-Cu versus 2G(n), 2G(n)-Cu, and 3G(n), 3G(n)-Cu. A direct relationship between the growth inhibitory effect and the number of terminal moieties or the amount of copper complexed to the dendrimer was observed in copper-complexed 1 series and noncomplexed 1 series. These data clearly suggested that cytotoxicity increased with the number of terminal moieties available and was boosted by the presence of complexed Cu atoms. Importantly, no cytotoxic effect was observed with CuCl2 at the same concentrations. Finally, 1G3 and 1G3-Cu have been selected for antiproliferative studies against a panel of tumor cell lines: 1G3 and 1G3-Cu demonstrated potent antiproliferative activities with IC50 values ranging 0.3-1.6 μM. Interestingly, the complexation of the terminal ligands of 1G3 dendrimers by copper(II) metal strongly increased the IC50 values in noncancer cells lines referred to as "safety" cell lines.


Retargeting of human regulatory T cells by single-chain bispecific antibodies.
Bispecific Abs hold great potential for immunotherapy of malignant diseases. Because the first components of this new drug class are now entering clinical trials, all aspects of their mode of action should be well understood. Several studies proved that CD8(+) and CD4(+) effector T cells can be successfully redirected and activated against tumor cells by bispecific Abs both in vitro and in vivo. To our knowledge, this study provides the first evidence that bispecific Abs can also redirect and activate regulatory T cells against a surface Ag, independently of their TCR specificity. After cross-linking, via a bispecific Ab, redirected regulatory T cells upregulate the activation markers CD69 and CD25, as well as regulatory T cell-associated markers, like CTLA-4 and FOXP3. The activated regulatory T cells secrete the immunosuppressive cytokine IL-10, but, in contrast to CD8(+) and CD4(+) effector T cells, almost no inflammatory cytokines. In addition, the redirected regulatory T cells are able to suppress effector functions of activated autologous CD4(+) T cells both in vitro and in vivo. Therefore, the potential risk for activation of regulatory T cells should be taken into consideration when bispecific Abs are applied for the treatment of malignant diseases. In contrast, an Ag/tissue-specific redirection of regulatory T cells with bispecific Abs holds great potential for the treatment of autoimmune diseases and graft rejection.


The effects of lower body passive heating combined with mixed-method cooling during half-time on second-half intermittent sprint performance in the heat.
This study examined the effects of combined cooling and lower body heat maintenance during half-time on second-half intermittent sprint performances. In a repeated measures design, nine males completed four intermittent cycling trials (32.1 ± 0.3 °C and 55.3 ± 3.7% relative humidity), with either one of the following half-time recovery interventions; mixed-method cooling (ice vest, ice slushy and hand cooling; COOL), lower body passive heating (HEAT), combined HEAT and COOL (COMB) and control (CON). Peak and mean power output (PPO and MPO), rectal (Tre), estimated muscle (Tes-Mus) and skin (TSK) temperatures were monitored throughout exercise. During half-time, the decrease in Tre was substantially greater in COOL and COMB compared with CON and HEAT, whereas declines in Tes-Mus within HEAT and COMB were substantially attenuated compared with CON and COOL. The decrease in TSK was most pronounced in COOL compared with CON, HEAT and COMB. During second-half, COMB and HEAT resulted in a larger decrease in PPO and MPO during the initial stages of the second-half when compared to CON. In addition, COOL resulted in an attenuated decrease in PPO and MPO compared to COMB in the latter stages of second-half. The maintenance of Tes-Mus following half-time was detrimental to prolonged intermittent sprint performance in the heat, even when used together with cooling.


Hormonal influence on utero-ovarian blood flow distribution in the mid-luteal pseudopregnant rat.
Uterine and ovarian blood flows and ovarian fractional perfusion were measured in adult rats anaesthetized with sodium pentobarbital i.p. on day 6 of pseudopregnancy, by using radioactive microspheres. Fifty i.u. human chorionic gonadotropin (hCG) was found to increase the fractional perfusion of the ovary, and this was at least partially accomplished at the expense of the uterine blood flow, since this was reduced. Noradrenaline (NA) infusion (2 nmol min-1) reduced ovarian as well as uterine blood flow acutely. After 20 min of NA infusion (2 nmol min-1), however, ovarian blood flow and fractional perfusion were increased, while no significant effect was seen for uterine blood flow. Antidiuretic hormone (ADH) in a 20-min infusion (1 nmol min-1) markedly reduced ovarian and uterine blood flows.


Electron-density-dependent fused-sphere surfaces derived from pseudopotential calculations.
Fused-sphere surfaces can be used to mimic a molecular "boundary" associated with a constant value of the electron density. The simplest of such fused-sphere models are constructed by using the atomic radii for the spherical isodensity surfaces of individual atoms. In this work, we discuss the extension of this model to molecules containing atoms beyond the second row. In these many-electron systems, the computation of electron densities is usually simplified by adopting a pseudopotential (or effective-core potential) approach. Here, we discuss the performance of large- and small-core pseudo-potential calculations as a tool to derive atomic radii. Our results provide an optimum set of variable radii that can be used to build fused-sphere surfaces. This continuum of surfaces provides a simple approximation to the low-electron-density regions around molecules with heavy atoms.


[Role of contrast-enhanced ultrasound in management of splenic trauma].
To study the role of contrast-enhanced ultrasound (CEUS) in the management of splenic trauma Forty patients with splenic trauma underwent CEUS examination and then different management were provided according to the examination results and the general status of patients: conservative treatment (group I, n = 19); CEUS-guided injective therapy (group II, n = 6); and surgery (group III, n = 15). Eighteen patients were cured in group I and one patient experienced rehaemorrhagia. The haemostatic effect of CEUS-guided injective therapy was obvious in all six patients in group II. Among them, one patient experienced arteriovenous fistula, which was resolved after one week of injective therapy. Fifteen patients in Group III underwent surgery and were cured. CEUS can provide reliable information for therapy mode selection in patients with splenic trauma and can be used to guide injective therapy.


Safe and efficient Boc-SPPS for the synthesis of glycopeptide-α-thioesters.
Boc-solid phase peptide synthesis is useful for the preparation of peptide-α-thioesters. However, this strategy usually employs hydrogen fluoride for the final deprotection step. These strongly acidic conditions cannot be applied for the synthesis of acid-labile glycopeptide-α-thioesters. The protocol presented here is a modified in situ neutralization Boc-solid phase peptide synthesis employing 10% sulfuric acid/dioxane conditions for intermediate Boc removal and TfOH for the final side-chain deprotection step. These conditions were found to be applicable for the synthesis of acid-labile glycopeptide-α-thioesters. In this protocol, a glycopeptide is synthesized as α- thioester on a thiol linker, and the product glycopeptide-α-thioester is released from the resin by thiolysis after side-chain deprotection step with an acid cocktail containing TfOH instead of hydrogen fluoride.


Laparoscopy in zoological medicine.
The technique of laparoscopy was adapted and utilized in zoological medicine for various mammals, birds, and reptiles for reproductive and diagnostic studies as well as clinically related research. It was concluded that since anesthesia was routinely required for most manipulative procedures in zoo animals, and since laparoscopy adds little additional risk, the use of this technique provides an additional diagnostic aid when indicated. Laparoscopy was found to be effective for evaluating reproductive status, particularly ovarian anatomy and function, direct visual biopsy of internal organs, sex determination in selected birds, and as a surgical means of fertility control.


Spatial and Temporal Resolution of the Visual System of the Anna's Hummingbird (Calypte anna) Relative to Other Birds.
Hummingbirds are an emerging model for studies of the visual guidance of flight. However, basic properties of their visual systems, such as spatial and temporal visual resolution, have not been characterized. We measured both the spatial and temporal visual resolution of Anna's hummingbirds using behavioral experiments and anatomical estimates. Spatial visual resolution was determined behaviorally using the optocollic reflex and anatomically using peak retinal ganglion cell densities from retinal whole mounts and eye size. Anna's hummingbirds have a spatial visual resolution of 5-6 cycles per degree when measured behaviorally, which matches anatomical estimates (fovea: 6.26 ± 0.12 cycles per degree; area temporalis: 5.59 ± 0.15 cycles per degree; and whole eye average: 4.64 ± 0.08 ). To determine temporal visual resolution, we used an operant conditioning paradigm wherein hummingbirds were trained to use a flickering light to find a food reward. The limits of temporal visual resolution were estimated as 70-80 Hz. To compare Anna's hummingbirds with other bird species, we used a phylogenetically controlled analysis of previously published data on avian visual resolutions and body size. Our measurements for Anna's hummingbird vision fall close to and below predictions based on body size for spatial visual resolution and temporal visual resolution, respectively. These results indicate that the enhanced flight performance and foraging behaviors of hummingbirds do not require enhanced spatial or temporal visual resolution. This finding is important for interpreting flight control studies and contributes to a growing understanding of avian vision.


Clinical tools designed to assess motor abilities in children with cerebral palsy.
This systematic review aimed to list the tools used by rehabilitation professionals to test motor abilities in children with cerebral palsy (CP), to determine if these tools have psychometric properties specifically measured for CP, and to identify the main characteristics of these tools. Web of Science, PEDro, PubMed/MEDLINE, Science Direct, and SciELO databases were searched to identify the tools. PubMed/MEDLINE was then searched to identify the studies assessing those tools' psychometric properties. The agreement-based standards for the selection of health measurement tools and the Terwee criteria were used to assess the quality and the results of each included study, respectively. Eighteen tools were identified. The psychometric properties of many of the tools used with children with CP have not been evaluated for this population. The psychometric properties evaluated often have a poor methodological quality of measurement. Overall, we suggest the tools with most empirical support to evaluate children with CP.


Delta opioid receptor enhancement of mu opioid receptor-induced antinociception in spinal cord.
Although the mu selective agonist [D-Ala2-MePhe4-Gly-ol5]enkephalin (DAMGO) and the delta selective agonist [D-Pen2,D-Pen5]enkephalin (DPDPE) are both antinociceptive when administered directly into the spinal cord of mice, 50% of antinociceptive dose (AD50) of DAMGO is about 2 orders of magnitude lower than the AD50 of DPDPE. In contrast, the two ligands show similar affinities for their respective receptors in in vitro binding assays. One possible explanation for this discrepancy is that DPDPE antinociception in the spinal cord is mediated through not delta but mu receptors, for which it has an several hundred-fold lower affinity than DAMGO. In support of this, we found that DPDPE-mediated antinociception was blocked by the mu selective antagonist D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (CTAP). The pA2 value of CTAP for DPDPE was virtually identical with that for DAMGO. However, because its action also was blocked by naltrindole, an antagonist selective for delta receptors, the latter must also play a role in antinociception. When DAMGO and DPDPE were administered i.t. together at ratios ranging from 1:200 to 1:500, the AD50 of DAMGO was lowered as much as 10-fold relative to its AD50 when given alone. Thus DPDPE had a potentiating effect on DAMGO, although the reverse was not observed. This potentiation was lost in animals made tolerant to systemic morphine. The loss of potentiation seemed to be caused by changes in the delta receptors, because a) the AD50 of DAMGO (i.t.) given alone to tolerant animals was virtually the same as for naive animals, whereas the AD50 of DPDPE given alone increased by 4-fold; and b) the AD50 of DPDPE given alone in the tolerant animal was increased only slightly by naltrindole, whereas CTAP was still a very potent antagonist. We conclude that DPDPE, a selective delta agonist, mediates antinociception in the spinal cord through mu receptors, consistent with results of recent studies of "knock-out" mice lacking mu receptors. At the same time, however, the delta agonist acting through delta receptors can potentiate the mu receptor-mediated antinociceptive action of either mu or delta agonists. This potentiating effect, like the synergistic effect observed between mu receptors at spinal and supraspinal sites, is lost during tolerance.


Water fluoridation information found on the World Wide Web.
There is much controversy over the fluoridation of community water supplies. This study surveyed the informational content of internet sites on the World Wide Web (WWW) regarding water fluoridation. One hundred websites were identified and 59 were evaluated with a 6 point scoring system using predetermined criteria. Of these 59 sites, 54% recommend water fluoridation as compared to 31% that oppose it. The informational content of WWW sites may range from factual, to unsubstantiated opinions, to frank fraud. However, this information is presented to the public indiscriminately. The informational content of the WWW contains varied views and recommendations for community water supply fluoridation. Public health officials and practicing physicians should be aware that the internet WWW may influence public opinion.


Peculiarities of the binding of restriction endonuclease EcoRII to synthetic DNA duplexes.
The binding of restriction endonuclease EcoRII to synthetic oligodeoxyribonucleotide substrates 11 to 30 bp long was investigated by gradient polyacrylamide gel electrophoresis under nondenaturing conditions in the absence of Mg2+ ions. Irrespective of the substrate's length, two types of specific DNA-protein complexes were shown to be formed. Their mobility in gel was close to that of the monomer and the dimer of the marker ovalbumin. The number of such complexes in solution depended on the ratio of the molar concentrations of restriction endonuclease EcoRII and the DNA duplex. The possible structure of the complexes is discussed.


Inhibitors of the proteasome block the degradation of most cell proteins and the generation of peptides presented on MHC class I molecules.
Reagents that inhibit the ubiquitin-proteasome proteolytic pathway in cells have not been available. Peptide aldehydes that inhibit major peptidase activities of the 20S and 26S proteasomes are shown to reduce the degradation of protein and ubiquitinated protein substrates by 26S particles. Unlike inhibitors of lysosomal proteolysis, these compounds inhibit the degradation of not only abnormal and short-lived polypeptides but also long-lived proteins in intact cells. We used these agents to test the importance of the proteasome in antigen presentation. When ovalbumin is introduced into the cytosol of lymphoblasts, these inhibitors block the presentation on MHC class I molecules of an ovalbumin-derived peptide by preventing its proteolytic generation. By preventing peptide production from cell proteins, these inhibitors block the assembly of class I molecules. Therefore, the proteasome catalyzes the degradation of the vast majority of cell proteins and generates most peptides presented on MHC class I molecules.


Philosophy of medicine: should it be teleologically or socially construed?
This response to Kevin Wildes's article in the previous issue of the Kennedy Institute of Ethics Journal addresses several major points of disagreement between Pellegrino and Wildes regarding the nature and scope of a philosophy of medicine, in particular how it is derived and by what method of philosophical enquiry it is best pursued.


Cellular transport of lutein is greater from uncooked rather than cooked spinach irrespective of whether it is fresh, frozen, or canned.
Lutein, a carotenoid found in significant levels in spinach, has attracted a great deal of attention owing to its reported function as a shield against the photooxidative effects of blue light. Therefore, the rationale of this study was to examine the effects of various processing and cooking methods on lutein bioavailability from spinach (Spinacia oleracea) using an in vitro digestion procedure coupled with the use of a human intestinal Caco-2 cell model. Fresh, frozen, and canned spinach were analyzed uncooked and after boiling or microwave cooking. Lutein content from the uncooked and cooked digested food (digestate) and appropriate micelles was determined. Micellarized lutein from the spinach samples was adjusted to 0.1 micromol/L and added to Caco-2 cells. Cellular uptake and secretion (cellular transport) of lutein were determined. Our results showed that digestate obtained from uncooked canned spinach had greater lutein content (P < .05) than uncooked fresh or frozen spinach. Microwave cooking, but not boiling, significantly lowered the lutein content of canned spinach digestate and micelles compared with their uncooked counterparts. Interestingly, there were no differences in the micellarization of lutein between the cooking and processing methods. Cellular transport of lutein was greater from uncooked spinach micelles compared with boiled or microwave-cooked spinach. To conclude, although the lutein content of digesta and micelles may have been modified, its micellarization was not significantly affected by any of the cooking or processing methods tested. In general, cellular transport of lutein was greatest in uncooked spinach irrespective of whether the spinach was fresh, frozen, or canned.


Mobile Lower Body Negative Pressure Suit as an Integrative Countermeasure for Spaceflight.
BACKGROUND: Persistent headward fluid shift and mechanical unloading cause neuro-ocular, cardiovascular, and musculoskeletal deconditioning during long-term spaceflight. Lower body negative pressure (LBNP) reintroduces footward fluid shift and mechanical loading.METHODS: We designed, built, and tested a wearable, mobile, and flexible LBNP device (GravitySuit) consisting of pressurized trousers with built-in shoes to support ground reaction forces (GRF) and a thoracic vest to distribute load to the entire axial length of the body. In eight healthy subjects we recorded GRF under the feet and over the shoulders (Tekscan) while assessing cardiovascular response (Nexfin) and footward fluid shift from internal jugular venous cross-sectional area (IJVa) using ultrasound (Terason).RESULTS: Relative to normal bodyweight (BW) when standing upright, increments of 10 mmHg LBNP from 0 to 40 mmHg while supine induced axial loading corresponding to 0%, 13 ± 3%, 41 ± 5%, 75 ± 11%, and 125 ± 22% BW, respectively. Furthermore, LBNP reduced IJVa from 1.12 ± 0.3 cm² to 0.67 ± 0.2, 0.50 ± 0.1, 0.35 ± 0.1, and 0.31 ± 0.1 cm², respectively. LBNP of 30 and 40 mmHg reduced cardiac stroke volume and increased heart rate while cardiac output and mean arterial pressure were unaffected. During 2 h of supine rest at 20 mmHg LBNP, temperature and humidity inside the suit were unchanged (23 ± 1°C; 47 ± 3%, respectively).DISCUSSION: The flexible GravitySuit at 20 mmHg LBNP comfortably induced mechanical loading and desired fluid displacement while maintaining the mobility of hips and knee joints. The GravitySuit may provide a feasible method to apply low-level, long-term LBNP without interfering with daily activity during spaceflight to provide an integrative countermeasure.Petersen LG, Hargens A, Bird EM, Ashari N, Saalfeld J, Petersen JCG. Mobile lower body negative pressure suit as an integrative countermeasure for spaceflight. Aerosp Med Hum Perform. 2019; 90(12):993-999.


Comparison of angiotensinogen and tetradecapeptide as substrates for human renin. Substrate dependence of the mode of inhibition of renin by a statine-containing hexapeptide.
The kinetic properties of two different substrates for human renin, a synthetic tetradecapeptide and the natural substrate human angiotensinogen, have been compared. While the Vmax was similar for the two substrates, the Km values differed by a factor of 10, i.e., 11.7 +/- 0.7 microM (tetradecapeptide) and 1.0 +/- 0.1 microM (angiotensinogen). The mode of inhibition of renin by a statine (Sta)-containing hexapeptide, BW897C, that is a close structural analog of residues 8-13 of human angiotensinogen (Phe-His-Sta-Val-Ile-His-OMe), was determined for the two substrates. Competitive inhibition was observed when tetradecapeptide was the substrate (Ki = 2.0 +/- 0.2 microM), but a more complex mixed inhibition mode (Ki = 1.7 +/- 0.1 microM, Ki' = 3.0 +/- 0.23 microM) was found with angiotensinogen as substrate. This mixed inhibition probably results from the formation of an enzyme-inhibitor-substrate or enzyme-inhibitor-product complex and reflects the more extensive interactions that the protein angiotensinogen, as opposed to the small tetradecapeptide substrate, can make with renin. We conclude that the mixed inhibition observed when angiotensinogen is used as renin substrate could be important in the clinical application of renin inhibitors because it is less readily reversed by increased concentrations of substrate than is simple competitive inhibition.


Cost-neutral policies to increase Social Security benefits for widows: a simulation for 1992.
Among older women, widows are more likely to live in poverty than married women. Thus, increasing Social Security benefits to widows seems desirable. Shifting some Social Security benefits from the period when women live as part of a couple to the period when they are widows could reduce poverty. This article uses the 1991 Survey of Income and Program Participation exactly matched to the Social Security Administration's record of benefits to evaluate the effect on poverty rates of four cost-neutral proposals that transfer Social Security benefits from married couples to surviving widows. The policies would moderately decrease poverty rates among older women by reducing the rate for widows more than the slight increase in the rate for couples. The evaluated proposals include a proposal supported by the majority of the 1994-96 Advisory Council on Social Security that would calculate the survivor's benefit as 75 percent of the couple's benefit, reduce the spouse's benefit from 50 to 33 percent of the husband's benefit, and reduce benefits by 1.5 percent.


Somatic cell mapping of the genes for anti-müllerian hormone and osteonectin in cattle: identification of a new bovine syntenic group.
DNA probes from the bovine anti-Müllerian hormone and osteonectin genes were hybridized onto Southern blots containing DNAs from cow-hamster and cow-mouse hybrid somatic cell lines segregating bovine chromosomes. Bovine anti-Müllerian hormone and osteonectin loci were fully concordant with each other in 96 hybrid somatic cell lines, but were not concordant with any other bovine syntenic group described to date. As such, these two genes represent another syntenic group in cattle, bringing to 27 the number of autosomal syntenic groups identified thus far.


PFT interpretive strategies: American Thoracic Society/ European Respiratory Society 2005 guideline gaps.
All pulmonologists, including those recently completing training, should be competent in critically evaluating and interpreting pulmonary function tests (PFTs). In addition, some authorities recommend that respiratory therapists learn to provide preliminary PFT interpretations for the medical directors of PFT labs. The 2005 American Thoracic Society/European Respiratory Society guidelines for interpreting PFTs lack recommendations for the best reference equations for lung volumes and diffusing capacity of the lung for carbon monoxide (D(LCO)), and lack reference equations for non-whites. The pre-test probability of lung disease should be determined using a short questionnaire. The "nonspecific pattern" occurs in about 15% of patients referred to a PFT lab, but it has many clinical correlates and the course is usually benign. Less common PFT patterns and those resulting from comorbid conditions (such as obesity, respiratory muscle weakness, or heart failure) are not discussed by the guidelines. More than half of patients with interstitial lung disease have a normal ratio of D(LCO)/V(A) (alveolar volume), and many have a normal total lung capacity.


An American Board of Orthodontics case report.
A case report is presented of an Angle Class I dental malocclusion, significant bimaxillary dentoalveolar protrusion, anterior open bite, and spacing of the maxillary and mandibular incisors. [This case was presented to the American Board of Orthodontics in partial fulfillment of the requirements for the certification process conducted by the Board.]


Hemilabyrinthectomy causes both an increase and a decrease in corticotropin releasing factor mRNA in rat inferior olive.
It was previously shown [NeuroReport, 3 (1992) 829-832] that unilateral labyrinthectomy (UL) induces Fos expression in several brainstem regions, including the beta subnucleus of the inferior olive. Using isotopic 33P in situ hybridization, the present results demonstrate significant changes in oligonucleotide-probed mRNA levels for corticotropin-releasing factor (CRF) in the rat inferior olivary nucleus 4 days following unilateral labyrinthectomy (UL). In the medulla of normal animals there was strong CRF mRNA labeling in the inferior olivary nucleus, and weaker labeling in the vestibular nuclei and prepositus hypoglossi. Following unilateral labyrinthectomy, the contralateral olivary beta subnucleus showed a significant increase in CRF message, similar to the contralateral Fos labeling observed after hemilabyrinthectomy [NeuroReport, 3 (1992) 829-832]. In addition, the contralateral A and B subnuclei (IOA/B) of the inferior olive showed a strong increase in CRF labeling, while the ipsilateral dorsal cap of Kooy (IOK) showed a decrease. This novel bidirectional alteration in CRF message in different subdivisions of the same nuclear group indicates the existence of both up and down regulatory mechanisms controlling CRF peptide expression, and reflects the dynamic neurochemical alterations occurring during vestibular compensation.


Study of RNA synthesis in the livers of aging mice by means of electron microscopic radioautography.
The application of 3H-uridine radioautography results in labeling of the liver cells in which RNA is synthesized at various ages of the mouse. Quantitative changes of RNA synthesis in the hepatocytes of aging mice were studied by electron microscopic radioautography. The silver grains were mainly located in the nucleoli and nuclei and a few in the mitochondria and rough surfaced endoplasmic reticulum of almost all of the cell populations at various ages. The number of silver grains in the hepatocyte gradually increased after birth, reached the maximum at 14 days of postnatal age, then decreased to 24 months with aging. The number of silver grains of the euchromatin was more than those of the heterochromatin of the hepatocyte nuclei at various ages. The number of silver grains of the granular components was more than those of the fibrillar components of the hepatocyte nucleoli at various ages. However, the ratio of silver grains among euchromatin, heterochromatin, granular components and fibrillar components remained approximately constant.


Asymmetric subcellular distribution of glucose transporters in the endothelium of small contractile arteries.
The authors have recently reported the presence and asymmetric distribution of the glucose transporters GLUT-1 to -5 and SGLT-1 in the endothelium of rat coronary artery (Gaudreault et al. 2004, Diabetologica, 47, 2081-2092). In the present study the authors investigate and compare the presence and subcellular distribution of the classic glucose transporter isoforms in endothelial cells of cerebral, renal, and mesenteric arteries. The GLUTs and SGLT-1 were examined with immunohistochemistry and wide-field fluorescence microscopy coupled to deconvolution in en face preparation of intact artery. We identified GLUT-1 to -5 and SGLT-1 in the endothelial cells of all three vascular beds. The relative level of expression for each isoform was found comparable amongst arteries. Clusters of the glucose transporter isoforms were found at a high density in proximity to the cell-to-cell junctions. In addition, a consistent asymmetric distribution of GLUT-1 to -5 was found, predominantly located on the abluminal side of the endothelium in all three vascular beds examined (ranging from 68% to 91%, p<.05). The authors conclude that the expression and subcellular distribution of glucose transporters are similar in endothelial cells from vascular beds of comparable diameter and suggest that their subcellular organization may facilitate transendothelial transport of glucose in small contractile arteries.


Coarse-graining intermittent intracellular transport: Two- and three-dimensional models.
Viruses and other cellular cargo that lack locomotion must rely on diffusion and cellular transport systems to navigate through a biological cell. Indeed, advances in single particle tracking have revealed that viral motion alternates between (a) diffusion in the cytoplasm and (b) active transport along microtubules. This intermittency makes quantitative analysis of trajectories difficult. Therefore, the purpose of this paper is to construct mathematical methods to approximate intermittent dynamics by effective stochastic differential equations. The coarse-graining method that we develop is more accurate than existing techniques and applicable to a wide range of intermittent transport models. In particular, we apply our method to two- and three-dimensional cell geometries (disk, sphere, and cylinder) and demonstrate its accuracy. In addition to these specific applications, we also explain our method in full generality for use on future intermittent models.


Anti-candida antibodies in human serum. Detection and definition of Ig classes by a new elution technique.
The Partigen Elution Test (P.E.T.), a partially quantitative method for the determination of Ig classes to which the antibodies against superficial Candida antigens belong, has been assessed. The sensitivity of the test is comparable to that of the Ouchterlony test. The P.E.T. has the advantage of simplicity, great reproducibility and absolute specificity. With P.E.T. the level of single class antibodies can be quantitated on the basis of the diameter of immunoprecipitation ring. It seems possible to establish a correlation between the number of precipitation lines in the Ouchterlony test and the diameter of the precipitation ring in LC-Partigen plates. In addition as association among the width of the ring diameter, the Ig classes implicated and the severity of Candida affection is apparent.


[Comparative characteristics of microbial proteases by the level of hydrolysis of protein substrates].
Screening of enzyme preparations displaying a maximum proteolytic activity at pH 4.0-5.5 and effecting deep proteolysis of plant proteins was performed. Amyloprotooryzin prepared from Aspergillus oryzae 387 containing a complex of proteolytic enzymes was the most effective. The amino acid composition of the hydrolysates obtained was studied. Amyloprotooryzin increased the contents of amino acids by 108-227%, depending on the substrate used. The enzymatic complex of amyloprotooryzin was studied; in addition, proteases, alpha-amylase, exo-beta-glucanase, and xylanase were detected in the complex.


One- and two-dimensional NMR relaxation studies of dynamics and structure in bile salt-phosphatidylcholine mixed micelles.
A series of one- and two-dimensional 1H-NMR relaxation measurements has been conducted on simple and mixed micellar aggregates of taurocholate, diphenylvaleroylphosphatidylcholine (diPVPC) and egg yolk phosphatidylcholine (egg PC). The results are interpreted to provide structural and dynamic comparisons between micelles and vesicles, between phospholipids of varying chain length, and between different lipid components within the same micellar aggregate. Both chemical shift changes and two-dimensional nuclear Overhauser effect cross peaks suggest direct interaction of taurocholate and PC chemical sites, although the latter observations may also be accounted for by PC-PC interactions. These experiments demonstrate the promise of NMR relaxation techniques for investigations of molecular organization in model substrate for lipolytic enzymes.


The role of M cells in the protection of mucosal membranes.
The mucosa-associated lymphoid tissues continuously take up antigenic matter from the lumen to generate immune responses or to maintain immune tolerance. This antigen sampling is performed by highly specialised epithelial cells, the membranous (M) cells of the dome epithelia. M cells possess a unique ultrastructure and lie in close contact to lymphoid cells. They endocytose soluble and solid substances, including entire microorganisms, at their apical membrane, transport these in vesicles to their basolateral membrane and exocytose them to the intercellular space. This review summarises the structural and functional peculiarities of M cells in different species and at the different sites of lymphoid tissue along the digestive and respiratory tracts. Specialisations of M cells for antigen uptake and transport comprise the composition of their apical membrane and its glycocalyx, a modified cytoskeleton as compared to enterocytes and a pocket-like invagination of the basolateral membrane populated by lymphocytes and macrophages. Besides ultrastructural characteristics, histochemical markers are listed that are currently available for detecting M cells by light microscopy. The origin, differentiation and distribution of M cells and other epithelial cell types of the dome epithelium are outlined. As M cells are used as entry sites by various pathogens and, in the future, might be employed for the oral application of vaccines and drugs, the clinical relevance of M cells in health and disease is discussed.


Molecular Clock as Insight to Estimate the Evolutionary History and Times of Divergence for 10 Nominal Astyanax Species (Characiformes, Characidae): An Evolutionary Approach in Species with 2n = 36, 46, 48, and 50 Chromosomes.
Astyanax is a genus with a wide distribution ranging from the south United States to north of Patagonia (Argentina). The available cytogenetic data on Astyanax indicate a high karyotypic diversity, with diploid number of 36-52 chromosomes, presence of B chromosomes, heterochromatin polymorphism, and variations with respect to the number and localization of nucleolar organizer regions (NORs) and 18S and 5S ribosomal DNA sites. In the present study, we estimated the evolutionary history and times of divergence for 10 nominal Astyanax species from the South and Central/North American (Cna) continents, which present distinct chromosomal characteristics, based on molecular clocks inferred from mitochondrial DNA sequence. The molecular clock results indicate the origin of three distinct clades (Humeral dark spot [Hds]; Diffuse humeral spot [Dhs]; Cna group) during the late Miocene about 11.2 million years ago (Mya). Thus, Astyanax mexicanus (Cna) represent a species that diverged a long time ago (∼8.6 Mya) from the Hds group, and Astyanax schubarti is the oldest species (∼6.5 Mya) among the Dhs species.


Influence of streptavidin on the absorption and fluorescence properties of cyanine dyes.
Cyanine dyes have become widely used fluorescence labels in clinical and biological chemistry. In particular, cyanine dyes with excitation wavelengths lambda(ex) > 600 nm are often used in biological applications. However, aggregation behavior and matrix effects on cyanine fluorescence are not fully understood yet and interfere with the data interpretation. In this study, we analyzed the spectroscopic characteristics of a model system consisting of the biotinylated cyanine dyes DY-635 and DY-647 and their streptavidin conjugates. On the basis of the spectroscopic data, the interaction processes between cyanine dye molecules and proteins are discussed. Binding to streptavidin had a significant influence on both fluorescence and anisotropy decays of the cyanine dyes investigated. In particular, the fluorescence anisotropy was significantly altered, making it a promising detection parameter for bioanalytical applications in connection with the cyanine dyes used in the present study. In order to evaluate the time-resolved anisotropy, the introduction of a sophisticated kinetic model was required to describe the contributions from different fluorescing species properly. The rotational motion of streptavidin-bound dyes was analyzed using the associated anisotropy model, which allowed discrimination between contributions from different microenvironments. The anisotropy decay times increased by a factor of up to 20 due to protein binding.


Angiotensin II stimulates an endogenous response in Xenopus laevis ovarian follicles.
While responses to angiotensin II have previously been induced in Xenopus laevis oocytes after injection of messenger RNA extracted from mammalian tissue, no endogenous responses of ovarian tissue to this hormone have been reported. Here we describe such an endogenous dose-dependent response to angiotensin II, detected by conventional electrophysiological techniques, in follicular oocytes. The ED50 of the response was estimated to be 0.15 +/- 0.07 microM (S.E.M.). Maximal depolarization, obtained at 1 microM angiotensin II, was 18.3 +/- 1.4 mV (n = 18, three experiments using oocytes from two toads, mean resting membrane potential = -42 +/- 2 mV). The response was absent from collagenase-treated oocytes or follicular oocytes treated with octanol, suggesting that the receptors are predominantly in the follicular layer surrounding the oocytes.


Source and nature of inhaled atmospheric dust from trace element analyses of human bronchial fluids.
Rapid volcanic eruptions quickly ejecting large amounts of dust provoke the accumulation of heavy metals in people living in surrounding areas. Analyses of bronchoalveolar lavage samples (BAL) collected from people exposed to the paroxysmal 2001 Etna eruption revealed a strong enrichment of many toxic heavy metals. Comparing the BAL to the dust composition of southeastern Sicily, we found that only V, Cr, Mn, Fe, Co, and U enrichment could be related to the volcanic event, whereas Ni, Cu, Cd, and Pb contents come from the dissolution of particles of anthropogenic origin. Furthermore, the nature of these inhaled anthropogenic particles was revealed by anomalous La and partially Ce concentrations in BAL that were consistent with a mixture of road dust and petroleum refinery emissions. Our results indicate that trace element distribution in BAL is a suitable tracer of human exposure to different sources of inhaled atmospheric particulates, allowing investigations into the origin of source materials inhaled by people subjected to atmospheric fallout.


Control of cardiovascular function and its evolution in Crustacea.
Work in the last decade has shown that crustacean open circulatory systems are highly efficient and controlled in a complex manner. Control occurs at several levels. Myocardial contraction is initiated in the cardiac ganglion but constantly modulated by the central nervous system, both directly via the cardioregulatory nerves and indirectly via the neurohormonal system. Heart rate and stroke volume can be controlled independently and measurements of both are needed to assess cardiac output accurately. Haemolymph outflow from many arthropod hearts is via a complex multiarterial distribution system, and the regional distribution of cardiac output is tightly controlled via cardioarterial valves at the base of each artery. These valves contain innervated muscle, and differential contraction serves to regulate the efflux of oxygenated haemolymph into a particular system. The major influence on both the evolution and control of arthropod open blood vascular systems is efficiency of oxygen uptake and delivery. This influence is illustrated by reference to a variety of crustacean and other arthropod types.


Visualization of sporopollenin-containing pathogenic green micro-alga Prototheca wickerhamii by fluorescent in situ hybridization (FISH).
Fluorescent in situ hybridization (FISH) using taxon-specific, rRNA-targeted oligonucleotide probes is one of the most powerful tools for the rapid identification of harmful microorganisms. However, eukaryotic algal cells do not always allow FISH probes to permeate over their cell walls. Members of the pathogenic micro-algal genus Prototheca are characterized by their distinctive cell-wall component, sporopollenin, an extremely tough biopolymer that resists acid and alkaline hydrolysis, enzyme attack, and acetolysis. To our knowledge, there has been no report of the successful permeation by the oligonucleotide probes over the cell walls of unicellular green micro-algae, which contain sporopollenin. The DNA probes passed through the cell wall of Prototheca wickerhamii after treating the algal cells with cetyltrimethylammonium bromide (CTAB). Most cells in the middle logarithmic growth phase culture fluoresced when hybridized with the rRNA-targeted universal probe for eukaryotes, though individual cells included in this culture differed in the level of cell-wall vulnerability to attack by the polysaccharide-degrading enzyme, thus reflecting the different stages of the life cycle. This is the first report regarding the visualization of sporopollenin-containing, green micro-algal cells by FISH.