Immunonutrition: Role in Wound Healing and Tissue Regeneration.
Significance: The role of immunonutrition in wound healing has been an area of both interest and controversy for many years. Although deficiencies in certain nutrients have long been known to impair healing, supplementation of specific immune modulating nutrients has not consistently yielded improvements in wound healing. Still, the prospect of optimizing nutrition to assist the immune system in wound repair bears great significance in both medical and surgical fields, as the costs of wound care and repair cannot be ignored. Recent Advances: Recent studies have rekindled efforts to elucidate the roles of specific immunonutrients, and we now have a better understanding of the conditionally essential role of various nutrients such as arginine, which becomes essential in certain clinical situations such as for the trauma patient or patients at high risk for malnutrition. Immunonutrition in its current formulation usually includes supplementation with arginine, glutamine, omega-3 fatty acids, vitamins, and trace minerals, and its use has often been associated with decreased infectious complications and sometimes with improvements in wound healing. Critical Issues: A key to understanding the role of immunonutrition in wound healing is recognizing the distinct contributions and importance of the various elements utilized. Future Directions: Critical areas for future study include identifying the specific populations, timing, and ideal composition of immunomodulating diets in order to optimize the wound healing process.


[Cause of mortality in schizophrenic patients: prospective study of years of a cohort of 150 chronic schizophrenic patients].
Overmortality in schizophrenic patients in comparison to the reference population has been found. At the present time this over mortality is mainly due to suicide or certain natural causes such as respiratory, cardio-vascular and cerebro-vascular diseases. In France there are not psychiatric cas registers that could allow us to study the mortality of psychiatric patients. The aim of the study was first to determine the standardized mortality ratio (SMR) in a group of 150 chronic schizophrenics followed during 8 years and secondly to detect the variables that could predict this mortality. The subjects filled out the RDC criteria for definite chronic schizophrenia and were included from 1991 to 1995. The subjects were inpatients or outpatients and their evaluation was made by psychiatrist. The subjects were selected from the different departments of two psychiatric hospitals corresponding to two French geographic areas (the Somme and Oise, two French "département"). At the initial assessment socio-demographic, clinical and psychometrical variables were collected: sex, age, educative level, number of hospitalizations, mean duration of the illness, scores on the Physical Anhedonia Scale, Brief Psychiatric Rating Scale (BPRS), Positive and Negative Syndrome Scale (PANSS). For the BPRS and PANSS, negative, positive and general subscales were extracted. In May 1999 all the subjects were contacted in order to know if they are alive or not and if they are death to know the date and the causes of their death. For the subjects that were still alive we used either direct assessment by interview of their psychiatrist or general practioner or indirect assessment by interview of their family. For the deceased subjects, we obtained informations about the date and the causes of the death by their psychiatrist or general practioner. If the patients were lost sight of we send a letter to the city of their place of birth in order to know if they are alive or not and if they are dead to know the date of their death. Moreover demographic data concerning the French and the Somme populations as well as the corresponding data concerning the mortality according to age and gender were obtained. A comparison of global mortality between patients and the French general or the Somme populations was made by the SMR. Moreover the deceased subjects and the survivors were compared by unidimensional statistical tests (chi 2 analyses for qualitative variables or Student's t test for quantitative variables) for the sociodemographic, clinical or psychometric variables. For each significant difference at p level < or = 0.05, the corresponding variable was retained for a multivariate step by step discriminant analysis. We found 13 deaths (10 males, 3 females): 3 suicides, 3 cardiac diseases, 2 cancers, 1 respiratory disease, 1 car crash, 1 homicide, 1 infectious disease, 1 respiratory arrest. The mortality rate (without correction for age) were 1.08% for both sexes, 1.44% and 0.6% for males and females respectively. The mortality rates (corrected for age) were 2.47% in our cohort and 0.988% in the Somme population. The corresponding SMR was 2.5. (chi 2 = 3.15, df = 1, p < 0.01). The overmortality was found only for males (chi 2 = 2.57, df = 1, p < 0.01) and not for females (chi 2 = 0.034, df = 1, p > 0.05). Concerning the comparisons between the deceased subjects and the survivors, there were five significant differences: gender, age, duration of the illness, neuroleptic dosage, negative symptoms (BPRS negative subscale). The deceased subjects were older, there was more men, the duration of the illness and the neuroleptic dosage were higher and the BPRS negative subscale was lower. These five variables were introduced in the discriminant analysis to explore notably their respecting weight. The corresponding power of the five variables were in decreasing order: neuroleptic dosage, negative symptoms, age, gender, duration of the illness. Our study confirm the overmortality in schizophrenic patients, this overmortality was especially explained by natural and non natural causes of death. The overmortality concerned only schizophrenic males patients whereas schizophrenic females did not have an overmortality. This negative result could be explain by a bias selection, the males being overrepresented in our cohort. Among the variables that were linked to the overmortality, the low level of negative symptomatology confirmed previous studies that have shown a low suicide rate in deficit schizophrenic. Moreover a high level of positive symptomatology could lead to high risk behaviors (suicide attempts, sexual disinhibition...). The neuroleptic dosage was the variable whom discriminate power was the highest. At least two explanations can be proposed. (ABSTRACT TRUNCATED)


Responses of regulatory and effector T-cells to low-dose interleukin-2 differ depending on the immune environment after allogeneic stem cell transplantation.
CD4+Foxp3+ regulatory T cells (Tregs) play a central role in the maintenance of immune tolerance after allogeneic hematopoietic stem cell transplantation (HSCT). Tregs promptly respond to low concentrations of IL-2 through the constitutive expression of high-affinity IL-2 receptors. It has been reported that low-dose IL-2 therapy increased circulating Tregs and improved clinical symptoms of chronic GVHD. Clinical studies of IL-2 therapy so far have mainly targeted patients in the chronic phase of transplantation when acute immune responses has subsided. However, the biological and clinical effects of exogenous IL-2 in an acute immune environment have not been well investigated. In the current study, we investigated the impact of exogenous IL-2 therapy on the post-transplant homeostasis of T cell subsets which influence the balance between GVHD and GVL in the acute phase, by setting the various immune environments early after HSCT in murine model. We initially found that 5,000 IU of IL-2 was enough to induce the active proliferation of Treg without influencing other conventional T cells (Tcons) when administered to normal mice. However, activated Tcons showed the response to the same dose of IL-2 in recipients after allogeneic HSCT. In a mild inflammatory environment within a threshold, exogenous IL-2 could effectively modulate Treg homeostasis with just limited influence to activated T cells, which resulted in an efficient GVHD suppression. In contrast, in a severely inflammatory environment, exogenous IL-2 enhanced activated T cells rather than Tregs, which resulted in the exacerbation of GVHD. Of interest, in an immune-tolerant state after transplant, exogenous IL-2 triggered effector T-cells to exert an anti-tumor effect with maintaining GVHD suppression. These data suggested that the responses of Tregs and effector T cells to exogenous IL-2 differ depending on the immune environment in the host, and the mutual balance of the response to IL-2 between T-cell subsets modulates GVHD and GVL after HSCT. Our findings may provide useful information in the optimization of IL-2 therapy, which may be personalized for each patient having different immune status.


Hyperbaric oxygen exposures at 3 and 4 atmospheres absolute pressure for experimental gas gangrene: succinate protection against oxygen toxicity.
The concurrent effect of succinate administration to protect against oxygen toxicity and of hyperbaric oxygen (HBO) exposures to treat model gas gangrene in mice was tested to determine whether succinate would interfere with the therapeutic efficacy of HBO. HBO (seven 90-min exposures) at 3 atmospheres absolute pressure (ATA) had been shown to reduce significantly the mortality of mice injected with Clostridium perfringens suspended in 10 mug of Adrenalin. When succinate was tested with this system, mortality of HBO-exposed infected animals was again significantly reduced (79% control mortality versus 17% HBO-exposed mortality), indicating that succinate does not interfere with the action of HBO. Exposures to 4 ATA of O(2) were evaluated in the same model clostridial infection with succinate used to prevent oxygen toxicity. Five 30-min exposures at 4 ATA reduced the mortality of infected animals (62% control versus 6% HBO-exposed mortality). Intraperitoneal succinate injections (10 mmoles/kg) were given 20 to 25 min prior to four of the seven 3-ATA exposures and before three of the five 4-ATA exposures. The intermittent succinate injections gave significant protection against the development of oxygen toxicity in infected and noninfected mice at both O(2) pressures. These studies support the potential clinical use of succinate or other oxygen-protective agents (i) to shorten HBO exposure times by using higher pressures to deliver the necessary O(2) dose, (ii) to increase the O(2) dose for difficult clinical situations by using maximal exposures at 4 ATA or more prolonged exposures at 2 to 3 ATA, and (iii) to continue HBO exposures in patients who require treatment but develop symptoms of oxygen toxicity.


Variables affecting the competency maintenance behaviors of occupational therapists.
The purpose of this study was to examine the relationship between work environment and the competency maintenance activities of occupational therapists. A survey of 121 occupational therapists in three states and interviews with 8 occupational therapists in Ohio were used. Analysis of survey data indicated that workplace support and the degree of competency monitoring are significant determinants of competency maintenance behavior. State continuing education requirements had no impact on reported levels of competency maintenance. Personal motivation of the therapist emerged as a potentially important moderator of the relationship between environment and competency maintenance. Although environmental factors enhance competency maintenance activities, active participation in competency maintenance is also related to the personal commitment of the individual therapist. Strategies that reduce barriers to effective competency maintenance and promote a culture of continuing competency in the profession merit further investigation.


CuFe2O4 magnetic nanocrystal clusters as a matrix for the analysis of small molecules by negative-ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
CuFe2O4 magnetic nanocrystal clusters (CuFe2O4 MNCs) were proposed as a new matrix for small molecule analysis by negative ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the first time. We demonstrated its advantages over conventional organic matrices in the detection of small molecules such as amino acids, peptides, nucleobases, fatty acids, and steroid hormones. A systematic comparison of CuFe2O4 MNCs with different ionization modes revealed that MS spectra obtained for the CuFe2O4 MNC matrix in the negative ion mode was only featured by deprotonated ion peaks with a free matrix background, which was different from the complicated alkali metal adducts produced in the positive ion mode. The developed method was found relatively tolerant to salt contamination and exhibited good reproducibility. A detection limit down to the subpicomolar level was achieved when testosterone was analyzed. In addition, by comparison of the MS spectra obtained from bare Fe3O4 and MFe2O4 MNC (M = Co, Ni, Cu, Zn) matrices, two main factors of MFe2O4 MNC matrices were revealed to play a vital role in assisting the negative ion desorption/ionization (D/I) process: doping transition metals into ferrite nanocrystals favoring laser absorption and energy transfer and a good match between the UV absorption of MFe2O4 MNCs and the excitation of nitrogen laser source facilitating LDI efficiency. This work creates a new branch of application for MFe2O4 MNCs and provides an alternative solution for small molecule analysis.


[Role of arthroscopy in the diagnosis of chronic monoarthritis of the knee joint].
The author analyzes the arthroscopy findings of 70 knee joints in 68 patients (35 men and 33 women) suffering from chronic monoarthritis of the knee joints with a disease standing from 3 months to 12 years. According to the x-ray data the overwhelming majority of the joints (54) showed stage 0-1, 15 stage II and 1 stage III of the pathological process. In accordance with the preliminary diagnoses 32 patients had synovitis of unclear genesis, 4 rheumatoid arthritis, 3 gonarthrosis deformans, 2 psoriatic arthritis, 5 patients were afflicted with Bechterew's disease, 1 with chondromatosis, and in 21 patients the diagnosis was doubtful. Arthroscopy promoted considerable improvement of the diagnosis, since rheumatoid arthritis including juvenile one was recognized in 12 patients, psoriatic arthritis in 8, reactive synovitis due to gonarthrosis in 16, Bechterew's disease in 12, Reiter's disease in 2, chondromatosis in 4, and pigmented villous-nodular synovitis in 3. The arthroscopic appearance of the disease remained unclear in 7 patients. Therefore, arthroscopy proved a highly effectual method that permits one to differentiate between reactive synovitis and genuine inflammatory diseases and to differentiate with a great probability between rheumatoid synovitis, psoriatic arthropathy and Bechterew's disease.


Shear bond strength of permanent 3D-printed resin and milled zirconia to primary teeth using different luting agents.
To evaluate the shear bond strengths (SBS) of permanent 3D-printed resin (PR) to primary dentin using different luting agents. 90 primary teeth were prepared. 45 cylinders (3 x 3 mm) were printed using PR, and 45 cylinders were milled using a Z block (to control). The cylinders were bonded to primary dentin by using three types of luting agent [glass-ionomer cement (GIC), resin-modified glass-ionomer cement (RMGIC), and self-adhesive resin cement (SRC)]. The SBS values of the specimens were calculated, and the fracture modes were examined. There was a statistically significant difference between the three different luting agents that were used to lute the PR to primary dentin (P< 0.001). Changing the material (PR or Z) did not affect the SBS values of the luting agents (P> 0.05). The adhesive failure between cement and dentin in the PR-SRC group was significantly higher than the other groups (P< 0.001). The SBS values of the newly developed PR to primary dentin with RMGIC and SRC were similar, but GIC showed lower values than the others. This laboratory study suggests that bond strength of the permanent 3D-printed resin can be like that of zirconia. As the resin-modified glass-ionomer cement and self-adhesive resin cement showed higher bond strength to primary teeth making the 3D-printed resin a treatment option.


Chronic Stressors and Adolescents' Externalizing Problems: Genetic Moderation by Dopamine Receptor D4. The TRAILS Study.
The existing literature does not provide consistent evidence that carriers of the Dopamine D4 Receptor 7-repeat allele are more sensitive to adverse environmental influences, resulting in enhanced externalizing problems, compared to noncarriers. One explanation is that the adverse influences examined in prior studies were not severe, chronic, or distressing enough to reveal individual differences in sensitivity reflected by DRD4-7R. This study examined whether the 7-repeat allele moderated the association between chronic stressors capturing multiple stressful aspects of individuals' lives and externalizing problems in adolescence. We expected that chronic stressor levels would be associated with externalizing levels only in 7-repeat carriers. Using Linear Mixed Models, we analyzed data from 1621 Dutch adolescents (52.2% boys), obtained in three measurement waves (mean age approximately 11, 13.5, and 16 years) from the TRacking Adolescents' Individual Lives Survey (TRAILS) population-based birth cohort and the parallel clinic-referred cohort. Across informants, we found that higher levels of chronic stressors were related to higher externalizing levels in 7-repeat carriers but not in noncarriers, as hypothesized. Although previous studies on the 7-repeat allele as a moderator of environmental influences on adolescents' externalizing problems have not convincingly demonstrated individual differences in sensitivity to adverse environmental influences, our findings suggest that adolescent carriers of the Dopamine D4 Receptor 7-repeat allele are more sensitive to chronic, multi-context stressors than noncarriers.


Refractive index of human articular cartilage varies with tissue structure and composition.
Optical properties of biological tissues, such as refractive index, are fundamental properties, intrinsically linked to a tissue's composition and structure. This study aims to investigate the variation of refractive index (RI) of human articular cartilage along the tissue depth (via collagen fibril orientation and optical density) and integrity (based on Mankin and Osteoarthritis Research Society International (OARSI) scores). The results show the relationship between RI and PG content (p=0.042), collagen orientation (p=0.037), and OARSI score (p=0.072). When taken into account, the outcome of this study suggests that the RI of healthy cartilage differs from that of pathological cartilage (p=0.072). This could potentially provide knowledge on how progressive tissue degeneration, such as osteoarthritis, affects changes in cartilage RI, which can, in turn, be used as a potential optical biomarker of tissue pathology.


Phase transition and compressibility in silicon nanowires.
Silicon nanowires (Si NWs), one-dimensional single crystalline, have recently drawn extensive attention, thanks to their robust applications in electrical and optical devices as well as in the strengthening of diamond/SiC superhard composites. Here, we conducted high-pressure synchrotron diffraction experiments in a diamond anvil cell to study phase transitions and compressibility of Si NWs. Our results revealed that the onset pressure for the Si I-II transformation in Si NWs is approximately 2.0 GPa lower than previously determined values for bulk Si, a trend that is consistent with the analysis of misfit in strain energy. The bulk modulus of Si-I NWs derived from the pressure-volume measurements is 123 GPa, which is comparable to that of Si-V NWs but 25% larger than the reported values for bulk silicon. The reduced compressibility in Si NWs indicates that the unique wire-like structure in nanoscale plays vital roles in the elastic behavior of condensed matter.


Differential gene expression of thyroid hormone receptor alpha and beta in fish development.
To understand the mechanisms of flounder metamorphosis, which is controlled by thyroid hormone, gene expression of the thyroid hormone receptors (TR alpha A, TR alpha B, TR beta 1, and TR beta 2) was studied in developing flounder larvae, using quantitative reverse-transcriptase polymerase chain reaction and in situ hybridization. TR gene transcripts were found at very low levels in fertilized flounder eggs. Substantial amounts of TR mRNAs were present in premetamorphic larvae, except for TR alpha B mRNA, which was low throughout larval development. TR alpha A gene transcripts increased rapidly in metamorphic climax and decreased rapidly postclimax. The expression level of TR beta s increased in climax, reached its peak postclimax, and remained high in metamorphosed juveniles. In situ hybridization confirmed the decrease in TR alpha transcripts in most tissues postclimax and further revealed the ubiquitous expression of TR genes and distinct tissue specificity of alpha and beta subtypes in the overall fish body. These results suggest that thyroid hormone exerts effects directly on each tissue during fish metamorphosis and that gene expression of TR subtypes is differentially regulated both temporally and regionally. Thus, the results suggest that the development of each tissue of the flounder by thyroid hormone is further controlled at the receptor level by the differential expression of TRs.


Plasma insulin levels in weaned pigs fed protein or energy restricted diets.
Two experiments were conducted with pigs to investigate changes in plasma insulin levels during protein-energy malnutrition. Three or four week old pigs were fed a control diet (18% protein), an energy-restricted diet (19% protein) or a low protein diet (6% protein) for 8 weeks. Energy restriction was achieved by feeding the control diet in amounts that allowed some growth, but only to an extent equal to the low protein diet. At the end of the restriction period, all pigs were fed the control diet for another 8 weeks. Blood samples from the superior vena cava were collected at intervals throughout the experimental period. Plasma samples were analyzed for insulin by radioimmunoassay. Protein restriction after weaning resulted in persistently low insulin levels during depletion and rehabilitation periods, while high levels of plasma insulin were observed in energy-restricted pigs only during the depletion period compared to pigs fed the control diet.


Multiple neurilemmomas of the cauda equina, cavernous hemangioma of the spinal cord, and degeneration of the lateral corticospinal tracts in a man with the clinical diagnosis of multiple sclerosis.
Clinical and neuropathological findings of a 72-year-old man are reported who was thought to have had multiple sclerosis for 44 years. The main clinical symptom was a slowly progressive spastic tetraparesis, but during the last years he also developed bilateral neurogenic deafness, dysfunction of the urinary bladder and loss of vibratory and positional sensation in the legs. Autopsy revealed a unique combination of neuropathological findings, namely 1) multiple neurilemmomas of the cauda equina with loss of nerve fibers in dorsal roots and Goll's tracts, 2) symmetric fiber loss in the lateral corticospinal tracts throughout the spinal cord, and 3) a cavernous hemangioma of the lower thoracic spinal cord. No changes of multiple sclerosis were found. It is postulated that the patient may have had both neurofibromatosis and a sporadic or familial form of primary lateral sclerosis. The hemangioma in the spinal cord is probably unrelated to these two conditions, although it cannot be excluded that it represents a manifestation of von Recklinghausen's disease.


ZmaR, a novel and widespread antibiotic resistance determinant that acetylates zwittermicin A.
ZmaR is a resistance determinant of unusual abundance in the environment and confers on gram-positive and gram-negative bacteria resistance to zwittermicin A, a novel broad-spectrum antibiotic produced by species of Bacillus. The ZmaR protein has no sequence similarity to proteins of known function; thus, the purpose of the present study was to determine the function of ZmaR in vitro. Cell extracts of E. coli containing zmaR inactivated zwittermicin A by covalent modification. Chemical analysis of inactivated zwittermicin A by 1H NMR, 13C NMR, and high- and low-resolution mass spectrometry demonstrated that the inactivated zwittermicin A was acetylated. Purified ZmaR protein inactivated zwittermicin A, and biochemical assays for acetyltransferase activity with [14C]acetyl coenzyme A demonstrated that ZmaR catalyzes the acetylation of zwittermicin A with acetyl coenzyme A as a donor group, suggesting that ZmaR may constitute a new class of acetyltransferases. Our results allow us to assign a biochemical function to a resistance protein that has no sequence similarity to proteins of known function, contributing fundamental knowledge to the fields of antibiotic resistance and protein function.


Clinical and histologic outcome in a dog surviving massive hepatic necrosis.
This report describes the clinical and histologic recovery of a 2-year-old mixed-breed dog presented with hypovolemic shock, markedly increased serum alanine amino transferase activity, and hemoabdomen. Emergency exploratory surgery revealed a friable liver with multiple capsule hemorrhages necessitating removal of the left lateral lobe. Histologic evaluation showed acute massive hepatic necrosis with centrilobular and midzonal distribution. The dog survived, and all monitored laboratory values normalized within 7 weeks. A liver biopsy taken 8 weeks after presentation revealed normal hepatic architecture with a few, randomly distributed neutrophilic foci. Follow-up included intermittent determination of liver variables including liver function tests for a period of 7 years. The dog's health status, and all test results remained normal during this time. Complete recovery and good long-term quality of life after life-threatening acute liver failure secondary to massive hepatic necrosis is possible in dogs.


Fixed and mobile-bearing total ankle prostheses: Effect on tibial bone strain.
Total ankle replacement is associated to a high revision rate. To improve implant survival, the potential advantage of prostheses with fixed bearing compared to mobile bearing is unclear. The objective of this study was to test the hypothesis that fixed and mobile bearing prostheses are associated with different biomechanical quantities typically associated to implant failure. With a validated finite element model, we compared three cases: a prosthesis with a fixed bearing, a prosthesis with a mobile bearing in a centered position, and a prosthesis with mobile bearing in an eccentric position. Both prostheses were obtained from the same manufacturer. They were tested on seven tibias with maximum axial compression force during walking. We tested the hypothesis that there was a difference of bone strain, bone-implant interfacial stress, and bone support between the three cases. We also evaluated, for the three cases, the correlations between bone support, bone strain and bone-implant interfacial stress. There were no statistically significant differences between the three cases. Overall, bone support was mainly trabecular, and less effective in the posterior side. Bone strain and bone-implant interfacial stress were strongly correlated to bone support. Even if slight differences are observed between fixed and mobile bearing, it is not enough to put forward the superiority of one of these implants regarding their reaction to axial compression. When associated to the published clinical results, our study provides no argument to warn surgeons against the use of two-components fixed bearing implants.


Myelodysplastic syndromes in childhood: report of 49 patients from a French multicentre study. French Society of Paediatric Haematology and Immunology.
We describe the clinical, cytological and cytogenetic features of 49 cases of myelodysplastic syndromes (MDS) in childhood. Three children had received prior cytotoxic treatment (group 1); all of these had cytogenetic abnormalities and died shortly after diagnosis. 22 children had constitutional anomalies (group 2). The remaining 24 MDS were considered as 'primary' (group 3). Hypoplastic marrow was found in nine cases, and only 53% of the MDS fitted the adult FAB classification. Transformation to AML occurred in 11 cases, development of aplastic anaemia in three cases, and spontaneous remission in one case each of RA and RAEB. Differences were observed between groups 2 and 3 in terms of mean age at diagnosis (11.1 months v 5 years), rate of cytogenetic anomalies (15% v 38%) and rate of progression towards acute leukaemia (13% v 29%). In group 2, all the fur girls studied exhibited a polyclonal pattern of X-inactivation, which suggests that MDS may be only the haematological expression of an embryological defect with different target tissues. This study suggests that some MDS in childhood can exhibit particular features such as congenital anomalies associated with MDS, bone marrow hypoplasia, polyclonality, and spontaneous remission. It emphasizes that the FAB classification is not adequate for children and addresses the question of whether these MDS are always malignant diseases.


Therapeutic Effect of Gypenosides on Nonalcoholic Steatohepatitis via Regulating Hepatic Lipogenesis and Fatty Acid Oxidation.
Nonalcoholic steatohepatitis (NASH) is the most frequent cause of liver dysfunction and a common global problem. Gypenosides can decrease pathological modifications of high-fat diet-induced rat atherosclerosis; however, its effect and mechanism on NASH remain unclear. In this study, rats were randomly divided into normal control and model groups. Model rats were fed with a high-fat diet and treated with gypenosides, rosiglitazone, or water for 6 weeks. We found that liver tissues showed significant hepatic steatosis and vacuolar degeneration with significantly higher triglyceride (TG), free fatty acid (FFA) and malonyl CoA, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT) activities in model group versus normal control group (p<0.01). Liver tissue mRNA and protein levels of sterol regulatory element binding protein-1c (SREBP-1c), carbohydrate response element binding protein (ChREBP), acetyl-CoA carboxylase (ACCase), and stearoyl CoA desaturase enzyme 1 (SCD1) were significantly increased, while the carnitine palmitoyl transferase-1 (CPT-1) level was significantly decreased in the model group versus the normal control group (p<0.01). Pathological changes of hepatic steatosis; body weight and liver wet weight; liver tissue TG, FFA and malonyl CoA concentrations; serum ALT, AST and GGT activities; liver tissue mRNA and protein levels of SREBP-1c, ChREBP, ACCase, and SCD-1 were significantly decreased; protein and mRNA levels of CPT-1 were significantly increased in the gypenosides group versus model group (p<0.01). In conclusion, gypenosides has therapeutic effect on NASH through regulating key transcriptional factors and lipogenic enzymes involved in fatty acid oxidation during hepatic lipogenesis.


Retrovirus-mediated gene transfer of 6-pyruvoyl-tetrahydropterin synthase corrects tetrahydrobiopterin deficiency in fibroblasts from hyperphenylalaninemic patients.
Tetrahydrobiopterin (BH4) deficiency, a variant form of hyperphenylalaninemia with progressive neurological dysfunction, is primarily caused by autosomal recessive mutations in the gene encoding the 6-pyruvoyl-tetrahydropterin synthase (PTPS). PTPS is a biosynthetic enzyme for the BH4 co-factor, and its deficiency is associated with a malfunction of the phenylalanine catabolism in the liver and a lack of biogenic amine neurotransmitters dopamine and serotonin in the brain. We have previously isolated the wild-type PTPS cDNA and identified several mutations responsible for a decreased enzyme in patients. This study reports the in vitro correction of BH4 deficiency by using retrovirus mediated transfer of the PTPS cDNA into primary fibroblast cultures established from different patients. The Bing packaging cell line was used for amphotropic virus production. Following PTPS gene transfer, stimulation with cytokines restored biosynthesis of BH4 in originally defective cells to values comparable to those of heterozygous fibroblasts from clinically healthy subjects. These results not only provide a direct proof that the mutations in PTPS were causative for the mutant phenotype, but they are also the first step toward gene therapy as a potential alternative approach to treat BH4 deficiency.


The Update of NGAL in Acute Kidney Injury.
Acute kidney injury (AKI) is one of the most common complications of various serious conditions, and early diagnosis is therefore critical for the treatment of AKI. Increase of the classic AKI biochemical markers such as serum creatinine is not evident until renal function is irreversibly damaged, which adds to the difficulties of early identification of AKI and results in an increase of the mortality rate. In order to improve the prevention, diagnosis, treatment, and prognosis prediction of AKI, novel early markers of AKI are required. Recent evidence demonstrates that neutrophil gelatinase- associated lipocalin (NGAL) is closely associated with AKI. Several experimental and clinical studies have shown that the expression of urine and serum NGAL increases significantly in AKI. In particular, the urine NGAL level is closely associated with the severity of kidney injury, and could be detected earlier than other AKI markers. Therefore, NGAL shows potential to be a new effective early biochemical marker of AKI. Further studies are needed to confirm the significant advantages of NGAL in the diagnosis of early AKI and its value in clinical applications.


Evaluation of bacterial interference and beta-lactamase production in management of experimental infection with group A beta-hemolytic streptococci.
The in vivo effects of penicillin and cefprozil therapy on the interaction between organisms commonly recovered from inflamed tonsils were studied by using a subcutaneous abscess model in mice. These organisms were group A beta-hemolytic streptococci (GABHS), Streptococcus salivarius (which is capable of interfering with GABHS), and Staphylococcus aureus. In mice infected with GABHS and S. salivarius alone or in combination, penicillin eliminated both organisms and cefprozil eliminated GABHS and S. aureus but not S. salivarius. Penicillin did not, however, reduce the number of GABHS or S. salivarius in the presence of S. aureus. The present study demonstrated the ability of beta-lactamase-producing S. aureus to protect GABHS from penicillin. However, no such protection was present following the administration of cefprozil. Furthermore, the preservation of S. salivarius that interferes with GABHS growth may provide protection from reinfection with GABHS. This study supports and provides an explanation for the increased efficacies of cephalosporins administered orally over that of penicillin when treating patients with acute GABHS pharyngitis or tonsillitis.


Association of caffeine to MDMA does not increase antinociception but potentiates adverse effects of this recreational drug.
Ecstasy (MDMA) street tablets often contain several other compounds in addition to MDMA, particularly caffeine. Then, it becomes necessary to study the consequences of caffeine plus MDMA combination. MDMA (1 mg/kg) elicited an analgesic response both at the spinal and supraspinal levels. However, when associated, MDMA and caffeine did not show any synergistic interaction. When caffeine was administered prior to MDMA, a potentiation of locomotor activity was observed, which consisted in an increase in maximal values and in a prolonged time of activity. In the neurotoxicity studies, a hyperthermic effect of MDMA was observed. Although caffeine alone failed to alter body temperature, it potentiated MDMA-induced hyperthermia. This association also significantly increased MDMA lethality (from 22% to 34%). Following administration of MDMA to rats, there was a persistent decrease in the number of serotonin transporter sites in the cortex, striatum and hippocampus, which was potentiated by caffeine co-treatment. This MDMA toxicity in rats was accompanied by a transient dopaminergic impairment in the striatum, measured as decreased [(3)H]WIN35428 binding sites, by 31% 3 days after treatment, which was not modified by caffeine. A transient down-regulation of 5-HT(2) receptors occurred in the cortex of MDMA-treated rats, whose recovery was slowed by co-treatment with caffeine. In conclusion, the association of MDMA with caffeine does not generate any beneficial effects at the antinociceptive level. The acute effects stemming from this association, in tandem with the final potentiation of serotonergic terminals injury, provide evidence of the potentially greater long-term adverse effects of this particular recreational drug combination.


Depression, diabetes, and chronic disease risk factors among US women of reproductive age.
Depression and chronic disease have implications for women's overall health and future pregnancies. The objective of this study was to estimate the prevalence and predictors of diabetes and chronic disease risk factors among reproductive-age women with depression. We used population-based data from the 2006, 2008, and 2010 Behavioral Risk Factor Surveillance System to examine prevalence of diabetes and prediabetes, binge and heavy drinking, smoking, overweight and obesity, and physical inactivity among 69,043 women aged 18 to 44 years with current major or minor depression, a past depression diagnosis, or no depression. In a multivariable logistic regression model, we calculated adjusted odds ratios (AORs) and 95% confidence intervals (CIs) of 1, 2, and 3 or more chronic disease risk factors by depression status. We found that 12.8% of reproductive-aged women experienced both current depression and 1 or more chronic disease risk factors. Compared to women with no depression, currently depressed women and those with a past diagnosis had higher prevalence of diabetes, smoking, binge or heavy drinking, obesity, and physical inactivity (P < .001 for all). Odds of 3 or more chronic conditions and risk factors were elevated among women with major (AOR, 5.7; 95% CI, 4.3-7.7), minor (AOR, 4.7; 95% CI, 3.7-6.1), and past diagnosis of depression (AOR, 2.8; 95% CI, 2.4-3.4). Depressed women of reproductive age have high rates of chronic disease risk factors, which may affect their overall health and future pregnancies.


Effects of Cryopreserved Amniotic Membrane-Umbilical Cord Allograft on Total Ankle Arthroplasty Wound Healing.
Relatively high rates of wound healing complications continue to be reported with a total ankle arthroplasty (TAA) anterior incision. The amniotic membrane-umbilical cord (AM-UC) allograft is a regenerative orthobiologic adjunct that modulates wound healing by down-regulating inflammation, enhancing local healing and antimicrobial factors, and reducing scar formation. The purpose of this study was to determine whether local application of a cryopreserved AM-UC allograft enhances soft tissue healing after TAA. A total of 104 patients with symptomatic ankle arthritis who failed conservative management underwent standard TAA. At skin closure, patients were allocated to either the treatment (local application of AM-UC) or control (no allograft) group. Demographic data, patient comorbidities, and radiographic findings were collected. The primary outcome was a major complication necessitating reoperation. Secondary outcomes were time to healing, minor complications (i.e., skin dehiscence, local wound care, use of antibiotics), and patient scar assessment. Local application of an AM-UC allograft significantly decreased the overall time to skin healing (28.5 days vs 40 days; p = .03). Two patients required a reoperation for soft tissue wound complications, with no difference (p = 1.00) between the groups. No statistically significant difference was detected in terms of skin dehiscence, local wound care, or antibiotic prescriptions in the 2 groups. Regenerative technology using local application of a cryopreserved AM-UC allograft may enhance TAA outcomes by decreasing the time to healing. Larger randomized controlled trials are needed to determine whether an AM-UC allograft enhances soft tissue wound healing and ultimately reduces the incidence of devastating soft tissue complications.


[Specificity of inhibition between Salmonella strains].
A phenomenon of inhibition among Salmonella organisms colonizing the gut of young animals can be demonstrated which is important in connection with immunization using live Salmonella vaccines. For practical utilization of this inhibition phenomenon, research into its specificity is very important. In vitro broth culture and animal experiments have shown inhibition to be genus-specific, i.e. there is no inhibition between strains of different genera of Enterobacteriaceae in most cases. The strongest inhibition occurs between isogenic (identical genotype) strains (strain-specific inhibition). This applies to Salmonella and other genera of Enterobacteriaceae. There was a close correlation between the degree of the inhibitory effect and the genetic relatedness of the strains, as has been demonstrated in Salmonella strains of different epidemiological classification. The inhibitory capacity was most pronounced among closely related strains. Clonal strains of Salmonella Enteritidis phage type 4 fully inhibited caecal colonization by Salmonella Enteritidis phage type 4 strains in 65% of the combinations tested in chicks. In broth culture experiments, the share of combinations with complete inhibition was above 90%. A significant inhibitory competence of a selected strongly inhibitory Salmonella Enteritidis phage type 4 strain against the majority of Salmonella Enteritidis field isolates can be expected. However, Salmonella Enteritidis strains of all phase types showed almost no inhibitory competence against other Salmonella serotypes.


Isotropic component trace analysis.
A new method for analyzing diffusion tensor imaging (DTI) of the brain, based on a recently introduced algorithm, lambda chart analysis (LCA), is presented. Pre-treatment of a given DTI data set with LCA, which effectively segregates isotropic and anisotropic components, allows for total removal of the anisotropic component from the DTI data set. The remaining pure isotropic component can therefore be subjected to further analysis similar to that applied in the trace histogram method. Deconvolution of the trace function yielded 3 Gaussian elements. Remapping of these 3 deconvoluted isotropic elements back onto the 2-dimensional image plane provided anatomical correlates of each element. The algorithm, referred to here as isotropic component trace analysis, can be used as a pictorial analytic tool, as well as a numerical analytical tool, for the noninvasive assessment of isotropic parenchymal components. The presented method provides quantitative indices of certain parenchymal parameters with better clarity than currently available methods. A ready-to-use program, EZ-LCA, for this powerful method is provided (available at http://coe.bri.niigata-u.ac.jp).


Robust Time-of-Arrival Location Estimation Algorithms for Wildlife Tracking.
Time-of-arrival transmitter localization systems, which use measurements from an array of sensors to estimate the location of a radio or acoustic emitter, are now widely used for tracking wildlife. Outlier measurements can severely corrupt estimated locations. This article describes a new suite of location estimation algorithms for such systems. The new algorithms detect and discard outlier time-of-arrival observations, which can be caused by non-line-of-sight propagation, radio interference, clock glitches, or an overestimation of the signal-to-noise ratio. The new algorithms also detect cases in which two locations are equally consistent with measurements and can usually select the correct one. The new algorithms can also infer approximate altitude information from a digital elevation map to improve location estimates close to one of the sensors. Finally, the new algorithms approximate the covariance matrix of location estimates in a simpler and more reliable way than the baseline algorithm. Extensive testing on real-world data involving mobile transmitters attached to wild animals demonstrates the efficacy of the new algorithms. Performance testing also shows that the new algorithms are fast and that they can easily cope with high-throughput real-time loads.


Radiation dose and image quality in pediatric chest CT: effects of iterative reconstruction in normal weight and overweight children.
New CT reconstruction techniques may help reduce the burden of ionizing radiation. To quantify radiation dose reduction when performing pediatric chest CT using a low-dose protocol and 50% adaptive statistical iterative reconstruction (ASIR) compared with age/gender-matched chest CT using a conventional dose protocol and reconstructed with filtered back projection (control group) and to determine its effect on image quality in normal weight and overweight children. We retrospectively reviewed 40 pediatric chest CT (M:F = 21:19; range: 0.1-17 years) in both groups. Radiation dose was compared between the two groups using paired Student's t-test. Image quality including noise, sharpness, artifacts and diagnostic acceptability was subjectively assessed by three pediatric radiologists using a four-point scale (superior, average, suboptimal, unacceptable). Eight children in the ASIR group and seven in the control group were overweight. All radiation dose parameters were significantly lower in the ASIR group (P < 0.01) with a greater than 57% dose reduction in overweight children. Image noise was higher in the ASIR group in both normal weight and overweight children. Only one scan in the ASIR group (1/40, 2.5%) was rated as diagnostically suboptimal and there was no unacceptable study. In both normal weight and overweight children, the ASIR technique is associated with a greater than 57% mean dose reduction, without significantly impacting diagnostic image quality in pediatric chest CT examinations. However, CT scans in overweight children may have a greater noise level, even when using the ASIR technique.


From land cover change to land function dynamics: a major challenge to improve land characterization.
Land cover change has always had a central role in land change science. This central role is largely the result of the possibilities to map and characterize land cover based on observations and remote sensing. This paper argues that more attention should be given to land use and land functions and linkages between these. Consideration of land functions that provide a wide range of goods and services makes more integrated assessments of land change possible. The increasing attention to multifunctional land use is another incentive to develop methods to assess changes in land functions. A number of methods to quantify and map the spatial extent of land use and land functions are discussed and the implications for modeling are identified based on recent model approaches in land change science. The mixed use of land cover, land use and land function in maps and models leads to inconsistencies in land change assessments. Explicit attention to the non-linear relations between land cover, land use and land function is essential to consistently address land change. New methods to map and quantify land function dynamics will enhance our ability to understand and model land system change and adequately inform policies and planning.


Proliferative Fasciitis in Childhood: A Review of Clinical Data Apropos of a Case.
Proliferative Fasciitis (PF) is a benign lesion with histologic and clinical features overlapping with those of malignant soft tissue tumours. Its occurrence in children is considered very rare. We present a case of PF appearing as a painful, red, gradually increasing in size lesion, during a period of a few weeks, on the finger of a five-year-old boy compromising the dermis and subcutaneous tissue. We were able to locate literature on 20 paediatric PF cases, which we review. Only five of these focus on the clinical data, the rest describing mainly histological findings. It is the first reported paediatric case appearing on the finger.


Caretaker-adolescent communication on sexual and reproductive health: a cross-sectional study in Unguja-Tanzania Zanzibar.
Caretakers/parents/caregivers/guardians play important roles in improving Sexual and Reproductive Health (SRH) of adolescents. Caretaker-adolescent sexual communication suggested to influence young people's sexual behaviours. Despite this significance, the communication is believed to be low in Unguja due to the increase of risky sexual behaviours among adolescents. This study assessed the pattern of such communication using IMB model as a framework. This is a cross-sectional study targeted caretakers of adolescents aged 15-19. One thousand caretakers of adolescents were interviewed using structured questionnaire. Comparison between male and female caretakers on discussing different SRH topics to both sexes of adolescents was made. The mean-score difference of overall communication was examined using Univariate analysis of variance (ANOVA). Bivariate correlation and simple path analysis via regression was conducted to determine the association of IMB variables in relation to communication practice. This study finds 40.7% of caretakers had ever communicated with their adolescents on SRH matters and 9.2% reported to have had communicated in the past 30 days. The weighted topic measure revealed only 26.5% of caretakers communicated with their adolescents. Both caretakers communicated more with their female adolescents. The communication was more common between same sex and between caretakers and their biological adolescents (p < 0.000). Both male and female caretakers mostly discussed sexual abstinence to female adolescents while to male adolescents, HIV/STIs was mostly discussed by female caretakers and pregnancy by male caretakers. The least discussed topics to both sexes are safer sex and other contraceptives use. The bivariate correlations suggested that IMB constructs were inter-related and associated with communication practice. Caretakers-adolescents communication on SRH in Unguja is low and it is not comprehensive. Caretakers fail to communicate with their adolescents on sensitive issues but do so on less sensitive ones. The pattern of communication found to vary across gender of caretaker and that of adolescent and depends on the nature of relationship between caretaker and adolescent. There is gender differences in selecting SRH topics of discussion. Interventions programmes have to include strategies that enhance caretaker's information, motivation and skills so as to improve SRH communication between caretakers and adolescent.


[Streptococcus suis meningitis in pig farmers: report of first two cases in Chile].
Human infection by Streptococcus suis is a zoonosis with a known occupational risk. Meningitis is its most frequent clinical manifestation. We present the first two cases in Chile. 54-year-old female patient, pig-farmer. She presented headache, vomiting, confusion and meningismus. She presented septic shock. Second case: 48-year-old male patient, also pig farmer, presented headache, vomiting and meningismus. A Gram's staining of cerebrospinal fluid (CSF) showed gram-positive cocci in both cases. Ceftriaxone and dexamethasone treatment was administered. The CSF cultures were positive for Streptococcus suis serotype 2. The patients experienced a good outcome, without neurological sequelae at the time of discharge. It is considerable to evaluate epidemiologic factors in order to suspect this etiological agent in cases of meningitis. These cases enhance the need of heighten awareness of potential for occupational exposure and infection by this emerging human pathogen. Educating population at risk about simple preventive measures must be considered.


A pediatric patient with interstitial pneumonia due to enterovirus D68.
Enterovirus D68 (EV-D68) infection is associated with upper and lower respiratory tract symptoms such as fever, cough, and wheezing. Pediatric patients with EV-D68 infection easily develop more severe respiratory complications compared to patients infected with other species of enterovirus, and consequently, have a higher rate of hospitalization and admission to intensive care units. Therefore, the clinical picture of respiratory complications associated with EV-D68 infection needs to be elucidated. Here, we report a 4-year-old girl of EV-D68 infection that required artificial respiration management within 24 h from the onset of cold symptoms. The patient was diagnosed with interstitial pneumonia on the basis of chest imaging findings with patchy, funicular and frosted glassy shadows, increased blood markers of surfactant protein-A, surfactant protein-D and sialylated carbohydrate antigen KL-6, and increased neutrophils and lymphocytes in the bronchoalveolar lavage. Steroids showed a remarkable effect in her treatment. Further investigations are needed to confirm the efficacy of steroids for interstitial pneumonia due to EV-D68 infection. As rapid deterioration of respiratory status is observed in EV-D68 infection, the possibility of interstitial pneumonia may be considered.


A Nonlinear Reduced-Order Model of the Corpus Callosum Under Planar Coronal Excitation.
Traumatic brain injury (TBI) is often associated with microstructural tissue damage in the brain, which results from its complex biomechanical behavior. Recent studies have shown that the deep white matter (WM) region of the human brain is susceptible to being damaged due to strain localization in that region. Motivated by these studies, in this paper, we propose a geometrically nonlinear dynamical reduced order model (ROM) to model and study the dynamics of the deep WM region of the human brain under coronal excitation. In this model, the brain hemispheres were modeled as lumped masses connected via viscoelastic links, resembling the geometry of the corpus callosum (CC). Employing system identification techniques, we determined the unknown parameters of the ROM, and ensured the accuracy of the ROM by comparing its response against the response of an advanced finite element (FE) model. Next, utilizing modal analysis techniques, we determined the energy distribution among the governing modes of vibration of the ROM and concluded that the demonstrated nonlinear behavior of the FE model might be predominantly due to the special geometry of the brain deep WM region. Furthermore, we observed that, for sufficiently high input energies, high frequency harmonics at approximately 45 Hz, were generated in the response of the CC, which, in turn, are associated with high-frequency oscillations of the CC. Such harmonics might potentially lead to strain localization in the CC. This work is a step toward understanding the brain dynamics during traumatic injury.


Migration systems in Europe: evidence from harmonized flow data.
Empirical tests of migration systems theory require consistent and complete data on international migration flows. Publicly available data, however, represent an inconsistent and incomplete set of measurements obtained from a variety of national data collection systems. We overcome these obstacles by standardizing the available migration reports of sending and receiving countries in the European Union and Norway each year from 2003-2007 and by estimating the remaining missing flows. The resulting harmonized estimates are then used to test migration systems theory. First, locating thresholds in the size of flows over time, we identify three migration systems within the European Union and Norway. Second, examining the key determinants of flows with respect to the predictions of migration systems theory, our results highlight the importance of shared experiences of nation-state formation, geography, and accession status in the European Union. Our findings lend support to migration systems theory and demonstrate that knowledge of migration systems may improve the accuracy of migration forecasts toward managing the impacts of migration as a source of social change in Europe.


Sensitivity of a clinical scale of balance and gait in frail nursing home residents.
The purpose of this paper was to describe this application, and to assess the sensitivity of the application when evaluating clinical interventions for improving balance and gait. The records of fifteen consecutive patients referred to physical therapy for mobility problems or recent falls were reviewed for this study. The subjects were evaluated upon initial referral to physical therapy and after 30 days of treatment. Treatment was developed around the problems noted on initial evaluation and applied 5 days/week for 20 sessions. A Wilcoxon signed-rank comparison of the initial and 30 day re-evaluation of the balance and the gait assessment indicated that significant improvement had occurred in the balance scores (Z = -3.20, p = 0.001) and the gait scores (Z = -2.82, p = 0.005) in this group. These assessments are sensitive to clinical improvements in mobility among frail elders.


PSA3, a Protein on the Stromal Face of the Thylakoid Membrane, Promotes Photosystem I Accumulation in Cooperation with the Assembly Factor PYG7.
PSI is a large protein-pigment complex located in the thylakoid membrane in cyanobacteria, plants, and algae. Although the structure and components of PSI are well characterized, mechanisms that orchestrate its assembly are poorly understood. In this study, we discovered a novel nucleus-encoded protein, Photosystem I Assembly3 (PSA3), that is required for PSI accumulation. PSA3 is conserved among green photosynthetic eukaryotes but is lacking in cyanobacteria. Mutations in the psa3 gene cause the specific loss of PSI in Arabidopsis (Arabidopsis thaliana) and maize (Zea mays). Ribosome profiling and pulse-labeling analyses showed that chloroplast- encoded PSI subunits are synthesized at normal rates in psa3 mutants, indicating that PSA3 is involved in the biogenesis of PSI at a posttranslational step. PSA3 resides on the stromal face of the thylakoid membrane, where it is found in a complex that is slightly smaller than PSI. Structural predictions suggest that PSA3 binds a basic peptide in a manner that is sensitive to the oxidation state of Cys pairs flanking the predicted peptide binding groove. PSA3 and the previously described PSI biogenesis factor PYG7 interact in yeast two-hybrid and bimolecular fluorescence complementation assays, and they are found in thylakoid membrane complexes of similar size. These and other results indicate that PSA3 cooperates with PYG7 to promote the stable assembly of PSI, and that the PsaC subunit is likely to be the primary target of their action.


Ultrasound lung comets induced by repeated breath-hold diving, a study in underwater fishermen.
Pulmonary edema has been reported in breath-hold divers during fish-catching diving activity. The present study was designed to detect possible increases in extravascular lung water (EVLW) in underwater fishermen after a competition. Thirty healthy subjects were studied. They participated in two different 5-h fish-catching diving competitions: one organized in the winter (10 subjects) and one organized in the autumn (20 subjects). A questionnaire was used to record underwater activity and note respiratory problems. An increase in EVLW was investigated from the detection of ultrasound lung comets (ULC) by chest ultrasonography. Complementary investigations included echocardiography and pulmonary function testing. An increase in EVLW was detected in three out of 30 underwater fishermen after the competition. No signs of cardiovascular dysfunction were found in the entire population and in divers with an increase in the ULC score. Two divers with raised ULC presented respiratory disorders such as cough or shortness of breath. Impairment in spirometric parameters was recorded in these subjects. An increase in EVLW could be observed after a fish-catching diving competition in three out of 30 underwater fishermen. In two subjects, it was related to respiratory disorders and impairment in pulmonary flow.


X-ray microanalysis of mitochondrial calcium.
X-ray microanalytical techniques using wavelength dispersive (WDS) and energy dispersive spectrometry (EDS) indicate that Ca2+ is present in intramitochondrial granules which occur in myocardial cells following irreversible ischemic injury. Preliminary results using tissue prepared for routine electron microscopy suggest that the degree of calcium binding that occurs in the mitochondria increases with increased duration of ischemia. Free ions are leached out of the tissue during processing; hence, the role of ion redistribution in producing myocardial necrosis cannot be elicited from a study of this nature. However, with continued progress in the development of techniques of tissue preparation x-ray spectrometry may provide a means of assessing quantitative ion alterations that occur during ischemia.


The Subtype Selectivity in Search of Potent Hypotensive Agents among 5,5-Dimethylhydantoin Derived α1-Adrenoceptors Antagonists.
In order to find new hypotensive drugs possessing higher activity and better selectivity, a new series of fifteen 5,5-dimethylhydantoin derivatives (1-15) was designed. Three-step syntheses, consisting of N-alkylations using standard procedures as well as microwaves, were carried out. Crystal structures were determined for compounds 7-9. All of the synthesized 5,5-dimethylhydantoins were tested for their affinity to α1-adrenergic receptors (α1-AR) using both in vitro and in silico methods. Most of them displayed higher affinity (Ki < 127.9 nM) to α1-adrenoceptor than urapidil in radioligand binding assay. Docking to two subtypes of adrenergic receptors, α1A and α1B, was conducted. Selected compounds were tested for their activity towards two α1-AR subtypes. All of them showed intrinsic antagonistic activity. Moreover, for two compounds (1 and 5), which possess o-methoxyphenylpiperazine fragments, strong activity (IC50 < 100 nM) was observed. Some representatives (3 and 5), which contain alkyl linker, proved selectivity towards α1A-AR, while two compounds with 2-hydroxypropyl linker (11 and 13) to α1B-AR. Finally, hypotensive activity was examined in rats. The most active compound (5) proved not only a lower effective dose than urapidil but also a stronger effect than prazosin.


Cardiac rupture following blunt trauma.
A case of cardiac rupture following blunt trauma with no additional injuries is described. This can be a difficult diagnosis because of the presence of additional injuries and the lack of clinical symptoms. Here, there was a rupture of the pericardium, all chambers of the heart and the thoracic aorta.


Neuronal HuD gene encoding a mRNA stability regulator is transcriptionally repressed by thyroid hormone.
Many genes governed by thyroid hormone (T3) lack binding sites for its receptor (TR) and are thought to be post-transcriptionally regulated by T3. Here we demonstrate that the HuD gene, which encodes a neurone-specific protein that binds to mRNA and modulates its stability, is regulated by T3. HuD RNA and protein expression were strongly up-regulated in specific areas of the hypothyroid rat brain, and reduced by T3 in rat PC12 and mouse N2a cells containing appropriate TR levels. Furthermore, T3 inhibited the transcription of HuD in run-on assays. Finally, HuD protein bound with high affinity to two sequences in acetylcholinesterase mRNA, and ectopic HuD expression increased its abundance in N2a cells. This is the first report of a gene encoding an mRNA stability regulator that is under T3 control. The results suggest that HuD may mediate some T3 effects by altering the half-life of mRNAs for acetylcholinesterase and other genes.


Adaptive Nonparametric Kinematic Modeling of Concentric Tube Robots.
Concentric tube robots comprise telescopic precurved elastic tubes. The robot's tip and shape are controlled via relative tube motions, i.e. tube rotations and translations. Non-linear interactions between the tubes, e.g. friction and torsion, as well as uncertainty in the physical properties of the tubes themselves, e.g. the Young's modulus, curvature, or stiffness, hinder accurate kinematic modelling. In this paper, we present a machine-learning-based methodology for kinematic modelling of concentric tube robots and in situ model adaptation. Our approach is based on Locally Weighted Projection Regression (LWPR). The model comprises an ensemble of linear models, each of which locally approximates the original complex kinematic relation. LWPR can accommodate for model deviations by adjusting the respective local models at run-time, resulting in an adaptive kinematics framework. We evaluated our approach on data gathered from a three-tube robot, and report high accuracy across the robot's configuration space.


Cytogenetics of Triticum x Dasypyrum hybrids and derived lines.
Genomic in situ hybridization was used to study Triticum x Dasypyrum wide hybrids and derived lines. A cytogenetic investigation was carried out in progenies of (i) amphiploids derived from T. turgidum var. durum (T. durum; 2n = 14; genomes AABB) x D. villosum (2n = 14; genome VV), (ii) three-parental hybrids (T. durum x D. villosum) x T. aestivum (2n = 42, genomes A'A'B'B'D'D'), and (iii) T. aestivum aneuploid lines carrying D. villosum chromosomes or chromatin. The amphiploids derived from T. durum x D. villosum showed a stable chromosomal constitution, made up of 14 V chromosomes, 14 chromosomes carrying the wheat A genome and 14 chromosomes carrying the B genome. High karyological instability was observed in the progenies of three-parental hybrids ([T. durum x D. villosum] x T. aestivum). Plants having the expected 14 A chromosomes, 14 B chromosomes, 7 D chromosomes, and 7 V chromosomes were rather rare (4.5%). Many progeny plants (45.5%) had the hexaploid wheat genome with 42 chromosomes and lacked any detectable D. villosum chromatin. Other plants (50%) had 14 A chromosomes and 14 B chromosomes, plus variable numbers of D and V chromosomes, the former being better retained than the latter in most cases. Some T. aestivum lines carrying D. villosum chromosomes or chromatin, as the result of addition, substitution, or recombination events or even a combination of these karyological events, were found to be stable. Other lines were unstable, and these lines carried 1V, 3V, or 5V chromosomes or their portions. Substitution or recombination events where 1V chromosomes were involved could concern the homeologous counterparts in both the A and B and D genomes of wheat. No line could be recovered where the shorter arm of 3V chromosomes was present. Changes in the morphology and banding pattern of V chromosomes were observed in hybrids that did not carry the entire D. villosum complement. By comparing the results of our cytogenetic analyses with certain phenotypic characteristics of the lines studied, genes for discrete traits could be assigned to specific V chromosomes or V chromosome arms. From the frequency of V chromosomes that were involved in chromatin exchanges with or substituted for one of their homeologous counterparts in the A, B, and D wheat genomes, it was inferred that D. villosum belongs to the same phyletic lineage as T. urartu (donor of the A genome of wheat) and Aegilops speltoides (B genome), and that Ae. squarrosa (D genome) diverged earlier from D. villosum.


Low-grade meningioma showing nearly equal density with spinal fluid on radiographic images.
A 61-year-old woman had an intracranial tumour that was located on the falx. Meningioma was suspected and the tumour rapidly grew over 1 year. It showed nearly equal density with spinal fluid showing almost no enhancement on radiographic images, like microcystic meningioma. Successful removal of the tumour was achieved. Histopathologically, the tumour was diagnosed as low-grade meningioma. The meningioma had variable sized microcysts and the appearance of solid area was meningothelial meningioma. This is a rare radiographic image for meningothelial meningioma.


Telomere length in prospective and retrospective cancer case-control studies.
Previous studies have reported that shorter mean telomere length in lymphocytes was associated with increased susceptibility to common diseases of aging, and may be predictive of cancer risk. However, most analyses have examined retrospectively collected case-control studies. Mean telomere length was measured using high-throughput quantitative real-time PCR. Blood for DNA extraction was collected after cancer diagnosis in the East Anglian SEARCH Breast (2,243 cases and 2,181 controls) and SEARCH Colorectal (2,249 cases and 2,161 controls) studies. Prospective case-control studies were conducted for breast cancer (199 cases) and colorectal cancer (185 cases), nested within the EPIC-Norfolk cohort. Blood was collected at least 6 months prior to diagnosis, and was matched to DNA from two cancer-free controls per case. In the retrospective SEARCH studies, the age-adjusted odds ratios for shortest (Q4) versus longest (Q1) quartile of mean telomere length was 15.5 [95% confidence intervals (CI), 11.6-20.8; p-het = 5.7 x 10(-75)], with a "per quartile" P-trend = 2.1 x 10(-80) for breast cancer; and 2.14 (95% CI, 1.77-2.59; p-het = 7.3 x 10(-15)), with a per quartile P-trend = 1.8 x 10(-13) for colorectal cancer. In the prospective EPIC study, the comparable odds ratios (Q4 versus Q1) were 1.58 (95% CI, 0.75-3.31; p-het = 0.23) for breast cancer and 1.13 (95% CI, 0.54-2.36; p-het = 0.75) for colorectal cancer risk. Mean telomere length was shorter in retrospectively collected cases than in controls but the equivalent association was markedly weaker in the prospective studies. This suggests that telomere shortening largely occurs after diagnosis, and therefore, might not be of value in cancer prediction.


RENEWAL OF MULTIPLE PRECIPITIN PRODUCTION ON INJECTION OF ONE ANTIGEN IN RABBITS SUCCESSIVELY IMMUNIZED WITH MANY ANTIGENS.
Multiple precipitin formation may be induced in the rabbit by the successive introduction of different antigens, and many precipitins may exist simultaneously in the blood for some time. Precipitins no longer demonstrable in the blood of a rabbit subjected to multiple successive immunization may reappear on the injection of only one of the antigens previously injected. Whether an unused antigen would have the same effect has not been determined.


Structural neuroimaging findings in migraine patients with restless legs syndrome.
One out of three migraine patients might have accompanying restless legs syndrome (RLS). In our study, we aimed to compare the volumes of the brain structures of migraineurs with and without RLS. We had 37 female patients with migraine and 17 females as the control group. Nineteen migraineurs had no RLS (RLS0) and 18 migraineurs had comorbidity of RLS (RLS1). The volumes of the brain structures were obtained by manual measurements, volBrain, and voxel-based morphometry (VBM). Manually, we measured caudate and putamen volumes. We used age, years of education, depression, anxiety scores, and total intracranial volume as covariates. According to VBM analyses, the volumes of the left superior occipital gyrus and precuneus were increased, and the substantia nigra and cuneus were decreased in the RLS1 group compared with the RLS0 group. RLS1 patients had larger superior temporal gyrus, Brodmann area 38, and left insula, and RLS0 patients had larger Brodmann area 22, right superior temporal gyrus, and Heschl gyrus compared with controls. Migraine and RLS0 patients had a smaller corpus callosum anteriorly, whereas RLS1 patients had a smaller splenium. Caudate volumes were larger in migraine patients via the three techniques. There was a positive relation between the caudate and putamen volumes and attack frequency. Comorbidity of RLS might be a confounding factor in structural neuroimaging studies in migraine. Deficits in the visual network seem to be related to accompanying RLS; deficits in the auditory network are particularly related to migraine.


Bosentan inhibits transient receptor potential channel expression in pulmonary vascular myocytes.
Bosentan, a dual endothelin receptor blocker, has been used clinically to treat idiopathic pulmonary arterial hypertension (IPAH). However, the mechanism of its antiproliferative effect on pulmonary artery smooth muscle cells (PASMCs) remains unclear. A rise in cytoplasmic Ca2+ stimulates PASMC proliferation and the canonical transient receptor potential (TRPC) channels are an important pathway for Ca2+ entry during PASMC proliferation. Bosentan (20-50 microM) significantly inhibited endothelin-1- or platelet-derived growth factor (PDGF)-mediated PASMC growth and [3H]thymidine uptake. In PASMCs, endothelin-1 (1 microM) and PDGF (10 ng/ml) both upregulated protein expression of TRPC6, whereas bosentan markedly downregulated TRPC6 protein levels. Furthermore, TRPC6 expression in PASMCs from patients with IPAH was greater than in normal PASMCs, and the antiproliferative effect of bosentan was significantly enhanced in IPAH-PASMCs in comparison with normal PASMCs. These observations demonstrate that the antiproliferative effect of bosentan on PASMCs involves the downregulation of TRPC6 channels via a mechanism possibly independent of endothelin receptor blockade. The greater effect of bosentan on IPAH-PASMCs than on normal PASMCs suggests that increased TRPC6 expression and function may be involved in the overgrowth of PASMCs in patients with IPAH.


Photodamage: cause, clinical manifestations, and prevention.
It is now recognized that intrinsic aging changes are distinct from those caused by habitual exposure to the sun (photoaging). Not only are these sun-induced alterations unsightly, but they also serve as a marker for patients at risk for developing skin cancer. Because of increased leisure time and affluence in our society, and an emphasis on sunbathing and other outdoor recreational activities during the last few decades, photoaging and skin cancer are becoming an ever-increasing problem and source of concern for health care professionals. The purpose of this article is to review the cause, clinical manifestations, and implications of photodamage. Photoprotection and other preventive measures to minimize photoaging and the risk of skin cancer will be discussed.


Clinical management of fusion in primary mandibular incisors: a systematic literature review.
Objective: Dental anomalies occurring in deciduous teeth can affect the eruption of the permanent dentition and the occlusion stability. The occurrence of dental anomalies such as double teeth during the primary dentition in the daily practice might be frequent. The study aimed to qualitatively summarize the therapeutic management of double teeth in primary incisors.Material and Methods: A systematic review regarding the therapy of primary fused incisors in the mandible was performed and the obtained data were assessed according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. The following electronic databases were screened from 1st January, 1996 until 30th July, 2019: PubMed, Scopus, EBSCO and the archives of paediatric dental journals. The search terms were grouped in anatomic entity: (tooth OR teeth OR incisor), pathological condition: (fused OR fusion OR geminated OR double), intervention: (treatment OR intervention OR therapy OR prevention OR control OR management OR restoration), observed parameters: (primary dentition OR primary tooth OR primary teeth).Results: Ten articles met all inclusion criteria. The data disclosed the occurrence of double teeth in mandibular incisors. The main management of this clinical condition is either preventive or surgical involving the extraction of fused teeth, based on the deciduous nature of the teeth, the degree of caries and malocclusion development risk.Conclusion: An early diagnosis of dental anomalies is fundamental for the application of proper preventive strategies to avoid a potential malocclusion in permanent dentition and to maintain these teeth sound and caries-free until the eruption of the permanent dentition.


High-resolution mapping reveals links of HP1 with active and inactive chromatin components.
Heterochromatin protein 1 (HP1) is commonly seen as a key factor of repressive heterochromatin, even though a few genes are known to require HP1-chromatin for their expression. To obtain insight into the targeting of HP1 and its interplay with other chromatin components, we have mapped HP1-binding sites on Chromosomes 2 and 4 in Drosophila Kc cells using high-density oligonucleotide arrays and the DNA adenine methyltransferase identification (DamID) technique. The resulting high-resolution maps show that HP1 forms large domains in pericentric regions, but is targeted to single genes on chromosome arms. Intriguingly, HP1 shows a striking preference for exon-dense genes on chromosome arms. Furthermore, HP1 binds along entire transcription units, except for 5' regions. Comparison with expression data shows that most of these genes are actively transcribed. HP1 target genes are also marked by the histone variant H3.3 and dimethylated histone 3 lysine 4 (H3K4me2), which are both typical of active chromatin. Interestingly, H3.3 deposition, which is usually observed along entire transcription units, is limited to the 5' ends of HP1-bound genes. Thus, H3.3 and HP1 are mutually exclusive marks on active chromatin. Additionally, we observed that HP1-chromatin and Polycomb-chromatin are nonoverlapping, but often closely juxtaposed, suggesting an interplay between both types of chromatin. These results demonstrate that HP1-chromatin is transcriptionally active and has extensive links with several other chromatin components.


Cut it out! Thoracic surgeon's approach to pulmonary mucormycosis and the role of surgical resection in survival.
Mucormycosis portends a poor prognosis with mortality rates ranging from 50% to 70% in pulmonary mucormycosis (PM) and up to 95% in disseminated disease. However, detailed outcomes data have been lacking. It remains unknown how to identify patients who would benefit from surgical resection. We present our experience with patients undergoing surgical resection for PM, including an analysis of factors affecting postoperative survival. We also describe a thoracic surgeon's approach through illustrative cases. We conducted a single-centre retrospective study of all adult patients with PM who received antifungal therapy and underwent surgical resection or who received antifungal therapy alone at Stanford between January 2004 and June 2018. Twelve patients received antifungal therapy and underwent surgical resection and 13 patients received antifungal therapy alone. From infection onset to death (or right-censoring if still alive), patients who underwent surgical resection had a median survival of 406 days (mean, 561.3; range, 22-2510), and patients who received antifungal therapy alone had a median survival of 28 days (mean, 66.7; range, 8-447). In patients who underwent surgical resection, median postoperative survival time was 154 days (range, 11-2495), in-hospital mortality was 16.7%, and 1-year mortality was 50.0%. Age, primary disease, ASA status, extrapulmonary dissemination, laterality, multilobar involvement, number of lesions, largest lesion size, platelet count, surgical approach, type of resection or extent of resection were not significantly associated with postoperative survival. Surgical resection significantly increases survival and should be strongly considered for selected patients with PM.


Analogue transformations in physics and their application to acoustics.
Transformation optics has shaped up a revolutionary electromagnetic design paradigm, enabling scientists to build astonishing devices such as invisibility cloaks. Unfortunately, the application of transformation techniques to other branches of physics is often constrained by the structure of the field equations. We develop here a complete transformation method using the idea of analogue spacetimes. The method is general and could be considered as a new paradigm for controlling waves in different branches of physics, from acoustics in quantum fluids to graphene electronics. As an application, we derive an "analogue transformation acoustics" formalism that naturally allows the use of transformations mixing space and time or involving moving fluids, both of which were impossible with the standard approach. To demonstrate the power of our method, we give explicit designs of a dynamic compressor, a spacetime cloak for acoustic waves and a carpet cloak for a moving aircraft.


Oral anticoagulants for preventing stroke in patients with non-valvular atrial fibrillation and no previous history of stroke or transient ischemic attacks.
Non-valvular atrial fibrillation (AF) is associated with an increased risk of stroke. The objective of this review was to characterize the efficacy and safety of oral anticoagulation (OAC) with vitamin K antagonists for the primary prevention of stroke in patients with chronic AF. We searched the Cochrane Stroke Group Specialised Register of Trials (June 1999), MEDLINE database, and the database of the Antithrombotic Trialists Collaboration, as well as reference lists of relevant articles. All randomized controlled trials comparing the value of OAC versus control in patients with non-valvular chronic atrial fibrillation and no history of transient ischemic attack (TIA) or stroke. Trials for inclusion were independently selected by two reviewers who also extracted each outcome and double-checked the data. The Peto method was used for combining odds ratios. All analysis were, as far as possible, "intention-to-treat". Since the published results of four trials included 3-8% of participants with prior stroke or TIA, unpublished results excluding these participants were obtained from the Atrial Fibrillation Investigators. Of 2313 participants without prior cerebral ischemia from five trials, about half (n = 1154) were randomized to adjusted-dose OAC with an estimated mean INRs ranging between 2.0-2.6 during 1.5 years/participant average follow-up. Participant features and study quality were similar between trials. OAC was associated with large, highly statistically significant reductions in ischemic stroke (OR = 0.34, 95% CI 0.23 - 0.52), all stroke (OR = 0.39, 95% CI 0.26 - 0. 59), all disabling or fatal stroke (OR = 0.47, 95% CI 0.28 - 0.80), and the combined endpoint of all stroke, MI or vascular death (OR = 0.56, 95% CI 0.42 - 0.76). The observed rates of intracranial and extracranial hemorrhage not significantly increased by OAC therapy, but confidence intervals were wide. Adjusted-dose OAC (achieved INRs between 2-3) reduces stroke as well as disabling/fatal stroke for patients with nonvalvular AF, and these benefits were not substantially offset by increased bleeding among participants in randomized clinical trials. Limitations include relatively short follow-up and imprecise estimates of bleeding risks from these selected participants. For primary prevention in AF patients who have an average stroke rate of 4%/year, about 25 strokes and about 12 disabling fatal strokes would be prevented yearly for every 1000 given OAC.


Isomorphic emissive GTP surrogate facilitates initiation and elongation of in vitro transcription reactions.
The fastidious behavior of T7 RNA polymerase limits the incorporation of synthetic nucleosides into RNA transcripts, particularly at or near the promoter. The practically exclusive use of GTP for transcription initiation further compounds this challenge, and reactions with GTP analogs, where the heterocyclic nucleus has been altered, have not, to our knowledge, been demonstrated. The enzymatic incorporation of (th)GTP, a newly synthesized isomorphic fluorescent nucleotide with a thieno[3,4-d]pyrimidine core, is explored. The modified nucleotide can initiate and maintain transcription reactions, leading to the formation of fully modified and highly emissive RNA transcripts with (th)G replacing all guanosine residues. Short and long modified transcripts are synthesized in comparable yields to their natural counterparts. To assess proper folding and function, transcripts were used to assemble a hammerhead ribozyme with all permutations of natural and modified enzyme and substrate strands. The (th)G modified substrate was effectively cleaved by the natural RNA enzyme, demonstrating the isomorphic features of the nucleoside and its ability to replace G residues while retaining proper folding. In contrast, the (th)G modified enzyme showed little cleavage ability, suggesting the modifications likely disrupted the catalytic center, illustrating the significance of the Hoogsteen face in mediating appropriate contacts. Importantly, the ribozyme cleavage reaction of the emissive fluorescent transcripts could be followed in real time by fluorescence spectroscopy. Beyond their utility as fluorescent probes in biophysical and discovery assays, the results reported point to the potential utility of such isomorphic nucleosides in probing specific mechanistic questions in RNA catalysis and RNA structural analysis.


Kinetics and regulation of a polarized Na(+)-H+ exchanger from Caco-2 cells, a human intestinal cell line.
The kinetics and regulation of Na(+)-H+ exchange were studied using BCECF to measure pHi in Caco-2 cells grown on membrane filters. Na(+)-H+ exchange was defined as a Na(+)-dependent H+ efflux in response to an acid load imposed by an NH4Cl prepulse in the absence of added CO2. Na(+)-H+ exchange was present exclusively on the basolateral membrane, had a Kt (Na+) of 21 +/- 2 mM, and an ID50 for amiloride dependent on medium [Na+] with an apparent Ki for amiloride of 3 microM. Na(+)-H+ exchange rates had a greater than first-order dependence on intracellular [H+], suggesting the presence of an internal proton modifier site. Results also suggest that Na(+)-H+ exchange is kinetically inactivated at resting pHi (7.35 +/- 0.02), since neither removal of Na+ nor addition of amiloride affected resting pHi, although monensin alkalinized cells to pHi 7.6. To evaluate regulation of Na(+)-H+ exchange, cells were exposed to either forskolin, 1,9-dideoxyforskolin (a noncyclase-activating forskolin derivative), 8-BrcAMP, E. coli STa toxin, ionomycin, phorbol dibutyrate, or cellular shrinkage in hypertonic medium. Only forskolin and 1,9-dideoxyforskolin caused a significant change (inhibition) in Na(+)-H+ exchange rate. Experiments performed with the Ussing chamber-voltage clamp technique verified that forskolin, 8-BrcAMP, E. coli STa toxin, ionomycin, and phorbol dibutyrate increased transepithelial Isc, verifying that all the regulatory pathways tested were functional and responsive to agonists. Results suggested that the Isc was due to Cl- secretion, since no net transcellular Na+ or Cl- flux was detected in basal conditions, and the Isc response to forskolin was abolished by omission of serosal Cl-. Because forskolin, but not 1,9-dideoxyforskolin, increased both cellular cAMP and Isc, the inhibition of Na(+)-H+ exchange by forskolin derivatives was mediated by a mechanism not involving activation of adenylyl cyclase. In conclusion, Caco-2 cells use a basolateral Na(+)-H+ exchanger to regulate pHi, but this exchanger is not affected by cell shrinkage or second messenger pathways that regulate Na(+)-H+ exchangers in other cell systems.


Characterization of GLUT4 and calpain expression in healthy human skeletal muscle during fasting and refeeding.
Calpain-10 and calpain-3 and the diabetes ankyrin repeat protein (DARP) have all been linked to insulin resistance and type 2 diabetes. We set out to measure the expression of these genes in human skeletal muscle and relate them to functional measurements of insulin action during fasting (which induces insulin resistance) and refeeding (which reverses it). Ten healthy male volunteers underwent 48 h of starvation followed by 24 h of high carbohydrate refeeding. On three occasions, before and after starvation and after refeeding, subjects underwent a 16 min insulin tolerance test to quantify insulin sensitivity. Muscle biopsies were obtained before and after fasting and after refeeding for the analysis of calpain-10 and calpain-3, GLUT4 and DARP expression by Western blotting and real-time PCR. Fasting led to a marked reduction in whole body insulin sensitivity by approx. 45% (P<0.01) and skeletal muscle GLUT4 gene expression by approx. 40% (P<0.05). However, fasting had no effect on calpain-10 and calpain-3 mRNA or protein levels, or DARP mRNA expression. Refeeding only partly restored insulin sensitivity and GLUT4 gene expression to their pre-fast values, but did not effect the expression of calpain-10, calpain-3 or DARP. These findings demonstrate that in healthy non-diabetic humans induction of insulin resistance by fasting and its reversal by refeeding with a high CHO diet is mirrored by changes in skeletal muscle GLUT4 but not calpain-10 and calpain-3 expression.


First-In-Class Inhibitors Targeting the Interaction between Bacterial RNA Polymerase and Sigma Initiation Factor Affect the Viability and Toxin Release of Streptococcus pneumoniae.
Novel antimicrobial classes are in desperate need for clinical management of infections caused by increasingly prevalent multi-drug resistant pathogens. The protein-protein interaction between bacterial RNA polymerase (RNAP) and the housekeeping sigma initiation factor is essential to transcription and bacterial viability. It also presents a potential target for antimicrobial discovery, for which a hit compound (C3) was previously identified from a pharmacophore model-based in silico screen. In this study, the hit compound was experimentally assessed with some rationally designed derivatives for the antimicrobial activities, in particular against Streptococcus pneumoniae and other pathogens. One compound, C3-005, shows dramatically improved activity against pneumococci compared to C3. C3-005 also attenuates S. pneumoniae toxin production more strongly than existing classes of antibiotics tested. Here we demonstrate a newly validated antimicrobial agent to address an overlooked target in the hit-to-lead process, which may pave the way for further antimicrobial development.


Salmonella enterica isolated from infections in Australian livestock remain susceptible to critical antimicrobials.
Salmonella enterica is a zoonotic pathogen causing a variety of diseases in humans and animals. Many countries are reporting an increase in the prevalence of multidrug-resistant (MDR) S. enterica in food animals. The aim of this study was to determine whether S. enterica isolated from livestock in New South Wales, Australia, have similar resistance traits to those reported internationally. Salmonella enterica (n=165) from clinical infections in food animals between 2007 and 2011 were serotyped and tested for susceptibility to 18 antimicrobials. Also, 22 antimicrobial resistance genes (ARGs), 3 integrons and 18 plasmid replicon types were screened for using PCR. Most isolates (66.1%) remained susceptible to all antimicrobials; 8.5% of the isolates were resistant to four or more antimicrobials. Antimicrobials with the highest prevalence of resistance were sulfafurazole (28.5%), ampicillin (17.0%), tetracycline (15.8%) and trimethoprim (8.5%). There was no resistance to fluoroquinolones or third-generation cephalosporins. The most common ARGs were blaTEM (15.2%), sul2 (10.3%), tetB (9.1%), tetA (5.5%), aphA1 (4.8%) and dhfrV (4.8%). Class 1 integrons (7.9%) and IncFIIA (69.7%) were the most commonly detected integron and plasmid replicon types, respectively. Class 1 integrons were positively associated with MDR phenotypes and ARG carriage (P≤0.001). Internationally prominent MDR serovars associated with severe disease in humans (e.g. AmpC-positive Salmonella Newport) were not detected. Overall, the comparatively favourable resistance status of S. enterica in Australian livestock represents minimal public health risk associated with MDR strains and supports a conservative approach to the registration of antimicrobial drug classes in food-producing animals.


Cloning, sequencing and characterization of the 5'-flanking region of the human collagenase-3 gene.
Collagenase-3 (matrix metalloprotease-13) is a recently discovered human collagenase produced in normal articular cartilage chondrocytes and thought to be involved in the pathological process of osteoarthritis. We have sequenced and characterized 1.6 kb of the human collagenase-3 gene 5'-flanking region. The transcription start site was located 22 bp upstream from the ATG start codon. Sequence analysis of the 5'-flanking region revealed the presence of the consensus recognition sites for the TATA and CCAAT DNA-binding proteins, activator protein-1 and E26 transformation specific/polyoma virus enhancer, as well as three core motifs of hormone response elements. Transient transfection assays demonstrated that a small fragment of 133 bp, containing the activator protein-1 and E26 transformation specific/polyoma virus enhancer sites promoted transcription in normal and osteoarthritic human chondrocytes with significantly higher activity than the original 1.6 kb fragment. Nucleotide sequence comparison of the promoter region of human collagenase-3 revealed a stronger similarity to the mouse collagenase-1 promoter than to the human collagenase-1 promoter.


Environmental swabs as a tool in norovirus outbreak investigation, including outbreaks on cruise ships.
In this study, we investigated whether environmental swabs can be used to demonstrate the presence of norovirus in outbreak settings. First, a procedure was set up based on viral RNA extraction using guanidium isothiocyanate buffer and binding of nucleic acids to silica. Subsequently, environmental swabs were taken at 23 Dutch restaurants and four cruise ships involved in outbreaks of gastroenteritis. Outbreaks were selected based on clinical symptoms consistent with viral gastroenteritis and time between consumption of suspected food and onset of clinical symptoms (>12 h). Norovirus RNA was demonstrated by real-time reverse transcriptase PCR in 51 of 86 (59%) clinical specimens from 12 of 14 outbreaks (86%), in 13 of 90 (14%) food specimens from 4 of 18 outbreaks (22%), and in 48 of 119 (40%) swab specimens taken from 14 of 27 outbreaks (52%). Positive swab samples agreed with positive clinical samples in seven outbreaks, showing identical sequences. Furthermore, norovirus was detected on swabs taken from kitchen and bathroom surfaces in five outbreaks in which no clinical samples were collected and two outbreaks with negative fecal samples. The detection rate was highest for outbreaks associated with catered meals and lowest for restaurant-associated outbreaks. The use of environmental swabs may be a useful tool in addition to testing of food and clinical specimens, particularlywhen viral RNA is detected on surfaces used for food preparation.


Identification of a dimeric KDG aldolase from Agrobacterium tumefaciens.
Agrobacterium tumefaciens is a Gram-negative bacterium and causative agent of Crown Gall disease that infects a variety of economically important plants. The annotated A. tumefaciens genome contains 10 putative dapA genes, which code for dihydrodipicolinate synthase (DHDPS). However, we have recently demonstrated that only one of these genes (dapA7) encodes a functional DHDPS. The function of the other nine putative dapA genes is yet to be determined. Here, we demonstrate using bioinformatics that the product of the dapA5 gene (DapA5) possesses all the catalytic residues canonical to 2-keto-3-deoxygluconate (KDG) aldolase, which is a class I aldolase involved in glucose metabolism. We therefore expressed, purified, and characterized recombinant DapA5 using mass spectrometry, circular dichroism spectroscopy, analytical ultracentrifugation, and enzyme kinetics. The results show that DapA5 (1) adopts an α/β structure consistent with the TIM-barrel fold of KDG aldolases, (2) possesses KDG aldolase enzyme activity, and (3) exists as a tight dimer in solution. This study shows for the first time that dapA5 from A. tumefaciens encodes a functional dimeric KDG aldolase.


Reduced cortical gamma-aminobutyric acid levels in depressed patients determined by proton magnetic resonance spectroscopy.
Several lines of emerging evidence suggest that dysfunction of gamma-aminobutyric acid (GABA) systems is associated with major depression. However, investigation of this hypothesis is limited by difficulty obtaining noninvasive in vivo measures of brain GABA levels. In this study we used in vivo proton magnetic resonance spectroscopy to investigate the hypothesis that abnormalities in the GABA neurotransmitter system are associated with the neurobiologic processes of depression. The GABA levels were measured in the occipital cortex of medication-free depressed patients meeting DSM-IV criteria (n = 14) and healthy control subjects with no history of mental illness (n = 18) using a localized difference editing proton magnetic resonance spectroscopy protocol. An analysis of covariance was employed to examine the effects of depression, sex, and age. The depressed patients demonstrated a highly significant (52%) reduction in occipital cortex GABA levels compared with the group of healthy subjects. While there were significant age and sex effects, there was no interaction of diagnosis with either age or sex. This study provides the first evidence of abnormally low cortical GABA concentrations in the brains of depressed patients.