[Gestational diabetes. Diagnosis in early stages of the pregnancy].
To determine whether women with early diagnosis of gestational diabetes have an increase of the rate of gestational hypertensive disease, metabolic imbalance, higher insulin dosage requirement, perinatal morbidity and perinatal death than those in whom diabetes developed after 24th week. All pregnant women with gestational diabetes were studied. Maternal age, pregestational body mass indexes, total weight gain during pregnancy, gestational age at diagnosis, and the timing of delivery were recorded. Two groups were studied: women with gestational diabetes diagnosed before 24 gestation weeks and those in which diabetes was diagnosed at 24 weeks or later. Total insulin dosage, rate of gestational hypertension, perinatal morbidity, perinatal death, and percentages of women with good metabolic control were analysed. In the whole group 28% were diagnosed before 24 weeks of gestation. Women that were diagnosed earlier had grater pregestational body mass indexes (30 +/- 6 kg/m2 vs 27 +/- 6 kg/m2) and total weight gain during pregnancy was lower during over all gestation (8 +/- 5 kg vs 10 +/- 6 kg). Insulin requirements were higher in women diagnosed earlier and there were no differences in the rates of gestational hypertension, and perinatal death. Perinatal morbidity was significantly higher in patients who were diagnosed later Early diagnosed and treatment may result in decreased perinatal morbidity. Women with early diagnosed diabetes required higher insulin dose, but they did not represent a high risk subgroup.


Intramuscular heat shock protein 72 and heme oxygenase-1 mRNA are reduced in patients with type 2 diabetes: evidence that insulin resistance is associated with a disturbed antioxidant defense mechanism.
To examine whether genes associated with cellular defense against oxidative stress are associated with insulin sensitivity, patients with type 2 diabetes (n = 7) and age-matched (n = 5) and young (n = 9) control subjects underwent a euglycemic-hyperinsulinemic clamp for 120 min. Muscle samples were obtained before and after the clamp and analyzed for heat shock protein (HSP)72 and heme oxygenase (HO)-1 mRNA, intramuscular triglyceride content, and the maximal activities of beta-hydroxyacyl-CoA dehydrogenase (beta-HAD) and citrate synthase (CS). Basal expression of both HSP72 and HO-1 mRNA were lower (P < 0.05) by 33 and 55%, respectively, when comparing diabetic patients with age-matched and young control subjects, with no differences between the latter groups. Both basal HSP72 (r = 0.75, P < 0.001) and HO-1 (r = 0.50, P < 0.05) mRNA expression correlated with the glucose infusion rate during the clamp. Significant correlations were also observed between HSP72 mRNA and both beta-HAD (r = 0.61, P < 0.01) and CS (r = 0.65, P < 0.01). HSP72 mRNA was induced (P < 0.05) by the clamp in all groups. Although HO-1 mRNA was unaffected by the clamp in both the young and age-matched control subjects, it was increased (P < 0.05) approximately 70-fold in the diabetic patients after the clamp. These data demonstrate that genes involved in providing cellular protection against oxidative stress are defective in patients with type 2 diabetes and correlate with insulin-stimulated glucose disposal and markers of muscle oxidative capacity. The data provide new evidence that the pathogenesis of type 2 diabetes involves perturbations to the antioxidant defense mechanism within skeletal muscle.


Acalculous adenomyomatosis of the gallbladder.
The course of acalculous adenomyomatosis of the gallbladder in six patients is described. It is suggested that, even in the absence of gallstones, cholecystectomy should be advised when this condition is demonstrated radiographically in symptomatic subjects. The cause of the pain is unknown but it is probably related to excessive neuromuscular activity of the hyperplastic gallbladder wall. One of the patients was found also to have an adenomatous polyp containing areas of adenocarcinoma. Although this polyp was not situated within an area of adenomyomatosis, it is possible that, as in most other patients with carcinoma of the gallbladder, previous disease may have predisposed to malignant change.


Scoliosis associated with osteogenesis imperfecta.
A survey was conducted to document the results of bracing and spinal fusion for scoliosis associated with osteogenesis imperfecta. Observations were made of 121 patients who underwent treatment by bracing or spinal fusion and who had been treated by 51 orthopaedic surgeons in 14 countries. The average curve before bracing measured 43 degrees. The braces were ineffective in stopping progression even in small curves. We were unable to determine whether braces slowed the rate of progression of curvature. The average age at fusion was 15 years 7 months, the average curve before operation measured 74 degrees, and the average correction was 36 per cent. The high incidence of complications was related to the size of the curve before spinal fusion, the use of Harrington instrumentation, and the presence of associated kyphosis. In the absence of pseudarthrosis or kyphosis, late bending of the fused spine did not seem to occur.


Management of chronic spontaneous urticaria: practical parameters.
Chronic urticaria (CU) is a skin disorder characterized by transient, pruritic wheals persisting for longer than 6 weeks. According to the European Academy of Allergy and Clinical Immunology (EAACI) guidelines, CU can be categorized into two main types: chronic spontaneous urticaria (CSU), in which the wheals appear spontaneously, and inducible urticaria, that is triggered by physical agents. CSU may be due to triggering factors such as food allergens or infections, but in at least 40% of cases it is autoimmune in origin, caused by circulating autoantibodies anti-FcεR1 or anti-IgE, or autoreactive. In the present paper, re-evaluating the EAACI guidelines, we have developed a document containing some practical indications which are useful for diagnosis and management of CSU in the context of the Italian situation. Concerning CSU treatment, second generation antihistamines are the first-line treatment; these drugs can be used, as second-line treatment, at a higher than licensed dose in patients who do not respond adequately at licensed doses. The third-line treatment includes leukotriene receptor antagonists which, however, do not have a specific indication for the treatment of CSU, cyclosporine, whose use in this disease is still off-label, and omalizumab. The latter is a recombinant monoclonal IgG antibody that binds free IgE, down regulates mast cell function and induces eosinophil apoptosis. Recently, it has emerged as an effective and safe treatment for antihistamine-unresponsive CSU of both autoimmune/autoreactive and non-autoimmune/non-autoreactive, and has been officially approved for use against this disease.


[Characteristics of the NK cells in Syrian hamsters carrying tumors with varying metastatic and resistance-suppressing activity].
Experiments in vivo and in vitro were made to study the effects of HETR-MLN-8 and HETR tumor cells differing in metastatic ability and inhibition of the natural host resistance to tumor on cytotoxic activity of NK from Syrian hamsters. Marked inhibition of cytotoxicity and ability for interferon activation was detected in NK isolated from tumors (as compared with blood), with that inhibition being far more pronounced in highly malignant HETR-MLN-8 tumors. This may indicate a direct inhibitory action of the tumor or its products on NK cytotoxicity. The in-vitro competition inhibition test yielded results showing that HETR-MLN-8 cells capable of in-vivo inhibition of the natural host resistance to the tumor also display much more demonstrable ability for in-vitro inhibition of NK cytotoxicity as compared to HETR cells.


The missense mutation W290R in Fgfr2 causes developmental defects from aberrant IIIb and IIIc signaling.
Missense mutations in the Fibroblast Growth Factor Receptor 2 (FGFR2) have been identified in human craniosynostotic syndromes such as Crouzon (CS) and Pfeiffer (PS). FGFR2 has two major isoforms, IIIb and IIIc, generated through alternative splicing with their own temporal, spatial, and ligand-binding specificities. In this study, we report the identification and characterization of a missense mutation in codon 290 of murine Fgfr2 (W290R). The defects in W290R mutants are suggestive of disruption of signalling in both IIIb and IIIc isoforms of the Fgfr2 gene. Heterozygous mutants presented with features resembling those found in patients with CS. Fgfr2(W290R) homozygotes displayed constitutive FGFR2 activation with increased, but correct tissue-specific, expression of the IIIb and IIIc isoforms in many of the defective organs. Our Fgfr2(W290R) mouse model thus represents an excellent mouse model of CS to probe the many questions around the pathogenesis of craniosynostotic birth defects consequent to defects in FGF signaling.


Synthesis and antimicrobial activity of N-analogous corollosporines.
Corollosporine is an antimicrobial metabolite, which was isolated from the marine fungus Corollospora maritima. Owing to its basic 4-hydroxyphthalic acid anhydride structure, it has become an attractive target for a structure/activity relationship modelling of derived compounds with potential antibiotic activity. In this regard we report on the straightforward synthesis of hetero analogous corollosporines, which were easily prepared by a three-step reaction sequence, taking advantage of a novel multicomponent reaction (AAD-reaction) and a subsequent aromatization/Grignard reaction protocol. Furthermore, the obtained products were tested in several biological assays to evaluate their antimicrobial activity.


[Changes in the levels of nitric oxide in the mononuclear cells and neutrophils of patients with tuberculosis with varying courses of pulmonary tuberculosis].
A hundred and forty four patients with different types of the course of pulmonary tuberculosis were examined. Before and 20 hours after incubation with BCG-killed cultures, mononuclear cells and neutrophils were used to estimate the level of generation of nitric oxide (NO) from the content of its stabile metabolites--nitrites determined by Griess reagent. Crucially active pulmonary tuberculosis was found to follow the increased leukocytic generation of NO. The most profound shifts occurred in the mononuclear cells. During a favorable involutional process, the mononuclear cells increased their baseline generation of NO and, moreover, preserved their potential for a responsive "nitric oxide burst" when they met a specific causative agent. In patients with acute pulmonary tuberculosis, the basal level of NO in the mononuclear cells was high, but their response to BCG proved to be decreased. In the neutrophils, the basal level of NO exhibited a significant scatter and their higher response to BCG was noted only during the involutional course of tuberculosis. Changes in the leukocytic levels of NO were related to the magnitude of a systemic inflammatory response, to the severity of respiratory failure, and to the rate of cellular lipid peroxidation. A clear relationship was observed between the nature of baseline changes in the values of NA and the efficiency of subsequent treatment.


Chemo Checker: Transforming Care and Increasing Patient Safety by Reducing Nurse Distractions.
On a high-volume academic ambulatory chemotherapy unit, a medication error triggered an evidence-based practice nursing work group to initiate a pilot "chemo checker" role as part of a new chemotherapy verification process. The group implemented two interventions to minimize nurse distractions and a 17-point chemotherapy verification label. An independent verification role was created to perform the first check for chemotherapy. Six months into the pilot project, medication administration errors and nurses' distractions during the chemotherapy verification process decreased, and nurses' perception of safety during the process increased (N = 22).


Biomaterials in the clinical treatment of lymphedema-a systematic review.
Lymphedema is a chronic condition caused by impaired lymphatic fluid drainage, resulting in progressive edema. The current mainstay of lymphedema therapy consists of conservative therapy and surgical therapy. In this systematic review, we investigated the novel role of biomaterials in clinical lymphedema therapy and assessed their objective outcomes and the complication rate associated with their use. Studies were identified through systematic review using the Embase and PubMed/MEDLINE databases. Only original articles reporting the use of biomaterials for clinical lymphedema therapy were included. The primary outcome measure was the objective reduction in limb volume after biomaterial use. The secondary outcome measure was the assessment of biomaterial safety. A total of 354 articles were identified in the first search, of which 10 met our inclusion criteria. These articles described the use of two biomaterials, nanofibrillar collagen scaffolds (NCSs) and silicone tubes (STs), for the treatment of lymphedema. NCS implantation showed an average excess limb volume reduction of 1% to 10.7% and clear evidence of lymphangiogenesis on imaging. No complications were 7documented after NCS implantation. ST implantation showed an average limb volume reduction of 700 to 887 mL and limb circumference reduction of 3.1 to 8 cm in patients with advanced stage lymphedema. Of 177 patients treated with ST implantation, only 11 (6.2%) developed local inflammation. Both NCS and ST implantation showed promising limb volume reduction; however, with the scarce literature available, additional research is needed to determine their effectiveness. Both demonstrated good safety profiles, with no complications after NCS implantation and a complication rate equivalent to other similar implants for ST implantation.


Modulation of leukocyte infiltration and phenotype in microporous tissue engineering scaffolds via vector induced IL-10 expression.
Biomaterial scaffolds are central to many tissue engineering strategies as they create a space for tissue growth and provide a support for cell adhesion and migration. However, biomaterial implantation results in unavoidable injury resulting in an inflammatory response, which can impair integration with the host and tissue regeneration. Toward the goal of reducing inflammation, we investigated the hypothesis that a lentiviral gene therapy-based approach to localized and sustained IL-10 expression at a scaffold could modulate the number, relative proportions, and cytokine production of infiltrating leukocyte populations. Flow cytometry was used to quantify infiltration of six leukocyte populations for 21 days following implantation of PLG scaffolds into intraperitoneal fat. Leukocytes with innate immune functions (i.e., macrophages, dendritic cells, neutrophils) were most prevalent at early time points, while T lymphocytes became prevalent by day 14. Reporter gene delivery indicated that transgene expression persisted at the scaffold for up to 28 days and macrophages were the most common leukocyte transduced, while transduced dendritic cells expressed the greatest levels of transgene. IL-10 delivery decreased leukocyte infiltration by 50% relative to controls, increased macrophage IL-10 expression, and decreased macrophage, dendritic cell, and CD4 T cell IFN-γ expression. Thus, IL-10 gene delivery significantly decreased inflammation following scaffold implant into the intraperitoneal fat, in part by modulating cytokine expression of infiltrating leukocytes.


Simple tests of homogeneity of controls in matched studies.
Consider matched case-control studies with a pair of distinguishable controls, say neighbourhood and hospital, wherein one of the controls may be missing. Liang and Stewart derive an efficient test for the possible difference in controls for the complete data case. They suggest inefficient tests for the situation where some triplets are incomplete. Levin, and Risch and Tibshirani, derive efficient tests for the incomplete triplet case by the methods of maximum likelihood estimator (Wald) tests and likelihood ratio tests, respectively. All these tests require the use of compute algorithms. Using score theory, we derive a simple efficient test for complete triplets which only requires solving a quadratic equation. Furthermore, we show that the usual normal approximation to McNemar's test is equivalent to the score test and is thus fully efficient as well. For incomplete triplets, score theory leads to a fully efficient test which only requires the solution of a cubic equation. We propose a nearly efficient test which is even more simply computed. The example of Liang and Stewart illustrates application of the results.


Plantar Ulcers and Neuropathic Arthropathies: Associated Diseases, Polyneuropathy Correlates, and Risk Covariates.
To evaluate the associated diseases, polyneuropathy correlates, and risk covariates of neuropathic plantar ulcers (PUs) and neuropathic arthropathies (NAs). The authors conducted a retrospective, observational study over 3.5 years of 69 patients with neuropathy, NA, or PU seen in a wound clinic who also had a comprehensive neurologic evaluation and neurophysiologic testing. Comparisons were made to a population representative cohort of patients with diabetes mellitus (DM; n = 259). Of the 69 wound clinic patients, 32 had PUs, 14 had NAs, and 23 had both. Of the 61 adequately assessed patients, 37 (61%) had DM, 22 (36%) had no known associated disease, and 2 (3%) had hereditary sensory and autonomic neuropathy. Of the 37 patients with DM, 35 had distal polyneuropathy, and 2 did not. In 22 patients with chronic idiopathic axonal polyneuropathy, 20 had distal polyneuropathy. Although DM was the disease most commonly associated with PUs and NAs, chronic hyperglycemia may not have been the major underlying risk factor. The major risk covariates are sensation loss from polyneuropathy, old age, obesity, repetitive foot injury, and inadequate foot care or treatment. Physicians and other healthcare providers can help by identifying patients at risk and instituting measures such as adequate foot care to decrease these risks.


Rural southeast Texas air quality measurements during the 2006 Texas Air Quality Study.
The authors conducted air quality measurements of the criteria pollutants carbon monoxide, nitrogen oxides, and ozone together with meteorological measurements at a park site southeast of College Station, TX, during the 2006 Texas Air Quality Study II (TexAQS). Ozone, a primary focus of the measurements, was above 80 ppb during 3 days and above 75 ppb during additional 8 days in summer 2006, suggestive of possible violations of the ozone National Ambient Air Quality Standard (NAAQS) in this area. In concordance with other air quality measurements during the TexAQS II, elevated ozone mixing ratios coincided with northerly flows during days after cold front passages. Ozone background during these days was as high as 80 ppb, whereas southerly air flows generally provided for an ozone background lower than 40 ppb. Back trajectory analysis shows that local ozone mixing ratios can also be strongly affected by the Houston urban pollution plume, leading to late afternoon ozone increases of as high as 50 ppb above background under favorable transport conditions. The trajectory analysis also shows that ozone background increases steadily the longer a southern air mass resides over Texas after entering from the Gulf of Mexico. In light of these and other TexAQS findings, it appears that ozone air quality is affected throughout east Texas by both long-range and regional ozone transport, and that improvements therefore will require at least a regionally oriented instead of the current locally oriented ozone precursor reduction policies.


Agreement between contemporaneously recorded and subsequently recalled time spent outdoors: implications for environmental exposure studies.
The aim of the study is to evaluate the agreement between contemporaneously recorded and subsequently recalled time spent outdoors during 1 week among members of an occupational cohort. One hundred twenty-five radiologic technologists from northern and southern geographic areas in the United States recorded time spent outdoors for 7 consecutive days in a daily diary. Six months later, study participants completed a mailed self-administered questionnaire of the number of outdoor hours during the same 7-day period. We tested the agreement between questionnaire responses and diary entries. Logistic regression models were used to identify variables significantly affecting agreement. Time spent outdoors comprised one fifth of the total time recorded in the diaries. Agreement (weighted kappa [kappa(w)]) between reported outdoor time during weekdays (kappa(w) = 0.49; 95% confidence interval [CI], 0.39-0.59) was significantly (p < 0.05) higher than for weekends (kappa(w) = 0.23; 95% CI, 0.12-0.34). Similarly, agreement was lower for weekends compared with weekdays in multivariate analyses, reaching statistical significance (p = 0.05) in only the southern regions. Although our investigation was carried out among volunteers from the US radiologic technologist cohort, we believe retrospective questionnaires may be more accurate in reporting time spent outdoors for weekdays compared with weekends in any group of indoor workers. These differences have implications for the wording in future questionnaires about time spent outdoors and level and sources of uncertainty characterizing estimated time spent outdoors on weekdays versus weekend days.


The impact of the current epidemiology of pediatric musculoskeletal infection on evaluation and treatment guidelines.
Methicillin-resistant Staphylococcus aureus is thought to have led to an increase in the incidence of severe musculoskeletal infection in children. Our purpose was (1) to compare the current epidemiology of musculoskeletal infection with historical data at the same institution 20 years prior and (2) to evaluate the spectrum of the severity of this disease process within the current epidemiology. Children with musculoskeletal infection, treated between January 2002 and December 2004, were studied retrospectively. Diagnoses of osteomyelitis, septic arthritis, pyomyositis, and abscess were established for each child based on overall clinical impression, laboratory indices, culture results, radiology studies, and intraoperative findings. Comparison was made with the experience reported at the same institution in 1982. Children within each diagnostic category were compared with respect to mean values of C-reactive protein and erythrocyte sedimentation rate at admission, number of surgical procedures, intensive care unit admissions, identification of deep venous thrombosis, and length of hospitalization. Five hundred fifty-four children were studied (osteomyelitis, n = 212; septic arthritis, n = 118; pyomyositis, n = 20; and deep abscess, n = 204). The annualized per capita incidence of osteomyelitis increased 2.8-fold, whereas that of septic arthritis was unchanged when compared with historical data from 20 years prior. Methicillin-resistant Staphylococcus aureus was isolated as the causative organism in 30% of the children. We identified increasing severity of illness according to a hierarchy of tissue involvement (osteomyelitis > septic arthritis > pyomyositis > abscess) and according to the identification of contiguous infections within in each primary diagnostic category. The incidence of musculoskeletal infection appears to have increased within our community. We found that a more comprehensive diagnostic classification of this disease is useful in understanding the spectrum of the severity of illness and identifying those who require the greatest amount of resources. Magnetic resonance imaging is useful early in the diagnostic process to enable a more detailed disease classification and to expedite surgical decisions. The recognition of the incidence of methicillin-resistant Staphylococcus aureus within our community has also led to a change in empirical antibiotic selection.


[Three cases of hereditary pancreatitis in two households in the same family associated with R122H mutation in cationic trypsinogen gene].
Hereditary pancreatitis is a rare, autosomal dominant, inherited disease characterized by recurrent attacks of acute pancreatitis with the development of chronic pancreatitis and an increased risk of pancreatic cancer. R122H or N29I mutation in cationic trypsinogen (protease serine 1, PRSS1) gene causes hereditary pancreatitis. R122H mutation is the most common mutation that causes pancreatitis by preventing deactivation of trypsin within the pancreas and prolonging its action. Three members of the family, the patient, her elder son, and her niece experienced recurrent attacks of pancreatitis. We analyzed five exons of the PRSS1 gene in DNA samples of five family members including her husband and younger son who were asymptomatic. We found out that four members of the family, the patient, her two sons, and her niece, had R122H mutation in the exon 3 of PRSS1 gene. Finally, we diagnosed hereditary pancreatitis in two households in the same family.


Characterization of a mammalian peroxiredoxin that contains one conserved cysteine.
A new type of peroxidase enzyme, named thioredoxin peroxidase (TPx), that reduces H2O2 with the use of electrons from thioredoxin and contains two essential cysteines was recently identified. TPx homologs, termed peroxiredoxin (Prx), have also been identified and include several proteins, designated 1-Cys Prx, that contain only one conserved cysteine. Recombinant human 1-Cys Prx expressed in and purified from Escherichia coli has now been shown to reduce H2O2 with electrons provided by dithiothreitol. Furthermore, human 1-Cys Prx transiently expressed in NIH 3T3 cells was able to remove intracellular H2O2 generated in response either to the addition of exogenous H2O2 or to treatment with platelet-derived growth factor. The conserved Cys47-SH group was shown to be the site of oxidation by H2O2. Thus, mutation of Cys47 to serine abolished peroxidase activity. Moreover, the oxidized intermediate appears to be Cys-SOH. In contrast to TPx, in which one of the two conserved cysteines is oxidized to Cys-SOH and then immediately reacts with the second conserved cysteine of the second subunit of the enzyme homodimer to form an intermolecular disulfide, the Cys-SOH of 1-Cys Prx does not form a disulfide. Neither thioredoxin, which reduces the disulfide of TPx, nor glutathione, which reduces the Cys-SeOH of oxidized glutathione peroxidase, was able to reduce the Cys-SOH of 1-Cys Prx and consequently could not support peroxidase activity. Human 1-Cys Prx was previously shown to exhibit a low level of phospholipase A2 activity at an acidic pH; the enzyme was thus proposed to be lysosomal, and Ser32 was proposed to be critical for lipase function. However, the mutation of Ser32 or Cys47 has now been shown to have no effect on the lipase activity of 1-Cys Prx, which was also shown to be a cytosolic protein. Thus, the primary cellular function of 1-Cys Prx appears to be to reduce peroxides with the use of electrons provided by an as yet unidentified source; the enzyme therefore represents a new type of peroxidase.


Automated fluorescent analysis for drug-induced cytotoxicity assays.
The human tumor clonogenic assay has been reported to predict for sensitivity of human tumors to a variety of drugs. However, this assay requires large numbers of viable cells, is time-consuming, and takes at least 2 weeks before results are available. To circumvent these problems, Weisenthal developed a microscope-based dye exclusion assay. Because this method is also time-consuming and subject to observer error, we have developed an automated method of quantitating drug cytotoxicity using a flow cytometric cell sorter (FCM). After incubation of drug-exposed tumor cells, acetaldehyde-fixed duck red blood cells (DRBC) are added. Dead tumor cells and the fixed DRBC are stained by the fluorescent dye propidium iodide, which penetrates dead cell membranes. A two-parameter analysis (cell size as measured by narrow angle light scatter vs propidium iodide fluorescence) enables determination of the live tumor cell:DRBC ratio. There was a strong correlation between the FCM method and manual counting (r = 0.958 for cell lines, r = 0.831 for fresh leukemic cells, P less than 0.0001 in both cases). We conclude that the automatized FCM method gives compatible results to the manual dye exclusion assay and increases efficiency.


Sensitivity of negative subsequent memory and task-negative effects to age and associative memory performance.
The present fMRI experiment employed associative recognition to investigate the relationships between age and encoding-related negative subsequent memory effects and task-negative effects. Young, middle-aged and older adults (total n=136) were scanned while they made relational judgments on visually presented word pairs. In a later memory test, the participants made associative recognition judgments on studied, rearranged (items studied on different trials) and new pairs. Several regions, mostly localized to the default mode network, demonstrated negative subsequent memory effects in an across age-group analysis. All but one of these regions also demonstrated task-negative effects, although there was no correlation between the size of the respective effects. Whereas negative subsequent memory effects demonstrated a graded attenuation with age, task-negative effects declined markedly between the young and the middle-aged group, but showed no further reduction in the older group. Negative subsequent memory effects did not correlate with memory performance within any age group. By contrast, in the older group only, task-negative effects predicted later memory performance. The findings demonstrate that negative subsequent memory and task-negative effects depend on dissociable neural mechanisms and likely reflect distinct cognitive processes. The relationship between task-negative effects and memory performance in the older group might reflect the sensitivity of these effects to variations in amount of age-related neuropathology. This article is part of a Special Issue entitled SI: Memory.


A Generic Approach to Lung Field Segmentation From Chest Radiographs Using Deep Space and Shape Learning.
Computer-aided diagnosis (CAD) techniques for lung field segmentation from chest radiographs (CXR) have been proposed for adult cohorts, but rarely for pediatric subjects. Statistical shape models (SSMs), the workhorse of most state-of-the-art CXR-based lung field segmentation methods, do not efficiently accommodate shape variation of the lung field during the pediatric developmental stages. The main contributions of our work are: 1) a generic lung field segmentation framework from CXR accommodating large shape variation for adult and pediatric cohorts; 2) a deep representation learning detection mechanism, ensemble space learning, for robust object localization; and 3) marginal shape deep learning for the shape deformation parameter estimation. Unlike the iterative approach of conventional SSMs, the proposed shape learning mechanism transforms the parameter space into marginal subspaces that are solvable efficiently using the recursive representation learning mechanism. Furthermore, our method is the first to include the challenging retro-cardiac region in the CXR-based lung segmentation for accurate lung capacity estimation. The framework is evaluated on 668 CXRs of patients between 3 month to 89 year of age. We obtain a mean Dice similarity coefficient of 0.96 ±0.03 (including the retro-cardiac region). For a given accuracy, the proposed approach is also found to be faster than conventional SSM-based iterative segmentation methods. The computational simplicity of the proposed generic framework could be similarly applied to the fast segmentation of other deformable objects.


Two-dimensional photoacoustic imaging by use of Fourier-transform image reconstruction and a detector with an anisotropic response.
Theoretical and experimental aspects of two-dimensional (2D) biomedical photoacoustic imaging have been investigated. A 2D Fourier-transform-based reconstruction algorithm that is significantly faster and produces fewer artifacts than simple radial backprojection methods is described. The image-reconstruction time for a 208 x 482 pixel image is approximately 1 s. For the practical implementation of 2D photoacoustic imaging, a rectangular detector geometry was used to obtain an anisotropic detection sensitivity in order to reject out-of-plane signals, thereby permitting a tomographic image slice to be reconstructed. This approach was investigated by the numerical modeling of the broadband directional response of a rectangular detector and imaging of various spatially calibrated absorbing targets immersed in a turbid phantom. The experimental setup was based on a Q-switched Nd:YAG excitation laser source and a mechanically line-scanned Fabry-Perot polymer-film ultrasound sensor. For a 800 microm x 200 microm rectangular detector, the reconstructed image slice thickness was 0.8 mm up to a vertical distance of z = 3.5 mm from the detector, increasing thereafter to 2 mm at z = 10 mm. Horizontal and vertical spatial resolutions within the reconstructed slice were approximately 200 and 60 microm, respectively.


Unpredictable chronic stress-induced reproductive suppression associated with the decrease of kisspeptin immunoreactivity in male mice.
Environmental stress affects various parts of mammals typically through the circulation of stress hormones. It has been identified as one of the possible reasons for male reproductive difficulties, but the complex mechanisms responsible for stress-induced reproductive suppression are poorly understood. Here, we examined the relationship between chronic environmental stress and hypothalamic kisspeptin, a recently discovered upstream regulator of the reproductive endocrine feedback system. We studied male mice under an unpredictable chronic stress procedure to replicate the situation of animals under chronic stress. Histological and immunohistochemical analyses were performed focusing on kisspeptin neurons in the arcuate hypothalamic nucleus (ARC) and DNA fragmented cells in seminiferous tubules. Although the ARC was not morphologically altered in either the stressed or non-stressed group, granular kisspeptin immunoreactivities decreased slightly in the stress group. In the testes of the stress group, several signs of testicular degeneration were observed, including increased numbers of ssDNA-positive cells per seminiferous tubule, thinning, vacuoled seminiferous epithelia and multinucleated giant cells. The decreases in kisspeptin in the stress group might be due to other hypothalamic peptides, such as corticotropin-releasing hormone and leptin, whose receptors are known to coexpress in the ARC. In addition, environmental stress directly and indirectly affects testicular function through stress hormones and gonadotropins. In summary, our findings enhance the understanding of stress-induced reproductive suppression possibly mediated by kisspeptin in the ARC.


[Reduction of methemoglobin with photoradicals of benzophenone-3,3',4,4'-tetracarbonic acid].
Reaction ability of partially reduced human methemoglobin was estimated by the rate of protein addition of carbon monoxide immediately after reduction. Impulse of photoexcitation allowed two variants of these experiments to be carried out: immediately after partial reduction of methemoglobin with photoradicals of benzophenone 3,3',4,4'-tetracarbon acid and after photodissociation of the carboxy-complex obtained in the first case. It has been shown that at protein reduction "p" less than or equal to 0.03 under these conditions the initially obtained R-form of protein is partially transformed into the less reactive T-form.


Design of polyamine-based therapeutic agents: new targets and new directions.
Enzymes in the biosynthetic and catabolic polyamine pathway have long been considered targets for drug development, and early drug discovery efforts in the polyamine area focused on the design and development of specific inhibitors of the biosynthetic pathway, or polyamine analogues that specifically bind DNA. More recently, it has become clear that the natural polyamines are involved in numerous known and unknown cellular processes, and disruption of polyamine functions at their effector sites can potentially produce beneficial therapeutic effects. As new targets for polyamine drug discovery continue to evolve, the rational design of polyamine analogues will result in more structurally diverse agents. In addition, the physical linkage of polyamine-like structures to putative drug molecules can have beneficial effects resulting from increases in DNA affinity and selective cellular uptake. The present chapter will summarize recent advances in the development of alkylpolyamine analogues as antitumour agents, and describe subsequent advances that have resulted from incorporating polyamine character into more diverse drug molecules. Specifically, new polyamine analogues, and the role of polyamine fragments in the design of antiparasitic agents, antitumour metal complexes, histone deacetylase inhibitors and lysine-specific demethylase 1 inhibitors, will be described.


Mutual capture of dipolar molecules at low and very low energies. II. Numerical study.
The low-energy rate coefficients of capture of two identical dipolar polarizable rigid rotors in their lowest nonresonant (j(1) = 0 and j(2) = 0) and resonant (j(1) = 0, 1 and j(2) = 1, 0) states are calculated accurately within the close-coupling (CC) approach. The convergence of the quantum rate coefficients to their quantum-classical counterparts is studied. A comparison of the present accurate numerical with approximate analytical results (Nikitin, E. E.; Troe, J. J. Phys. Chem. A 2010, 114, 9762) indicates a good performance of the previous approach which was based on the interpolation between s-wave fly wheel quantal and all-wave classical adiabatic channel limits. The results obtained apply as well to the formation of transient molecular species in the encounter of two atoms at very low collision energy interacting via resonance dipole-dipole interaction.


Update: Ebola virus disease epidemic - West Africa, February 2015.
CDC is assisting ministries of health and working with other organizations to end the ongoing epidemic of Ebola virus disease (Ebola) in West Africa. The updated data in this report were compiled from situation reports from the Guinea Interministerial Committee for Response Against the Ebola Virus, the Liberia Ministry of Health and Social Welfare, the Sierra Leone Ministry of Health and Sanitation, and the World Health Organization.


Glycocalyx volume: a critical review of tracer dilution methods for its measurement.
A clinical measure of endothelial glycocalyx structure would have great potential importance, because lesions of the glycocalyx may be the first changes to occur in diabetes and in a wide range of vascular diseases. A method recently described by Nieuwdorp et al. for estimating the volume of the luminal glycocalyx of the entire human vascular system would seem to be the first attempt to develop a measure of this kind. It is based on the tracer dilution principle, and this review considers the principles and conditions that underlie this method and the extent to which the conditions appear to have been fulfilled in this case. Our analysis raises two questions about 1) the estimation of the concentration of the tracer (dextran 40) at zero time and 2) the estimation of plasma volume, both of which can be answered by changes in experimental protocol. A third question, concerning the partition coefficient of the tracer between plasma and the fluid within the glycocalyx, cannot be answered at the present time, and until it has been resolved, glycocalyx volume cannot be estimated from the dilution of a macromolecular tracer.


Differential endothelial injury caused by vascular clamps and vessel loops. I. Normal vessels.
Silicone rubber looped tourniquets were compared with standard crossmembered metallic peripheral vascular occlusive clamps in a canine aorto-iliac vessel model to compare the degree and severity of endothelial injury as demonstrated by scanning electron microscopy. The moderate-to-severe injury incurred by the standard clamps was not seen whatsoever with the looped tourniquets. Selective and preferential use of silastic rubber vessel tourniquet loops for vascular occlusion and control is thus advocated.


Comparing systemic defence-related gene expression changes upon migratory and sedentary nematode attack in rice.
Complex defence signalling pathways, controlled by different hormones, are known to be involved in the reaction of plants to a wide range of biotic and abiotic stress factors. Here, we studied the differential expression of genes involved in stress and defence responses in systemic tissue of rice infected with the root knot nematode (RKN) Meloidogyne graminicola and the migratory root rot nematode Hirschmanniella oryzae, two agronomically important rice pathogens with very different lifestyles. qRT-PCR revealed that all investigated systemic tissues had significantly lower expression of isochorismate synthase, a key enzyme for salicylic acid production involved in basal defence and systemic acquired resistance. The systemic defence response upon migratory nematode infection was remarkably similar to fungal rice blast infection. Almost all investigated defence-related genes were up-regulated in rice shoots 3 days after root rot nematode attack, including the phenylpropanoid pathway, ethylene pathway and PR genes, but many of which were suppressed at 7 dpi. Systemic shoot tissue of RKN-infected plants showed similar attenuation of expression of almost all studied genes already at 3 dpi, with clear attenuation of the ethylene pathway and methyl jasmonate biosynthesis. These results provide an interesting starting point for further studies to elucidate how nematodes are able to suppress systemic plant defence mechanisms and the effect in multitrophic interactions.


SEPP: SATé-enabled phylogenetic placement.
We address the problem of Phylogenetic Placement, in which the objective is to insert short molecular sequences (called query sequences) into an existing phylogenetic tree and alignment on full-length sequences for the same gene. Phylogenetic placement has the potential to provide information beyond pure "species identification" (i.e., the association of metagenomic reads to existing species), because it can also give information about the evolutionary relationships between these query sequences and to known species. Approaches for phylogenetic placement have been developed that operate in two steps: first, an alignment is estimated for each query sequence to the alignment of the full-length sequences, and then that alignment is used to find the optimal location in the phylogenetic tree for the query sequence. Recent methods of this type include HMMALIGN+EPA, HMMALIGN+pplacer, and PaPaRa+EPA.We report on a study evaluating phylogenetic placement methods on biological and simulated data. This study shows that these methods have extremely good accuracy and computational tractability under conditions where the input contains a highly accurate alignment and tree for the full-length sequences, and the set of full-length sequences is sufficiently small and not too evolutionarily diverse; however, we also show that under other conditions accuracy declines and the computational requirements for memory and time exceed acceptable limits. We present SEPP, a general "boosting" technique to improve the accuracy and/or speed of phylogenetic placement techniques. The key algorithmic aspect of this booster is a dataset decomposition technique in SATé, a method that utilizes an iterative divide-and-conquer technique to co-estimate alignments and trees on large molecular sequence datasets. We show that SATé-boosting improves HMMALIGN+pplacer, placing short sequences more accurately when the set of input sequences has a large evolutionary diameter and produces placements of comparable accuracy in a fraction of the time for easier cases. SEPP software and the datasets used in this study are all available for free at http://www.cs.utexas.edu/users/phylo/software/sepp/submission.


Emerging therapeutic targets in esophageal adenocarcinoma.
The incidence of gastro-esophageal disease and associated rate of esophageal adenocarcinoma (EAC) is rising at an exponential rate in the United States. However, research targeting EAC is lagging behind, and much research is needed in the field to identify ways to diagnose EAC early as well as to improve the rate of pathologic complete response (pCR) to systemic therapies. Esophagectomy with subsequent reconstruction is known to be a morbid procedure that significantly impacts a patient's quality of life. If indeed the pCR rate of patients can be improved and those patients destined to be pCR can be identified ahead of time, they may be able to avoid this life-altering procedure. While cancer-specific biological pathways have been thoroughly investigated in other solid malignancies, much remains unexplored in EAC. In this review, we will highlight some of the latest research in the field in regards with EAC, along with new therapeutic targets that are currently being explored. After reviewing conventional treatment and current changes in medical therapy for EAC, we will focus on unchartered grounds such as cancer stem cells, genetics and epigenetics, immunotherapy, and chemoradio-resistant pathways as we simultaneously propose some investigational possibilities that could be applicable to EAC.


Dynamics of a microbial community associated with manure hot spots as revealed by phospholipid fatty acid analyses.
Microbial community dynamics associated with manure hot spots were studied by using a model system consisting of a gel-stabilized mixture of soil and manure, placed between layers of soil, during a 3-week incubation period. The microbial biomass, measured as the total amount of phospholipid fatty acids (PLFA), had doubled within a 2-mm distance from the soil-manure interface after 3 days. Principal-component analyses demonstrated that this increase was accompanied by reproducible changes in the composition of PLFA, indicating changes in the microbial community structure. The effect of the manure was strongest in the 2-mm-thick soil layer closest to the interface, in which the PLFA composition was statistically significantly different (P < 0.05) from that of the unaffected soil layers throughout the incubation period. An effect was also observed in the soil layer 2 to 4 mm from the interface. The changes in microbial biomass and community structure were mainly attributed to the diffusion of dissolved organic carbon from the manure. During the initial period of microbial growth, PLFA, which were already more abundant in the manure than in the soil, increased in the manure core and in the 2-mm soil layer closest to the interface. After day 3, the PLFA composition of these layers gradually became more similar to that of the soil. The dynamics of individual PLFA suggested that both taxonomic and physiological changes occurred during growth. Examples of the latter were decreases in the ratios of 16:1 omega 7t to 16:1 omega 7c and of cyclopropyl fatty acids to their respective precursors, indicating a more active bacterial community. An inverse relationship between bacterial PLFA and the eucaryotic 20:4 PLFA (arachidonic acid) suggested that grazing was important.


Bifidobacterium mechanisms of immune modulation and tolerance.
Bifidobacterium is a widely distributed commensal bacterial genus that displays beneficial pro-homeostatic and anti-inflammatory immunomodulatory properties. Depletion or absence of Bifidobacterium in humans and model organisms is associated with autoimmune responses and impaired immune homeostasis. At the cellular level, Bifidobacterium upregulates suppressive regulatory T cells, maintains intestinal barrier function, modulates dendritic cell and macrophage activity, and dampens intestinal Th2 and Th17 programs. While there has been a large volume of literature characterizing the probiotic properties of various Bifidobacterial species, the likely multifactorial mechanisms underlying these effects remain elusive, in particular, its immune tolerogenic effect. However, recent work has shed light on Bifidobacterium surface structural polysaccharide and protein elements, as well as its metabolic products, as commensal mediators of immune homeostasis. This review aims to discuss several mechanisms Bifidobacterium utilizes for immune modulation as well as their indirect impact on the regulation of gut microbiome structure and function, from structural molecules to produced metabolites. These mechanisms are pertinent to an increasingly networked understanding of immune tolerance and homeostasis in health and disease.


Use of reciprocal pool specific activities to model leucine metabolism in humans.
To investigate in humans whether interconversion of leucine and its alpha-keto acid, alpha-ketoisocaproate (KIC), is rate limiting for leucine metabolism, normal volunteers were infused with independent 3H and 14C labels of leucine and KIC. Total leucine carbon (leucine + KIC) entry into the plasma space determined by using both plasma specific activities (SA) of the infused isotopes or by using either plasma SA of the leucine moieties reciprocal to those being infused (e.g., [3H]KIC and [14C]leucine during [3H]leucine and [14C]KIC infusion) were nearly identical. When [3H]leucine and [14C]KIC were infused, the ratio of 3H/14C radioactivity of leucine in plasma proteins was similar both to the ratio of plasma [3H]KIC SA/[14C]leucine SA and to the ratio of the rate of [3H]leucine/[14C]KIC infused. Estimates of leucine oxidation using expired 14CO2 and the plasma SA of the 14C moiety reciprocal to infused [14C]leucine or [14C]KIC were similar. We conclude that the plasma SA of the leucine moiety reciprocal to infused labeled leucine or KIC may reflect more accurately whole-body leucine metabolism in postabsorptive humans.


Ascorbic Acid: a promising memory-enhancer in mice.
Alzheimer's disease is a progressive neurodegenerative disorder characterized by a gradual decline in memory. The occurrence of Alzheimer's disease is commonplace among the Asian population, particularly among senior citizens. The present study was undertaken to assess the potential of ascorbic acid as a memory-enhancer. Swiss mice of either sex were employed in the present investigation. Elevated plus-maze and passive-avoidance apparatus served as the exteroceptive behavioral models, and diazepam-, scopolamine-, and aging-induced amnesia served as the interoceptive behavioral models. Ascorbic acid (60, 120 mg/kg) injected for 3 and 8 consecutive days improved learning and memory of aged mice as indicated by decreased transfer-latency and increased step-down latency. Furthermore, ascorbic acid provided protection to the young animals from scopolamine- and diazepam-induced impairment of memory. Ascorbic acid was found to be more potent than piracetam as reflected by the smaller dose, more pronounced effect, and quicker onset of action. Ascorbic acid has shown promise as a powerful memory-improving agent particularly effective in aged animals. Hence, ascorbic acid might prove to be a useful memory-restorative agent in the treatment of dementia seen in elderly individuals. The underlying mechanism of action of ascorbic acid may be attributed to its antioxidant property.


Orifice locating with a microscope.
The purpose of this study was to determine whether the use of the dental operating microscope (DOM) could increase the number of root canal orifices located in mandibular molars. Ninety-three first and 111 second extracted mandibular molars were used. With the naked eye, all access cavities were prepared and the number of canals in each root was recorded. Using a DOM with x8-x13 magnification, all teeth had the access cavity preparations again examined. With the naked eye, a total of 641 canals were seen in all teeth. After the DOM examination, 50 more canals could be visualized, representing a 7.8% increase in the total number of located canals. From these canals, 35 were located in the first molars and 15 in the second molars. The use of the DOM increases the number of root canal orifices located.


Endothelial CCR6 expression due to FLI1 deficiency contributes to vasculopathy associated with systemic sclerosis.
We have recently demonstrated that serum CCL20 levels positively correlate with mean pulmonary arterial pressure in patients with systemic sclerosis (SSc). Considering a proangiogenic effect of CCL20 on endothelial cells via CCR6, the CCL20/CCR6 axis may contribute to the development of SSc vasculopathy. Therefore, we explored this hypothesis using clinical samples, cultured cells, and murine SSc models. The expression levels of CCL20 and CCR6 in the skin, mRNA levels of target genes, and the binding of transcription factor FLI1 to the target gene promoter were evaluated by immunostaining, quantitative reverse transcription PCR, and chromatin immunoprecipitation, respectively. Vascular permeability was evaluated by Evans blue dye injection in bleomycin-treated mice. Angiogenic activity of endothelial cells was assessed by in vitro angiogenesis assay. CCL20 expression was significantly elevated in dermal fibroblasts of patients with early diffuse cutaneous SSc, while CCR6 was significantly up-regulated in dermal small vessels of SSc patients irrespective of disease subtypes and disease duration. In human dermal microvascular endothelial cells, FLI1 siRNA induced the expression of CCR6, but not CCL20, and FLI1 bound to the CCR6 promoter. Importantly, vascular permeability, a representative SSc-like vascular feature of bleomycin-treated mice, was attenuated by Ccr6 siRNA treatment, and CCR6 siRNA suppressed the angiogenic activity of human dermal microvascular endothelial cells assayed by in vitro tube formation. The increased expression of endothelial CCR6 due to FLI1 deficiency may contribute to the development of SSc vasculopathy.


Aberrant alternative splicing is another hallmark of cancer.
The vast majority of human genes are alternatively spliced. Not surprisingly, aberrant alternative splicing is increasingly linked to cancer. Splice isoforms often encode proteins that have distinct and even antagonistic properties. The abnormal expression of splice factors and splice factor kinases in cancer changes the alternative splicing of critically important pre-mRNAs. Aberrant alternative splicing should be added to the growing list of cancer hallmarks.


Training primary health care physicians in Saudi Arabia to recognize psychiatric illness.
In order to evaluate the effect of training primary health care (PHC) physicians to recognize mental disorders, an intervention randomized controlled trial was conducted. Three categories of PHC physicians were selected: the index group (n = 12 doctors), control group 1 (n = 12) and control group 2 (n = 4). For 1 week randomly selected patients from all trial centres were asked to complete the General Health Questionnaire (GHQ-28), and the three groups of doctors were asked to identify the presence of and rate the severity of any mental disorder in these patients. The same process was repeated for all three groups after a 6-month training course which was provided for the index group only. The doctors' ability to detect mental disorders was measured by calculating Spearman's correlation coefficient between their severity rating and the GHQ scores of their patients. Training produced a noticeable improvement in the diagnostic accuracy of the index group, whereas there was minimal improvement in control group 1 and deterioration in control group 2. Recommendations to improve the ability of PHC physicians to detect mental disorders are discussed.


Giving birth in the hospital: childbirth experiences of Thai women in northern Thailand.
In this article, I examine how Thai women perceive and experience childbirth in hospitals. The article is based on in-depth interviews with 30 women living in Chiang Mai, Thailand. The women's narratives reveal that childbirth was managed within the medical system. The women believed that safety was the primary reason for their choice of birth in the hospital. Women's embodied experiences with hospital birth reveal the "passivity" discourse; women accord total trust to their doctors and very rarely question the many routine procedures in hospitals. It seems that in northern Thai hospitals the involvement of women's partners or their significant others is kept to a minimum. Of interest among postpartum care provided in Thai hospitals in the north is the use of a spotlight to help heal the episiotomy wound. This is an adaptation of Thai traditional confinement practices in the era of modernity. The use of a spotlight in hospital not only provides the women with symbolic ritual but also is believed to assist them in the healing process. Women in general were satisfied with postpartum care received during their hospital stay, except for rooming-in practice. The data suggest some differences between rural poor and urban middle-class women in terms of hospitals of birth, the opportunity to have a family member at birth, and so on. It is clear that middle-class educated women are able to exercise their choices and control over their childbirth experiences much more than rural poor women. I argue that care provided to women during birth needs to take into account women's emotional and subjective experience so that sensitive birthing care can be achieved. This will only make childbirth of many women a more positive one.


Molecular cloning of C4 gene and identification of the class III complement region in the shark MHC.
To clarify the evolutionary origin of the linkage of the MHC class III complement genes with the MHC class I and II genes, we isolated C4 cDNA from the banded hound shark (Triakis scyllium). Upon phylogenetic tree analysis, shark C4 formed a well-supported cluster with C4 of higher vertebrates, indicating that the C3/C4 gene duplication predated the divergence of cartilaginous fish from the main line of vertebrate evolution. The deduced amino acid sequence predicted the typical C4 three-subunits chain structure, but without the histidine residue catalytic for the thioester bond, suggesting the human C4A-like specificity. The linkage analysis of the complement genes, one C4 and two factor B (Bf) genes, to the shark MHC was performed using 56 siblings from two typing panels of T. scyllium and Ginglymostoma cirratum. The C4 and one of two Bf genes showed a perfect cosegregation with the class I and II genes, whereas two recombinants were identified for the other Bf gene. These results indicate that the linkage between the complement C4 and Bf genes, as well as the linkage between these complement genes and the MHC class I and II genes were established before the emergence of cartilaginous fish >460 million years ago.


Balance improvement by strength training for the elderly.
[Purpose] Aging is associated with a progressive decline in overall muscle strength. Loss of lower limb strength leads to an increased risk of falls and a sedentary lifestyle. The purpose of this study was to investigate whether lower limb strengthening exercise leads to improved lower limb strength and balance function for the elderly. [Subjects] From a total of 74 respondents, 50 subjects were randomly assigned to either a training group (n = 30) or a control group (n = 20). The subjects ranged in age from 65 to 82 years. A randomized controlled trial compared the effects of strengthening exercise and balance function. [Methods] Leg extension and lower curl exercises were performed during the 12-week study. [Results] After training, the lower limb strength and balance of the individuals in the training group had significantly improved compared to the baseline. [Conclusion] Improvement in lower limb strength may lead to balance enhancement in neurologically intact older persons.


Application of adaptive kinetic modelling for bias propagation reduction in direct 4D image reconstruction.
Parametric imaging in thoracic and abdominal PET can provide additional parameters more relevant to the pathophysiology of the system under study. However, dynamic data in the body are noisy due to the limiting counting statistics leading to suboptimal kinetic parameter estimates. Direct 4D image reconstruction algorithms can potentially improve kinetic parameter precision and accuracy in dynamic PET body imaging. However, construction of a common kinetic model is not always feasible and in contrast to post-reconstruction kinetic analysis, errors in poorly modelled regions may spatially propagate to regions which are well modelled. To reduce error propagation from erroneous model fits, we implement and evaluate a new approach to direct parameter estimation by incorporating a recently proposed kinetic modelling strategy within a direct 4D image reconstruction framework. The algorithm uses a secondary more general model to allow a less constrained model fit in regions where the kinetic model does not accurately describe the underlying kinetics. A portion of the residuals then is adaptively included back into the image whilst preserving the primary model characteristics in other well modelled regions using a penalty term that trades off the models. Using fully 4D simulations based on dynamic [(15)O]H2O datasets, we demonstrate reduction in propagation-related bias for all kinetic parameters. Under noisy conditions, reductions in bias due to propagation are obtained at the cost of increased noise, which in turn results in increased bias and variance of the kinetic parameters. This trade-off reflects the challenge of separating the residuals arising from poor kinetic modelling fits from the residuals arising purely from noise. Nonetheless, the overall root mean square error is reduced in most regions and parameters. Using the adaptive 4D image reconstruction improved model fits can be obtained in poorly modelled regions, leading to reduced errors potentially propagating to regions of interest which the primary biologic model accurately describes. The proposed methodology, however, depends on the secondary model and choosing an optimal model on the residual space is critical in improving model fits.


Democratizing the Assessment of Thermal Robustness of Metal-Organic Frameworks.
With the pressing need of having reliable materials for carbon dioxide capture, metal-organic frameworks (MOFs) have shown promising performance due to their flexible sign and tunable functionality by applying reticular chemistry principles. One of the main characteristics of practical MOFs is to design thermally robust candidates for sustainable functionality. Here, we introduce a comprehensive methodology for examining the thermal stability of MOFs by combining theoretical calculations and affordable experimental methods to fully describe their performance under thermal variations. We chose the prototypical MOF, HKUST-1, to assess the methodology by performing density functional theory and classical molecular dynamics simulations and validating with experiments such as in situ powder X-ray diffraction, differential scanning calorimetry, and thermogravimetric analysis. HKUST-1 shows thermal robustness until a temperature of 240 °C at different atmospheric gases with a reversible breathing trend with temperature. This methodology is affordable as it uses minimal experimental testing and can be applied to any MOF materials to explore its suitability for practical applications.


Thiamine Acquisition Strategies Impact Metabolism and Competition in the Gut Microbe Bacteroides thetaiotaomicron.
Thiamine (vitamin B1) is an essential cofactor for all organisms. Humans primarily acquire thiamine through their diet, and thiamine deficiencies have adverse neurological effects. However, the role gut microbes play in modulating thiamine availability is poorly understood, and little is known about how thiamine impacts the stability of microbial gut communities. To investigate thiamine's role in the gut, we utilized the model gut microbe Bacteroides thetaiotaomicron. Transcriptome sequencing (RNA-seq) revealed a global downregulation of thiamine and amino acid biosynthesis, glycolysis, and purine metabolism when thiamine was present. Using genetic mutants with thiamine biosynthesis and transport locus mutations, we determined both systems were critical for growth in thiamine-deficient medium. The defect in the double transport mutant suggests an uncharacterized feedback mechanism between thiamine transport and biosynthesis in B. thetaiotaomicron. Mutant phenotypes were recapitulated during pairwise competitions, reinforcing the importance of encoding versatile thiamine acquisition mechanisms when thiamine concentrations are variable. In addition, liquid chromatography-mass spectrometry (LC-MS) analyses corroborate that exogenous thiamine levels affect the internal thiamine pool of B. thetaiotaomicron. Furthermore, we computationally examined the ability of other gut microbes to acquire thiamine and identified lineage-specific differences in thiamine acquisition strategies. Among the Bacteroidetes, the capacities for both thiamine transport and biosynthesis are common. Together, these data show that thiamine acquisition mechanisms used by B. thetaiotaomicron not only are critical for its physiology and fitness but also provide the opportunity to model how other gut microbes may respond to the shifting availability of thiamine in the gut. IMPORTANCE Variation in the ability of gut microbes to transport, synthesize, and compete for vitamin B1 (thiamine) is expected to impact the structure and stability of the microbiota, and ultimately this variation may have both direct and indirect effects on human health. Our study identifies the diverse strategies employed by gut Bacteroidetes to acquire thiamine. We demonstrate how the presence or absence of thiamine biosynthesis or transport dramatically affects the abundance of B. thetaiotaomicron in a competitive environment. This study adds further evidence that altering the presence or concentrations of water-soluble vitamins such as thiamine may be an effective method for manipulating gut community composition. In turn, targeted thiamine delivery could be used therapeutically to alter dysbiotic communities linked to disease. Author Video: An author video summary of this article is available.


The importance of histopathology in the diagnosis of isolated renal sarcoidosis: a case report.
Sarcoidosis is a systemic inflammatory disease of unknown etiology, characterized by the presence of non-caseating granulomas in several organs; renal impairment alone is a rare condition. When it affects the kidneys, the most prevalent manifestations are hypercalcemia and hypercalciuria. This paper aims to address the topic of renal sarcoidosis, by means of a case report, and reinstate the importance of histopathology in its diagnosis. The data came from an observational clinical study with a qualitative approach, through an interview with the renal sarcoidosis patient and data from her medical records. Patient D.M.S., 50 years old, Caucasian, presented with reddish eyes and body pains lasting for fifteen days as first manifestations of the disease. Upon kidney ultrasound scan, we found renal parenchymal nephropathy. Serial renal function and metabolic tests reported anemia and progressive urea and creatinine changes, as well as hypercalcemia and hypercalciuria, confirming acute kidney failure (AKF). A histopathological examination suggested the diagnosis, which was confirmed by clinical, laboratory and histopathological data. There was therapeutic resolution after steroid therapy. The symptomatology of sarcoidosis is diverse and often non-specific. Renal manifestation, which usually occurs after organ involvement, is present in less than 5% of patients, and about 1% to 2% of these patients may develop AKF. The use of histopathology together with clinical and laboratory data to diagnose isolated renal sarcoidosis, rule out other etiologies and introduce early treatment is of paramount importance.


Treatment of severe acute ulcerative colitis in SARS-CoV-2 infected patients: report of three cases and discussion of treatment options.
In the wake of the coronavirus disease 2019 (COVID-19) pandemic, it is unclear how asymptomatic severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-infected patients who present with acute severe ulcerative colitis (UC) can be treated effectively and safely. Standard treatment regimens consist of steroids, immunomodulatory drugs, and biological therapies, but therapeutic decision-making becomes challenging as there are uncertainties about how to deal with these drugs in patients with COVID-19 and active UC. Importantly, guidelines for this particular group of patients with UC are still lacking. To inform therapeutic decision-making, we describe three consecutive cases of patients with active UC and COVID-19 and discuss their treatments based on theoretical knowledge, currently available evidence and clinical observations. Three patients were identified through our national inflammatory bowel disease network [Initiative on Crohn's and Colitis (ICC)] for whom diagnosis of SARS-CoV-2-infection was established by reverse transcription-polymerase chain reaction (RT-PCR) testing in nasopharynx, stools, and/or biopsies. Acute severe UC was diagnosed by clinical parameters, endoscopy, and histopathology. Clinical guidelines for SARS-CoV-2-negative patients advocate the use of steroids, calcineurin inhibitors, or tumor necrosis factor alpha (TNF-α)-antagonists as induction therapy, and experiences from the current three cases show that steroids and TNF-α-antagonists could also be used in patients with COVID-19. This could potentially be followed by TNF-α-antagonists, vedolizumab, or ustekinumab as maintenance therapy in these patients. Future research is warranted to investigate if, and which, immunomodulatory drugs should be used for COVID-19 patients that present with active UC. To answer this question, it is of utmost importance that future cases of patients with UC and COVID-19 are documented carefully in international registries, such as the SECURE-IBD registry.


Defense cells profile of cervical mucous during follicular and luteal phases of estrus cycle in river buffalo.
The aim of this study was to evaluate the defense cells changes of cervical mucous during follicular and luteal phases of estrus cycle in river buffalo. Reproductive organs of the adult and apparently healthy female buffaloes were collected from the slaughterhouse. By visual investigation of both the ovaries for presence of corpus luteum and growing follicles, the luteal and follicular phase of each buffalo was specified. Cervical discharge samples were collected by sterile swabs and then spread over the glass slides, dried and fixed with methanol. The specimens were undergone Giemsa staining. The percentage of lymphocytes, neutrophils, monocytes (macrophages), eosinophils and basophils in each case (for both the follicular and luteal phases) were obtained at 20 microscopic fields. The percentage of lymphocytes, neutrophils and basophils in luteal phase were higher than the follicular phase. The percentage of eosinophils in follicular phase was higher than the luteal phase. The percentage of monocytes (macrophages) in luteal and follicular phases was nearly equal. The statistical analysis showed that the differences of all cells between follicular and luteal phase were not significant (P > 0.05). The most defense cells in discharges of external os of cervix (both follicular and luteal phases) were neutrophils and lymphocytes.


Modeling approaches for qualitative and semi-quantitative analysis of cellular signaling networks.
A central goal of systems biology is the construction of predictive models of bio-molecular networks. Cellular networks of moderate size have been modeled successfully in a quantitative way based on differential equations. However, in large-scale networks, knowledge of mechanistic details and kinetic parameters is often too limited to allow for the set-up of predictive quantitative models.Here, we review methodologies for qualitative and semi-quantitative modeling of cellular signal transduction networks. In particular, we focus on three different but related formalisms facilitating modeling of signaling processes with different levels of detail: interaction graphs, logical/Boolean networks, and logic-based ordinary differential equations (ODEs). Albeit the simplest models possible, interaction graphs allow the identification of important network properties such as signaling paths, feedback loops, or global interdependencies. Logical or Boolean models can be derived from interaction graphs by constraining the logical combination of edges. Logical models can be used to study the basic input-output behavior of the system under investigation and to analyze its qualitative dynamic properties by discrete simulations. They also provide a suitable framework to identify proper intervention strategies enforcing or repressing certain behaviors. Finally, as a third formalism, Boolean networks can be transformed into logic-based ODEs enabling studies on essential quantitative and dynamic features of a signaling network, where time and states are continuous.We describe and illustrate key methods and applications of the different modeling formalisms and discuss their relationships. In particular, as one important aspect for model reuse, we will show how these three modeling approaches can be combined to a modeling pipeline (or model hierarchy) allowing one to start with the simplest representation of a signaling network (interaction graph), which can later be refined to logical and eventually to logic-based ODE models. Importantly, systems and network properties determined in the rougher representation are conserved during these transformations.


Complete Genome Sequence of Staphylococcus aureus Z172, a Vancomycin-Intermediate and Daptomycin-Nonsusceptible Methicillin-Resistant Strain Isolated in Taiwan.
We report the complete genome sequence of Z172, a representative strain of sequence type 239-staphylococcal cassette chromosome mec type III (ST239-SCCmec type III) hospital-associated methicillin-resistant Staphylococcus aureus in Taiwan. Strain Z172 also exhibits a vancomycin-intermediate and daptomycin-nonsusceptible phenotype.


Pushing the limits: an instrument for hard X-ray imaging below 20 nm.
Hard X-ray microscopy is a prominent tool suitable for nanoscale-resolution non-destructive imaging of various materials used in different areas of science and technology. With an ongoing effort to push the 2D/3D imaging resolution down to 10 nm in the hard X-ray regime, both the fabrication of nano-focusing optics and the stability of the microscope using those optics become extremely challenging. In this work a microscopy system designed and constructed to accommodate multilayer Laue lenses as nanofocusing optics is presented. The developed apparatus has been thoroughly characterized in terms of resolution and stability followed by imaging experiments at a synchrotron facility. Drift rates of ∼2 nm h(-1) accompanied by 13 nm × 33 nm imaging resolution at 11.8 keV are reported.


Alpha-fetoprotein elevation in NUT midline carcinoma: a case report.
Nuclear protein in testis (NUT) midline carcinoma is a rarely diagnosed and potentially under-recognized type of squamous carcinoma that is considered one of the most aggressive human solid tumors. Alpha-fetoprotein elevation has been associated with chronic liver diseases and a limited number of cancers. In particular, in presence of a mediastinal mass in a young man, alpha-fetoprotein elevation is considered nearly pathognomonic of a non-seminoma germ-cell tumor. A 22-year old man without any comorbidity was diagnosed with a large mediastinal mass with skeletal and lymph node metastases. The clinical picture was dominated by a life-threatening superior vena cava syndrome with elevated alpha-fetoprotein and lactate dehydrogenase that supported the diagnostic suspicion of mediastinal germ-cell tumor. However, a biopsy showed a poorly-differentiated and diffusely necrotic carcinoma. We eventually reached the diagnosis of the peculiar entity of NUT midline carcinoma, but the differential diagnosis was quite challenging also because alpha-fetoprotein is not reported as a marker of NUT midline carcinoma. Notably, alpha-fetoprotein levels correlated with disease course. The life-threatening aggressiveness of NUT midline carcinoma mandates to reach the right diagnosis in the shortest possible time. In this regard, poorly differentiated carcinomas lacking glandular differentiation mandate testing for NUT expression by immunohistochemistry. Awareness of a potentially misleading tumor marker elevation can help to broaden the differential diagnosis and establish the most appropriate treatment.


Clinical signs of impending death in cancer patients.
The physical signs of impending death have not been well characterized in cancer patients. A better understanding of these signs may improve the ability of clinicians to diagnose impending death. We examined the frequency and onset of 10 bedside physical signs and their diagnostic performance for impending death. We systematically documented 10 physical signs every 12 hours from admission to death or discharge in 357 consecutive patients with advanced cancer admitted to two acute palliative care units. We examined the frequency and median onset of each sign from death backward and calculated their likelihood ratios (LRs) associated with death within 3 days. In total, 203 of 357 patients (52 of 151 in the U.S., 151 of 206 in Brazil) died. Decreased level of consciousness, Palliative Performance Scale ≤20%, and dysphagia of liquids appeared at high frequency and >3 days before death and had low specificity (<90%) and positive LR (<5) for impending death. In contrast, apnea periods, Cheyne-Stokes breathing, death rattle, peripheral cyanosis, pulselessness of radial artery, respiration with mandibular movement, and decreased urine output occurred mostly in the last 3 days of life and at lower frequency. Five of these signs had high specificity (>95%) and positive LRs for death within 3 days, including pulselessness of radial artery (positive LR: 15.6; 95% confidence interval [CI]: 13.7-17.4), respiration with mandibular movement (positive LR: 10; 95% CI: 9.1-10.9), decreased urine output (positive LR: 15.2; 95% CI: 13.4-17.1), Cheyne-Stokes breathing (positive LR: 12.4; 95% CI: 10.8-13.9), and death rattle (positive LR: 9; 95% CI: 8.1-9.8). We identified highly specific physical signs associated with death within 3 days among cancer patients.


Family-of-origin maltreatment, posttraumatic stress disorder symptoms, social information processing deficits, and relationship abuse perpetration.
In this study, the authors examined the interrelations among family-of-origin maltreatment variables, posttraumatic stress disorder (PTSD) symptoms, social information processing deficits, and male-to-female psychological and physical intimate relationship abuse perpetration in adulthood among a community sample of 164 men and their partners. In bivariate analyses, higher family-of-origin childhood parental rejection was associated with the perpetration of psychological and physical abuse in adulthood, and childhood exposure to interparental violence was also associated with adult psychological abuse perpetration. Structural equation modeling analyses indicated that when childhood variables and other study variables were considered together, only childhood parental rejection was associated with the abuse perpetration outcomes, and these effects were indirect through PTSD symptoms and social information processing deficits. Results indicate a need for further investigation into the mechanisms accounting for the impact of early maltreatment on the development of abusive intimate relationship behavior.


Long-term clinical outcome after Lyme neuroborreliosis in childhood.
To determine long-term clinical outcome in children with confirmed Lyme neuroborreliosis (LNB) and to evaluate persistent subjective symptoms compared with a control group. After a median of 5 years, 84 children with confirmed LNB underwent a neurologic re-examination, including a questionnaire. Medical records were analyzed, and a control group (n = 84) was included. The total recovery rate was 73% (n = 61). Objective neurologic findings, defined as "definite sequelae," were found in 16 patients (19%). The majority of these children had persistent facial nerve palsy (n = 11), but other motor or sensory deficits occurred (n = 5). Neurologic signs and/or symptoms defined as "possible sequelae" were found in another 7 patients (8%), mainly of sensory character. Nonspecific subjective symptoms were reported by 35 patients (42%) and 32 controls (38%) (nonsignificant). Affected daily activities or school performance were reported to the same extent in both groups (23% vs 20%, nonsignificant). The long-term clinical recovery rate was 73% in children with confirmed LNB. Persistent facial nerve palsy occurred in 13%, whereas other motor or sensory deficits were found in another 14%. Neurologic deficits did not affect daily activities or school performance more often among patients than controls and should be considered as mild. Furthermore, nonspecific subjective symptoms such as headache, fatigue, or memory or concentration problems were reported as often among patients as controls and should not be considered as sequelae after LNB.


Clinical presentation, course, and prognostic factors in lymphocyte-predominant Hodgkin's disease and lymphocyte-rich classical Hodgkin's disease: report from the European Task Force on Lymphoma Project on Lymphocyte-Predominant Hodgkin's Disease.
Recent studies have suggested that lymphocyte-predominant Hodgkin's disease (LPHD) is both clinically and pathologically distinct from other forms of Hodgkin's disease, including classical Hodgkin's disease (CHD). However, large-scale clinical studies were lacking. This multicenter, retrospective study investigated the clinical characteristics and course of LPHD patients and lymphocyte-rich classical Hodgkin's disease (LRCHD) patients classified according to morphologic and immunophenotypic criteria. Clinical data and biopsy material of all available cases initially submitted as LPHD were collected from 17 European and American centers, stained, and reclassified by expert pathologists. The 426 assessable cases were reclassified as LPHD (51%), LRCHD (27%), CHD (5%), non-Hodgkin's lymphoma (3%), and reactive lesion (3%); 11% of cases were not assessable. Patients with LPHD and LRCHD were predominantly male, with early-stage disease and few risk factors. Patients with LRCHD were significantly older. Survival and failure-free survival rates with adequate therapy were similar for patients with LPHD and LRCHD, and were stage-dependent and not significantly better than stage-comparable results for CHD (German trial data). Twenty-seven percent of relapsing LPHD patients had multiple relapses, which is significantly more than the 5% of relapsing LRCHD patients who had multiple relapses. Lymphocyte-predominant Hodgkin's disease patients had significantly superior survival after relapse compared with LRCHD or CHD patients; however, this was partly due to the younger average age of LPHD patients. The two subgroups of LPHD and LRCHD bore a close clinical resemblance that was distinct from CHD; the course was similar to that of comparable nodular sclerosis and mixed cellularity patients. Thorough staging is necessary to detect advanced disease in LPHD and LRCHD patients. The question of how to treat such patients, either by reducing treatment intensity or following a "watch and wait" approach, remains unanswered.


The improved resistance of germinated spores to ultraviolet irradiation: Comparison with chlorine.
Fungi outbreaks in water will include a series of processes, including spore aggregation, germination, biofilm, and finally present in a mixed state in the aquatic environment. More attention is paid to the control of dispersed fungal spores, however, there was little knowledge of the control of germinated spores. This study investigated the inactivation kinetics and mechanism of ultraviolet (UV) treatment for fungal spores with different germination percentages compared with dormant spores. The results indicated that the inactivation rate constants (k) of spores with 5%-45% germination were 0.0278-0.0299 cm2/mJ for Aspergillus niger and 0.0588-0.0647 cm2/mJ for Penicillium polonicum, which were lower than those of dormant spores. It suggested that germinated spores were more tolerant to UV irradiation than dormant spores, and it may be due to the defensive barrier (upregulated pigments) and some reductive substance (upregulated enoyl reductase) by absorbing UV or reacting with reactive oxygen species according to transcriptome analysis. Compared to dormant spores, the k-UV of germinated spores decreased by 18.17%-26.56% for Aspergillus niger, which was less than k-chlorine (62.33%-69.74%). A slighter decrease in k-UV showed UV irradiation can efficiently control fungi contamination, especially when dormant spores and germinated spores coexisted in actual water systems. This study indicates that more attention should be paid to germinated spores.


[Statistical analysis for count data: use of healthcare services applications].
To describe some of the statistical models for the study of count variables in the context of the use of health services. We used the Seguro Popular Evaluation Survey to estimate the effect of Seguro Popular on the frequency of use of outpatient health services, using Poisson regression models and negative binomial, zero-inflated negative binomial and the hurdle negative binomial models. We used the Akaike Information Criterion (AIC) to define the best model. Results show that the best statistical approach to model the use of health services is the hurdle model, taking into account both the main theoretical assumptions and the statistical results of the AIC. The modelling of count data requires the application of statistical models to model data dispersion; in the presence of an excess of zeros, the hurdle model is an appropriate statistical option.


[Expression of p21 in hepatocellular carcinoma and liver cirrhosis and its relation with HBV infection].
Determination of p21, a product of Ha-ras oncogene, and HBsAg in hepatocellular carcinoma (HCC) and liver cirrhosis etc. was carried out with ABC method. The results showed that HCC tissues exhibited enhancement of p21 expression with a positive rate of 72.4%, which was obviously higher than the expression of p21 in tissues from liver cirrhosis, chronic hepatitis and hepatoblastoma. The p21 positive rate of regenerative cirrhosis nodules close to the HCC was 87.2%. The p21 expression level in HBsAg positive regenerative nodules of cirrhosis close to the HCC was significantly high, and its positive rate reached 93.9%. The expression level of p21 protein in well-differentiated HCC was higher than that of poorly differentiated and undifferentiated HCC. Therefore, the result suggests that the expression level of p21 in liver cirrhosis is related to persistent infection of HBV. The elevated expression of p21 plays an important role in the development of regenerative nodules in liver cirrhosis towards HCC, and it is also an important factor in the early stage of HCC development.


Impact of glycemic variability and hypoglycemia on adverse hospital outcomes in non-critically ill patients.
To determine if glycemic variability is associated with hospitalization outcomes in non-critically ill patients, and if this association remains after controlling for hypoglycemia. A retrospective review was performed on 1276 medical admissions (801 patients) in which insulin was given, ≥6 point of care glucose (POCG) measurements and length of stay (LOS) 2-30 days. Coefficient of variation (%CV) was used to measure glycemic variability. Outcomes included LOS and a composite outcome based on ICU transfer, hospital acquired infections, and acute renal failure (ARF). There were a median of 18.5 POCG measurements per admission with a mean %CV 34.2 ± 11.1. Hypoglycemia (POCG ≤70 mg/dl [3.9 mmol/l]) occurred in 35.0% of admissions. ICU transfer occurred in 3.3%, hospital acquired infections 4.8%, ARF 8.3%, and composite outcome 13.5%. Adjusting for age, sex, race and Charlson score, every 10 unit increase in %CV was associated with an increase in LOS of 0.27 days (p=0.004), while there was no association between %CV and the composite outcome. For LOS, there was a significant interaction between %CV and hypoglycemia (p=0.07). While there was a non-significant correlation in patients without hypoglycemia, LOS correlated negatively with %CV in patients with hypoglycemia. When considered simultaneously with %CV, hypoglycemia was associated with increased odds of the composite outcome [OR 2.03 (95% CI 1.36-3.01), p=<0.001] and an increase of 2 days in LOS for those with average %CV. Hypoglycemia, compared to glycemic variability, is more strongly associated with adverse outcomes in hospitalized, non-critically ill patients.


Synthesis and hydrolysis of ATP and the phosphate-ATP exchange reaction in soluble mitochondrial F1 in the presence of dimethylsulfoxide.
In medium containing 40% dimethylsulfoxide, soluble F1 catalyzes the hydrolysis of ATP introduced at concentrations lower than that of the enzyme [Al-Shawi, M.K. & Senior, A.E. (1992), Biochemistry 31, 886-891]. At this concentration of dimethylsulfoxide, soluble F1 also catalyzes the spontaneous synthesis of a tightly bound ATP to a level of approximately 0.15 mol per mol F1 [Gómez-Puyou, A., Tuena de Gómez-Puyou, M. & de Meis, L. (1986) Eur. J. Biochem. 159, 133-140]. The mechanisms that allow soluble F1 to carry out these apparently opposing reactions were studied. The rate of hydrolysis of ATP bound to F1 under uni-site conditions and that of synthesis of ATP were markedly similar, indicating that the two ATP molecules lie in equivalent high affinity catalytic sites. The number of enzyme molecules that have ATP at the high affinity catalytic site under conditions of synthesis or uni-site hydrolysis is less than the total number of enzyme molecules. Therefore, it was hypothesized that when the enzyme was treated with dimethylsulfoxide, a fraction of the F1 population carried out synthesis and another hydrolysis. Indeed, measurements of the two reactions under identical conditions showed that different fractions of the F1 population carried out simultaneously synthesis and hydrolysis of ATP. The reactions continued until an equilibrium level between F1.ADP + Pi <--> F1.ATP was established. At equilibrium, about 15% of the enzyme population was in the form F1.ATP. The DeltaG degrees of the reaction with 0.54 microM F1, 2 mM Pi and 10 mM Mg2+ at pH 6.8 was -2.7 kcal.mol-1 in favor of F1.ATP. The DeltaG degrees of the reaction did not exhibit important variations with Pi concentration; thus, the reaction was in thermodynamic equilibrium. In contrast, DeltaG degrees became significantly less negative as the concentration of dimethylsulfoxide was decreased. In water, the reaction was far to the left. The equilibrium constant of the reaction diminished linearly with an increase in water activity. The effect of solvent is fully reversible. In comparison to other enzymes, F1 seems unique in that solvent controls the equilibrium that exists within an enzyme population. This results from the effect of solvent on the partition of Pi between the catalytic site and the medium, and the large energetic barrier that prevents release of ATP from the catalytic site. In the presence of dimethylsulfoxide and Pi, ATP is continuously hydrolyzed and synthesized with formation and uptake of Pi from the medium. This process is essentially an exchange reaction analogous to the phosphate-ATP exchange reaction that is catalyzed by the ATP synthase in coupled energy transducing membranes.


A 3D "sandwich" co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro.
Various methods for ex utero culture systems have been explored. However, limitations remain regarding the in vitro culture platforms used before implanting mouse embryos and the normal development of mouse blastocysts in vitro. Furthermore, vascular niche support during mouse embryo development from embryonic day (E) 3.5 to E7.5 is unknown in vitro. This study established a three-dimensional (3D) "sandwich" vascular niche culture system with in vitro culture medium (IVCM) using human placenta perivascular stem cells (hPPSCs) and human umbilical vein endothelial cells (hUVECs) as supportive cells (which were seeded into the bottom layer of Matrigel) to test mouse embryos from E3.5 to E7.5 in vitro. The development rates and greatest diameters of mouse embryos from E3.5 to E7.5 were quantitatively determined using SPSS software statistics. Pluripotent markers and embryo transplantation were used to monitor mouse embryo quality and function in vivo. Embryos in the IVCM + Cells (hPPSCs + hUVECs) group showed higher development rates and greater diameters at each stage than those in the IVCM group. Embryos in the IVCM + Cells group cultured to E5.5 morphologically resembled natural egg cylinders and expressed specific embryonic cell markers, including Oct4 and Nanog. These features were similar to those of embryos developed in vivo. After transplantation, the embryos were re-implanted in the internal uterus and continued to develop to a particular stage. The 3D in vitro culture system enabled embryo development from E3.5 to E7.5, and the vascularization microenvironment constructed by Matrigel, hPPSCs, and hUVECs significantly promoted the development of implanted embryos. This system allowed us to further study the physical and molecular mechanisms of embryo implantation in vitro.


Designing a Rational Follow-Up Schedule for Patients with Extremity Soft Tissue Sarcoma.
The risk of tumor recurrence after resection of soft tissue sarcoma (STS) necessitates surveillance in follow-up. The objective of this study was to determine the frequency/timing of metastasis and local recurrence following treatment for soft tissue sarcoma, and to use these data to justify an evidence-based follow-up schedule. Utilizing a prospective database, a retrospective single center review was performed of all patients with minimum 2-year follow-up after resection of a localized extremity STS. Kaplan-Meier estimates were used to calculate the incidence of local recurrence and metastases on an annual basis for 10 years. We identified a total of 230 low-grade, 626 intermediate-grade and 940 high-grade extremity STS and a total of 721 events, 150 local recurrences and 571 metastases. Based on tumor size and grade, follow-up cohorts were developed that had similar metastatic risk. Using pre-determined thresholds for metastatic event, a follow-up schedule was established for each cohort. Based on our results we recommend that patients with small low-grade tumors undergo annual follow-up for 5 years following definitive local treatment. Patients with large low-grade tumors, small intermediate-grade and small high-grade tumors should have follow-up every 6 months for the first 2 years, then yearly to 10 years. Only patients with large intermediate- or high-grade tumors require follow-up every 3 months for the first 2 years, then every 6 months for years 3-5, followed by annually until 10 years.


Hemophagocytic lymphohistiocytosis associated with visceral leishmaniasis.
Visceral leishmaniasis (VL), is a systemic disease caused by the dissemination of protozoan parasite Leishmania throughout the reticuloendothelial system. It may mimic or lead to several types of hematological disorders including hemophagocytosis. Infection associated hemophagocytic syndrome implicating Leishmania is very rare and often difficult to diagnose. Here, we describe a child with hemophagocytic lymphohistiocytosis (HLH) associated with VL.


West Nile virus transmission via organ transplantation and blood transfusion - Louisiana, 2008.
Three years after the introduction and spread of West Nile virus (WNV) in the United States, transmission through blood transfusion and solid organ transplantation was documented in 2002. Within a year, these findings led to nationwide screening of blood donors for WNV. Although screening is extremely sensitive, current methods still do not detect all WNV-infected blood donations, and organ donors are not screened routinely. In October 2008, the Louisiana Department of Health (LDH) was notified of a heart transplant recipient with suspected West Nile neuroinvasive disease (WNND). LDH launched an investigation to confirm the diagnosis and determine whether the organ recipient's infection was derived from the organ donor or blood products the donor received before organ donation. The investigation concluded that two cases of probable transfusion-transmitted WNV resulted from a common blood donor; one infection resulted in WNND via an organ donor, and the other resulted in asymptomatic WNV infection via blood transfusion directly. This investigation also found that criteria used by the blood-screening laboratory to screen the implicated blood donation for WNV were less stringent than criteria used by other blood collection centers in the area. Use of the more stringent screening criteria might have detected the WNV and prevented the blood donation from being used. To increase the likelihood of detecting WNV-positive donations, blood centers should use the most sensitive screening criteria feasible and communicate frequently with nearby blood centers on screening results during times of high WNV activity in their geographic area. In addition, health-care providers should consider WNND as a possible cause of neurologic complications in patients after blood transfusion or organ transplantation.