[Gastrointestinal involvement in progressive systemic sclerosis].
25 patients with clinical, radiological and manometrical features of PSS in the gastrointestinal tract were reviewed, looking mainly for the esophageal involvement. All of the data obtained in our serie agreed with those of most of the authors. Outlining: The lack of relationship between the evolution of the skin involvement and GI tract involvement. The high incidence of esophageal involvement, especially functional alterations even in the absence of clinical and/or radiological symptomatology. The usefulness of manometric method in the diagnosis of motor involvement of esophages, especially for the evaluation of lower esophageal esphincter. Although the esophageal and intestinal involvement are more frequent and well known, any area of the GI tract may be damaged during the course of this disease. Since up to now, an ethiological therapy to stop the course of the disease is not known, it's important to search for earlier alterations in order to start with a pathophysiological and symptomatic treatment to avoid complications.


Summation of forces from multiple motor units in the cat soleus muscle.
Nearly all muscle models and most motor control concepts assume that forces from individual muscle fibers and motor units sum in an additive manner once effects of in-series tendon compliance are taken into account. Due to the numerous mechanical linkages between individual fibers, though, it is unclear whether this assumption is warranted. This work examined motor unit force summation over a wide range of muscle forces in the cat soleus. Nonadditive summation implies a nonlinear summation of motor unit forces. Summation nonlinearities were quantified during interactions of 10 individual motor units and 4 motor unit bundles containing approximately 10 units each. These protocols allowed motor unit force summation to be examined from approximately 0 to 25% of tetanic muscle force. Nonlinear summation was assessed by comparing the actual forces to the algebraic sum of individual units and bundles stimulated in isolation. Superadditive summation meant that the actual force exceeded the algebraic sum, whereas subadditive summation meant that the actual force was smaller than the algebraic sum. Experiments tested the hypothesis that superadditive summation occurs at low force levels when few motor units are recruited, whereas subadditive summation prevails above 10% of tetanic force. Results were consistent with this hypothesis. As in previous studies, nonlinear summation in the soleus was modest, but a clear transition from predominately superadditive to predominantly subadditive summation occurred in the range of 6-8% of tetanic force. The largest nonlinearities were transient and appeared at the onset of recruitment and derecruitment of groups of motor units. The results are discussed in terms of the mechanical properties of the connective tissue forming the tendon and linking muscle fibers.


The listening cube: a three dimensional auditory training program.
Here we present the Listening Cube, an auditory training program for children and adults receiving cochlear implants, developed during the clinical practice at the KIDS Royal Institute for the Deaf in Belgium. We provide information on the content of the program as well as guidance as to how to use it. The Listening Cube is a three-dimensional auditory training model that takes the following into consideration: the sequence of auditory listening skills to be trained, the variety of materials to be used, and the range of listening environments to be considered. During auditory therapy, it is important to develop training protocols and materials to provide rapid improvement over a relatively short time period. Moreover, effectiveness and the general real-life applicability of these protocols to various users should be determined. Because this publication is not a research article, but comes out of good daily practice, we cannot state the main results of this study. We can only say that this auditory training model is very successful. Since the first report was published in the Dutch language in 2003, more than 200 therapists in Belgium and the Netherlands followed a training course elected to implement the Listening Cube in their daily practice with children and adults with a hearing loss, especially in those wearing cochlear implants. The Listening Cube is a tool to aid in planning therapeutic sessions created to meet individual needs, which is often challenging. The three dimensions of the cube are levels of perception, practice material, and practice conditions. These dimensions can serve as a visual reminder of the task analysis and of other considerations that play a role in structuring therapy sessions.


Comparison of Total Hip Arthroplasty Outcomes after Failed Femoral Wedge or Curved Varus Osteotomy.
Differences in clinical and radiographic results following total hip arthroplasty between failed wedge and curved varus osteotomy are unclear. To investigate differences in clinical and radiographic results following total hip arthroplasty in patients who exhibited failed wedge or curved varus osteotomy. We performed 18 total hip arthroplasties after failed femoral varus osteotomy. Hips were divided into two groups: 14 had failed wedge varus osteotomy and four had failed curved varus osteotomy. Average ages at osteotomy and total hip arthroplasty were 27 years old (range, 10-46 years old) and 56 years old (range, 25-74 years old), respectively. The average duration of follow-up monitoring was 72.2 months (range, 54-91 months). Clinical and radiographic evaluations were completed for each group. The Japanese Orthopaedic Association hip score of total hip arthroplasty after failed varus osteotomy significantly improved at the last follow-up in both groups. However, hip score at the last follow-up was significantly higher after failed curved varus osteotomy than after failed wedge varus osteotomy (p<0.01). Four hips that failed wedge varus osteotomy underwent subtrochanteric corrective osteotomy with total hip arthroplasty. Radiographic evaluation showed that three stems for total hip arthroplasty after failed wedge varus osteotomy were inserted in malposition, and all stems in total hip arthroplasty after failed curved varus osteotomy were inserted in the normal position. Surgeons performing femoral varus osteotomy should consider possible future conversion to total hip arthroplasty. Curved varus osteotomy is more suitable than wedge varus osteotomy for future conversion to total hip arthroplasty.


An Intracellular Model of Hepatitis B Viral Infection: An In Silico Platform for Comparing Therapeutic Strategies.
Hepatitis B virus (HBV) is a major focus of antiviral research worldwide. The International Coalition to Eliminate HBV, together with the World Health Organisation (WHO), have prioritised the search for a cure, with the goal of eliminating deaths from viral hepatitis by 2030. We present here a comprehensive model of intracellular HBV infection dynamics that includes all molecular processes currently targeted by drugs and agrees well with the observed outcomes of several clinical studies. The model reveals previously unsuspected kinetic behaviour in the formation of sub-viral particles, which could lead to a better understanding of the immune responses to infection. It also enables rapid comparative assessment of the impact of different treatment options and their potential synergies as combination therapies. A comparison of available and currently developed treatment options reveals that combinations of multiple capsid assembly inhibitors perform best.


Mineralization capacity of bacteria and fungi from the rhizosphere-rhizoplane of a semiarid grassland.
A radiotracer glucose mineralization assay was used with streptomycin and actidione to monitor the relative seasonal contributions of bacteria and fungi to mineralization processes in soils derived from the rhizosphere-rhizoplane zone of plants from a shortgrass prairie ecosystem. Bacteria played a major role in glucose mineralization in both the rhizosphere and rhizoplane. These results indicate that the bacteria may play a greater role in glucose mineralization processes in the rhizosphere and rhizoplane zones of a semiarid grassland than would be assumed, based on available biomass estimates. This technique appears to be valuable for determining bacterial versus fungal contributions to glucose mineralization in the rhizosphere and rhizoplane and may be useful for measuring the decomposition of other more complex substances in this zone of intense microbial activity.


The characteristics of superficial siderosis and convexity subarachnoid hemorrhage and clinical relevance in suspected cerebral amyloid angiopathy.
Systematic studies of superficial siderosis (SS) and convexity subarachnoid hemorrhage (cSAH) in patients with suspected cerebral amyloid angiopathy (CAA) without lobar intracerebral hemorrhage (ICH) are lacking. We sought to determine the potential anatomic correlation between SS/cSAH and transient focal neurological episodes (TFNE) and whether SS/cSAH is predictor of future cerebral hemorrhagic events in these patients. We enrolled 90 consecutive patients with suspected CAA (due to the presence of strictly lobar microbleeds (CMBs) and/or SS/cSAH) but without the history of symptomatic lobar ICH who underwent brain MRI including T2*-weighted, diffusion-weighted imaging and fluid-attenuated inversion recovery sequences from an ongoing single center CAA cohort from 1998 to 2012. Evaluation of SS, cSAH and CMBs was performed. Medical records and follow-up information were obtained from prospective databases and medical charts. TFNE was defined according to published criteria and electroencephalogram reports were reviewed. Forty-one patients (46%) presented with SS and/or cSAH. The prevalence of TFNE was significantly higher in those with SS/cSAH (61 vs. 10%; p < 0.001) and anatomically correlated with the location of cSAH, but not SS. The majority of TFNE in patients with SS/cSAH presented with spreading sensory symptoms. Intermittent focal slowing on electroencephalogram was present in the same area as SS/cSAH in 6 patients, but no epileptiform activity was found in any patients. Among those with available clinical follow-up (76/90 patients, 84%), ten patients with SS/cSAH (29%, median time from the scan for all patients with SS/cSAH: 21 months) had a symptomatic cerebral bleeding event on follow up (average time to events: 34 months) compared with only 1 event (2.4%, 25 months from the scan) in patients without SS/cSAH (time to event: 25 months) (p = 0.001). The location of hemorrhages on follow-up scan was not in the same location of previously noted SS/cSAH in 9 of 10 patients. Follow-up imaging was obtained in 9 of 17 patients with cSAH and showed evidence of SS in the same location as initial cSAH in all these 9 cases. SS/cSAH is common in patients with suspected CAA without lobar intracerebral hemorrhage and may have a significantly higher risk of future cerebral bleeding events, regardless of the severity of the baseline CMB burden. The findings further highlight a precise anatomical correlation between TFNE and cSAH, but not SS. Distinct from transient ischemic attack or seizure, the majority of TFNE caused by SS/cSAH appear to present with spreading sensory symptoms.


Changes in concentrations of estradiol nuclear receptors in the preoptic area, medial basal hypothalamus, amygdala, and pituitary gland of middle-aged and old cycling rats.
This study determined whether the biochemical characteristics of the estradiol (E2) nuclear receptor located in three different brain areas and the pituitary gland (PIT) change in aging rats undergoing declining reproductive function. To measure maximal E2 nuclear receptor concentrations, groups of young (3-4 months old), middle-aged (8-11 months old), and old (16-18 months old) cycling rats were injected iv with E2 to translocate maximally cytosolic receptors into the nucleus. One hour later they were killed and nuclear extracts were prepared. Maximal E2 nuclear receptor concentrations and dissociation constants were assessed in the preoptic area (POA), medial basal hypothalamus (MBH), amygdala (AMYG), and PIT using an in vitro exchange assay and covariance analyses of double reciprocal plots (Lineweaver-Burk). Middle-aged cycling rats exhibited decreased E2 nuclear receptor concentrations in the POA, but no change in the MBH, AMYG, or PIT. Old rats exhibited decreased E2 nuclear receptor concentrations in the POA, MBH, and PIT and an unexpected increase in the AMYG. There was no change in the affinity of the receptor with age, although an apparent decrease in the pituitary E2 nuclear receptor of middle-aged rats was observed. This difference was not detected when saturation analyses were performed using varying dilutions of the pituitary nuclear extract. These results demonstrate that changes occur in maximal numbers of E2 nuclear receptors in the POA of middle-aged rats. As animals age further, the changes encompass a larger brain area and include the PIT. The data suggest that changes in E2 nuclear receptor concentrations may contribute to the age-related decline in reproductive function.


BACH1 controls hepatic insulin signaling and glucose homeostasis in mice.
Hepatic insulin resistance is central to the metabolic syndrome. Here we investigate the role of BTB and CNC homology 1 (BACH1) in hepatic insulin signaling. BACH1 is elevated in the hepatocytes of individuals with obesity and patients with non-alcoholic fatty liver disease (NAFLD). Hepatocyte-specific Bach1 deletion in male mice on a high-fat diet (HFD) ameliorates hyperglycemia and insulin resistance, improves glucose homeostasis, and protects against steatosis, whereas hepatic overexpression of Bach1 in male mice leads to the opposite phenotype. BACH1 directly interacts with the protein-tyrosine phosphatase 1B (PTP1B) and the insulin receptor β (IR-β), and loss of BACH1 reduces the interaction between PTP1B and IR-β upon insulin stimulation and enhances insulin signaling in hepatocytes. Inhibition of PTP1B significantly attenuates BACH1-mediated suppression of insulin signaling in HFD-fed male mice. Hepatic BACH1 knockdown ameliorates hyperglycemia and improves insulin sensitivity in diabetic male mice. These results demonstrate a critical function for hepatic BACH1 in the regulation of insulin signaling and glucose homeostasis.


Bone marrow fibrosis in patients with primary myelodysplastic syndromes has prognostic value using current therapies and new risk stratification systems.
Bone marrow fibrosis has recently been recognized as an adverse histological feature in patients with primary myelodysplastic syndromes. In this study, we assessed the prognostic impact of bone marrow fibrosis in patients with primary myelodysplastic syndromes under the recently revised new risk stratification systems: the New Comprehensive Cytogenetic Scoring System and the Revised International Prognostic Scoring System. From 2002 to 2012, a total of 79 (13%) patients with primary myelodysplastic syndromes and moderate/severe bone marrow fibrosis were identified; and these patients were compared with a control group of 166 patients with myelodysplastic syndromes but no significant fibrosis. Bone marrow fibrosis predicted an inferior overall survival and leukemia event-free survival for patients who received no hematopoietic stem cell transplant in univariate and multivariate analysis. Eleven patients with bone marrow fibrosis and 32 control group patients underwent hematopoietic stem cell transplant; and bone marrow fibrosis was an independent risk for an inferior overall survival but not leukemia-free survival. In addition, 17 (4%) patients developed bone marrow fibrosis during the course of myelodysplastic syndromes, which was accompanied by clinical and cytogenetic evidence of disease progression. JAK2 V617F mutations were detected in 6 of the 28 patients with bone marrow fibrosis presenting at the time of diagnosis and 2 of the 7 patients with bone marrow fibrosis developing in the course of disease, significantly higher than the control group patients. We conclude that bone marrow fibrosis is an adverse risk feature in primary myelodysplastic syndromes in the current therapeutic era, and this risk feature is not captured by newly revised risk stratification systems. Inclusion of bone marrow fibrosis in patient assessment may further aid in risk-adapted therapeutic decisions.


[Ocular side-effects associated with amiodarone therapy].
Amiodarone, one of the most effective anti-arrhythmic drugs, is also known for its ability to accumulate lipid-pharmakon complexes in the lysosomes of different tissues. In the eye the lysosomal storage leads to typical side-effects. Whorl-like epithelial, reversible corneal inclusions occur in about 70 to 100 % of the patients on amiodarone therapy. Tiny lens opacities without visual impairment have been reported in 50 % of patients who had been treated with amiodarone. At present the most severe complication of amiodarone is optic neuropathy with an incidence of 1.3 to 1.8 %. The optic neuropathy, as the rule, is only reversible approximately in (1/2) of the patients after discontinuing the drug. The fundoscopic picture of amiodarone neuropathy is similar to classic AION. Retinal involvement has also been reported; however, a relationship with amiodarone has not been proven yet.


[Formula: see text] Does training working memory or inhibitory control produce far-transfer improvements in set shifting for children with ADHD? A randomized controlled trial.
Children with ADHD show impairments in set shifting task performance. However, the limited available evidence suggests that directly training shifting may not improve shifting performance in this population. We hypothesized that this incongruence may be because impairments exhibited by children with ADHD during shifting tasks are due to deficits in other executive functions, as shifting tasks also engage children's working memory and/or inhibitory control abilities. This randomized controlled trial examined the extent to which neurocognitive training of working memory vs. inhibitory control can produce downstream (far-transfer) improvements in set shifting task performance. Children with ADHD ages 8-12 (M = 10.41, SD = 1.46; 12 girls; 74% White/Non-Hispanic) were randomized to either central executive training (CET; n = 25) or inhibitory control training (ICT; n = 29), two next-generation digital therapeutics previously shown to improve their intended neurocognitive targets. Two criterion set shifting tests were administered at pre- and post-treatment. Results indicated that ICT was superior to CET for improving shifting accuracy (treatmentxtime: p = .03, BF10 = 3.01, η2 = .09, d = 0.63). ICT was also superior to CET for improving shifting speed, albeit on only one of the two outcome tasks (p = .02, BF10 = 4.53, η2 = .08, d = 0.59). CET did not produce improvements in shifting speed or accuracy on either task (p > .52, BF01 > 2.62), but showed evidence for more general (non-shifting-specific) improvement in response times on one of the outcome tasks (shift trials, d = 0.70; non-shift trials, d = 0.68). Taken together, these findings confirm that inhibitory control is important for successful performance on shifting tests, and suggest that training inhibitory control may reflect a method for improving set shifting difficulties in children with ADHD.


Hepatitis B Virus and DNA Stimulation Trigger a Rapid Innate Immune Response through NF-κB.
Cell-intrinsic innate immunity provides a rapid first line of defense to thwart invading viral pathogens through the production of antiviral and inflammatory genes. However, the presence of many of these signaling pathways in the liver and their role in hepatitis B virus (HBV) pathogenesis is unknown. Recent identification of intracellular DNA-sensing pathways and involvement in numerous diverse disease processes including viral pathogenesis and carcinogenesis suggest a role for these processes in HBV infection. To characterize HBV-intrinsic innate immune responses and the role of DNA- and RNA-sensing pathways in the liver, we used in vivo and in vitro models including analysis of gene expression in liver biopsies from HBV-infected patients. In addition, mRNA and protein expression were measured in HBV-stimulated and DNA-treated hepatoma cell lines and primary human hepatocytes. In this article, we report that HBV and foreign DNA stimulation results in innate immune responses characterized by the production of inflammatory chemokines in hepatocytes. Analysis of liver biopsies from HBV-infected patients supported a correlation among hepatic expression of specific chemokines. In addition, HBV elicits a much broader range of gene expression alterations. The induction of chemokines, including CXCL10, is mediated by melanoma differentiation-associated gene 5 and NF-κB-dependent pathways after HBV stimulation. In conclusion, HBV-stimulated pathways predominantly activate an inflammatory response that would promote the development of hepatitis. Understanding the mechanism underlying these virus-host interactions may provide new strategies to trigger noncytopathic clearance of covalently closed circular DNA to ultimately cure patients with HBV infection.


[Biomechanic results in impingement syndrome of the shoulder].
Forces underneath the coraco-acromial vault during elevation of the arm were evaluated with a dynamic shoulder model. The deltoid muscle and the rotator cuff muscles were simulated with a hydrodynamic device, in ten autopsy specimens. Controlled cycles of glenohumeral joint motion were initiated with computerised regulation. An ultrasonic device measured the position of the arm in all spatial orientations. Capacitive sensors recorded forces underneath the coraco-acromial vault. The mean force during one cycle of elevation averaged 13.9 Newton +/- 12.5 Newton underneath the coraco-acromial ligament and 3.44 Newton +/- 4.37 Newton underneath the coracoid process. The peak force averaged 37.8 Newton +/- 33.2 Newton underneath the acromion, 3.03 Newton +/- 2.62 Newton underneath the coraco-acromial ligament and 6.93 Newton +/- 7.38 Newton underneath the coracoid process. The force markedly increased at the final stage of arm elevation and during early reverse-elevation in most specimens, corresponding to the painful arc sign. In some specimens, the force under the coracoid process exceeded the force under the acromion. Osteophytes protruding into the subacromial space may lead to a concentration of force and to high regional pressures.


Role of DNA polymerase beta in the excision step of long patch mammalian base excision repair.
The two base excision repair (BER) subpathways in mammalian cells are characterized by the number of nucleotides synthesized into the excision patch. They are the "single-nucleotide" BER pathway and the "long patch" (several nucleotides incorporated) BER pathway. Both of these subpathways involve excision of a damaged base and/or nearby nucleotides and DNA synthesis to fill the excision gap. Whereas DNA polymerase beta (pol beta) is known to participate in the single-nucleotide BER pathway, the identity of polymerases involved in long patch BER has remained unclear. By analyzing products of long patch excision generated during BER of a uracil-containing DNA substrate in mammalian cell extracts we find that long patch excision depends on pol beta. We show that the excision of the characteristic 5'-deoxyribose phosphate containing oligonucleotide (dRP-oligo) is deficient in extracts from pol beta null cells and is rescued by addition of purified pol beta. Also, pol beta-neutralizing antibody inhibits release of the dRP-oligo in wild-type cell extracts, and the addition of pol beta after inhibition with antibody completely restores the excision reaction. The results indicate that pol beta plays an essential role in long patch BER by conducting strand displacement synthesis and controlling the size of the excised flap.


Application of sub-regional analysis to bone mineral density of the lower limb from whole body DXA scans.
Bone mineral density at spine and hip is widely used to diagnose osteoporosis. Certain conditions cause changes in bone density at other sites, particularly in the lower limb, with fractures occurring in non-classical locations. Bone density changes at these sites would be of interest for diagnosis and treatment. We describe an application, based on an existing software option for Hologic scanners, which allows reproducible measurement of bone density at six lower limb sites (upper femur, mid-femur, lower femur; upper leg, mid-leg, lower leg). In 30 unselected subjects, referred for bone density, precision (CV%) measured on 2 occasions, separated by repositioning, ranged from 1.7% (mid-femur) to 4.5% at the lowest leg site. Intra-operator precision, measured by three operators on ten subjects on three occasions, was between 1.0% and 2.9%, whilst inter-operator precision was between 1.0% and 3.6%, according to region. These values compare well with those at the spine and upper femur, and in the literature. There was no evidence that this operator agreement improved between occasions 1 and 3. This technique promises to be useful for assessing bone changes at vulnerable sites in the lower limb, in diverse pathological states and in assessing response to treatment.


Estimating heterogeneous transmission with multiple infectives using MCMC methods.
We developed a general procedure for estimating the transmission probability adjusting for covariates when susceptibles are exposed to several infectives concurrently and taking correlation within transmission units into account. The procedure is motivated by a study estimating efficacy of pertussis vaccination based on the secondary attack rate in a rural sub-Saharan community (Niakhar, Senegal) and illustrated with simulations. The procedure is also appropriate to estimate the pairwise transmission probability in transmission studies of live vaccine virus in a collection of transmission units, such as day-care centres or retirement centres. Previously, analyses either excluded transmission units with multiple infectives or ignored co-infectives. Excluding transmission units with multiple infectives is statistically less efficient and ignoring co-infectives can lead to biased estimation. Modelling is carried out by regressing the latent pairwise transmission probability from each infective to a susceptible on covariates and specifying a transmission linkage function linking the latent pairwise transmission probability to the overall transmission probability. Parameters are estimated using Markov chain Monte Carlo methods.


Territorial bell miners and other birds affecting populations of insect prey.
Bell miners (Manorina melanophrys) feed primarily on the lerps and nymphs of psyllids, and they defend communal territories against other bird species. As bell miners were removed exprimentally from a psyllid-infested patch of eucalypt forest, birds of 11 other species moved in to feed on psyllids and within 4 months they eradicated the infestation. This shows the ability of other birds to control these insects in the absence of bell miners and the value of interspecific territoriality to bell miners.


Successful pregnancy in maple syrup urine disease: a case report and review of the literature.
Maple syrup urine disease (MSUD) is an autosomal recessive disorder of branched-chain amino acid metabolism. Patients with MSUD are at risk of life-threatening metabolic decompensations with ketoacidosis and encephalopathy. These episodes are often triggered by physiological stress. Only few cases of pregnancies in MSUD mothers have been reported so far. We present the favorable outcome of a pregnancy in a woman with classical MSUD. She presented in the metabolic outpatient clinic in week 7 of gestation. Branched-chain amino acid concentrations were measured at least weekly to adjust dietary leucine intake. Despite excellent compliance, leucine concentrations frequently exceeded the target value of < 300 μmol/L during the first trimester. From the second trimester until delivery, protein and leucine intake increased continuously to about threefold compared to pre-pregnancy values. To maximize patient safety during delivery and the postpartum period, a detailed plan including peripartal infusion therapy, dietary recommendations and monitoring parameters was developed. Primary Caesarean section was performed in week 38 of gestation, and the patient gave birth to a healthy girl. Lactation was successfully implemented. Leucine levels were maintained within the target range throughout the complete postpartum period. In addition to our case, we give an overview about all cases of pregnancies in MSUD mothers published so far. Management of pregnancy, delivery, postpartum period and lactation may be challenging in patients with MSUD. Careful monitoring and interdisciplinary collaboration is essential to minimize the risk of metabolic crisis, especially after delivery.


The use of concurrent hormonotherapy and radiotherapy does not deteriorate radiation-induced cardiac toxicity.
Postmenopausal patients with breast cancer have two options for adjuvant endocrine therapy, tamoxifen and aromatase inhibitors (AIs) as well as radiotherapy (RT) and chemotherapy. However, there is limited data regarding the optimal sequencing of RT and tamoxifen/AIs. Thus, we aimed to evaluate the effects of tamoxifen and AIs on radiation-induced cardiotoxicity. Eighty ovariectomized rats were divided into eight groups (G). G1 was defined as a control group; G2, G3, G4, and G5 were RT, tamoxifen, anastrozle, and letrozole groups, respectively; G6, G7, and G8 were RT plus tamoxifen, anastrozle, and letrozole groups, respectively. Drugs were started 1 week before RT and continued until the animals were killed 16 weeks after RT. The heart tissues were then dissected and examined with light microscopy to determine endocardial thickness and cardiac fibrosis. The endocardial thickness scores of both RT alone and the tamoxifen groups as well as the cardiac fibrosis score of RT alone were higher than that the control group ( p < 0.05 for all). There was no difference in the endocardial thickness and cardiac fibrosis scores of the RT-only group and the RT plus hormonotherapy groups ( p > 0.05 for all). Concurrent administration of RT and hormonal therapy with either tamoxifen or AIs did not further amplify radiation-induced cardiac toxicity. This issue warrants further study.


Survival of children after liver transplantation for hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) in childhood differs from adult HCC because it is often associated with inherited liver disease. It is, however, unclear whether liver transplantation (LT) for HCC in childhood with or without associated inherited disease has a comparable outcome to adult HCC. On the basis of data from the European Liver Transplant Registry (ELTR), we aimed to investigate if there are differences in patient and graft survival after LT for HCC between children and adults and between patients with underlying inherited versus noninherited liver disease, respectively. We included all 175 children who underwent LT for HCC and were enrolled in ELTR between 1985 and 2012. Of these, 38 had an associated inherited liver disease. Adult HCC patients with (n = 79) and without (n = 316, matched by age, sex, and LT date) inherited liver disease served as an adult comparison population. We used multivariable piecewise Cox regression models with shared frailty terms (for LT center) to compare patient and graft survival between the different HCC groups. Survival analyses demonstrated a superior longterm survival of children with inherited liver disease when compared with children with HCC without inherited liver disease (hazard ratio [HR], 0.29; 95% CI, 0.10-0.90; P = 0.03) and adults with HCC with inherited liver disease (HR, 0.27; 95% CI, 0.06-1.25; P = 0.09). There was no survival difference between adults with and without inherited disease (HR, 1.05; 95% CI, 0.66-1.66; P = 0.84). In conclusion, the potential survival advantage of children with an HCC based on inherited disease should be acknowledged when considering transplantation and prioritization for these patients. Further prospective studies accounting for tumor size and extension at LT are necessary to fully interpret our findings. Liver Transplantation 24 246-255 2018 AASLD.


Evaluation of a potential clinical interaction between ceftriaxone and calcium.
In April 2009, the FDA retracted a warning asserting that ceftriaxone and intravenous calcium products should not be coadministered to any patient to prevent precipitation events leading to end-organ damage. Following that announcement, we sought to evaluate if the retraction was justified. A search of the FDA Adverse Event Reporting System was conducted to identify any ceftriaxone-calcium interactions that resulted in serious adverse drug events. Ceftazidime-calcium was used as a comparator agent. One hundred four events with ceftriaxone-calcium and 99 events with ceftazidime-calcium were identified. Adverse drug events were recorded according to the listed description of drug involvement (primary or secondary suspect) and were interpreted as probable, possible, unlikely, or unrelated. For ceftriaxone-calcium-related adverse events, 7.7% and 20.2% of the events were classified as probable and possible for embolism, respectively. Ceftazidime-calcium resulted in fewer probable embolic events (4%) but more possible embolic events (30.3%). Among cases that considered ceftriaxone or ceftazidime and calcium as the primary or secondary drug, one case was classified as a probable embolic event. That patient received ceftriaxone-calcium and died, although an attribution of causality was not possible. Our analysis suggests a lack of support for the occurrence of ceftriaxone-calcium precipitation events in adults. The results of the current analysis reinforce the revised FDA recommendations suggesting that patients >28 days old may receive ceftriaxone and calcium sequentially and provide a transparent and reproducible methodology for such evaluations.


Stimulation and inhibition of 1,3-bis(2-chloroethyl)-1-nitrosourea-induced strand breaks and interstrand cross-linking in Col E1 plasmid deoxyribonucleic acid by polyamines and inorganic cations.
The influence of various polyamines and metallic cations on 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU)-induced DNA single-strand breaks and DNA interstrand cross-linking was in Col E1 plasmid using electrophoretic techniques. Spermidine and spermine (0.4 to 1.5 mM concentration range) markedly stimulated BCNU-induced DNA nicking, whereas putrescine had no effect on the nicking process. In contrast to the polyamines, BCNU-induced DNA nicking was decreased by the three inorganic cations, Na+ (100 and 200 mM), Mg2+ (0.5 and 1.5 mM), and Co3+ (NH3)6 (0.2 and 0.4 mM), with the trivalent hexamminecobalt ions being most inhibitory. When the monofunctional N-methyl-N-nitrosourea (MNU) was used (instead of the bifunctionally active BCNU) to alkylate Col E1 DNA, nicking of the DNA was inhibited by spermidine. Furthermore, the ability of chloroethylated Col E1 DNA to form interstrand cross-links after treatment with BCU was inhibited by 0.5 mM spermidine and 0.5 mM spermine, both concentrations within the intracellular range. Putrescine at 3-6 mM only marginally stimulated DNA cross-linking. In comparison, the inorganic cations all enhanced Col E1 DNA cross-linking by BCNU, with the rank order of cross-link stimulation being Mg2+, Na+, and Co3+ (NH3)6. These results provide evidence that polyamines can interact with DNA to modulate chloroethylnitrosourea-induced DNA damage and that the interaction is not only a function of the charge on the polyamine molecule but also of the chemical structure of the polyamine.


Prolonged Survival of a Patient with Chronic Myeloid Leukemia in Accelerated Phase with Recurrent Isolated Central Nervous System Blast Crisis.
BACKGROUND Chronic myeloid leukemia (CML) is usually a tri-phasic myeloproliferative neoplasm, characterized by the presence of the BCR-ABL1 fusion gene, derived from a balanced translocation, t(9;22)(q34;q11). BCR-ABL tyrosine kinase inhibitors (TKI) are used to treat patients with CML. The addition of pegylated interferon-alpha2b to imatinib or dasatinib results in promising deep molecular responses. Because imatinib shows poor penetration into the central nervous system (CNS), the CNS may become a sanctuary site in patients on prolonged imatinib therapy for CML. It is extremely rare for the blast phase in patients with chronic phase CML to affect the CNS without concomitant bone marrow involvement. CASE REPORT This report describes a 57-year-old woman who was diagnosed with accelerated phase (AP) CML and failed high dose imatinib therapy. Despite achieving an excellent molecular response to dasatinib in 6 months, she developed recurrent isolated CNS blast crisis. Survival was prolonged after treatment with intrathecal chemotherapy and whole-brain radiation therapy combined with dasatinib. After achieving long and deep molecular remission with dasatinib and a few months of pegylated interferon-alpha2a, she lived for 18 months in treatment-free-remission (TFR). At age 65 years, she died of progressive rectal carcinoma with septic shock, cancer-related venous thromboembolism, and a possible autoimmune disorder. CONCLUSIONS This patient with accelerated phase CML and 2 isolated CNS blast crises died in TFR 8.5 years after her initial diagnosis and 7.5 years after her first isolated CNS blast crisis. Survival resulted from tailoring of therapies around her comorbidities.


[Lesions of the coronary bed in patients with ischemic heart disease and ECG changes at rest].
The data of ECG at rest and of coronary angiography have been analyzed in 184 patients. Correlations between various ECG changes and the coronary bed involvement have been revealed. The authors have calculated the sensitivities and specificities of some ECG symptoms for the prediction of an atherosclerotic involvement of the coronary arteries. A sufficiently high specificity of ECG examination (89.1%) makes it useful for active detection of coronary patients, whereas its low specificity (54.2%) necessitates employment of supplementary methods of investigation.


End-stage renal disease in India and Pakistan: incidence, causes, and management.
Chronic renal failure is a devastating medical, social, and economic problem for patients and their families in India and Pakistan. Reliable data on the true incidence and prevalence of end-stage renal disease (ESRD) in India and Pakistan are lacking because no national registries exist. Among reported cases, chronic glomerulonephritis is the most common cause, accounting for more than one third of patients, while diabetic nephropathy accounts for approximately 20% of all patients in India. Delayed diagnosis and failure to institute measures to slow the progression of renal failure have resulted in a predominantly young ESRD population, with a median age of 44 years. Because of financial constraints, less than one third of all patients referred to a tertiary care center receive any kind of renal replacement therapy. Most hemodialysis patients who stop treatment and die do so because of cost constraints within the first three months, and approximately 5% of patients are started on ambulatory peritoneal dialysis. Renal transplantation is the cheapest option, but < 10% of all patients with ESRD have a transplant. Cyclosporine, azathioprine, and prednisolone continue to be the backbone of post-transplant immunosuppression, but cyclosporine is stopped in a significant proportion of patients at one year post-transplant to cut down costs. Living related donor transplants constitute 70% of all transplants; two thirds of the donors are females, while more than three fourths of all recipients are males. Spouses account for 20% of donors from within families. Almost 30% of transplanted kidneys are donated by living unrelated donors, while cadaver donors account for only 2%. More resources must be mobilized to care for these patients; early detection of renal disease must be facilitated, and measures to delay ESRD must be implemented.


Nitrocatechol Derivatives of Chalcone as Inhibitors of Monoamine Oxidase and Catechol-O-Methyltransferase.
The efficacy of L-dopa in the treatment of Parkinson's disease depends on its metabolic conversion to dopamine in the brain, however extensive peripheral metabolism of L-dopa diminishes its availability for uptake into the brain. L-Dopa is extensively decarboxylated in the gastrointestinal tract and peripheral tissues by Aromatic L-Amino Acid Decarboxylase (AADC), and AADC inhibitors are thus frequently combined with L-dopa therapy. When AADC is inhibited, 3-Omethylation Catalysed by Catechol-O-Methyltransferase (COMT) becomes a dominant metabolic pathway for L-dopa, and COMT inhibitors may thus also be used as adjuncts to L-dopa in Parkinson's disease. Monoamine Oxidase (MAO), in turn, metabolises dopamine in the brain, and MAO-B inhibitors may exert a dopamine sparing effect in the brain. Based on the roles of COMT and MAO in the metabolism of L-dopa and dopamine, the present study attempts to discover novel dual inhibitors of these enzymes. For this purpose, nitrocatechol derivatives of chalcone were synthesised and evaluated as inhibitors of COMT and MAO. The chalcone class of compounds is well known to potently inhibit MAO-B, while nitrocatechol derivatives (e.g. tolcapone and entacapone) are clinically used COMT inhibitors. The results document that all of the derivatives are high potency in vitro inhibitors of rat liver COMT with IC50 values ranging from 0.07 to 0.29 μM. Under these experimental conditions, tolcapone and entacapone display IC50 values of 0.26 µM and 0.25 µM, respectively. The chalcones are less potent as inhibitors of MAO with the most potent inhibitor possessing a Ki of 4.6 µM for the in vitro inhibition of human MAO-B. This study shows that nitrocatechol derivatives of chalcone may act as COMT and MAO-B inhibitors, and proposes a general strategy for further enhancing MAO-B inhibition while retaining the potent COMT inhibition activity of this class.


Creatine metabolism and safety profiles after six-week oral guanidinoacetic acid administration in healthy humans.
Guanidinoacetic acid (GAA) is a natural precursor of creatine, yet the potential use of GAA as a nutritional additive for restoring creatine availability in humans has been limited by unclear efficacy and safety after exogenous GAA administration. The present study evaluated the effects of orally administered GAA on serum and urinary GAA, creatine and creatinine concentration, and on the occurrence of adverse events in healthy humans. Twenty-four healthy volunteers were randomized in a double-blind design to receive either GAA (2.4 grams daily) or placebo (PLA) by oral administration for 6 weeks. www.clinicaltrials.gov, identification number NCT01133899. Serum creatine and creatinine increased significantly from before to after administration in GAA-supplemented participants (P < 0.05). The proportion of participants who reported minor side effects was 58.3% in the GAA group and 45.5% in the placebo group (P = 0.68). A few participants experienced serum creatine levels above 70 µmol/L. Exogenous GAA is metabolized to creatine, resulting in a significant increase of fasting serum creatine after intervention. GAA had an acceptable side-effects profile with a low incidence of biochemical abnormalities.


Attitude and practices of parents toward their children's oral health care during COVID-19 pandemic.
The impact of COVID-19 has affected billions of people affecting nearly all domains of life, including patient's healthcare needs. Due to constrained dental services during lockdown, pediatric dental needs were severely compromised therefore, parental attitude and practices of maintaining child's oral hygiene impose significant impact on child's oral hygiene. This study aims to assess the attitude and practices of parents toward their children's oral health care during COVID-19 pandemic. The present cross-sectional study was conducted with a sample of 120 parents selected randomly from the outpatient department after obtaining informed consent. A structured questionnaire was used to assess attitude and practice of parents toward their child's oral hygiene during lockdown in COVID-19 pandemic. The overall attitude and practice score of parents in maintaining their child's oral hygiene was low, with mean value of 2.36 ± 1.36 and 2.51 ± 1.50, respectively. 60.8% of parents reported the need of dental treatment for their child during the lockdown period whereas 50.8% parents contacted the dentist during this period; majority (37.5%) of which reported due to toothache. 68.3% of parents would like to visit nearby dental hospital for emergencies during lockdown period. Only 33.3% of parents took extra efforts to maintain their child's oral hygiene while 45% made dietary changes in their child's diet to prevent tooth decay. The overall attitude and practices of the parents were poor toward the oral health of their children during the lockdown period in COVID-19 pandemic. There is urgent need of increasing awareness among parents regarding the importance of maintaining their child's oral health. The inability to see the doctor should at least encourage better home practice measures for future. The concept of tele-dentistry is very new and will be the need of the coming modern times.


[Hazards and complications of combined use of anti-arrhythmia drugs in ambulatory conditions].
To prevent paroxysms of atrial fibrillation many patients resort to long-term use of various antiarrhythmic agents. In this case a question arises: how to recover a sinus rhythm if the agent's effect is not over. Two clinical cases have been described when the patients themselves tried to arrest arrhythmia by increasing the dose of the main drug and using additionally other antiarrhythmics, which led to severe complications.


Stress and Androgen Activity During Fetal Development.
Prenatal stress is known to alter hypothalamic-pituitary-adrenal axis activity, and more recent evidence suggests that it may also affect androgen activity. In animal models, prenatal stress disrupts the normal surge of testosterone in the developing male, whereas in females, associations differ by species. In humans, studies show that (1) associations between prenatal stress and child outcomes are often sex-dependent, (2) prenatal stress predicts several disorders with notable sex differences in prevalence, and (3) prenatal exposure to stressful life events may be associated with masculinized reproductive tract development and play behavior in girls. In this minireview, we examine the existing literature on prenatal stress and androgenic activity and present new, preliminary data indicating that prenatal stress may also modify associations between prenatal exposure to diethylhexyl phthalate, (a synthetic, antiandrogenic chemical) and reproductive development in infant boys. Taken together, these data support the hypothesis that prenatal exposure to both chemical and nonchemical stressors may alter sex steroid pathways in the maternal-placental-fetal unit and ultimately alter hormone-dependent developmental endpoints.


Job stress interventions and the organization of work.
Interventions that aim at improving health by changing the organization of work-in terms of task characteristics, work conditions, and social aspects-have shown their potential, but results are mixed, and many studies do not use their methodological potential. It is proposed that interventions at the organizational level are likely to have a more diverse effect than at the individual level, as the number of subsystems, with potentially diverging interests, is larger. Even well-implemented interventions are not likely to lead to improvements in all parameters for all participants, and trade-offs have to be considered. Methodological improvement is necessary but should not only focus on design issues, but also on careful documentation and subgroup analyses. A combination of person-focused and organization-focused approaches is the most promising. Finally, evidence points to the limited utility of economic arguments for the acceptance of health promotion projects; the necessity of professional trust is therefore emphasized.


Clinical Decision-Making when Treating Diabetic Macular Edema Patients with Dexamethasone Intravitreal Implants.
Diabetes mellitus (DM) is a metabolic disease frequently associated with comorbidities that include diabetic macular edema (DME). The current medical approach to treating DME involves intravitreal injections with either anti-vascular endothelial growth factors or steroids. However, the burden associated with intravitreal injections and DM-derived complications is high, underlining the need to find optimal treatment regimens. In this article we describe the considerations we apply when treating DME patients with dexamethasone intravitreal implants (Ozurdex®), particularly those that influence the clinical decision-making process during the follow-up period. These considerations are based both on the available medical literature and on our clinical experience following the use of these implants in this type of patient, the goal being to optimize the number of injections and the clinical outcome of this therapy. We also provide a general overview of the pathophysiology of DME, highlighting the inflammatory component as a rationale to use steroids in these patients.


Uptake and toxicity of glyphosate in the lichen Xanthoria parietina (L.) Th. Fr.
This study investigated if treatment of the lichen Xanthoria parietina (L.) Th. Fr. with glyphosate caused uptake of this herbicide as well as physiological alterations. Samples were treated with Glifene SL®, a common commercial glyphosate-based herbicide, at the lowest recommended doses (3.6g/L) as well as with doses slightly higher than the highest suggested (36 g/L). The results clearly showed glyphosate uptake in X. parietina proportionally to the dose provided. Adverse physiological effects were evident on the photosynthetic apparatus (photosynthetic efficiency, chlorophyll a content, chlorophyll degradation) as well as on the fungal respiration rates and cell membrane integrity (ergosterol content, dehydrogenase activity) already after 24h from treatment, also at the low application dose. It is concluded that lichens are suitable organisms for monitoring unwanted biological effects from the application of glyphosate-based herbicides, as well as for detecting the accumulation of this compound in the biota, thus screening for its environmental fate.


Synthesis and biological evaluation of a fluorescent analogue of folic acid.
A fluorescein derivative of the lysine analogue of folic acid, N alpha-pteroyl-N epilson-(4'-fluoresceinthiocarbamoyl)-L-lysine (PLF), was synthesized as a probe for dihydrofolate reductase (DHFR) and a membrane folate binding protein (m-FBP). Excitation of PLF at 282 nm and at 497 nm gave a fluorescence emission maximum at 518 nm. Binding of PLF to human DHFR or human placental m-FBP results in approximately a 20-fold enhancement in the magnitude of the fluorescence emission, suggesting that the ligand interacts with a hydrophobic region on these proteins. Additional evidence suggests that an energy transfer may occur between the pteridine and the fluorescein moieties. PLF binds to the active site of human DHFR since methotrexate (MTX) competes stoichiometrically and the denatured enzyme in the presence of PLF did not exhibit fluorescent enhancement. The dissociation constant for the fluorescein derivative with respect to human DHFR is 115 nM as compared to 111 nM for folic acid. The Ki value for the competitive inhibition of human DHFR by the fluorescent analogue of folic acid is 2.0 microM compared to 0.48 microM for folic acid. PLF was reduced to N alpha-(7,8-dihydropteroyl)-N epilson-(4'-fluoresceinthiocarbamoyl)-L-lysine (H2PLF) and assayed by the enzymatic conversion to the tetrahydro derivative. The Km value for human DHFR for the dihydrofolate analogue is 2.0 microM. The KD value for H2PLF to human DHFR is 47 nM as compared to 44 nM for dihydrofolate. The KD values for both H2PLF and PLF indicate that the fluorescein moiety does not significantly affect folate binding in enzyme binary complexes.(ABSTRACT TRUNCATED AT 250 WORDS)


Post-surgical inflammatory neuropathy.
Post-surgical neuropathies are usually attributed to mechanical factors, such as compression, stretch, contusion or transection. The role of inflammatory mechanisms in neuropathies occurring after surgeries is poorly appreciated and not well characterized, and may provide a rationale for immunotherapy. A total of 23 selected patients with post-surgical neuropathies received nerve biopsies, of which 21 demonstrated increased inflammation. Here we report the clinical features in these 21 cases of biopsy-confirmed and 12 cases of clinically suspected post-surgical inflammatory neuropathies, in whom no trauma to the nerves was documented. All neuropathies developed within 30 days of a surgical procedure. Of 33 patients, 20 were male and the median age was 65 years (range 24-83). Surgical procedures were orthopaedic (n=14), abdominal/pelvic (n=12), thoracic (n=5) and dental (n=2). Patients developed focal (n=12), multifocal (n=14) or diffuse (n=7) neuropathies. Focal and multifocal neuropathies typically presented with acute pain and weakness, and focal neuropathies often mimicked mechanical aetiologies. Detailed analyses, including clinical characteristics, electrophysiology, imaging and peripheral nerve pathology, were performed. Electrophysiology showed axonal damage. Magnetic resonance imaging of roots, plexuses and peripheral nerves was performed in 22 patients, and all patients had abnormally increased T(2) nerve signal, with 20 exhibiting mild (n=7), moderate (n=12) or severe (n=1) enlargement. A total of 21 patients had abnormal nerve biopsies that showed increased epineurial perivascular lymphocytic inflammation (nine small, five moderate and seven large), with 15 diagnostic or suggestive of microvasculitis. Evidence of ischaemic nerve injury was seen in 19 biopsies. Seventeen biopsies had increased axonal degeneration suggesting active neuropathy. Seventeen biopsied patients were treated with immunotherapy. In 13 cases with longitudinal follow-up (median 9 months, range 3-71 months), the median neuropathy impairment score improved from 30 to 24 at the time of last evaluation (P=0.001). (i) not all post-surgical neuropathies are mechanical, and inflammatory mechanisms can be causative, presenting as pain and weakness in a focal, multifocal or diffuse pattern; (ii) these inflammatory neuropathies may be recognized by their spatio-temporal separation from the site and time of surgery and by the characteristic magnetic resonance imaging features; (iii) occasionally post-surgical inflammatory and mechanical neuropathies are difficult to distinguish and nerve biopsy may be required to demonstrate an inflammatory mechanism, which in our cohort often, but not exclusively, exhibited pathological features of microvasculitis and ischaemia; and (iv) recognizing the role of inflammation in these patients' neuropathy led to rational immunotherapy, which may have resulted in the subsequent improvement of neurological symptoms and impairments.


Intrinsic disorder in proteins associated with oxidative stress-induced JNK signaling.
The c-Jun N-terminal kinase (JNK) signaling cascade is a mitogen-activated protein kinase (MAPK) signaling pathway that can be activated in response to a wide range of environmental stimuli. Based on the type, degree, and duration of the stimulus, the JNK signaling cascade dictates the fate of the cell by influencing gene expression through its substrate transcription factors. Oxidative stress is a result of a disturbance in the pro-oxidant/antioxidant homeostasis of the cell and is associated with a large number of diseases, such as neurodegenerative disorders, cancer, diabetes, cardiovascular diseases, and disorders of the immune system, where it activates the JNK signaling pathway. Among different biological roles ascribed to the intrinsically disordered proteins (IDPs) and hybrid proteins containing ordered domains and intrinsically disordered protein regions (IDPRs) are signaling hub functions, as intrinsic disorder allows proteins to undertake multiple interactions, each with a different consequence. In order to ensure precise signaling, the cellular abundance of IDPs is highly regulated, and mutations or changes in abundance of IDPs/IDPRs are often associated with disease. In this study, we have used a combination of six disorder predictors to evaluate the presence of intrinsic disorder in proteins of the oxidative stress-induced JNK signaling cascade, and as per our findings, none of the 18 proteins involved in this pathway are ordered. The highest level of intrinsic disorder was observed in the scaffold proteins, JIP1, JIP2, JIP3; dual specificity phosphatases, MKP5, MKP7; 14-3-3ζ and transcription factor c-Jun. The MAP3Ks, MAP2Ks, MAPKs, TRAFs, and thioredoxin were the proteins that were predicted to be moderately disordered. Furthermore, to characterize the predicted IDPs/IDPRs in the proteins of the JNK signaling cascade, we identified the molecular recognition features (MoRFs), posttranslational modification (PTM) sites, and short linear motifs (SLiMs) associated with the disordered regions. These findings will serve as a foundation for experimental characterization of disordered regions in these proteins, which represents a crucial step for a better understanding of the roles of IDPRs in diseases associated with this important pathway.


Does Napping Enhance the Consolidation of Clinically Relevant Information? A Comparison of Individuals with Low and Elevated Depressive Symptoms.
Sleep, both overnight and daytime naps, can facilitate the consolidation of declarative memories in healthy humans. However, it is unclear whether such beneficial effects of sleep occur in special populations, such as individuals with elevated neuropsychiatric symptoms, and if they apply to clinically relevant material that may have personal significance to those populations. We examined memory retention over a 60-minute interval of wakefulness or nap opportunity in participants with low or elevated scores (≤13 and ≥21, respectively) on the Beck Depression Inventory-II (BDI-II). Memory for depression-related information was assessed by (a) free-recall of a video depicting a personal experience narrative of the impact of depression on cognition and workplace performance; and (b) a paired-associates task linking depression-related cognitive symptoms to appropriate coping strategies. The results showed no overall difference in recall between the nap and waking condition. However, across the full sample of participants, there were significant positive correlations between total sleep time and paired associates recall, and slow wave sleep (SWS) percentage and story free recall performance. Unexpectedly, participants with elevated BDI-II scores exhibited better free-recall performance compared to those with low scores. These results suggest that sleep, specifically SWS, may stabilize memories for clinically relevant information in populations with low and elevated depressive symptoms. The superior recall in participants with elevated-BDI scores may be related to the personal significance and stronger encoding of depression-related information. These observations raise the possibility that mnemonic deficits in depressed patients may be, at least in part, related to the type of information used to assess memory performance.


Motor deficits following dorsal corticospinal tract transection in rats: voluntary versus skilled locomotion readouts.
Following spinal cord injury, severe deficits result from damages to ascending and descending tracts, such as the corticospinal tract (CST) which is highly relevant for the motor execution in humans. Unfortunately, no curative treatment is available and intensive efforts are deployed in animal models, such as the CST transection model, to identify interventions providing functional regeneration after spinal cord injury. The CatWalk XT is a system for multi-parameter gait analysis of voluntary locomotion. In this study, the performance of the CatWalk XT for monitoring of functional deficits associated with dorsal CST lesion in rats was compared to skilled locomotion tests. Motor deficits associated with dorsal CST transection could be reliably monitored over seven weeks based on skilled locomotion testing, i.e. Horizontal Ladder Walk and Grid Walk. The collateral lesion to the overlaying gracile and cuneate funiculi occurring during dorsal CST transection resulted in slight hyposensitivity and proprioceptive deficit, which likely contributed to the lowered performance in skilled locomotion. In contrast, parameters of voluntary locomotion were not significantly affected by dorsal CST transection. Finally, an abnormal adduction reflex was detected immediately after lesion of the CST and could be conveniently used to confirm successful CST lesion in rats of experimental groups. The functional relevance of the dorsal CST in locomotion of rats is not as prominent as compared to in humans and thus challenging the motor execution is mandatory to reliably investigate CST function. A detailed analysis of voluntary walking using the CatWalk XT is not adequate to detect deficits following dorsal CST lesion in rats.


Turning danger into opportunity: teaching psychiatric nursing in the aftermath of a disaster.
In the wake of a devastating flood that destroyed the ecology of a small town, undergraduates taking their first clinical course in psychiatric mental health nursing formed AGAST (Awesome Guerillas Against Situational Trauma) and, with their instructor, performed individual and group crisis intervention over an intensive 90-hour clinical rotation. Three theories were used to help guide the students assessments and interventions with survivors. The clinical experience required innovation, close connection with the host community, and multi-level support from the educational institution. Students rose to the occasion and blossomed in the rich clinical matrix of the experience. The example can be applied to the use of other naturally-occurring events as theaters for innovative clinical teaching.


[Socio-demographic impact of 15 years of family planning].
Familiar planning is an important branch of the preventive medicine that can have a great impact on the health of the humanity. The present study is an evaluation by a cross section of the effects obtained by the program of familiar planning during a period of 15 years in the Aguascalientes Delegation of the Mexican Institute of the Social Security, establishing therefore a diagnosis and identifying elements that can contribute in the suitable planning of strategies to improve the quality of the attention and to respond to social and health necessities of the population.


Subgroups of New York City children at high risk of PTSD after the September 11 attacks: a signal detection analysis.
Case finding is an important challenge in mental health programs responding to large-scale disasters. Most people who experience psychological symptoms after such events return to normal functioning within a few months. Yet a significant minority continues to experience enduring symptoms. This study demonstrated the use of signal detection analyses of community survey data to identify subgroups of children who were at highest risk of posttraumatic stress disorder (PTSD) after the September 11 attacks. This study reanalyzed results of a needs assessment survey conducted six months after the World Trade Center attacks on September 11, 2001, with a representative sample of 7,832 New York City public school students in grades 4 to 12. Receiver operating characteristic (ROC) analyses conducted on half the sample resulted in a decision tree for classifying children into groups at varying levels of risk of PTSD. These decision rules were subsequently retested on the second half of the sample. We could reliably classify children into groups with varying probabilities of screening positive on a PTSD screen. Nearly two-thirds of children in grades 4 to 12 who screened positive for probable PTSD were concentrated among 4th graders (35%) and among children who had a friend or family member directly exposed to the attacks (28%). Signal detection analysis of community needs assessment surveys can identify community subgroups most likely to screen positive for mental health problems after a disaster or terrorist attack. This information can help target screening and outreach efforts to community segments that have the highest need for services.


Activation and co-ordination of InsP3-mediated elementary Ca2+ events during global Ca2+ signals in Xenopus oocytes.
1. The activation of elementary calcium release events ('puffs') and their co-ordination to generate calcium waves was studied in Xenopus oocytes by confocal linescan imaging together with photorelease of inositol 1,4,5-trisphosphate (InsP3) from a caged precursor. 2. Weak photolysis flashes evoked no responses or isolated calcium puffs, whereas flashes of increasing strength evoked more frequent puffs, often occurring in flurries as abortive waves, and then a near-simultaneous calcium liberation originating at multiple sites. The numbers of sites activated increased initially as about the fourth power of photoreleased [InsP3]. 3. Following repeated, identical photolysis flashes, puffs arose after stochastically varying latencies of a few hundred milliseconds to several seconds. The cumulative number of events initially increased as about the third power of time. No rise in free [Ca2+] was detected preceding the puffs, suggesting that this co-operativity arises through binding of multiple InsP3 molecules, rather than through calcium feedback. 4. The mean latency to onset of calcium liberation shortened as about the square of the flash strength, and the dispersion in latencies between events reduced correspondingly. 5. Weak stimuli often evoked coupled puffs involving adjacent sites, and stronger flashes evoked saltatory calcium waves, propagating with non-constant velocity. During waves, [Ca2+] rose slowly between puff sites, but more abruptly at active sites following an initial diffusive rise in calcium. 6. Initial rates of rise of local [Ca2+] at release sites were similar during puffs and release induced by much (> 10-fold) greater [InsP3]. In contrast, macroscopic calcium measurements averaged over the scan line showed a graded dependence of rate of calcium liberation upon [InsP3], due to recruitment of additional sites and decreasing dispersion in activation latencies. 7. We conclude that the initiation of calcium liberation depends co-operatively upon [InsP3] whereas the subsequent regenerative increase in calcium flux depends upon local calcium feedback and is largely independent of [InsP3]. Wave propagation is consistent with the diffusive spread of calcium evoking regenerative liberation at heterogeneous discrete sites, the sensitivity of which is primed by InsP3.


Giant cell lesions of the jaws: does the level of vascularity and angiogenesis correlate with behavior?
To compare vascularity and angiogenic activity in aggressive and nonaggressive giant cell lesions (GCLs) of the jaws. This is a retrospective study of 14 GCLs treated at the University of California, San Francisco. Immunohistochemistry was used to determine of the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), CD34, and CD31. VEGF and bFGF expression in giant cells (GCs) and surrounding mononuclear stroma was classified into 1) high immunoreactivity (>50% staining) and 2) low immunoreactivity (<50% staining). CD31- and CD34-stained vessels were counted at 200× magnification. Clinical and radiographic records were reviewed to classify lesions as aggressive or nonaggressive. Of the lesions, 8 were aggressive and 6 were nonaggressive. High VEGF expression was found within the GCs in 4 of 8 aggressive lesions compared with 1 of 6 nonaggressive lesions. The stroma in both groups had low staining. High staining of the GCs for bFGF was found in 6 of 8 aggressive lesions compared with 3 of 6 nonaggressive lesions. The stroma of all aggressive cases showed high expression of bFGF compared with 3 of 6 nonaggressive cases. The aggressive group had a mean of 20.1 ± 5.4 vessels/high-powered field (hpf) stained for CD31 compared with 11.5 ± 5.6 vessels/hpf in the nonaggressive group. The aggressive group had 24.6 ± 7.0 vessels/hpf stained with CD34 compared with 18.5 ± 4.0 vessels/hpf in the nonaggressive group. The vascularity and level of angiogenesis within aggressive GCLs are higher than those in nonaggressive lesions.


At-hatch administration of probiotic to chickens can introduce beneficial changes in gut microbiota.
Recent advances in culture-free microbiological techniques bring new understanding of the role of intestinal microbiota in heath and performance. Intestinal microbial communities in chickens assume a near-stable state within the week which leaves a very small window for permanent microbiota remodelling. It is the first colonisers that determine the fate of microbial community in humans and birds alike, and after the microbiota has matured there are very small odds for permanent modification as stable community resists change. In this study we inoculated broiler chicks immediately post hatch, with 3 species of Lactobacillus, identified by sequencing of 16S rRNA and pheS genes as L. ingluviei, L. agilis and L. reuteri. The strains were isolated from the gut of healthy chickens as reproducibly persistent Lactobacillus strains among multiple flocks. Birds inoculated with the probiotic mix reached significantly higher weight by 28 days of age. Although each strain was able to colonise when administered alone, administering the probiotic mix at-hatch resulted in colonisation by only L. ingluviei. High initial abundance of L. ingluviei was slowly reducing, however, the effects of at-hatch administration of the Lactobacillus mix on modifying microbiota development and structure remained persistent. There was a tendency of promotion of beneficial and reduction in pathogenic taxa in the probiotic administered group.


The effects of acute work stress and appraisal on psychobiological stress responses in a group office environment.
The high prevalence of office stress and its detrimental health consequences are of concern to individuals, employers and society at large. Laboratory studies investigating office stress have mostly relied on data from participants that were tested individually on abstract tasks. In this study, we examined the effect of psychosocial office stress and work interruptions on the psychobiological stress response in a realistic but controlled group office environment. We also explored the role of cognitive stress appraisal as an underlying mechanism mediating the relationship between work stressors and the stress response. Ninety participants (44 female; mean age 23.11 ± 3.80) were randomly assigned to either a control condition or one of two experimental conditions in which they were exposed to psychosocial stress with or without prior work interruptions in a realistic multi-participant laboratory setting. To induce psychosocial stress, we adapted the Trier Social Stress Test for Groups to an office environment. Throughout the experiment, we continuously monitored heart rate and heart rate variability. Participants repeatedly reported on their current mood, calmness, wakefulness and perceived stress and gave saliva samples to assess changes in salivary cortisol and salivary alpha-amylase. Additionally, cognitive appraisal of the psychosocial stress test was evaluated. Our analyses revealed significant group differences for most outcomes during or immediately after the stress test (i.e., mood, calmness, perceived stress, salivary cortisol, heart rate, heart rate variability) and during recovery (i.e., salivary cortisol and heart rate). Interestingly, the condition that experienced work interruptions showed a higher increase of cortisol levels but appraised the stress test as less threatening than individuals that experienced only psychosocial stress. Exploratory mediation analyses revealed a blunted response in subjective measures of stress, which was partially explained by the differences in threat appraisal. The results showed that experimentally induced work stress led to significant responses of subjective measures of stress, the hypothalamic-pituitary-adrenal axis and the autonomic nervous system. However, there appears to be a discrepancy between the psychological and biological responses to preceding work interruptions. Appraising psychosocial stress as less threatening but still as challenging could be an adaptive way of coping and reflect a state of engagement and eustress.


Are the early childhood antecedents of men's external locus of control similar to those of their female partners?
Background: The concept of locus of control of reinforcement was introduced by Julian Rotter and has been the focus of intense research for nearly half a century. Surprisingly little research has been directed at clarifying antecedents of locus of control (LOC) orientations in adult men apart from a few small studies. We previously identified a number of independent antecedents associated with women's LOC, including features of their parents and early childhood. This raised the question as to whether these factors were also associated with the development of LOC in men. Methods: To identify antecedents of LOC orientations in a representative population of women we previously analysed information concerning characteristics of their parents and their own childhood experiences using pregnant women taking part in the Avon Longitudinal Study of Parents and Children (ALSPAC). Here we use the same design to determine whether their male partners have similar antecedents of LOC orientation. As previously, we use a hypothesis-free exposome technique using all available information on the parents and childhood of the individuals. Results: We show that men had many of the same antecedent characteristics as the women - in particular, their mother's year of birth and father's social group, being exposed to cigarette smoke prenatally, starting to smoke regularly before the age of 11, and having a friend die were all associated with being external. Associations of internality common to both were warm maternal care, being breast fed, being born in an area other than that where they currently live, attending boarding school and having a parent admitted to hospital. Conclusions: In general, the antecedents of male external and internal personalities have many similarities to those of women, thus providing some features to inform the possible theoretical background as to how LOC might develop over time.


Sixteen-year change in acoustic-admittance measures among older adults: data from a population-based study.
The primary purpose of this study was to measure the 16-year change in peak compensated static acoustic admittance (Peak Y(tm)) in a population-based cohort of older adults, and to determine whether age was associated with any observed change in Peak Y(tm). Other tympanometric measures also were taken and analyzed. Data from two examinations (1993-1995 and 2009-2010) of the population-based Epidemiology of Hearing Loss Study (n = 1,439 with data at both examinations, ages 48-84 years at baseline) were utilized to assess 16-year change. Tympanometric measures were taken using a 226-Hz probe tone, a positive-to-negative direction of pressure change, and a measured pump speed of 600/200 daPa/s. During the 16-year period, Peak Y(tm) declined an average of 0.009 mmho/year (0.009/year for women, 0.007/year for men). Among women, older baseline age was associated with greater decline in Peak Y(tm). Among men, baseline age was associated with change in Peak Y(tm), but in a nonlinear pattern. Other tympanometric measures demonstrated little change after 16 years. These results demonstrate a small degree of middle-ear stiffening after 16 years among these older adults, but not enough to affect function in a manner that would influence clinical decisions.


Modified double papillae flap technique: a new surgical approach for the treatment of isolated gingival recession defects. A case series.
There have been no modifications made to the original double papillae flap surgical technique introduced by Cohen and Ross in 1968. The aim of the present case series is to evaluate the effectiveness of a modified surgical approach to the double papillae flap design in the treatment of isolated gingival recession. A total of 12 healthy, young patients (age range 20 to 28 years) with isolated gingival recession were enrolled in this study. The gingival recession had the following characteristics: isolated defect, Miller Class I or II defect, visible cementoenamel junction (CEJ), and well represented interdental papillae. The following clinical measurements were taken before the surgery and at the 1-year follow-up examination: probing depth (PD) at the treated tooth, clinical attachment level (CAL), amount of keratinized tissue (KT) at the treated site, and recession depth (RD). All measurements were taken at baseline and after 12 months by means of a manual probe and were rounded up to the nearest millimeter. At the 1-year follow-up examination, the same clinical parameters taken at baseline were rerecorded. No statistical differences were noted for the PD (P=0.54). The mean increase for the CAL was statistically significant (P=0.04). Furthermore, the mean amount of KT increased by 2.5±0.4 mm (P<0.001). Finally, the mean RD decreased to 0.5±0.6 mm, corresponding to a root coverage of 3.8±0.8 mm (P<0.001). In terms of mean root coverage (MRC), 88.4% of exposed root surface was covered with soft tissue, and 9 of the 12 treated teeth (75%) showed complete coverage of the root surface. The results of the present case series show that the modified double papillae flap technique is effective in obtaining root coverage of isolated gingival recession defects. These rootcoverage outcomes were associated with clinically and statistically highly significant clinical attachment gain, with no noticeable change in either the PD values or the height of KT on the buccal aspect of the treated teeth.


Uptake of folic acid supplements before and during pregnancy: focus group analysis of women's views and experiences.
To reduce risk of neural tube defects, current guidance recommends that all women who could become pregnant should take a daily 400 μg folic acid supplement before conception and until the 12th week of pregnancy. It is recognised that compliance with this guidance is sub-optimal, although little is known about the reasons why. The present study aims to explore the rationale behind women's decision-making on folic acid supplement use to inform health communications. Women attending routine health visitor led baby clinics completed a questionnaire to establish their folic acid use in their most recent pregnancy. Participants were then invited to join focus group discussions to explore motivators and barriers to folic acid supplement use before and during pregnancy. Of 292 women approached, 211 (70%) provided information on supplement use. Of these, 67 (31%) reported having taken folic acid supplements as recommended; 118 (56%) only during pregnancy [22 (18%) only intermittently]; and 26 (12%) had not taken folic acid at all. Eight focus group discussions were held comprising 24 participants. Discussions indicated the rationale behind current recommendations was known. Participants often linked folic acid use with morning sickness, and invoked busy lives, competing priorities for concern, and poor memory in accounting for intermittent use. Building a 'lay evidence base' from their own experiences, many cited healthy pregnancy outcomes without supplement use and expressed scepticism about its preventive action. The findings of the present study highlight the importance of guidance on the importance of daily folic acid supplement use, the severity of neural tube defects and the provision of evidence on risk reduction.


The simplified HF E-800 high-frequency audiometer: clinical applications.
In a prior study (Gauz and Smith, J. Aud. Res., 1985, 25, 101-122) threshold tracings were collected at 8-20 kc/s on 108 Ss using a Bekesy-type Simplified High-Frequency Audiometer (SHF E-800) set in the fixed-frequency mode. Of these, 23 Ss did not meet screening criteria for a normative study; their data are reported here. Examples are given of HF audiometric results on Ss with one or more conditions affecting hearing thresholds, including age, conductive pathologies, noise exposure, cardiovascular disorders, hearing loss in the conventional audiometric frequencies, and a miscellaneous category. Results were obtained generally in line with expectations based on literature review. The present case studies provide information useful in the clinical application of HF audiometry.


Economics redefining the practice of psychiatry.
Major economic forces are dramatically affecting the practice of psychiatry. In the light of these changes, the author suggests how psychiatrists can have an expanded role in the emerging outcome-driven world of health care.


Functional disorganization of small-world brain networks in mild Alzheimer's Disease and amnestic Mild Cognitive Impairment: an EEG study using Relative Wavelet Entropy (RWE).
Previous neuroscientific findings have linked Alzheimer's Disease (AD) with less efficient information processing and brain network disorganization. However, pathological alterations of the brain networks during the preclinical phase of amnestic Mild Cognitive Impairment (aMCI) remain largely unknown. The present study aimed at comparing patterns of the detection of functional disorganization in MCI relative to Mild Dementia (MD). Participants consisted of 23 cognitively healthy adults, 17 aMCI and 24 mild AD patients who underwent electroencephalographic (EEG) data acquisition during a resting-state condition. Synchronization analysis through the Orthogonal Discrete Wavelet Transform (ODWT), and directional brain network analysis were applied on the EEG data. This computational model was performed for networks that have the same number of edges (N = 500, 600, 700, 800 edges) across all participants and groups (fixed density values). All groups exhibited a small-world (SW) brain architecture. However, we found a significant reduction in the SW brain architecture in both aMCI and MD patients relative to the group of Healthy controls. This functional disorganization was also correlated with the participant's generic cognitive status. The deterioration of the network's organization was caused mainly by deficient local information processing as quantified by the mean cluster coefficient value. Functional hubs were identified through the normalized betweenness centrality metric. Analysis of the local characteristics showed relative hub preservation even with statistically significant reduced strength. Compensatory phenomena were also evident through the formation of additional hubs on left frontal and parietal regions. Our results indicate a declined functional network organization even during the prodromal phase. Degeneration is evident even in the preclinical phase and coexists with transient network reorganization due to compensation.


Long-term consequences of intraoperative spillage of bile and gallstones during laparoscopic cholecystectomy.
Laparoscopic cholecystectomy is associated with a higher incidence of iatrogenic perforation of the gallbladder than open cholecystectomy. The long-term consequences of spilled bile and gallstones are unknown. Data were collected prospectively from 1059 consecutive patients undergoing laparoscopic cholecystectomy over a 3-year period. Details of the operative procedures and postoperative course of patients in whom gallbladder perforation occurred were reviewed. Long-term follow-up (range 24 to 59 months) was available for 92% of patients. Intraoperative perforation of the gallbladder occurred in 306 patients (29%); it was more common in men and was associated with increasing age, body weight, and the presence of omental adhesions (each P < 0.001). There was no increased risk in patients with acute cholecystitis (P = 0.13). Postoperatively pyrexia was more common in patients with spillage of gallbladder contents (18% vs. 9%; P < 0.001). Of the patients with long-term follow-up, intra- abdominal abscess developed in 1 (0.6%) of 177 with spillage of only bile, and in 3 (2.9%) of 103 patients with spillage of both bile and gallstones, whereas no intra- abdominal abscesses occurred in the 697 patients in whom the gallbladder was removed intact ( P < 0.001). Intraperitoneal spillage of gallbladder contents during laparoscopic cholecystectomy is associated with an increased risk of intra-abdominal abscess. Attempts should be made to irrigate the operative field to evacuate spilled bile and to retrieve all gallstones spilled during the operative procedure.


Leukocyte diapedesis in vivo induces transient loss of tight junction protein at the blood-retina barrier.
Although much is now understood about the molecular structure of tight junctions (TJs), little is known about the regulation of their function during neural inflammatory disease processes in vivo. The mechanisms by which leukocytes transmigrate the blood-retina barrier (BRB) without affecting endothelial permeability are controversial. Confocal immunofluorescence microscopy of ex vivo retinal wholemounts was used to study BRB integrity during leukocyte adhesion and migration during experimental autoimmune uveoretinitis (EAU). Western blot analysis was used to measure levels of TJ proteins in EAU retina and RPE and in normal retina or RPE cultured with cytokines or chemokines. No evidence for discontinuity or other weakness of the endothelial or epithelial barrier at tricellular corners was observed, and maximum disruption of TJ protein expression was focused in retinal venules correlating with sites of leukocyte extravasation. Areas of maximum TJ protein loss in vivo also correlated with redistribution or loss of ensheathing astrocyte processes on venules but not adjacent capillaries or arterioles. Exposure of normal choroidal and retinal explants ex vivo to cytokines and chemokines alone did not downregulate total occludin-1 or claudin-3 TJ protein expression. The data presented herein support an active role for leukocytes in TJ disruption and blood-retina barrier (BRB) breakdown during retinal inflammation and further implicate venule microenvironment as a key factor in leukocyte recruitment to retinal tissue in vivo.


[Improved diagnostics in invasive aspergillosis and systematic monitoring in patients at high risk of infection].
Invasive aspergillosis remains an important cause of morbidity and mortality in patients with prolonged and severe immune suppression such as following haematopoietic stem-cell transplantation. Consensus definitions, which allow categorisation of patients based on diagnostic criteria, are an important improvement in uniform registration of invasive mycoses in clinical trials. Prospective monitoring of high-risk patients for the circulating aspergillus cell-wall component galactomannan, results in earlier diagnosis in two-thirds of patients when compared with conventional diagnostic methods. High-resolution CT (HRCT) enables the lesions characteristic of invasive mycoses to be detected earlier and better than by chest radiograph. In addition, invasive mycoses cause characteristic lesions on the HRCT scan including the halo-sign and the air-crescent sign. The pre-emptive management strategy which combines monitoring of patients for surrogate markers with a HRCT scan appears to be a promising approach to the early identification and treatment of patients with invasive aspergillosis.


Intracellular Ca2+ silences L-type Ca2+ channels in mesenteric veins: mechanism of venous smooth muscle resistance to calcium channel blockers.
Calcium channel blockers (CCBs) exert their antihypertensive effect by reducing cardiac afterload but not preload, suggesting that Ca(2+) influx through L-type Ca(2+) channels (LTCC) mediates arterial but not venous tone. The object of this study was to resolve the mechanism of venous resistance to CCBs. We compared the sensitivity of depolarization (KCl)-induced constriction of rat small mesenteric arteries (MAs) and veins (MVs) to the dilator effect of CCBs. Initial findings confirmed that nifedipine progressively dilated depolarization-induced constrictions in MAs but not MVs. However, Western blots showed a similar expression of the alpha(1C) pore-forming subunit of the LTCC in both vessels. Patch-clamp studies revealed a similar density of whole-cell Ca(2+) channel current between single smooth muscle cells (SMCs) of MAs and MVs. Based on these findings, we hypothesized that LTCCs are expressed but "silenced" by intracellular Ca(2+) in venous SMCs. After depletion of intracellular Ca(2+) stores by the SERCA pump inhibitor thapsigargin, depolarization-induced constrictions in MVs were blocked 80% by nifedipine suggesting restoration of Ca(2+) influx through LTCCs. Similarly, KCl-induced constrictions were sensitive to block by nifedipine after depletion of intracellular Ca(2+) stores by caffeine, ryanodine, or 2-aminoethoxydiphenyl borate. Cell-attached patch recordings of unitary LTCC currents confirmed rare channel openings during depolarization of venous compared to arterial SMCs, but chelating intracellular Ca(2+) significantly increased the open-state probability of venous LTCCs. We report that intracellular Ca(2+) inactivates LTCCs in venous SMCs to confer venous resistance to CCB-induced dilation, a fundamental drug property that was previously unexplained.


Novel 3,6-Dihydroxypicolinic Acid Decarboxylase-Mediated Picolinic Acid Catabolism in Alcaligenes faecalis JQ135.
Picolinic acid (PA), a typical C-2-carboxylated pyridine derivative, is a metabolite of l-tryptophan and many other aromatic compounds in mammalian and microbial cells. Microorganisms can degrade and utilize PA for growth. However, the precise mechanism of PA metabolism remains unknown. Alcaligenes faecalis strain JQ135 utilizes PA as its carbon and nitrogen source for growth. In this study, we screened a 6-hydroxypicolinic acid (6HPA) degradation-deficient mutant through random transposon mutagenesis. The mutant hydroxylated 6HPA into an intermediate, identified as 3,6-dihydroxypicolinic acid (3,6DHPA), with no further degradation. A novel decarboxylase, PicC, was identified to be responsible for the decarboxylation of 3,6DHPA to 2,5-dihydroxypyridine. Although, PicC belonged to the amidohydrolase 2 family, it shows low similarity (<45%) compared to other reported amidohydrolase 2 family decarboxylases. Moreover, PicC was found to form a monophyletic group in the phylogenetic tree constructed using PicC and related proteins. Further, the genetic deletion and complementation results demonstrated that picC was essential for PA degradation. The PicC was Zn2+-dependent nonoxidative decarboxylase that can specifically catalyze the irreversible decarboxylation of 3,6DHPA to 2,5-dihydroxypyridine. The Km and kcat toward 3,6DHPA were observed to be 13.44 μM and 4.77 s-1, respectively. Site-directed mutagenesis showed that His163 and His216 were essential for PicC activity. This study provides new insights into the microbial metabolism of PA at molecular level.IMPORTANCE Picolinic acid is a natural toxic pyridine derived from l-tryptophan metabolism and other aromatic compounds in mammalian and microbial cells. Microorganisms can degrade and utilize picolinic acid for their growth, and thus a microbial degradation pathway of picolinic acid has been proposed. Picolinic acid is converted into 6-hydroxypicolinic acid, 3,6-dihydroxypicolinic acid, and 2,5-dihydroxypyridine in turn. However, there was no physiological and genetic validation for this pathway. This study demonstrated that 3,6-dihydroxypicolinic acid was an intermediate in picolinic acid catabolism and further identified and characterized a novel amidohydrolase 2 family decarboxylase PicC. PicC was also shown to catalyze the decarboxylation of 3,6-dihydroxypicolinic acid into 2,5-dihydroxypyridine. This study provides a basis for understanding picolinic acid degradation and its underlying molecular mechanism.


Applications of multiple reaction monitoring targeted proteomics assays in human plasma.
Introduction: Multiple (or selected) reaction monitoring-mass spectrometry (MRM/SRM) is a targeted proteomic method that can be used for relative and absolute quantification. Multiple reports exist supporting the potential of the approach in proteomic biomarker validation. Areas covered: To get an overview of the applications of MRM in protein quantification in plasma, a search in MedLine/PubMed was performed using the keywords: 'MRM/SRM plasma proteomic/proteomics/proteome'. The retrieved studies were further filtered to focus on disease biomarkers and the main results are summarized. Expert opinion: MRM is increasingly employed for the quantification of both well-established but also newly discovered putative biomarkers and occasionally their post-translationally modified forms in plasma. Fractionation is regularly required for the detection of low abundance proteins. Standardized procedures to facilitate assay establishment and marker quantification have been proposed and, in few cases, implemented. Nevertheless, in most cases, absolute quantification is not performed. To advance, multiple technical issues including the regular use of standard labeled peptides and appropriate quality controls to monitor assay performance should be considered. Additionally, clinical aspects involving careful study design to address biomarker clinical use should also be considered.


In vivo induction of tyrosylprotein sulfotransferase by ethanol: role of increased enzyme synthesis.
Tyrosine sulfation is a posttranslational modification involved in the synthesis, secretion, and biological activity of proteins and peptides. Our previous studies have demonstrated that the enzyme activity was induced by ethanol. In the present work, the induction was studied in detail. Initial experiments were conducted to examine the time course of tyrosylprotein sulfotransferase (TPST) induction in rats pair-fed liquid diets containing either ethanol or carbohydrate substitute (controls). Marked elevation of TPST activity (3-fold) was measured on day 10 in the liver and gastric mucosa of ethanol-fed rats. Ethanol-mediated enhancement was also noticed by Western-blot analysis with anti-TPST antibody in both the liver and gastric mucosa on days 5 and 10. We then determined the steady-state TPST protein turnover in ethanol-fed and control animals that were given 35S-methionine after 10 days of pair-feeding with liquid diet. The rates of TPST synthesis assessed by measuring initial rates of incorporation of 35S-methionine into TPST was increased in the liver and gastric mucosa of animals fed with ethanol. Monophasic exponential decay curves showed that TPST protein half-lives for liver (control: 34 hr, ethanol: 32 hr) and gastric mucosa (control: 52 hr, ethanol: 48 hr) did not differ between control and ethanol groups. Our overall results indicate that the in vivo induction of TPST by ethanol involves increased enzyme synthesis rather than decreased enzyme degradation.


What is the state of the stone analysis techniques in urolithiasis?
To compare and evaluate the mostly used methods of urinary stone analysis. We searched PubMed and Google Scholar for "urolithiasis, nephrolithiasis, renal stone, and kidney stone" combined with "stone analysis, spectroscopy, X-ray diffraction, chemical analysis, mass spectrometry, and laser-induced breakdown spectroscopy, review article, and quality control assessment". We identified 24 articles about reviews of the principles of stone analysis techniques and their quality control trials. Seven articles were not in English language; hence, were omitted from this review. The remaining 17 articles and their related references were studied thoroughly. There are various chemical and physical techniques available for urinary stone analysis. The correct stone analysis has to identify not only all stone components, but also the molecular structure and crystalline forms of them with the exact quantitative determination of each component. The knowledge of urinary stone composition is important for understanding pathophysiology, choice of treatment modality, and prevention of recurrences of urolithiasis, but up to now, a standard method has not been defined. Although there are many techniques available for identifying the urinary stone composition and structure, no single method can provide all the requiring information. Therefore, a combination of structural and morphological tests is needed for this purpose.


[Update: new forms of therapy for type-2-diabetes].
Update: new forms of therapy for type-2-diabetes Abstract. In the past few years medical treatment of type-2-diabetes experienced fundamental changes. New medications were approved which have no intrinsic risk of hypoglycemia and exert weight loss. Cardiovascular outcome trials demonstrated positive effects on cardiovascular morbidity and mortality for GLP-1-receptor agonists and SGLT-2-inhibitors, the latter showing also specific nephroprotective effects. The growing bulk of data leads to modified therapy strategies: Persons with established cardiovascular disease or high cardiovascular risk should be treated primary with these medications. This review starts with an overview on newer antidiabetic substances (DPPIV-inhibitors, GLP-1-receptor agonists, SGLT-2-inhibitors). Then practical aspects of treatment regimens according to actual national and international guidelines are discussed.