Assembly pathway of newly synthesized LamB protein an outer membrane protein of Escherichia coli K-12.
The assembly of newly induced LamB protein (phage lambda receptor) was investigated in an operon fusion strain of Escherichia coli, in which the lamB gene is expressed under lac promoter control. The induction kinetics both for total cellular and for cell surface-exposed LamB protein were studied by immunochemical detection methods, using two distinct antisera directed against detergent-solubilized LamB trimers and completely denatured LamB monomers, respectively. Anti-trimer antibodies recognized both monomers and trimers, whereas anti-monomer antibodies only reacted with monomers. Provided appropriate solubilization conditions were used, both antisera were able to immunoprecipitate intracellular mature LamB protein quantitatively. Following induction, the first LamB antigenic determinants were detected after 60 to 80 seconds; detection of the newly synthesized protein by anti-monomer antibodies slightly preceded that by anti-trimer antibodies, a finding that could be partly explained by the observation that anti-monomer antibodies recognized a larger fraction of nascent LamB than did anti-trimer antibodies. Exposure of antigenic determinants at the cell surface was delayed for 30 to 50 seconds with respect to their synthesis. Therefore, either translocation or conformational changes must be rate-limiting in the series of processes that eventually convert the newly synthesized protein into its mature outer membrane state. LamB protein was found to occur in at least three clearly distinguishable states. State I is the LamB monomer, state II corresponds to a metastable trimer that dissociates in sodium dodecyl sulphate above 60 degrees C, and state III is the state LamB trimer that dissociates in sodium dodecyl sulphate only at temperatures above 90 degrees C. The chase kinetics of these states showed that conversion of newly synthesized LamB monomers to stable LamB trimers occurred in two stages: state I monomers were chased into metastable state II trimers rapidly (t 1/2 = 20 s), whereas stabilization of state II trimers to state III trimers was a relatively slow (t 1/2 = 5.7 min) process. Based on our results, a timing sequence in the assembly of outer membrane LamB protein is proposed.


Protein synthesis in skeletal muscle of rats following starvation and refeeding.
Determination of protein synthesis in individual tissues is important to understand the changes in protein metabolism during catabolic states. Three methods based on different underlying assumptions were compared in assessing muscle protein synthesis during nutritional manipulation. Rats were nonstarved, starved for 1 or 3 days, or refed for 2 days after 3 days of starvation. The extensor digitorum longus (EDL) muscles from the two hindlegs were used for analysis. In one EDL muscle the concentration and size distribution of ribosomes as well as the incorporation of [14C]leucine into protein in a cell-free system were determined. The other EDL muscle was incubated as such and the incorporation of [14C]phenylalanine into protein was measured. The total ribosome concentration per milligram of DNA decreased to 65% on the third day of starvation and remained low after refeeding. The amount of polyribosomes in the percentage of total ribosomes fell to 90% on the first day of starvation, regained the initial level on the third day, and reached 110% upon refeeding. During refeeding amino acid incorporation into protein in a cell-free system decreased to 40% and that in intact muscle to 64% of the prestarvation level. Upon refeeding, the activity increased to or above the original values. The use of several different techniques in parallel to assess protein synthesis in skeletal muscle is recommended since it gives information about the factors involved in regulation of the translational process in intact mammalian tissues.


Excessive Seizure Clusters in an Otherwise Well-Controlled Epilepsy as a Possible Hallmark of Untreated Vitamin B6-Responsive Epilepsy due to a Homozygous PLPBP Missense Variant.
Recently, mutations in the PLPBP gene were described as a novel cause for vitamin B6-responsive epilepsy. We report the outcome in case of a male adolescent with a novel homozygous missense variant in PLPBP who was never treated with pyridoxine until the age of 16 years. He presented with only mild cognitive impairment and an early-onset, well-controlled epilepsy. In our patient, excessive seizure clusters and anxiety states occurred intermittently, suggesting that the combination might be a hallmark in untreated patients. Thus, mutations in PLPBP should be addressed even in adolescent patients with only mild learning disabilities and relatively good seizure control over the years.


Molecular bases of rod and cone differences.
In the vertebrate retina, rods and cones both detect light, but they differ in functional aspects such as light sensitivity and temporal resolution, and in some cell biological aspects. For functional aspects, both types of photoreceptors use a phototransduction cascade, consisting of a series of enzymatic reactions, to convert photon capture to an electrical signal. To understand the mechanisms underlying the functional differences between rods and cones at the molecular level, we compared biochemically, each of the reactions in the phototransduction cascades of rods and cones using the cells isolated and purified from carp retina. Although the cascade proteins are identical or are functionally similar between rods and cones, their activities together with their expression levels are mostly different. In general, reactions that generate a response are somewhat less effective in cones than in rods, but each of the reactions for termination and recovery of a response are much more effective in cones. These findings explain lower light sensitivity and briefer light responses in cones than in rods. In addition, our considerations suggest that a Ca2+-binding protein, S-modulin or recoverin, has a currently unnoticed role in shaping light responses. Upon comparison of the expression levels of proteins and/or mRNAs using purified cells, several proteins were found to be specifically or predominantly expressed in cones. These proteins will be of interest in future studies aimed at characterizing the differences between rods and cones.


A randomized, double-blind, placebo-controlled comparison of emedastine 0.05% ophthalmic solution with loratadine 10 mg and their combination in the human conjunctival allergen challenge model.
When selecting treatment for allergic conjunctivitis, a primary concern is whether to choose local or systemic therapy. This study compared the efficacy of topical emedastine 0.05% ophthalmic solution with that of oral loratadine 10 mg and their combination in the conjunctival allergen challenge model of allergic conjunctivitis. This was a single-center, randomized, double-masked, placebo-controlled, parallel-group study. At visit 1, eligible subjects underwent conjunctival allergen challenge to identify the dose required to elicit a positive allergic reaction. After 7 days, subjects returned for visit 2, at which the allergen dose was confirmed. At visit 3, which took place 2 weeks later, subjects were randomized to receive either emedastine plus placebo capsules, loratadine plus placebo eyedrops, or both emedastine and loratadine. One hour after receiving study drug, subjects were challenged with allergen in both eyes. Allergic signs and symptoms were graded using standardized 5-point scales. The primary efficacy variables were itching and conjunctival hyperemia. Secondary efficacy variables were ciliary and episcleral hyperemia, chemosis, lid swelling, and tearing. Itching was graded subjectively at 3, 5, and 10 minutes after challenge. All other variables were assessed at 5, 10, and 20 minutes after challenge. Eighty subjects (mean age, 43.68 years) were randomized to receive study treatment. Forty subjects (20 men, 20 women) received emedastine plus placebo capsules, 20 (7 men, 13 women) received loratadine plus placebo eyedrops, and 20 (12 men, 8 women) received both active treatments. In the between-group efficacy comparison at visit 3, the difference in itching and hyperemia scores between emedastine and loratadine was statistically significant at all time points (all, P < 0.05). Efficacy scores for the combination of emedastine and loratadine were significantly better than those for loratadine alone at 2 of 3 time points for itching and all time points for hyperemia (P < 0.05). The combination was significantly better than emedastine alone at I of 3 time points for itching and 6 of 9 time points for hyperemia (P < 0.05). In this study, emedastine was more efficacious than loratadine for reducing the itching and redness associated with allergic conjunctivitis in the human conjunctival allergen challenge model.


Sixteen-year follow-up of hyperopic laser in situ keratomileusis.
To assess the long-term efficacy and safety of hyperopic laser in situ keratomileusis (LASIK). Department of Ophthalmology, St. Thomas' Hospital, London, United Kingdom. Prospective cohort study. Hyperopic LASIK was performed using a mechanical microkeratome, an optical zone of 6.5 mm, and a blend zone of 1.5 mm. Simple hyperopia was treated. The mean patient age was 51.6 years (range 34 to 60 years). Preoperatively, the mean spherical equivalent (SE) was +3.74 diopters (D) (range +1.25 to +6.50 D). The mean attempted correction was +3.64 D (range +1.5 to +6.0 D). The mean follow-up was 16.5 years. The mean SE was +0.28 D (range -1.0 to +1.5 D) at 12 months, +0.84 D (range -0.75 to +3.35 D) at 5 years, and +1.74 D (range -0.75 to +4.13 D) at 16 years, representing an increase in hyperopia of +1.47 D ± 1.43 (SD) between 1 year and 16 years (P < .0001) and of +1.13 ± 0.8 D between 5 years and 16 years (P < .03). The uncorrected distance visual acuity improved at 16 years (P < .0001); corrected distance visual acuity (CDVA) was unchanged (P < .2). The efficacy index was 0.5 and the safety index, 1.09. One eye (3%) lost 2 lines of CDVA. Keratometry remained stable between 1 year and 16 years (P < 1.0). Four eyes (12%) had cataract surgery, and 2 (6%) had laser iridotomy. There was no ectasia. After hyperopic LASIK, an increase in hyperopia occurred between 1 and 5 years and 16 years. At 16 years, efficacy was limited but with no sight-threatening complications. Dr. Marshall was a consultant to Summit Technology, Inc. Dr. O'Brart holds a noncommercial research grant from Alcon Laboratories, Inc. None of the other authors has a financial or proprietary interest in any material or method mentioned.


Reducing Sedentary Behavior Versus Increasing Moderate-to-Vigorous Intensity Physical Activity in Older Adults.
To compare the effects of behavioral interventions targeting decreased sedentary behavior versus increased moderate-to-vigorous intensity physical activity (MVPA) in older adults. Inactive older adults ( N = 38, 68 ± 7 years old, 71% female) were randomized to 12-week interventions targeting decreased sedentary behavior ( Sit Less) or increased MVPA ( Get Active). The SenseWear armband was used to objectively assess activity in real time. Assessments included a blinded armband, the Community Health Activites Model Program for Senior (CHAMPS) questionnaire, 400-meter walk, and the Short Physical Performance Battery (SPPB). Objectively measured MVPA increased in Get Active (75 ± 22 min/week, p < .001); self-reported MVPA increased in both groups ( p < .05). Sedentary behavior did not change in either group (all p > .05). Only the Sit Less group improved the SPPB score (0.5 ± 0.3, p = .046). Targeting reduced sedentary behavior had a greater effect on physical function among inactive but high functioning older adults over 12 weeks. Future studies of longer duration and combining increased MVPA with reduced sedentary behavior are needed.


Diving into debate: Comparing discussion-based and single-presenter journal club formats in a large PM&R department.
Journal clubs in physical medicine and rehabilitation (PM&R) advance the educational mission by uniting colleagues to learn of literature updates, consider clinical applications, practice critical thinking, and engage in lively dialogue and community. Although discussion-based journal clubs have been shown to enhance quality, a model for their application across a large and clinically diverse department of PM&R has not been described, nor has one been evaluated in comparison to a single-speaker podium format. To develop a discussion-based PM&R department-wide journal club, present elements of the journal club model in a manner that would enable replication, and assess effectiveness as perceived by participants, compared to the prior (podium-based, single-speaker) format. It was hypothesized that a discussion-based journal club would more effectively achieve educational goals and would be perceived by participants to be of greater quality and value. Pre-post educational intervention study, using surveys of PM&R resident and faculty participants. Survey items used a 5-point Likert scale. Unpaired 2-tailed t-tests were used to compare the formats. A large academic PM&R department. PM&R faculty, residents, fellows: 26 respondents (preintervention) and 26 respondents (postintervention) out of a total of 94 and 98 people invited to participate, respectively. A discussion-based departmental journal club was designed and implemented, replacing the previous single-speaker, podium-based journal club. Pre- and post- intervention respondent ratings of (a) journal club quality and value, and (b) effectiveness in achieving specific educational goals. Compared to the traditional format, the discussion-based format more effectively met the educational objectives, was of higher quality and value as perceived by respondents, and increased desire to attend the activity. This discussion-based journal club format can serve as a model for academic PM&R programs looking to enhance the educational value of journal club.


Micro-computed tomography with iodine staining resolves the arrangement of muscle fibres.
We illustrate here microCT images in which contrast between muscle and connective tissue has been achieved by means of staining with iodine. Enhancement is shown to be dependent on the concentration of iodine solution (I(2)KI), time in solution and specimen size. Histological examination confirms that the arrangement of individual muscle fibres can be visualised on the enhanced microCT images, and that the iodine accumulates in the muscle fibres in preference to the surrounding connective tissues. We explore the application of this technique to describe the fibrous structure of skeletal muscle, and conclude that it has the potential to become a non-destructive and cost-effective method for investigating muscle fascicle architecture, particularly in comparative morphological studies.


Binding of calcium by organic anions, determined by perturbation of the equilibrium solubility of [14C]calcium oxalate.
Confirmation is needed of the reported binding of calcium ions (Ca2+) by bile salts, which is believed to decrease the activity of free calcium ions [Ca2+] available for precipitation of insoluble calcium salts of organic anions in pigment gallstones. We report a new method to determine the association constants (K'f) of calcium for organic anions, from the perturbation by the added anion of the equilibrium solubilization of calcium [14C]oxalate monohydrate crystals (CaOx*). CaOx* crystals were prepared by stepwise conversion of [14C]oxalic acid to its K+ and Ca2+ salts. Structure and purity were confirmed by X-ray diffraction of the crystals. CaOx* was incubated (37 degrees C, under N2) in 0.15 M NaCl in CO2-free deionized H2O at pH 6.3. Dissolution of CaOx*, estimated by radioassay of the 0.22-micron Millipore filtrate, attained equilibrium at 3 days, with K'sp = [Ca2+] * [Ox=] = 2.34 X 10(-8) M2, calculated using known affinity constants for the soluble complexes of NaOx- (K'NaOx = 3.215 M-1) and CaOx (K'CaOx = 195.0 M-1). Keeping total [Na+] = 0.15 M, we added anions that formed soluble complexes with Ca2+. This decreased free [Ca2+], causing more CaOx* to dissolve in amounts related to the concentration of added anion and its K'f for Ca2+. With this method, K'f values for citrate, malonate and malate were similar to the values we determined with the Ca2+ ion electrode, and to published values obtained with the Ca2+ ion electrode and other methods. The sensitivity of the CaOx method permits determination of K'f values with small quantities and low concentrations of the anions and calcium.


[Acute respiratory tract infections due to a human metapneumovirus in children: descriptive study and comparison with respiratory syncytial virus infections].
A new paramyxovirus, the human metapneumovirus was recently isolated. We report the first French cases collected between 2000 and 2002. Samples were obtained from nasopharyngeal aspirates from children hospitalised for acute respiratory tract infection in hospitals of Caen and Flers in Basse-Normandie. Human metapneumovirus was studied by polymerase chain reaction on negative samples for respiratory syncytial virus, influenza A and B virus, parainfluenza (1, 2 and 3) virus, adenovirus, coronavirus and rhinovirus. Comparison between metapneumovirus virus and respiratory syncytial virus infections was done after matching sex, age and infection month. Twenty-six human metapneumovirus infections were identified. A comparative study of a matched group of children infected by respiratory syncytial virus found no significative difference for hospitalisation motive, clinical criteria and treatment. The human metapneumovirus is responsible for typical acute bronchiolitis in children.


Dopamine abnormalities in the neocortex of patients with temporal lobe epilepsy.
Experiments were designed to evaluate different variables of the dopaminergic system in the temporal cortex of surgically treated patients with temporal lobe epilepsy (TLE) associated with mesial sclerosis (MTLE, n=12) or with cerebral tumor or lesion (n=8). In addition, we sought to identify dopaminergic abnormalities in those patients with epilepsy that had comorbid anxiety and depression. Specifically, we investigated changes in dopamine and its metabolites, D1 and D2 receptors, tyrosine hydroxylase (TH) and dopamine transporter. Results obtained from patients with epilepsy were compared with those found in experiments using autopsy material. The neocortex of patients with MTLE demonstrated high D1 expression (1680%, p<0.05) and binding (layers I-II, 31%, p<0.05; layers V-VI, 28%, p<0.05), and decreased D2 expression (77%, p<0.05). The neocortex of patients with TLE secondary to cerebral tumor or lesion showed high expression of D1 receptors (1100%, p<0.05), and D2-like induced activation of G proteins (layers I-II, 503%; layers III-IV, 557%; layers V-VI, 964%, p<0.05). Both epileptic groups presented elevated binding to the dopamine transporter and low tissue content of dopamine and its metabolites. Analysis revealed the following correlations: a) D1 receptor binding correlated negatively with seizure onset age and seizure frequency, and positively with duration of epilepsy; b) D2 receptor binding correlated positively with age of seizure onset and negatively with duration of epilepsy; c) dopamine transporter binding correlated positively with duration of epilepsy and frequency of seizures; d) D2-like induced activation of G proteins correlated positively with the age of patients. When compared with autopsies and patients with anxiety and depression, patients without neuropsychiatric disorders showed high D2-like induced activation of G proteins, an effect that correlated positively with age of patient and seizure onset age, and negatively with duration of epilepsy. The present study suggests that alterations of the dopaminergic system result from epileptic activity and could be involved in the physiopathology of TLE and the comorbid anxiety and depression.


Clinical features and prevalence of Klinefelter syndrome in transgender individuals: A systematic review.
Previous studies have suggested a higher prevalence of Klinefelter syndrome amongst transgender individuals. We undertook a systematic review to determine the prevalence of Klinefelter syndrome amongst transgender individuals presumed male at birth and summarize the clinical features and potential treatment implications for individuals with Klinefelter syndrome commencing gender-affirming hormone therapy. Using preferred reporting items for systematic review and meta-analysis guidelines, we searched EMBASE, MEDLINE and the Cochrane Central Register of Controlled Trials (CENTRAL) up to 31 December 2021. All studies reporting on the prevalence or clinical features of transgender individuals with Klinefelter syndrome were included. This study is registered with the International Prospective Register of Systematic Reviews, number CRD42021227916. Our search strategy retrieved 11 cohort studies comprising 1376 transgender individuals. In all, 14 of 1376 (1.02%) individuals were diagnosed with Klinefelter syndrome. Based on the seven studies in which karyotype was undertaken in all individuals, the prevalence is 9/1013 (0.88%; 95% CI, 0.41%-1.68%). Case reports highlight unique treatment considerations in this population, including azoospermia, venous thromboembolism, and monitoring of breast cancer and bone health. Compared to the general population, observational studies document a higher prevalence of Klinefelter syndrome amongst transgender individuals, though underdiagnosis in the general population limits conclusions. Routine karyotype in transgender people initiating gender-affirming hormone therapy is not supported unless clinical features of Klinefelter syndrome, such as small testicular volume, or hypergonadotropic hypogonadism are present. Transgender individuals with Klinefelter syndrome need to manage a unique risk profile if they desire feminizing gender-affirming hormone therapy.


Evidence for calcitonin-producing cells in human lingual thyroids.
The thyroid contains two types of cells, the thyroid follicular cells and the calcitonin-producing cells. The site of origin of the thyroid follicular cells is the median thyroid anlage, an endothelial diverticulum in the midline of the ventral pharynx between the first and the second pharyngeal pouches. The ultimobranchial bodies (UBB), a pair of transient embryonic structures evaginated from the fourth pharyngeal pouch and located symmetrically on the sides of the developing neck, are the source of calcitonin-producing cells. In human embryos, the thyroid bud starts its migration at embryonic day 24 and reaches its final location in front of the trachea at embryonic day 45-50. The UBB fuse with the primitive thyroid when thyroid migration is completed. Lingual thyroids result from the failure of the thyroid precursor cells to migrate from the primordial pharynx to the anterior part of the neck. Therefore, calcitonin-producing cells are not expected to be present in lingual thyroids. Our objective was to determine whether calcitonin-producing C cells are present in ectopic lingual thyroids. We performed calcitonin immunolabeling and transcript detection on four flash-frozen ectopic lingual thyroids. Additional calcitonin immunolabeling was performed on two other paraffin-embedded ectopic lingual thyroids. We report evidence of calcitonin-producing cells in six independent cases of ectopic lingual thyroids. The UBB are not the only source of calcitonin-producing cells in humans. Interactions between calcitonin-producing and thyroid follicular cells occur earlier than previously accepted.


Requirement for integration of phorbol 12-myristate 13-acetate and calcium pathways is preserved in the transactivation domain of NFAT1.
The transcription factor NFAT integrates signals from both calcium- and phorbol ester-stimulated signaling pathways. The calcium signal activates the calmodulin (CaM)-dependent phosphatase calcineurin, which dephosphorylates the regulatory domain of NFAT and promotes its nuclear import, while the phorbol ester signal results in synthesis and activation of Fos and Jun, transcription factors that bind cooperatively with the NFAT DNA-binding domain in the nucleus to mediate the transcription of many target genes. Here we show that transactivation by a GAL4 fusion protein containing the strong acidic N-terminal transactivation domain (TAD) of NFAT1 also requires both calcium and phorbol ester stimulation. The calcium requirement can be mimicked by coexpression of activated versions of two CaM-dependent enzymes, calcineurin and CaM kinase IV. Our data indicate that a 144-amino acid segment of NFAT1, containing the N-terminal TAD but lacking the DNA-binding and Fos/Jun interaction domains, resembles the full-length protein in requiring a combined input from two separate signaling pathways for optimal function in cells.


Cardiopulmonary bypass in sickle cell anaemia without exchange transfusion.
To minimize sickling during cardiopulmonary bypass, exchange transfusion is frequently recommended peri-operatively for patients with homozygous sickle cell anaemia to reduce the circulating concentration of HbS. We report the successful management of two children both aged 12 years with sickle cell anaemia (homozygous SS), several sickle cell crises and multiple blood transfusions who underwent cardiopulmonary bypass for mitral valve replacement. No pre- or intra-operative exchange transfusions were employed in the two cases. Furthermore, moderate hypothermia in both cases was not associated with adverse sequelae.


Remodeling dendritic spines in the rat pilocarpine model of temporal lobe epilepsy.
Dendritic degeneration is a common pathology in temporal lobe epilepsy and its animal models. However, little is known when and how the degeneration occurs. In the present study of the rat pilocarpine model, visualization of dendrites of the hippocampal dentate granule cells (DGCs) by biocytin revealed a generalized spine loss immediately after the acute seizure induced by pilocarpine. However, this generalized damage was followed by recovery and plastic changes in spine shape and density, which occurred 15-35 days after the initial acute seizure, i.e., during the period of establishing a chronic phase of this model with the induction of spontaneous seizures. The present finding suggests that initial acute seizures do not cause permanent damages in dendrites and spines of DGCs; instead, dendritic spines are dynamically maintained in the course of the establishment and maintenance of spontaneous seizures. Local dendritic spine degeneration, detected later in the chronic phase of epilepsy, is likely to have a separate cause from initial acute insults.


Ethanol-lock technique in the treatment of bloodstream infections in pediatric oncology patients with broviac catheter.
To assess the ethanol-lock technique as a means of treating central venous line infections. Bloodstream infections in patients with tunneled central venous catheters can lead to removal of the lines. Twenty-eight children and adolescents aged 2 to 18 years, with different types of cancer, had Broviac catheters and presented with positive blood culture and clinical signs of infection between January 2000 and December 2001. The ethanol-lock technique was performed 24 times in 18 patients in addition to empiric (initially) and specific (after antibiogram) intravenous antibiotic treatment. In another 15 cases, 13 children were treated with systemic antibiotics alone. Sixty-seven percent of the patients treated with ethanol locks had no infectious relapse of any kind within 4 weeks of treatment or during subsequent aplasia, compared with 47% treated with systemic antibiotics alone. In one boy the catheter infection could not be cleared with systemic antibiotics alone, but after one course of ethanol locks no more blood culture-positive infectious episodes were observed. No severe clinical side effects of ethanol flush were observed. Mild symptoms that occurred were tiredness, headaches, dizziness, nausea, and light-headedness. The ethanol-lock technique appears to be a safe, well tolerated, and effective way to treat central venous line infections, even in small children. A prospective randomized study should be designed to compare antibiotic-lock, ethanol-lock technique, and systemic antibiotics alone in the treatment of device-associated bloodstream infection.


Direct synthesis of bicyclic guanidines through unprecedented palladium(ii) catalysed diamination with copper chloride as oxidant.
Palladium catalysed intramolecular guanidine transfer to alkenes can be accomplished with copper chloride as the oxidant to give bicyclic guanidines with complete selectivity and in high yields.


Increased plasma triglyceride secretion in EFA-deficient rats fed diets with or without saturated fat.
Metabolic responses to essential fatty acid-deficiency in rats include an increased rate of triglyceride secretion into the plasma, a large reduction in the HDL1 plasma lipoprotein concentration, and increased concentrations of liver triacylglycerols and cholesteryl esters. Because of differences in the types of EFA-deficient diets used, it is not clear whether these responses were solely due to the absence of EFA from the diet or whether saturated fat, or differences in acyl group chain length in this fat, might be responsible. Therefore, we fed rats diets differing only in amounts and kinds of fat, and measured triacylglycerol secretion rates and liver concentrations of triacylglycerols and cholesteryl esters, for comparison with our earlier measurements of plasma high density lipoprotein subpopulations in rats fed exactly the same diets. The purified diets contained either no fat, 5% by weight hydrogenated coconut oil, 5% hydrogenated cottonseed oil, or each of these three diets supplemented with 1% safflower oil, or 5% corn oil. We also fed some rats a nonpurified stock diet for comparison with literature reports. The present results indicate that the metabolic responses to essential fatty acid deficiency described above are definitely due to essential fatty acid-deficiency and not to the presence or chain length of acyl groups in saturated fat in the diet.


c-Abl is required for the signaling transduction induced by L-selectin ligation.
Lymphocyte recruitment onto inflamed tissues requires cells tethering to and rolling on vascular surfaces under flow. L-selectin is constitutively expressed on leukocytes to mediate the leukocytes' initial capture and subsequent rolling along the vessel. Apart from its adhesive function, engagement of L-selectin also results in cell activation, which is related to the completed signaling transduction. Here we show that ligation of L-selectin with its mAb increases c-Abl kinase activity, and that the activated c-Abl kinase can be recruited to and phosphorylate the cytoplasmic domain of L-selectin. In addition, the activated c-Abl kinase can regulate Zap70 kinase by increasing the phosphorylation of the Y319 site of Zap70 kinase and connect with Zap70 kinase through its SH2 domain. These results indicate that c-Abl kinase plays an important role in accepting and transferring the upstream activation events induced by L-selectin ligation.


Does weight-adjusted anti-tumour necrosis factor treatment favour obese patients with Crohn's disease?
Adalimumab (ADA) is a subcutaneous anti-tumour necrosis factor (anti-TNF) agent, effective in inducing and maintaining remission in Crohn's disease (CD). Unlike Infliximab (IFX), ADA dosing is not weight adjusted and dose frequency is based on clinical response. To determine whether obesity is a risk factor for early loss of response (LOR) to anti-TNF treatment and whether weight-adjusted anti-TNF treatment is favourable. A hospital database of CD patients receiving anti-TNF treatment was analyzed retrospectively. The relationship between time to LOR and BMI was examined by Kaplan-Meier (KM) survival curves and a Cox proportional hazards model. ADA patients: Of the 54 patients (46 BMI<30 and 8 BMI≥30), KM estimation indicated a significantly shorter time to dose escalation in the BMI of at least 30 (χ=6.117, P=0.01). The Cox proportional hazards model showed that an increased hazard of LOR to ADA is related to increases in BMI (P=0.04). IFX patients: Of the 76 patients (62 BMI<30 and 14 BMI≥30), KM estimation showed that the differences in survival curves were not significant (χ=1.933, P=0.16) for the BMI groups. This was supported by the Cox proportional hazard model (P=0.36). BMI appears to be important in predicting ADA efficacy (LOR) in CD. IFX appears to overcome this reduction of efficacy in obese patients. A prospective study evaluating the effect of weight on anti-TNF drug response and serum drug levels is warranted.


Survivorship care for cancer patients in primary versus secondary care: a systematic review.
Cancer survivorship care is traditionally performed in secondary care. Primary care is often involved in cancer management and could therefore play a more prominent role. To assess outcomes of cancer survivorship care in primary versus secondary care. A systematic search of MEDLINE and EMBASE was performed. All original studies on cancer survivorship care in primary versus secondary care were included. A narrative synthesis was used for three distinctive outcomes: (1) clinical, (2) patient-reported, and (3) costs. Sixteen studies were included: 7 randomized trials and 9 observational studies. Meta-analyses were not feasible due to heterogeneity. Most studies reported on solid tumors, like breast (N = 7) and colorectal cancers (N = 3). Clinical outcomes were reported by 10 studies, patient-reported by 11, and costs by 4. No important differences were found on clinical and patient-reported outcomes when comparing primary- with secondary-based care. Some differences were seen relating to the content and quality of survivorship care, such as guideline adherence and follow-up tests, but there was no favorite strategy. Survivorship care in primary care was associated with lower societal costs. Overall, cancer survivorship care in primary care had similar effects on clinical and patient-reported outcomes compared with secondary care, while resulting in lower costs. Survivorship care in primary care seems feasible. However, since the design and outcomes of studies differed, conclusive evidence for the equivalence of survivorship care in primary care is still lacking. Ongoing studies will help provide better insights.


Associations between Chinese college students' anxiety and depression: A chain mediation analysis.
Anxiety and depression are great public health concerns among college students. The purpose of this study was to explore whether sleep quality and quality of life (QoL) play mediating roles in anxiety and depression among Chinese college students. A total of 2757 college students (mean age = 19.07; SD = 1.14) completed the questionnaires, including a brief demographic survey. The 2-item General Anxiety Disorder (GAD-2) and the 2-item Patient Health Questionnaire (PHQ-2) were used to assess the symptoms of anxiety and depression, respectively. And the Pittsburgh Sleep Quality Index (PSQI) and the Short-Form 36 Health Survey (SF-36) were used to evaluate college students' sleep quality and QoL, respectively. Mediation analyses were conducted by using PROCESS macro in the SPSS software. Anxiety had both direct and indirect effects on depression. Sleep quality and QoL were not only independent mediators in the relationship between anxiety and depression but also chain mediators. The results of the current study highlight the crucial role of early intervention for depression with a focus on college students with anxiety, more especially, on those with poorer sleep quality and lower QoL.


[Unexplained, subclinical chronically elevated transaminases].
Unexplained, subclinical chronically elevated transaminases is mainly a marker of non-alcoholic fatty liver disease, metabolic syndrome, alcoholism and diabetes, which are very common situations but viral hepatitis and iatrogenic origin must also be considered. Before looking for hepatic or genetic rare diseases, it is worth considering hypertransaminasemia as a clue for muscular disease, particularly in paediatric settings, and creatine phosphokinase is a specific marker. Then, patient history, examination and appropriate biologic requests can permit the identification of less frequent disorders where isolated hypertransaminasemia is possibly the unique marker of the disease for a long while: hemochromatosis, celiac disease, autoimmune hepatitis, Wilson's disease, α1-anti-trypsine deficiency, thyroid dysfunctions, Addison's disease. Liver biopsy should be performed only in patients with aspartate aminotransferases upper the normal range or alanine aminotransferases higher than twice the normal range after 6 months delay with dietetic corrections.


Novel mutation in the NRLP3 manifesting as an intermediate phenotype of cryopyrinopathies.
Cryopyrin-associated periodic syndromes (CAPS) are a group of autoinflammatory diseases associated with NLRP3 gain of function mutations. CAPS associated mutations are found predominantly in exon 3. The objective of this study is to describe a new variant on NRLP3 gene and its phenotype. Case report description of a new NRLP3 pathogenic variant and literature case-based search through INFEVERS database. A 21-year old male who presented multiple tonic-clonic seizures on his 3rd day of life. At age 2, he had recurrent central facial palsy, high fever (40 °C), painful and persistent oral ulcers, abdominal pain, nausea and vomiting, and delayed neuropsychomotor development, with polyarthritis in wrists and knees. Over the years, several symptoms were observed: livedo reticularis, Raynaud's phenomenon, positive pathergy test, heat allodynia, extremely painful genital ulcers, and sporadic conjunctivitis. Laboratory studies revealed persistently elevated inflammatory markers and serum amyloid protein A (30 μg/l). The genetic panel for autoinflammatory diseases revealed heterozygous mutation in the NLRP3, (c.2068G > C, p.E690Q) with 0% of frequency in the general population. The patient denies rash and did not have frontal bossing or patellar overgrowth. We found a positive familial history on mother and brother, who carried the same mutation. The patient was started on canakinumab which controlled his symptoms. Currently, 241 missense variants in the NLRP3 have been described. We presented a new mutation in exon 3 of the NRLP3 gene in a patient that fulfills clinical criteria for CAPS who had complete clinical response to Canakinumab, supporting the idea that this mutation is pathogenic.


Family intervention effects on co-occurring early childhood behavioral and emotional problems: a latent transition analysis approach.
This study used latent transition analysis (LTA) to examine changes in early emotional and behavioral problems in children age 2 to 4 years resulting from participation in a family-centered intervention. A sample of 731 economically disadvantaged families was recruited from among participants in a national food supplement and nutrition program. Families with toddlers between age 2 and 3 were randomized either to the Family Check-Up (FCU) or to a nonintervention control group. The FCU's linked interventions were tailored to each family's needs. Assessments occurred at age 2, 3, and 4. The FCU followed age 2 and age 3 assessments. Latent class analyses were conducted on mother reports of behavior and emotional problems from age 2 to 4 to study transitions among the following four groups: (a) externalizing only, (b) internalizing only, (c) comorbid internalizing and externalizing, and (d) normative. LTA results revealed that participation in the FCU increased the likelihood of transitioning from either the comorbid or the internalizing class into the normative class by age 4. These results suggest family interventions in early childhood can potentially disrupt the early emergence of both emotional and behavioral problems.


Effects of a cervical disc prosthesis on maintaining sagittal alignment of the functional spinal unit and overall sagittal balance of the cervical spine.
The object of this study is to review the early clinical results and radiographic outcomes following insertion of the Bryan Cervical Disc Prosthesis (Medtronic Sofamor Danek, Memphis, TN), together with its effect on maintaining sagittal alignment of the functional spinal unit (FSU) and overall sagittal balance of the cervical spine for the treatment of single-level or two-level symptomatic disc disease. Forty-seven patients with symptomatic single or two-level cervical disc disease who received the Bryan Cervical Artificial Disc were reviewed prospectively. A total of 55 Bryan disc were placed in 47 patients. A single-level procedure was performed in 39 patients and a two-level procedure in the other eight. Radiographic and clinical assessments were made preoperatively and at 1.5, 3, 6, 9, 12, and 18 and up to 33 months postoperatively. Mean follow-up duration was 24 months, ranging from 13 to 33 months. Periods were categorized as early follow up (1.5-3 months) and late follow up (6-33 months). The visual analogue scale (VAS), neck disability index(NDI), Odom's criteria were used to assess pain and clinical outcomes. Static and dynamic radiographs were measured by hand and computer to determine the range of motion (ROM), the angle of the functional segmental unit (FSU), and the overall cervical alignment (C2-7 Cobb angle). With all of these data, we evaluated the change of the preoperative lordosis (or kyphosis) of the FSU and Overall sagittal balance of the cervical spine during the follow-up period. There was a statistically significant improvement in the VAS score from 7.0 +/- 2.6 to 2.0 +/- 1.5 (paired-t test, P = 0.000), and in the NDI from 21.5 +/- 5.5 to 4.5 +/- 3.9 (paired-t test P = 0.000). All of the patients were satisfied with the surgical results by Odom's criteria. The postoperative ROM of the implanted level was preserved without significant difference from preoperative ROM of the operated level. Only 36% of patients with a preoperative lordotic sagittal orientation of the FSU were able to maintain lordosis following surgery. However, the overall sagittal alignment of the cervical spine was preserved in 86% of cases at the final follow up. Interestingly, preoperatively kyphotic FSU resulted in lordotic FSU in 13% of patients during the late follow-up, and preoperatively kyphotic overall cervical alignment resulted in lordosis in 33% of the patients postoperatively. Clinical results are encouraging, with significant improvement seen in the Bryan Cervical Artificial disc. The Bryan disc preserves motion of the FSU. Although the preoperative lordosis (or kyphosis) of the FSU could not always be maintained during the follow-up period, the overall sagittal balance of the cervical spine was usually preserved.


Autologous collagen vocal fold injection: a preliminary clinical study.
This preliminary study reports the first use of injectable autologous collagen for vocal fold augmentation. In previous studies, the authors showed cross-linked bovine collagen to be effective in more than 150 patients with glottic incompetence, particularly those with focal defects or scarred or atrophic vocal folds. However, concerns about possible adverse immunologic responses to the bovine material have limited its use. The authors studied eight patients with difficult vocal fold pathology, including sulcus vocalis, atrophy, and scarring secondary to trauma and cordectomy. Skin was harvested under local anesthesia, processed into a naturally cross-linked injectable form (Autologen), and injected using indirect laryngoscopy. Voice production was evaluated prior to the injection and at intervals after the injection, using subjective, perceptual, aerodynamic, acoustic, and videostroboscopic assessments. The results indicate that autologous collagen is comparable to injectable bovine collagen in the management of several difficult glottic insufficiency problems and that the likelihood of a hypersensitivity response is negligible. Unlike bovine collagen preparations, Autologen does not require breakdown of the natural collagen molecule, so it is anticipated that this material will be better tolerated and more stable over time.


Cardiovascular effects of B-vitamins and/or N-3 fatty acids: the SU.FOL.OM3 trial.
Mechanisms involved in coronary stenosis evolution are different than those involved in clinical events. Because of differential vascular effects, N-3 polyunsatured fatty acids (PUFA) and B vitamins could have differential effects on different types of cardiovascular clinical events in high-risk patients. We analyzed the effects of n-3 PUFA and of B vitamins on both coronary revascularization and on hard coronary events risks in a subgroup of the SU.FOL.OM3 trial, a randomized, double-blind, placebo-controlled secondary prevention trial. Data were analyzed according to the intention-to-treat principle, with the use of Cox proportional-hazards models. After a mean follow-up of 4.2 ± 1.0 years among the 1,863 participants with coronary heart disease, 163 coronary revascularizations were performed, and 95 patients experienced a hard coronary event. Neither treatment with n-3 PUFA, nor treatment with B vitamins was associated with any significant effect on the occurrence of hard coronary events. Allocation to n-3 PUFA was not associated with any significant effect on coronary revascularization. However, treatment with B vitamins was associated with a statistically significant 52% increase in the risk of coronary revascularization (multivariate HR: 1.52; 95% CI: [1.11-2.10]; p=0.01). Neither n-3 PUFA, nor B vitamins reduced the rates of hard coronary events and of coronary revascularization. Furthermore, B vitamins significantly increased the rate of coronary revascularization. Consistent with the findings of previous trials, our results do not support the routine use of dietary supplements containing n-3 PUFA and argue against using dietary supplements containing B-vitamins in coronary patients in secondary cardiovascular prevention.


Diagnostic significance of microRNAs as novel biomarkers for bladder cancer: a meta-analysis of ten articles.
Previous studies have revealed the importance of microRNAs' (miRNAs) function as biomarkers in diagnosing human bladder cancer (BC). However, the results are discordant. Consequently, the possibility of miRNAs to be BC biomarkers was summarized in this meta-analysis. In this study, the relevant articles were systematically searched from CBM, PubMed, EMBASE, and Chinese National Knowledge Infrastructure (CNKI). The bivariate model was used to calculate the pooled diagnostic parameters and summary receiver operator characteristic (SROC) curve in this meta-analysis, thereby estimating the whole predictive performance. STATA software was used during the whole analysis. Thirty-one studies from 10 articles, including 1556 cases and 1347 controls, were explored in this meta-analysis. In short, the pooled sensitivity, area under the SROC curve, specificity, positive likelihood ratio, diagnostic odds ratio, and negative likelihood ratio were 0.72 (95%CI 0.66-0.76), 0.80 (0.77-0.84), 0.76 (0.71-0.81), 3.0 (2.4-3.8), 8 (5.0-12.0), and 0.37 (0.30-0.46) respectively. Additionally, sub-group and meta-regression analyses revealed that there were significant differences between ethnicity, miRNA profiling, and specimen sub-groups. These results suggested that Asian population-based studies, multiple-miRNA profiling, and blood-based assays might yield a higher diagnostic accuracy than their counterparts. This meta-analysis demonstrated that miRNAs, particularly multiple miRNAs in the blood, might be novel, useful biomarkers with relatively high sensitivity and specificity and can be used for the diagnosis of BC. However, further prospective studies with more samples should be performed for further validation.


Measuring social orienting in preschoolers with autism spectrum disorder using cartoons stimuli.
Altered social orienting (SO) was proposed as the primary source of socio-communicative difficulties in autism spectrum disorder (ASD). Eye-tracking studies generally confirm a decreased SO in ASD population. However, SO has been scarcely investigated using minimally social stimuli such as cartoons. The extent to which SO might be decreased when watching cartoons is therefore unknown. Yet, it could allow for malleable and child-friendly paradigms that could be sensitive to early atypical visual preference. In this study, 90 preschoolers with ASD (age = 3.19 ± 0 .88) and 20 TD (age = 2.95 ± 1.26) watched two eye-tracking preference tasks. One Realistic task, displaying children dancing versus geometric shapes moving repetitively and a Cartoon task, displaying social and non-social cartoon stimuli with similar movements. We measured SO percentage along with refined visual exploration parameters and compared those of ASD children to TDs. In addition, we investigated their relations with behavioral measures such as symptom severity, developmental and adaptive levels. We evidenced a decreased SO percentage in ASD compared to TD children when watching the Realistic task but not the Cartoon task. We did not identify any other between groups differences. However, we identified several correlations between eye-tracking measures and developmental as well as adaptive measures within the Cartoon task. Together, our results support a preferential orientation of children with autism towards repetitively moving shapes but no decreased SO when measured with minimally social stimuli. Nonetheless, when investigating finer visual exploration parameters, even socially simple stimuli elicited atypical gaze patterns related to early developmental delay.


Calcium transients and intramembrane charge movement in skeletal muscle fibres.
Myoplasmic calcium transients in response to pulse depolarisations have been monitored using the dye antipyralazo III, which provides a linear measure of the change in cytoplasmic free calcium concentration. Intramembrane charge movements were recorded simultaneously with the calcium transients. Except for its initial phase, the calcium transient during each pulse closely followed the time course expected for calcium redistribution between three intracellular compartments with time-independent flux coefficients. The time at which the flux coefficients attained their steady values occurred just after the intramembrane charge movement had been completed. This is the required sequence if charge movement were to control one or more coefficients f or calcium fluxes across the sarcoplasmic reliculum membrane.


Combining thrombolysis with the platelet glycoprotein IIb/IIIa inhibitor lamifiban: results of the Platelet Aggregation Receptor Antagonist Dose Investigation and Reperfusion Gain in Myocardial Infarction (PARADIGM) trial.
The trial was designed to assess the safety, pharmacodynamics and effects on reperfusion of the platelet glycoprotein (GP) IIb/IIIa inhibitor lamifiban when given with thrombolysis to patients with ST segment elevation acute myocardial infarction. Studies of fibrinolytic agents in acute myocardial infarction have demonstrated a direct relationship between early complete reperfusion and survival. Blockade of the platelet GP IIb/IIIa receptor complex inhibits platelet aggregation and may speed reperfusion when given in conjunction with thrombolysis to patients with acute myocardial infarction. Patients with ST segment elevation presenting within 12 h of symptom onset who were treated with either tissue-plasminogen activator or streptokinase were enrolled in this three-part Phase II dose exploration study. In Part A, all patients received the GP IIb/IIIa inhibitor lamifiban in an open-label, dose escalation scheme. Parts B and C were a randomized, double-blind comparison of a bolus plus 24-h infusion of lamifiban versus placebo with patients randomized in a 2:1 ratio. The goal was to identify a dose(s) of lamifiban that provided >85% adenosine diphosphate (ADP)-induced platelet aggregation inhibition. A composite of angiographic, continuous electrocardiographic and clinical markers of reperfusion was the primary efficacy end point, and bleeding was the primary safety end point. Platelet aggregation was inhibited by lamifiban in a dose-dependent manner with the highest doses exceeding 85% ADP-induced platelet aggregation inhibition. There was more bleeding associated with lamifiban (transfusions in 16.1% lamifiban-treated vs. 10.3% placebo-treated patients). Lamifiban induced more rapid reperfusion as measured by all continuous electrocardiographic (ECG) parameters. Lamifiban given with thrombolytic therapy appears to be associated with more rapid and complete reperfusion than placebo. As expected in this small sample, there were no obvious clinical benefits to lamifiban over placebo. Reconciliation of ECG monitoring with clinical outcomes will require a larger, adequately powered clinical trial.


Is there a link between availability of food and beverage establishments and BMI in Mexican adults?
To study the association between density of stores (food and beverage stores, stores selling only fruits and vegetables, and supermarkets) and the BMI of adults aged ≥20 years in Mexico. A cross-sectional study was performed. Individual data came from the 2012 National Health and Nutrition Survey, while information on stores was taken from the National Institute of Geography and Statistics' National Statistics Directory of Economic Units. A weighted least-squares model was estimated to test the association between density of stores and BMI of adults adjusting for sex, age, education, presence of hypertension, diabetes or both, household assets index and marginality index at the municipality level. Mexico. An additional 1 sd in the density of fruit and vegetable stores was associated with a reduction of 0·24 (95 % CI -0·37, -0·12) kg/m2 in BMI when the densities of the other stores were at their mean values. For food and beverage store density, a difference of 1 sd was associated with an increase of 0·50 (95 % CI 0·33, 0·67) kg/m2 in BMI, while for supermarkets the corresponding association was a reduction of 0·48 (95 % CI -1·52, 0·56) kg/m2 in BMI. In places with a higher density of stores that offer unhealthy foods, the BMI of adults tends to be higher.


Exogenous proline induces regulation in 2-acetyl-1-pyrroline (2-AP) biosynthesis and quality characters in fragrant rice (Oryza sativa L.).
Proline is one of the precursors of the biosynthesis of 2-acetyl-1-pyrroline (2-AP) which is the key and characteristic volatile component of fragrant rice aroma. In order to study the effects of exogenous proline on 2-AP biosynthesis and other grain quality attributes in fragrant rice, two indica fragrant rice cultivars, "Meixiangzhan-2" and "Xiangyaxiangzhan", and one japonica fragrant rice, "Yunjingyou", were used in present study. At initial heading stage, proline solutions at 0 (CK), 0.10 (Pro1), 0.20 (Pro2) and 0.50 (Pro3) g L-1 were applied as foliar spray solution to fragrant rice plants. Compared with CK, Pro1, Pro2 and Pro3 treatments significantly increased the grain 2-AP content. The significant up-regulation effects due to proline treatments were observed in the contents of proline, △1-pyrrolidine-5-carboxylic acid (P5C) and △1-pyrroline which involved in 2-AP formation. Exogenous proline application also significantly decreased the grain γ-aminobutyric acid (GABA) content. Furthermore, proline treatments enhanced the activity of proline dehydrogenase (ProDH) as well as transcript level of gene PRODH. On the other hand, the transcript level of gene BADH2 and activity of betaine aldehyde dehydrogenase (BADH) decreased under proline treatments. Proline treatments (Pro2 and Pro3) also increased the grain protein content by 3.57-6.51%. Moreover, 32.03-34.25% lower chalky rice rate and 30.80-48.88% lower chalkiness were recorded in proline treatments (Pro2 and Pro3) for both Meixiangzhan and Xiangyaxiangzhan whilst for Yunjingyou, foliar application of proline had no significant effect on chalky rice rate and chalkiness. There was no remarkable difference observed in grain milled quality (brown rice rate, milled rice rate and head rice rate) and amylose content between CK and proline treatments. In conclusion, exogenous proline enhanced the 2-AP biosynthesis and promoted some grain quality characters of fragrant rice.


Randomized clinical trial comparing systemic interferon with diathermocoagulation in primary multiple and widespread anogenital condyloma.
Two hundred three patients (median age 26 years; range 18-45) with untreated multiple and widespread anogenital condyloma were randomly assigned to one of four study arms in order to compare the efficacy, toxicity, and tolerability of recombinant interferon alpha-2b with those of diathermocoagulation. Of 200 evaluable patients, 51 were treated intramuscularly (IM) with 3 x 10(6) U (3 MU)/m2 daily for 3 weeks (total dose 63 MU/m2), 50 received subcutaneous thrice-weekly injections of 3 MU/m2 for 4 weeks (total dose 36 MU/m2), 51 underwent diathermocoagulation, and 48 were not treated and were used as a control group. Six months after the end of treatment, the overall response rate (complete and partial responses) was 70%: 57 and 82% for patients receiving interferon alpha-2b (IM and subcutaneously) and diathermocoagulation, respectively, and 8% for the control group. After 6 months from therapy, no significant differences in complete response were found among the different types of treatment: 20, 20, and 35% for the two interferon groups and the diathermocoagulation group, respectively. Fifteen and two complete responders in the cauterization and interferon groups, respectively, experienced disease recurrence (P less than .01). All patients given interferon therapy complained of flu-like symptoms, which declined progressively after the first week of treatment. Fatigue, lasting as long as patients received interferon, was the most prevalent chronic side effect. We conclude that systemic recombinant interferon alpha-2b is active in treating patients with primary condyloma lesions and does so as well as cauterization.(ABSTRACT TRUNCATED AT 250 WORDS)


Decreased expression of gamma-aminobutyric acid type A/benzodiazepine receptor beta subunit mRNAs in brain of flurazepam-tolerant rats.
The expression of GABAA/benzodiazepine beta subunit mRNAs was studied in cerebral cortex, hippocampus, and cerebellum of flurazepam-treated rats. Immediately following 4 wk of treatment, beta 2 and beta 3 subunit mRNAs were significantly reduced in cerebellum and hippocampus, whereas only beta 2 was decreased in cortex. These decreases had largely reversed 48 h following flurazepam treatment. After 2 wk of treatment, both beta 2 and beta 3 mRNAs were reduced in cerebellum, and beta 3 mRNA was reduced in hippocampus, but neither was changed in cortex. Four hours after an acute flurazepam treatment, the only change was a decrease in beta 3 mRNA in hippocampus. These results indicate that the expression of GABAA receptor beta subunit mRNAs in different brain regions is differentially regulated during chronic flurazepam treatment, and some changes occur within hours after a single large dose.


Effects of N,N-dimethylformamide and sodium butyrate on enzymes of pyrimidine metabolism in cultured human tumor cells.
Effects of a 7-day treatment with the maturational agents DMF and sodium butyrate on enzymes of pyrimidine metabolism, growth rate and cell maturation were assessed in 5 human tumor cell lines, ARH-77 (myeloma), K-562 (chronic myeloid leukemia), KG-1 (myeloid leukemia), HL-60 (promyelocytic leukemia) and RWLy-1 (non-Hodgkin's lymphoma). DMF lengthened the doubling times of all five cell lines while sodium butyrate lengthened only those of K-562, HL-60 and RWLy-1. Full maturation was induced only in HL-60 by either agent and in K-562 by butyrate. Exposure resulted in a decreased activity of the anabolic enzyme orotate phosphoribosyltransferase (EC 220.127.116.11) and increased activities of the catabolic enzymes thymidine phosphorylase (EC 18.104.22.168) and dihydrouracil dehydrogenase (EC 18.104.22.168). Changes in the amphibolic enzyme, uridine phosphorylase (EC 126.96.36.199) did not follow any apparent pattern. This study indicates that the pattern of pyrimidine metabolism differs between the differentiated and slowly growing, and undifferentiated rapidly growing counterpart of several human tumors, suggesting that enzymes of pyrimidine metabolism can be used as markers for cellular growth and/or maturity.


Research on cholesterol, steroidal hormones and liposoluble vitamins by thin layer chromatography in some adipose organs of anuran amphibians. II. Liposoluble vitamins.
By means of thin layer chromatography and selective detection, vitamin A acetate, vitamin E and vitamin E acetate, vitamin D3 and cholesterol were identified in the prepericardic body, adrenal tissue, fat bodies and testis of Bubo bufo, as well as in the prepericardic body of Rana esculenta. On the contrary, liposoluble vitamins could not be isolated from the adipose tissue occurring in the thoracic cavity of Rana esculenta, or in Bufo bufo muscles. The results are discussed in correlation with the histological structure of the prepericardic body and the presumptive identification therein of steroid hormones. Lastly, the functional analogy between the prepericardic body and fat bodies in anuran amphibians and the brown adipose tissue of mammals is also postulated.


Detection of specific gene rearrangements by fluorescence in situ hybridization in 16 cases of clear cell sarcoma of soft tissue and 6 cases of clear cell sarcoma-like gastrointestinal tumor.
Clear cell sarcoma of soft tissue (CCSST) and clear cell sarcoma-like gastrointestinal tumor (CCSLGT) are malignant mesenchymal tumors that share some pathological features, but they also have several different characteristics. They are well known to express chimeric fusions of Ewing sarcoma breakpoint region 1 (EWSR1) and cAMP response element-binding protein (CREB) family members; namely, EWSR1-activating transcription factor 1 (ATF1) and EWSR1-CREB1. In addition, recent studies have suggested the presence of other fusions. We used fluorescence in situ hybridization to detect specific rearrangements including EWSR1, ATF1, CREB1, and cAMP response element modulator (CREM) in 16 CCSST and 6 CCSLGT cases. We also used reverse transcription polymerase chain reaction (RT-PCR) to detect specific chimeric fusions of EWSR1-ATF1 and EWSR1-CREB1 using fresh tumor samples in available cases. A total of 15 of 16 CCSST cases (93.8%) had EWSR1 rearrangement, of which 11 (68.8%) also had ATF1 rearrangement, suggestive of the presence of EWSR1-ATF1 fusions. One CCSST case (6.3%) was found to have EWSR1 and CREM rearrangements, and 4 of 6 CCSLGT cases (66.7%) had EWSR1 rearrangement, of which 2 (33.3%) showed ATF1 rearrangement and the other 2 cases (33.3%) showed CREB1 rearrangement. These cases most likely had EWSR1-ATF1 and EWSR1-CREB1 fusions, respectively. RT-PCR was performed in 8 available cases, including 6 CCSSTs and 2 CCSLGTs. All CCSSTs showed EWSR1-ATF1 fusions. Among the 2 CCSLGT cases, one had EWSR1-ATF1 fusion and the other had EWSR1-CREB1 fusion. Rearrangements of EWSR1 and ATF1 or EWSR1-ATF1 fusion were predominantly found in CCSST, whereas those of EWSR1 and CREB1 or EWSR1-CREB1 tended to be detected in CCSLGT. A novel CREM fusion was also detected in a few cases of CCSST and CCSLGT. The cases in which EWSR1 rearrangement was detected without definitive partner genes should be considered for the presence of CREM rearrangement.


Macular hole formation, progression, and surgical repair: case series of serial optical coherence tomography and time lapse morphing video study.
To use a new medium to dynamically visualize serial optical coherence tomography (OCT) scans in order to illustrate and elucidate the pathogenesis of idiopathic macular hole formation, progression, and surgical closure. Two patients at the onset of symptoms with early stage macular holes and one patient following repair were followed with serial OCTs. Images centered at the fovea and at the same orientation were digitally exported and morphed into an Audiovisual Interleaving (avi) movie format. Morphing videos from serial OCTs allowed the OCTs to be viewed dynamically. The videos supported anterior-posterior vitreofoveal traction as the initial event in macular hole formation. Progression of the macular hole occurred with increased cystic thickening of the fovea without evidence of further vitreofoveal traction. During cyst formation, the macular hole enlarged as the edges of the hole became elevated from the retinal pigment epithelium (RPE) with an increase in subretinal fluid. Surgical repair of a macular hole revealed initial closure of the macular hole with subsequent reabsorption of the sub-retinal fluid and restoration of the foveal contour. Morphing videos from serial OCTs are a useful tool and helped illustrate and support anterior-posterior vitreofoveal traction with subsequent retinal hydration as the pathogenesis of idiopathic macular holes.


TCR/CD3 mediated stop-signal is decoupled in T-cells from Ctla4 deficient mice.
CTLA-4 is a co-receptor that plays a pivotal role in regulating the threshold for T-cell activation. We recently reported that CTLA-4 ligation can over-ride the stop-signal induced by anti-CD3 ligation [Schneider H, Downey J, Smith A, Zinselmeyer BH, Rush C, Brewer JM, et al. Reversal of the TCR stop-signal by CTLA-4. Science 2006;313:1972]. While these studies compared CTLA-4 positive and negative T-cells from normal mice, little is known regarding the behaviour of T-cells from diseased Ctla4 deficient mice with auto-proliferative disease. In this study, we show that while activated wild-type and Ctla-4-/- T-cells have similar rates of motility, Ctla-4-/- T-cells show a marked resistance to the induction of a stop-signal by anti-CD3 ligation. By contrast, T-cells from normal mice and CD28 deficient mice underwent a normal slowing of motility in response to anti-CD3 ligation. Our findings identify a fundamental difference between normal versus CTLA-4-/- T-cells from diseased mice in the regulation of motility by anti-CD3 ligation. This dysregulation of motility may contribute to the tissue infiltration and the autoimmune disorder observed in Ctla-4-/- mice.


Bureaucratization and medical professionals' values: A cross-national analysis.
Understanding the impact of the bureaucratization of governance systems on the occupational values of medical professionals is a fundamental concern of the sociological research of healthcare professions. While previous studies have examined the impact of bureaucratized management, organizations, and healthcare fields on medical professionals' values, there is a lack of cross-national research on the normative impact of the bureaucratized systems of national governance. Using the European Social Survey data for 29 countries, this study examines the impact of the bureaucratization of national governance systems on the occupational values of medical professionals. The findings indicate that medical professionals who are employed in countries with the more bureaucratized systems of national governance are less concerned with openness to change values, that emphasize autonomy and creativity, and self-transcendence values, that emphasize common good. The findings also indicate that the negative effect of the bureaucratization of national governance on the openness to change values is stronger for medical professionals in more bureaucratized organizations with more rationalized administration systems.


Charge distribution within cell surface coats of single and interacting surfaces--a minimum free electrostatic energy approach. Conclusions for electrophoretic mobility measurements.
The fixed charge distribution in a cell surface coat model was calculated using a variational approach based on the minimum of electrostatic free energy. The influence of ionic strength on distribution was analyzed. It is shown that interaction with other coats or smooth surfaces changes the fixed charge distribution. The special effect of a possible membrane charge was calculated in addition. In this case there is a fixed, charge-free space near the membrane, depending on coat thickness, ionic strength and membrane charge/surface coat charge ratio. For several cases energies, electric potentials and fixed charge distributions are given. The general result is that fixed charge distribution is always composed of a constant space charge density over a part of or the whole surface coat and, with some exceptions, of a surface charge density at the border of the surface coat. Conclusions for electrophoretic mobility measurement interpretations are drawn.


The interaction between HIF-1 and AP-1 transcription factors in response to low oxygen.
Hypoxia-inducible factor-1 (HIF-1) is a critical regulator of the transcriptional response to low oxygen conditions (hypoxia/anoxia) experienced by mammalian cells in both physiological and pathophysiological circumstances. As our understanding of the biology and biochemistry of HIF-1 has grown, it has become apparent that cells adapt to signals generated by low oxygen through a network of stress responsive transcription factors or complexes, which are influenced by HIF-1 activity. This review summarizes our current understanding of the interaction of HIF-1 with AP-1, a classic example of a family of pleiotropic transcription factors that impact on diverse cellular processes and phenotypes, including the adaptation to low oxygen stress. The review focuses on experimental studies involving cultured cells exposed to hypoxia/anoxia, and describes both established and possible interactions between HIF-1 and AP-1 at different levels of cellular organization.


3-Deoxy-D-arabino-heptulosonate 7-phosphate synthase is regulated for the accumulation of polysaccharide-linked hydroxycinnamoyl esters in rice (Oryza sativa L.) internode cell walls.
Polysaccharide-linked hydroxycinnamoyl esters (PHEs) over-accumulate in the internodes of a rice (Oryza sativa L.) mutant, Fukei 71 (F71). This accumulation is accompanied by over-expression of phenylalanine ammonialyase (PAL). In this study, we show that only one member of the 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAHPS) family expresses in close correlation with PAL. Furthermore, substrate availability to DAHPS is promoted by down-regulating the expression of plastidic pyruvate kinase (PKp), a competitor of DAHPS. Since the over-production of PHEs is caused by D50 gene disruption, these results suggest that specific enzymes in the phenylpropanoid and shikimate pathways are coordinately up-regulated. In addition, the results indicate that carbon-flow into the shikimate pathway is modified for the synthesis of PHEs, and is probably controlled by D50.


Impaired vitamin D metabolism in CKD.
Vitamin D metabolism consists of both production and catabolism, which are enzymatically driven and highly regulated. Renal vitamin D metabolism requires filtration and tubular reabsorption of 25-hydroxyvitamin D and is regulated by parathyroid hormone, fibroblast growth factor-23, and 1,25-dihydroxyvitamin D. In chronic kidney disease, renal production of 1,25-dihydroxyvitamin D from 25-hydroxyvitamin D is reduced. In addition, pharmacokinetic studies and epidemiologic studies of 24,25-dihydroxyvitamin D, the most abundant product of 25-hydroxyvitamin D catabolism by CYP24A1, suggest that vitamin D catabolism also is reduced. New insights into the mechanisms and regulation of vitamin D metabolism may lead to novel approaches to assess and treat impaired vitamin D metabolism in chronic kidney disease.


Non-invasive electromechanical assessment during atrial fibrillation identifies underlying atrial myopathy alterations with early prognostic value.
Electromechanical characterization during atrial fibrillation (AF) remains a significant gap in the understanding of AF-related atrial myopathy. This study reports mechanistic insights into the electromechanical remodeling process associated with AF progression and further demonstrates its prognostic value in the clinic. In pigs, sequential electromechanical assessment during AF progression shows a progressive decrease in mechanical activity and early dissociation from its electrical counterpart. Atrial tissue samples from animals with AF reveal an abnormal increase in cardiomyocytes death and alterations in calcium handling proteins. High-throughput quantitative proteomics and immunoblotting analyses at different stages of AF progression identify downregulation of contractile proteins and progressive increase in atrial fibrosis. Moreover, advanced optical mapping techniques, applied to whole heart preparations during AF, demonstrate that AF-related remodeling decreases the frequency threshold for dissociation between transmembrane voltage signals and intracellular calcium transients compared to healthy controls. Single cell simulations of human atrial cardiomyocytes also confirm the experimental results. In patients, non-invasive assessment of the atrial electromechanical relationship further demonstrate that atrial electromechanical dissociation is an early prognostic indicator for acute and long-term rhythm control.


Progress on RNA-based therapeutics for genetic diseases.
RNA therapeutics inhibit the expression of specific proteins/RNAs by targeting complementary sequences of corresponding genes or encode proteins for the synthesis desired genes to treat genetic diseases. RNA-based therapeutics are categorized as oligonucleotide drugs (antisense oligonucleotides, small interfering RNA, RNA aptamers), and mRNA drugs. The antisense oligonucleotides and small interfering RNA for treatment of genetic diseases have been approved by the FDA in the United States, while RNA aptamers and mRNA drugs are still in clinical trials. Chemical modifications can be applied to RNA drugs, such as pseudouridine modification of mRNA, to reduce immunogenicity and improve the efficacy. The secure and effective delivery systems such as lipid-based nanoparticles, extracellular vesicles, and virus-like particles are under development to address stability, specificity, and safety issues of RNA drugs. This article provides an overview of the specific molecular mechanisms of eleven RNA drugs currently used for treating genetic diseases, and discusses the research progress of chemical modifications and delivery systems of RNA drugs.


A minimum core outcome dataset for the reporting of preclinical chemotherapeutic drug studies: Lessons learned from multiple discordant methodologies in the setting of colorectal cancer.
In vivo studies in animal models are critical tools necessary to study the fundamental complexity of carcinogenesis. A constant strive to improve animal models in cancer exists, especially those investigating the use of chemotherapeutic effectiveness. In the present systematic review, colorectal cancer (CRC) is used as an example to highlight and critically evaluate the range of reporting strategies used when investigating chemotherapeutic agents in the preclinical setting. A systematic review examining the methodology and reporting of preclinical chemotherapeutic drug studies using CRC murine models was conducted. A total of 45 studies were included in this systematic review. The literature was found to be highly heterogeneous with various cell lines, animal strains, animal ages and chemotherapeutic compounds/regimens tested, proving difficult to compare outcomes between similar studies or indeed gain any significant insight into which chemotherapeutic regimen caused adverse events. From this analysis we propose a minimum core outcome dataset that could be regarded as a standardised way of reporting results from in vivo experimentation.


Prins cyclizations in silyl additives with suppression of epimerization: versatile tool in the synthesis of the tetrahydropyran backbone of natural products.
[reactions: see text] A catalytic Prins cyclization reaction has been developed. The involvement of trimethylsilyl halides offers a versatile route to the formation of cis-4-halo-2,6-disubstituted tetrahydropyran rings. The problem of epimerization in Prins cyclization has also been addressed in the total synthesis of (-)-centrolobine using this methodology.


Anisotropic scattering profiles of charged gels in a deformed state.
The structure factor of weakly charged polymer gels under external mechanical strain is calculated considering both thermal and frozen concentration fluctuations as well as the screening of ionic interactions. The butterfly patterns are discussed based on our result.


Preservation of food products by irradiation.
The use of irradiation to preserve food has the potential to significantly enhance our capacity to maximize the quality and quantity of the food we consume. In a world in which distribution of food occurs across continents and in which malnourished populations are in dire need of basic food products, any safe, effective, and efficient means of preserving food is more than welcome. Irradiation, as a method for food preservation, has been studied for more than 30 years. This discussion focuses on this most recent method for the preservation of food with particular emphasis on its effects on the safety, nutritive, and aesthetic values of the food preserved by irradiation. The use of ionizing radiation as a method to preserve foods is one that has been demonstrated to be effective for a variety of food classes. Irradiation offers a means to decontaminate, disinfest, and retard the spoilage of the food supply. At the same time, it appears that the wholesomeness of these food products is maintained. Nutritive value can be sustained by use of effective doses of radiation. Concerns over the safety of irradiated food are rooted in questions regarding the potential induction of radioactivity, harmful radiolytic products, and pathogenic radiation-resistant or mutant strains of microorganisms. Research findings have allayed concerns over safety. However, more research is necessary to conclusively resolve these safety issues. Food irradiation is a promising technology that has and will contribute to our ability to feed the people of this world. This technology is but one of many available ways to preserve our greatest natural resource, the food supply. Enhancement of the ability to preserve food by irradiation will facilitate the distribution of food from fertile developed regions to the malnourished peoples of underdeveloped countries. It is in diminishing the problem of malnourishment and starvation that irradiation as a means to preserve food may find the greatest acceptance.


Prognostic value of pharmacological stress echocardiography in women with chest pain and unknown coronary artery disease.
In this study we sought to investigate the prognostic value of pharmacological stress echocardiography in women referred for chest pain, having unknown coronary artery disease. The noninvasive identification of a high-risk subgroup among women with chest pain and unknown coronary artery disease is an unresolved task to date. A total of 456 women (mean [+/-SD] age 63+/-10 years) underwent pharmacological stress echocardiography with either dipyridamole (n = 305) or dobutamine (n = 151) for evaluation of chest pain and were followed-up for 32+/-19 months. None of them had a previous diagnosis of coronary artery disease. No major complication occurred during stress testing. Five tests (1.1%) were prematurely interrupted because of the appearance of side effects. Echocardiographic positivity was identified in 51 patients. During the follow-up, 23 cardiac events occurred: 3 deaths, 10 infarctions and 10 cases of unstable angina; an additional 21 patients underwent coronary revascularization. At Cox analysis, the echocardiographic evidence of ischemia was found as the only independent predictor of hard cardiac events (death, infarction) (odds ratio [OR] = 27.5; 95% confidence interval [CI] = (6.5 to 115.5; p = 0.0000). When spontaneous cardiac events (death, infarction and unstable angina) were considered as endpoints, the positive echocardiographic result (OR = 23.9; 95% CI = 8.6 to 66.8; p = 0.0000) and family history of coronary artery disease (OR = 3.7; 95% CI = 1.5 to 9.1; p = 0.0037) were independently correlated with prognosis. By using an interactive stepwise procedure, the prognostic value of stress echocardiography was found to be incremental to that provided by clinical variables, both considering hard and spontaneous cardiac events as endpoints. The 3-year survival rate for the negative and the positive population was respectively, 99.5% and 69.5% (p = 0.0000) considering hard cardiac events, 99.2% and 50.6% (p = 0.0000) considering spontaneous cardiac events. Pharmacological stress echocardiography is safe, highly feasible and effective in risk stratification of women with chest pain and unknown coronary artery disease, also when hard endpoints are considered. Its use can have relevant implications in daily clinical practice for selection of patients needing further investigations.


Temporal profile and cell subtype distribution of activated caspase-3 following experimental traumatic brain injury.
This study investigated the temporal expression and cell subtype distribution of activated caspase-3 following cortical impact-induced traumatic brain injury in rats. The animals were killed and examined for protein expression of the proteolytically active subunit of caspase-3, p18, at intervals from 6 h to 14 days after injury. In addition, we also investigated the effect of caspase-3 activation on proteolysis of the cytoskeletal protein alpha-spectrin. Increased protein levels of p18 and the caspase-3-specific 120-kDa breakdown product to alpha-spectrin were seen in the cortex ipsilateral to the injury site from 6 to 72 h after the trauma. Immunohistological examinations revealed increased expression of p18 in neurons, astrocytes, and oligodendrocytes from 6 to 72 h following impact injury. In contrast, no evidence of caspase-3 activation was seen in microglia at all time points investigated. Quantitative analysis of caspase-3-positive cells revealed that the number of caspase-3-positive neurons exceeded the number of caspase-3-positive glia cells from 6 to 72 h after injury. Moreover, concurrent assessment of nuclear histopathology using hematoxylin identified p18-immunopositive cells exhibiting apoptotic-like morphological profiles in the cortex ipsilateral to the injury site. In contrast, no evidence of increased p18 expression or alpha-spectrin proteolysis was seen in the ipsilateral hippocampus, contralateral cortex, or hippocampus up to 14 days after the impact. Our results are the first to demonstrate the concurrent expression of activated caspase-3 in different CNS cells after traumatic brain injury in the rat. Our findings also suggest a contributory role of activated caspase-3 in neuronal and glial apoptotic degeneration after experimental TBI in vivo.


Specific sensitivity of patients with panic attacks to carbon dioxide inhalation.
One inhalation of 35% CO2 in oxygen was administered to 36 patients with anxiety disorders and 14 healthy controls. Eighteen patients had a diagnosis of panic disorder (PD) and 18 of obsessive-compulsive disorder (OCD). As a placebo control for CO2, compressed air was administered in a double-blind design. Immediately before and after the inhalation, levels of anxiety and DSM-III-R symptoms of panic were assessed. CO2 elicited high levels of subjective anxiety in the PD group. Patients with OCD were hardly affected by the inhalation, and did not differ from healthy controls. These results suggest that CO2 challenge should be considered as a specific probe for subjects with panic-anxiety. It is speculated that CO2 may trigger some as yet undefined mechanisms, possibly linked to ventilation control, which demarcate panic from other types of pathological anxiety.


IGF-1 promotes angiogenesis in endothelial cells/adipose-derived stem cells co-culture system with activation of PI3K/Akt signal pathway.
The aim of this study was to investigate the role of insulin-like growth factor-1 (IGF-1) and crosstalk between endothelial cells (ECs) and adipose-derived stem cells (ASCs) in the process of angiogenesis. A three-dimensional collagen gel used to culture mouse ASCs and mouse ECs in vitro was established. The effects of angiogenesis after exposure to IGF-1 were observed by confocal laser scanning microscopy. Western blotting and qPCR were performed to elucidate the underlying mechanisms. IGF-1 treatment promoted the formation of vessel-like structures and the recruitment of ASCs in the three-dimensional collagen gel. The angiogenic genes and proteins in ECs were up-regulated by IGF-1 and in co-culture. Similar changes in the genes and in the proteins were detected in ASCs after exposure to IGF-1 and co-culture. p-Akt expression levels were high in ECs and ASCs after exposure to IGF-1 and co-culture. IGF-1 and co-culture between cells facilitate the process of angiogenesis via the PI3-kinase/Akt signalling pathway. In ECs, IGF-1 stimulates the expression of angiogenesis-related growth factors with the activation of the PI3-kinase/Akt signalling pathway. Co-cultured ECs exposed to excess VEGF-A and other angiogenesis-related growth factors para-secreted from ASCs exhibit high expression of angiogenesis-related genes and proteins. In ASCs, IGF-1 induces the recruitment and function of ASCs by up-regulating the expression of PDGFB, MMPs and α-SMA. Crosstalk with ECs further facilitates changes in ASCs.


Total replacement of the ureter by a segment of the sigmoid colon in the treatment of giant hydronephrosis: calyco-sigmoido-cystostomy.
Two cases of giant hydronephrosis of a solitary kidney are reported. In both of them correctly performed pyelo-ureteroplasties had not assured satisfactory emptying of the pyelocalyceal system. Its dilatation and infection had not disappeared. In order to liquidate the nephrostomy, the ureters were replaced by a sigmoidal loop (calyco-sigmoido-cystostomy) with favorable results assessed 24 and 19 months after operation. The use of an isolated sigmoid segment for replacement of the ureter is discussed.


Versatile reporter systems show that transactivation by human T-cell leukemia virus type 1 Tax occurs independently of chromatin remodeling factor BRG1.
Potent activation of human T-cell leukemia virus type 1 (HTLV-1) gene expression is mediated by the virus-encoded transactivator protein Tax and three imperfect 21-bp repeats in the viral long terminal repeats. Each 21-bp repeat contains a cAMP-responsive-element core flanked by 5' G-rich and 3' C-rich sequences. Tax alone does not bind DNA. Rather, it interacts with basic domain-leucine zipper transcription factors CREB and ATF-1 to form ternary complexes with the 21-bp repeats. In the context of the ternary complexes, Tax contacts the G/C-rich sequences and recruits transcriptional coactivators CREB-binding protein (CBP)/p300 to effect potent transcriptional activation. Using an easily transduced and chromosomally integrated reporter system derived from a self-inactivating lentivirus vector, we showed in a BRG1- and BRM1-deficient adrenal carcinoma cell line, SW-13, that Tax- and 21-bp repeat-mediated transactivation does not require BRG1 or BRM1 and is not enhanced by BRG1. With a similar reporter system, we further demonstrated that Tax- and tumor necrosis factor alpha-induced NF-kappaB activation occurs readily in SW-13 cells in the absence of BRG1 and BRM1. These results suggest that the assembly of stable multiprotein complexes containing Tax, CREB/ATF-1, and CBP/p300 on the 21-bp repeats is the principal mechanism employed by Tax to preclude nucleosome formation at the HTLV-1 enhancer/promoter. This most likely bypasses the need for BRG1-containing chromatin-remodeling complexes. Likewise, recruitment of CBP/p300 by NF-kappaB may be sufficient to disrupt histone-DNA interaction for the initiation of transcription.


Doping reversed-phase media for improved peptide purification.
The purification of therapeutic peptides is most often performed using one or more reversed phase chromatography steps. This ensures high purities while keeping the costs of purification under control. In this paper, a doped reversed phase chromatographic material is tested and compared to traditional reversed phase materials. The doping consists of adding limited amounts of ion exchange ligands to the surface of the material to achieve orthogonal separation and increase the non-hydrophobic interactions with the surface. These ionic groups can either be attractive (opposite charge), or repulsive (same charge) to the peptide. The benefit of this new doped reversed phase material is shown through increases in selectivity in diluted conditions and yield and productivity in overloaded (i.e. industrial) conditions. It is the conjectured that all performance characteristics should increase using repulsive doping groups, whereas these characteristics should decrease when using attractive doping groups. This conjecture is shown to be true through several examples, including purifications of industrially relevant peptide crudes, in industrially relevant conditions. Moreover, the effect of ionic strength and organic modifier concentration was explored and shown to be in line with the expected behavior.


Epigenetic modulation of FOXM1-gene interacting network by BET inhibitors in breast cancer.
FOXM1 is a transcription factor that has been implicated in the genesis of several tumors by regulating the expression of genes involved in the mitotic process. Transcriptomic enrichment analysis was performed to evaluate deregulated pathways in breast cancer, and relapse-free survival associated with the upregulated genes of FOXM1 signature was explored using the KM Plotter online tool. Treatment with bromodomain and extraterminal (BET) inhibitor JQ1 was explored in breast cancer cell lines to evaluate FOXM1 by qPCR and proliferation by MTT colorimetric assays. The FOXM1 gene signature was clearly deregulated in breast cancer patients, particularly in the basal-like subgroup where it was linked with detrimental prognosis. Treatment with the BET inhibitor JQ1 reduced the expression of FOXM1, decreasing cell proliferation in a panel of cell lines, being more active in the basal-like subtype, MDA-MB-231 and HS-578T. Knockdown of FOXM1 or treatment with JQ1 reduced genes included in the FOXM1 signature. Similarly, genes downregulated by the FOXM1 small interfering RNA approach were associated with detrimental outcome in breast cancer patients. Finally, we observed that FOXM1 was amplified in the triple-negative breast cancer subtype in around 15% of patients. Our study demonstrates that activation of the FOXM1 pathway has a prognostic role in breast cancer. JQ1 can modulate the expression of the FOXM1-gene interacting network, opening the opportunity for the evaluation of this compound in breast cancer patients.


[Chemical Constituents from Dalbergia cochinchinensis].
To investigate the chemical constituents from the heartwood of Dalbergia cochinchinensis. Isolate and purify compounds by various column chromatographic methods. Spectral analysis were taken to identify the structures. Elev- en compounds were isolated and identified as dibutyl terephthalate (1), medicarpin (2), pterostilbene (3), 6-hydroxy-2-(2-hydroxy-4- methoxyphenyl)-benzofuran (4), pterocarpol (5), butyl isobutyl phthalate (6), pterolinus B (7), methyl 4-hydroxybenzoate (8), ethyl 4- hydroxybenzoate (9),2-(2'-methoxy-4'-hydroxy)-aryl-3-methyl-6-hydroxy-benzofuran (10) and 6α-hydroxycyclonerolidol (11). Compounds 1 and 6~10 are isolated from Dalbergia genus for the first time, and compounds 2, 4 and 11 are isolated from this plant for the first time.


Leprosy Diagnosed via Teledermatology in a U. S. Urban Academic Health Center Highlights Potential Benefits and a Pitfall of Existing Telemedicine Services.
Purpose: Teledermatology has emerged as an important tool to extend the reach of dermatologists. As utilization of teledermatology services increases, careful attention should be given to the design of the system for consulting dermatologists. Methodology: In this case study, we report how access to a patient's entire medical history in a teledermatology consultation was vital for an accurate diagnosis. Results: This case emphasizes the importance of full access to patients' medical records for improving diagnostic accuracy. We believe that full-access systems will lower the risk of misdiagnosis and will help to maximize the potential of teledermatology.


Constraint on the substrate cytochrome P-450 binding reaction in bovine adrenocortical microsomes at physiological temperature.
The addition of cholate to the microsomes at 37.5 degrees C resulted in a striking decrease in the apparent substrate dissociation constant (K's) and its temperature dependency. The microsomal membranes depleted of 80% of the lipids preserved the temperature dependency of the Ks and exhibited breaks in the Van't Hoff plot at the characteristic temperature of the lipids phase transition. The results indicate that the cytochrome P-450 is considerably restrained from expressing its maximum substrate binding potential at physiological temperature. In addition, the results indicate that the majority of the lipids apparently do not play a significant role in imposing constraint on the substrate-cytochrome -450 binding reaction and in the temperature dependency of the Ks.


Activation energy and force fields during topological transitions of fluid lipid vesicles.
Topological transitions of fluid lipid membranes are fundamental processes for cell life. For example, they are required for endo- and exocytosis or to enable neurotransmitters to cross the neural synapses. Here, inspired by the idea that fusion and fission proteins could have evolved in Nature in order to carry out a minimal work expenditure, we evaluate the minimal free energy pathway for the transition between two spherical large unilamellar vesicles and a dumbbell-shaped one. To address the problem, we propose and successfully use a Ginzburg-Landau type of free energy, which allows us to uniquely describe without interruption the whole, full-scale topological change. We also compute the force fields needed to overcome the involved energy barriers. The obtained forces are in excellent agreement, in terms of intensity, scale, and spatial localization with experimental data on typical fission protein systems, whereas they suggest the presence of additional features in fusion proteins.


[Palliative long-term urinary bladder drainage: the uro-geriatric point of view].
A serious voiding disorder or urinary incontinence represent indications for long-term catheterization of the urinary bladder. Treatment by a transurethral or suprapubic bladder catheter for life-long bladder drainage is accompanied by technical short-term complications and long-term sequelae. The mortality risk associated with inserting a suprapubic catheter is approximately 2% due to an incorrect bladder puncture. Long-term consequences of life-long bladder drainage are hematuria, infection, spasms, stone formation, obstruction, catheter loss, risk of falls and automanipulation especially in cognitively impaired persons. These constitute frequent reasons for emergency treatment or hospitalization. Further problem areas are dependency on caregivers to perform the catheter change and dermal problems in the area of the fistula stoma. To what extent this limits the quality of life of those affected remains unclear up to now. The acute complications and the long-term consequences of life-long bladder drainage make careful decision-making necessary after other therapeutic options have failed, are not appropriate or not desired.


Electrochemiluminescent immunoassay for neuron specific enolase by using amino-modified reduced graphene oxide loaded with N-doped carbon quantum dots.
An ultrasensitive electrochemiluminescence based sandwich immunoassay is presented for determination of neuron specific enolase. The method uses silver-cysteine nanowires as the capture probe and a composite made of amino-modified reduced graphene oxide and nitrogen-doped carbon quantum dots as the signal probe. It was synthesized by covalent coupling of amino-modified reduced graphene oxide to the carboxy groups of nitrogen-doped carbon quantum dots. The nanowires possess a large specific surface and abundant functional groups which facilitate immobilizing the primary antibody (Ab1). The amino-modified reduced graphene oxide is employed as a carrier for loading a large number of the quantum dots and secondary antibody (Ab2). This increases the electrochemiluminescence intensity of quantum dots. Response to neuron specific enolase is linear in the 0.55 fg·mL-1 to 5.5 ng·mL-1 concentration range. It has a detection limit of 0.18 fg·mL-1 (at S/N = 3). The relative standard deviation (for n = 6) is less than 2.9%. The assay is highly sensitive, reproducible, selective and stable. Graphical abstractA novel electrochemiluminescence immunosensor is described that uses amino-modified reduced graphene oxide (amino-rGO), nitrogen-doped carbon quantum dots (N-CQDs) and silver-cysteine nanowires (SCNWs). It was applied to the determination of neuron specific enolase (NSE). Bovine serum albumin: BSA;1-ethyl-3-(3-dimethylaminopropyl)carbodiimide: (EDC;, N-hydroxysuccinimide: NHS.


[Examination of stenotic bladder neck using the Kishev balloon stretch test with the Foley catheter (author's transl)].
A 2-year study with 125 patients who suffered from symptoms suggestive of stenotic bladder neck with or without overlapping symptoms of 'abacterial prostatitis' is described. More than 200 balloon stretch tests were carried out in order to choose 125 patients for operation. With a knife electrode designed by one of us, two deep furrows were cut along the 5 o'clock and the 7 o'clock positions, which begin at the trigonum and continue through the entire bladder neck and the floor of the prostatic part of the urethra to the apex of the verumontanum.