Sex differences in the myocardial inflammatory response to ischemia-reperfusion injury.
The myocardium generates inflammatory mediators during ischemia-reperfusion (I/R), and these mediators contribute to cardiac functional depression and apoptosis. The great majority of these data have been derived from male animals and humans. Sex has a profound effect over many inflammatory responses; however, it is unknown whether sex affects the cardiac inflammatory response to acute myocardial I/R. We hypothesized the existence of inherent sex differences in myocardial function, expression of inflammatory cytokines, and activation of the p38 mitogen-activated protein kinase (MAPK) signaling pathway after I/R. Isolated rat hearts from age-matched adult males and females were perfused (Langendorff), and myocardial contractile function was continuously recorded. After I/R, myocardium was assessed for expression of TNF-alpha, IL-1beta, and IL-6 (RT-PCR, ELISA); IL-1alpha and IL-10 mRNA (RT-PCR); and activation of p38 MAPK (Western blot). All indexes of postischemic myocardial function [left ventricular developed pressure, left ventricular end-diastolic pressure, and maximal positive (+dP/dt) and negative (-dP/dt) values of the first derivative of pressure] were significantly improved in females compared with males. Compared with males, females had decreased myocardial TNF-alpha, IL-1beta, and IL-6 (mRNA, protein) and decreased activation of p38 MAPK pathway. These data demonstrate that hearts from age-matched adult females are relatively protected against I/R injury, possibly due to a diminished inflammatory response.


Kinetic-energy-sensitive mass spectrometry for separation of different ions with the same m/z value.
A double-focusing mass spectrometer (MS) equipped with a superconducting-tunnel-junction (STJ) detector has been applied to measure relative ionization cross-sections for the production of ions that are accompanied by different ion species with the same mass-to-charge (m/z) value. The STJ detector fabricated for this study enables kinetic energy (E) measurement of incoming individual ions at a counting rate of up to approximately 100 k ions/s and an energy resolution (DeltaE/E) of 15%. Both high counting rate and high-energy resolution are necessary to independently determine both m and z and not the m/z value only in ion-counting MS experiments. Ions such as (14)N(2) (2+) and (14)N(+) with the same m/z value can be clearly discriminated using a kinetic-energy-sensitive MS. This fine discrimination capability allows direct determination of relative ionization cross-sections of the homonuclear diatomic ions (14)N(2) (2+)/(14)N(2) (+) and (16)O(2) (2+)/(16)O(2) (+), which are difficult to measure due to the strong interference by the signals of their dissociated atomic ions with noticeably large ionization cross-sections. The new instrument requires no low-abundance heteronuclear diatomic molecules of the forms (14)N(15)N or (16)O(17)O to carry out ionization studies and thus, is expected to be useful in fields such as atmospheric science, interstellar science, or plasma physics.


Characterization of the active site iron in tyrosine hydroxylase. Redox states of the iron.
Tyrosine hydroxylase is an iron-containing monooxygenase that uses a tetrahydropterin to catalyze the hydroxylation of tyrosine to dihydroxyphenylalanine in catecholamine biosynthesis. The role of the iron in this enzyme is not understood. Purification of recombinant rat tyrosine hydroxylase containing 0.5-0.7 iron atoms/subunit and lacking bound catecholamine has permitted studies of the redox states of the resting enzyme and the enzyme during catalysis. As isolated, the iron is in the ferric form. Dithionite or 6-methyltetrahydropterin can reduce the iron to the ferrous form. Reduction by 6-methyltetrahydropterin consumes 0.5 nmol/nmol of enzyme-bound iron, producing quinonoid 6-methyldihydropterin as the only detectable product. In the presence of oxygen, reoxidation to ferric iron occurs. During turnover the enzyme is in the ferrous form. However, a fraction is oxidized during turnover; this can be trapped by added catechol or by the dihydroxyphenylalanine formed during turnover.


Long-term outcomes of pediatric infections: from traditional infectious diseases to long Covid.
There is limited evidence available on the long-term impact of SARS-CoV-2 infection in children. In this article, the authors analyze the recent evidence on pediatric long Covid and lessons learnt from a pediatric post-Covid unit in Rome, Italy. To gain a better understanding of the concerns raised by parents and physicians in relation to the potential long-term consequences of this novel infection, it is important to recognize that long-term effect of a post-infectious disease is not a new phenomenon.


Prolonged length of stay in delayed cholecystectomy is not due to intraoperative or postoperative contributors.
Previous studies have reported that same-day laparoscopic cholecystectomy for acute cholecystitis is superior to delayed elective cholecystectomy. Although this practice is ideal, it requires significant hospital resources, particularly for an underprivileged inner-city population at a large, municipal hospital. We sought to evaluate the implementation of same-day laparoscopic cholecystectomy in a large, municipal hospital and assess the possible benefits of decreasing preoperative length of stay (LOS), particularly its effect on operative time and length of stay in patients with acute cholecystitis. This was a retrospective chart review of patients treated for symptomatic gallstone disease between September 2012 and November 2013. Medical records were reviewed, and relevant data points were collected. Univariate and multivariate regressions were performed to assess the correlation between time to operation (<36 h [no delay] or >36 h [delay]) and the main outcomes (operative time and total length of stay). Inclusion criteria were patients age ≥18 y who underwent same-admission cholecystectomy and had a diagnosis of cholecystitis on pathology. Eighty-eight patients met all inclusion criteria. The mean (standard deviation) preoperative LOS was 76.2 (±48.6) h, the mean operative time was 2.3 (±1.1) h, and the mean postoperative LOS was 60.3 (±60.1) h. The average total LOS was 136 (±79.8) h. Operative times and postoperative LOS were similar for patients in the delay and no delay groups. Patients with >36 h wait before surgery had a total length of stay twice as long as patients with <36 h wait (152 versus 83.3 h; P = 0.0005). These findings remained significant when adjusted for age, sex, radiologic findings, number of preoperative tests, and pathology. Increased preoperative LOS is not associated with a significant increase in operative time. However, it was associated with significantly increased length of stay. Further analysis is needed to explore the potential cost savings of decreasing preoperative LOS.


Neonatal outcomes in relation to timing of repeat cesarean delivery at term.
The increasing rates of repeat cesarean delivery necessitate, more than ever, the need to time deliveries to optimize neonatal outcomes. A recent large multicenter cohort study demonstrated that, contrary to current recommendations, a high proportion of elective cesarean deliveries in the USA are performed before 39 weeks' gestation. These early deliveries are associated with a significant increase in several adverse neonatal events, including respiratory complications, treated hypoglycemia, newborn sepsis and admission to the neonatal intensive care unit. This, together with results of previous studies, supports the recommendation to delay elective delivery to 39 weeks' gestation. From the current data, it is uncertain whether delivery before 39 weeks' gestation with documented fetal lung maturity will prevent the increased neonatal morbidity.


Achieving Food System Transformation: Insights From A Retrospective Review of Nutrition Policy (In)Action in High-Income Countries.
Comprehensive nutrition policies are required urgently to help transform food systems to more equitably deliver healthy, sustainable diets. Literature was searched systematically for nutrition policies of the then 34 Organisation for Economic Co-operation and Development (OECD) members as part of a scoping study. Recently, results were re-analysed, against the NOURISHING framework. Twenty-three nutrition policy documents were identified for 19 jurisdictions. Most policy actions focused on the behaviour change communication domain: all (100%) promoted consumption of 'healthy' choices. In the food environment domain, most policies included food labelling (84%), product reformulation (68%), providing healthy foods in public institutions (89%, mainly schools), and restricting food advertising (53%), largely through voluntary codes. Relatively few economic tools were being applied. There was very little focus on reducing consumption of 'unhealthy' food or drinks. Not all nutrition policy actions identified were covered by the NOURISHING framework. The NOURISHING framework could be expanded to more comprehensively encompass the health and sustainability dimensions of food systems, eg, by detailing optimum governance arrangements. As recently as seven years ago, half of the most developed economies globally did not have a publicly available nutrition policy. Existing policies were dominated by conventional nutrition education approaches, while policy actions targeting food environments, and regulatory and legislative reforms, were rare. This is consistent with a neo-liberal approach centring individual responsibility. No examples of the multi-strategy, inter-sectoral, coordinated, evidence-based policies required to drive systemic transformation were identified. Therefore, it is not surprising that rates of obesity and diet-related conditions have continued to rise in these jurisdictions, nor that governments are currently off-track to deliver the systemic transformation required to meet relevant global health and sustainable development goals.


HLA class II susceptibility to multiple sclerosis among Ashkenazi and non-Ashkenazi Jews.
To look for HLA class II alleles and haplotypes conferring susceptibility to multiple sclerosis (MS) in the Jewish population of Israel. Population-based cohort of clinically definite patients with MS tested prospectively over 7 years. Referral center in a neurology clinic at a university hospital in the greater Jerusalem area in Israel. A total of 162 consecutive patients with clinically definite MS from the 2 main ethnic Jewish groups in Israel: 104 Ashkenazi (80 with a relapsing remitting or secondary progressive and 24 with a primary chronic progressive course of the disease) and 58 non-Ashkenazi (36 with a relapsing remitting or secondary progressive course and 22 with a primary chronic progressive course of the disease), matched with 132 Ashkenazi and 120 non-Ashkenazi healthy controls. The relationship between the various HLA class II alleles and haplotypes and MS, as defined by the polymerase chain reaction and sequence-specific oligonucleotide probe hybridization, among the Ashkenazi and the non-Ashkenazi Jewish sections and with respect to the different clinical courses of the disease. The haplotype DRB1*1501, DQA1*0102, DQB1*0602 was found to be associated with MS among both Ashkenazi and non-Ashkenazi patients (P<.001 and P =.04, respectively). Among the non-Ashkenazi patients, a new association of haplotypes DRB1*1303, DQA1*05, and DQB1*030 with MS was detected (P = .03). The MS susceptibility alleles, DRB 1* 1501, DQA1*0102, and DQB1*0602 , were found in association with the Ashkenazi patients (P<.001, P=.02, and P=.01, respectively); DRB1*1501 and DRB1*1303 were more frequently observed among the non-Ashkenazi patients (P = .03, P = .04, respectively). On subdivision of the patients into clinical subgroups, associations of DRB1*0801, DQA1*0102, DQA1*0401, and DQB1*0602 with primary chronic progressive MS among the Ashkenazi patients were evident (P = .03, P = .04, P = .04 and P = .05, respectively), whereas DRB1* 1501, DRB1*03011, and DQB1*0602 were associated with relapsing remitting or secondary progressive among the non-Ashkenazi patients (P = .05, P = .05, and P = .03, respectively). This study, unlike previous ones, is the first to show a significant association between HLA class II alleles and MS in the Jewish population. The association with the HLA-DR2-related haplotype is similar to that among non-Jewish white patients with MS. Moreover, our data support the possibility that DRB1*1501 is the susceptibility allele responsible for the association between this haplotype and MS in the Jewish population. Our study also underscores differences in HLA profiles between Ashkenazi and non-Ashkenazi patients, and between the different clinical courses of the disease. The latter may indicate that the clinical courses of MS are influenced by the genetic background.


Impact of renin angiotensin system modulation on the hyperfiltration state in type 1 diabetes.
The initial stages of diabetic nephropathy are characterized by glomerular hyperfiltration and hypertension, processes that have been linked to initiation and progression of renal disease. Renin angiotensin system (RAS) blockade is commonly used to modify the hyperfiltration state and delay progression of renal disease. Despite this therapy, many patients progress to ESRD, suggesting heterogeneity in the response to RAS modulation. The role of the RAS in the hyperfiltration state in adolescents with uncomplicated type 1 diabetes was examined, segregated on the basis of the presence of hyperfiltration. Baseline renal hemodynamic function was characterized in 22 patients. Eleven patients exhibited glomerular hyperfiltration (GFR>or=135 ml/min), and in the remaining 11 patients, the GFR was <130 ml/min. Renal hemodynamic function was assessed in response to a graded angiotensin II (AngII) infusion during euglycemic conditions and again after 21 d of angiotensin-converting enzyme (ACE) inhibition with enalapril. AngII infusion under euglycemic conditions resulted in a significant decline in GFR and renal plasma flow in the hyperfiltration group but not in the normofiltration group. After ACE inhibition, GFR fell but did not normalize in the hyperfiltration group; the normofiltration group showed no change. These data show significant differences in renal hemodynamic function between hyperfiltering and normofiltering adolescents with type 1 diabetes at baseline, after AngII infusion and ACE inhibition. The response to ACE inhibition and AngII in hyperfiltering patients suggests that vasodilation may complement RAS activation in causing the hyperfiltration state. The interaction between glomerular vasoconstrictors and vasodilators requires examination in future studies.


Endothelium-dependent induction of vasorelaxation by the butanol extract of Phellinus igniarius in isolated rat aorta.
The butanol extract of Phellinus igniarius (BPI) induced relaxation of the phenylephrin e-precontracted rat aorta in a dose-dependent manner, and its effect was abolished by the removal of functional endothelium. Pretreatment of the aortic tissues with N(G)-nitro-L-arginine methyl ester (L-NAME), methylene blue, or 1H-[1,2,4]-oxadiazole-[4,3-alpha]-quinoxalin1-one (ODQ) inhibited the vascular relaxation induced by BPI. BPI-induced vascular relaxations were also markedly attenuated by the addition of verapamil or diltiazem, while the relaxant effect of BPI was not blocked by pretreatment with indomethacine, glibenclamide, tetraethylammonium (TEA), atropine, or propranolol. Incubation of endothelium-intact rat aorta with BPI increased the production of cGMP in a dose-dependent manner. These results suggest that BPI dilates vascular smooth muscle via endothelium-dependent nitric oxide-cGMP signaling pathway, with the possible involvement of L-type Ca(2+) channels.


The application of capillary electrophoresis to the analysis of vitamins in food and beverages.
Capillary electrophoretic (CE) methods have been used to separate and determine a wide range of water-soluble vitamins in pharmaceutical preparations, but has found limited application in determining vitamins at naturally occurring levels in food and beverages. CE has been used to determine vitamin C in fruits and beverages, niacin in a range of foods and thiamine in samples of meat and milk. The CE methodologies used to determine vitamins in pharmaceutical preparations and biological fluids are also included as examples of the effectiveness of CE in vitamin analysis.


Patterns of leisure activities of young adults with mild mental retardation.
Leisure activities were examined for young adults with mild mental retardation. Some had disappeared from mental retardation services after leaving school; others were in adult day services. Peers without mental retardation (comparison subjects) were also included. Various leisure activities were combined using cluster analysis. Some adults with mental retardation and some comparison subjects were found in each of the 13 clusters obtained but proportions varied. Leisure patterns were strongly influenced by gender and marital status as well as mental retardation, except for one cluster, Socially Isolated, Impoverished, where there were more adults with mental retardation, irrespective of gender and marital status.


Acoustic Classification and Optimization for Multi-Modal Rendering of Real-World Scenes.
We present a novel algorithm to generate virtual acoustic effects in captured 3D models of real-world scenes for multimodal augmented reality. We leverage recent advances in 3D scene reconstruction in order to automatically compute acoustic material properties. Our technique consists of a two-step procedure that first applies a convolutional neural network (CNN) to estimate the acoustic material properties, including frequency-dependent absorption coefficients, that are used for interactive sound propagation. In the second step, an iterative optimization algorithm is used to adjust the materials determined by the CNN until a virtual acoustic simulation converges to measured acoustic impulse responses. We have applied our algorithm to many reconstructed real-world indoor scenes and evaluated its fidelity for augmented reality applications.


Highly Sensitive and Selective Triethylamine Sensing through High-Entropy Alloy (Ti-Zr-Cr-V-Ni) Nanoparticle-Induced Fermi Energy Control of MoS2 Nanosheets.
A giant enhancement of nearly 100 times is seen in triethylamine response through Ti-Zr-Cr-V-Ni high-entropy alloy nanoparticle (HEA NP)-induced fermi energy control of two-dimensional molybdenum disulfide (MoS2) nanosheets. These Laves-phase HEA NP-decorated MoS2 samples are synthesized using cryomilling followed by 30 h of sonication. The prolonged sonication results in well-exfoliated MoS2 with fairly small (∼10-20 nm) HEA NPs anchored due to cryomilling confirmed by extensive microscopic and spectroscopic examinations. The presence of HEA NPs leads to reduction in edge oxidation of MoS2 as seen from X-ray photoelectron spectroscopy. Moreover, this edge state reduction causes strong Fermi level pinning, which is commonly observed in layered MoS2 with bulk metal electrodes. This leads to target gas-specific carrier-type response and selective oxidation of TEA vapors due to highly catalytically active metals. The resulting composite (MoS2 + NPs) exhibits high response (380% for 2000 ppm TEA vapors) along with selectivity toward TEA at 50 °C. The cross-sensitivity of the composite to other volatile organic compounds and NH3, CO, and H2 has been very minimal. Thus, the highly selective catalytic activity of metal alloy NPs and their Fermi energy control has been proposed as the prime factors for observed large sensitivity and selective response of MoS2 + NP nanocomposites.


Singlet exciton fission for solar cell applications: energy aspects of interchromophore coupling.
Singlet exciton fission, a process that converts one singlet exciton to a pair of triplet excitons, has the potential to enhance the efficiency of both bulk heterojunction and dye-sensitized solar cells and is understood in crystals but not well understood in molecules. Previous studies have identified promising building blocks for singlet fission in molecular systems, but little work has investigated how these individual chromophores should be combined to maximize triplet yield. We consider the effects of chemically connecting two chromophores to create a coupled chromophore pair and compute how various structural choices alter the thermodynamic and kinetic parameters likely to control singlet fission yield. We use density functional theory to compute the electron transfer matrix element and the thermodynamics of fission for several promising chromophore pairs and find a trade-off between the desire to maximize this element and the desire to keep the singlet fission process exoergic. We identify promising molecular systems for singlet fission and suggest future experiments.


Retrophylogenomics place tarsiers on the evolutionary branch of anthropoids.
One of the most disputed issues in primate evolution and thus of our own primate roots, is the phylogenetic position of the Southeast Asian tarsier. While much molecular data indicate a basal place in the primate tree shared with strepsirrhines (prosimian monophyly hypothesis), data also exist supporting either an earlier divergence in primates (tarsier-first hypothesis) or a close relationship with anthropoid primates (Haplorrhini hypothesis). The use of retroposon insertions embedded in the Tarsius genome afforded us the unique opportunity to directly test all three hypotheses via three pairwise genome alignments. From millions of retroposons, we found 104 perfect orthologous insertions in both tarsiers and anthropoids to the exclusion of strepsirrhines, providing conflict-free evidence for the Haplorrhini hypothesis, and none supporting either of the other two positions. Thus, tarsiers are clearly the sister group to anthropoids in the clade Haplorrhini.


Structural characterization of a novel GPVI-nanobody complex reveals a biologically active domain-swapped GPVI dimer.
Glycoprotein VI (GPVI) is the major signaling receptor for collagen on platelets. We have raised 54 nanobodies (Nb), grouped into 33 structural classes based on their complementary determining region 3 loops, against recombinant GPVI-Fc (dimeric GPVI) and have characterized their ability to bind recombinant GPVI, resting and activated platelets, and to inhibit platelet activation by collagen. Nbs from 6 different binding classes showed the strongest binding to recombinant GPVI-Fc, suggesting that there was not a single dominant class. The most potent 3, Nb2, 21, and 35, inhibited collagen-induced platelet aggregation with nanomolar half maximal inhibitory concentration (IC50) values and inhibited platelet aggregation under flow. The binding KD of the most potent Nb, Nb2, against recombinant monomeric and dimeric GPVI was 0.6 and 0.7 nM, respectively. The crystal structure of monomeric GPVI in complex with Nb2 revealed a binding epitope adjacent to the collagen-related peptide (CRP) binding groove within the D1 domain. In addition, a novel conformation of GPVI involving a domain swap between the D2 domains was observed. The domain swap is facilitated by the outward extension of the C-C' loop, which forms the domain swap hinge. The functional significance of this conformation was tested by truncating the hinge region so that the domain swap cannot occur. Nb2 was still able to displace collagen and CRP binding to the mutant, but signaling was abolished in a cell-based NFAT reporter assay. This demonstrates that the C-C' loop region is important for GPVI signaling but not ligand binding and suggests the domain-swapped structure may represent an active GPVI conformation.


Long-term efficacy and safety of tildrakizumab in Japanese patients with moderate to severe plaque psoriasis: Results from a 5-year extension of a phase 3 study (reSURFACE 1).
The three part, double-blind, randomized, controlled reSURFACE 1 trial and extension study (NCT01722331) evaluated efficacy and safety of tildrakizumab in adults with moderate to severe plaque psoriasis. Patients with ≥50% improvement from baseline in Psoriasis Area and Severity Index (PASI 50) following treatment with tildrakizumab 100 mg (TIL100) or 200 mg (TIL200) could enter the optional long-term extension study and continue treatment at the same dose for an additional 192 weeks. This subgroup analysis assessed the long-term efficacy and safety of tildrakizumab treatment for Japanese patients enrolled in reSURFACE 1 for up to 5 years of treatment. The primary efficacy outcomes were the proportions of patients who maintained PASI 75 and Physician Global Assessment (PGA) clear or minimal with ≥2-grade reduction from baseline (PGA 0/1) from base study week 64 to extension week 192. Secondary outcomes were the proportion of patients who maintained PASI 90/100 from base study week 64 to extension week 192. Adverse events (AEs) were monitored throughout the study and for up to 20 weeks after the last study visit. Of the 120 Japanese patients who entered the reSURFACE 1 extension study, 43 (79.6%) patients receiving tildrakizumab 100 mg and 58 (87.9%) patients receiving tildrakizumab 200 mg completed the extension study. Of all Japanese patients with PASI 75/90/100 and PGA 0/1 at week 64, 85%/88% receiving TIL100/TIL200 maintained PASI 75, 70%/96% maintained PASI 90, 63%/67% maintained PASI 100, and 68%/72% maintained PGA 0/1 at extension week 192. AEs led to discontinuation in 1.7 patients per 100 patient-years (P100PY) receiving tildrakizumab 100 mg and 0.8 P100PY receiving tildrakizumab 200 mg. Incidences of severe infections, malignancies, confirmed major adverse cardiac events, and hypersensitivity reactions were low in both treatment groups. Through 5 years of treatment, tildrakizumab maintained efficacy and was well tolerated with low rates of AEs of special interest.


Next generation sequencing yields the complete mitochondrial genome of the striped raphael catfish, Platydoras armatulus (Siluriformes: Doradidae).
Striped raphael catfish, Platydoras armatulus, belongs to catfish family Doradidae, which are endemic to South America and featured by a single row of thorny scutes along the lateral line. In this study, we determined the complete mitochondrial genome of P. armatulus by next generation sequencing of genomic DNA without prior sample processing or primer design. Bioinformatics analysis provided the full-length mitogenome sequence of 16,470 bp. The P. armatulus mitogenome is composed of 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and 1 control region, showing typical circular molecule structure as in other vertebrates. The whole genome base composition was estimated to be 30.9% A, 25.2% T, 15.9% G and 28.0% C, with A/T bias of 56.1%. The generation of P. armatulus mitogenome sequence enriched catfish molecular resources that are valuable for phylogenetic analysis and species identification studies.


A Microfluidic System for Stable and Continuous EEG Monitoring from Multiple Larval Zebrafish.
Along with the increasing popularity of larval zebrafish as an experimental animal in the fields of drug screening, neuroscience, genetics, and developmental biology, the need for tools to deal with multiple larvae has emerged. Microfluidic channels have been employed to handle multiple larvae simultaneously, even for sensing electroencephalogram (EEG). In this study, we developed a microfluidic chip capable of uniform and continuous drug infusion across all microfluidic channels during EEG recording. Owing to the modular design of the microfluidic channels, the number of animals under investigation can be easily increased. Using the optimized design of the microfluidic chip, liquids could be exchanged uniformly across all channels without physically affecting the larvae contained in the channels, which assured a stable environment maintained all the time during EEG recording, by eliminating environmental artifacts and leaving only biological effects to be seen. To demonstrate the usefulness of the developed system in drug screening, we continuously measured EEG from four larvae without and with pentylenetetrazole application, up to 60 min. In addition, we recorded EEG from valproic acid (VPA)-treated zebrafish and demonstrated the suppression of seizure by VPA. The developed microfluidic system could contribute to the mass screening of EEG for drug development to treat neurological disorders such as epilepsy in a short time, owing to its handy size, cheap fabrication cost, and the guaranteed uniform drug infusion across all channels with no environmentally induced artifacts.


A new experimental absolute nuclear magnetic shielding scale for oxygen based on the rotational hyperfine structure of H(2)(17)O.
The hyperfine structure in the rotational spectrum of water containing (17)O has been investigated experimentally and by means of quantum-chemical calculations. The Lamb-dip technique has been used to resolve the hyperfine structure due to spin-rotation as well as spin-spin interactions and allowed the determination of the corresponding hyperfine parameters with high accuracy. The experimental investigation and, in particular, the analysis of the spectra have been supported by quantum-chemical computations at the coupled-cluster level. The experimental (17)O isotropic spin-rotation constant of H(2)(17)O has been used in a further step for the determination of the paramagnetic part of the corresponding nuclear magnetic shielding constant, whereas the diamagnetic contribution as well as vibrational and temperature corrections have been obtained from quantum-chemical calculations. This joint procedure leads to a value of 325.3(3) ppm for the oxygen shielding in H(2)(17)O at 300 K, in good agreement with pure theoretical predictions, and in this way provides the basis for a new absolute oxygen shielding scale.


X-ray crystallographic evidence for the presence of the cysteine tryptophylquinone cofactor in L-lysine ε-oxidase from Marinomonas mediterranea.
We have determined the x-ray crystal structure of L-lysine ε-oxidase from Marinomonas mediterranea in its native and L-lysine-complex forms at 1.94- and 1.99-Å resolution, respectively. In the native enzyme, electron densities clearly indicate the presence of cysteine tryptophylquinone (CTQ) previously identified in quinohemoprotein amine dehydrogenase. In the L-lysine-complex, an electron density corresponding to the bound L-lysine shows that its ε-amino group is attached to the C6 carbonyl group of CTQ, suggesting the formation of a Schiff-base intermediate. Collectively, the present crystal structure provides the first example of an enzyme employing a tryptophylquinone cofactor in an amine oxidase.


Acute stress effects on GABA and glutamate levels in the prefrontal cortex: A 7T 1H magnetic resonance spectroscopy study.
There is ample evidence that the inhibitory GABA and the excitatory glutamate system are essential for an adequate response to stress. Both GABAergic and glutamatergic brain circuits modulate hypothalamus-pituitary-adrenal (HPA)-axis activity, and stress in turn affects glutamate and GABA levels in the rodent brain. However, studies examining stress-induced GABA and glutamate levels in the human brain are scarce. Therefore, we investigated the influence of acute psychosocial stress (using the Trier Social Stress Test) on glutamate and GABA levels in the medial prefrontal cortex of 29 healthy male individuals using 7 Tesla proton magnetic resonance spectroscopy. In vivo GABA and glutamate levels were measured before and 30 min after exposure to either the stress or the control condition. We found no associations between psychosocial stress or cortisol stress reactivity and changes over time in medial prefrontal glutamate and GABA levels. GABA and glutamate levels over time were significantly correlated in the control condition but not in the stress condition, suggesting that very subtle differential effects of stress on GABA and glutamate across individuals may occur. However, overall, acute psychosocial stress does not appear to affect in vivo medial prefrontal GABA and glutamate levels, at least this is not detectable with current practice 1H-MRS.


The burn registry program in Iran - First report.
Burn injury is still a major problem in Iran, resulting in many reports which are usually dispersed, uncoordinated and probably unreliable. We created a burn registry comprised of a data entry program with 222 variables for each admitted burn patient. This program was established in August 2010 at the Iran University of Medical Sciences, Motahari Burn Hospital. We had 14,277 burn patients from August 2010 to August 2011, 877 of whom were admitted to the hospital. Of the patients, 65.9% were male and 34.1% were female. The age was 28.85 years (SD = 19.77). The most prevalent cause of burn was flame 78.5% (pipe propane gas 57.2; kerosene 19.9%). The mean total body surface area (TBSA) involvement was 23%. A total of 77.8% of patients were discharged with partial recovery, while mortality was 8.9%. The mean hospital stay was 14.63 days (SD =11.07). The program is designed to help understand the scope of burn injury in Iran, providing information on patients, etiology, and course of treatment. It also highlights differences between various parts of the country in terms of the causes and frequency of burn injuries. Moreover, the burn registry provides a basis for further research and surveys for treatment and preventive programs. Our results showed that, although Emergency Medical Services (EMS) staff are highly capable and well-trained, their coverage seems to be less than 50% and needs to be increased. Marriage status was shown to have no influence on the occurrence of burns, and among our patients, 57.0 % were poorly educated.


Voltage-dependent potassium channels Kv1.3 and Kv1.5 in human fetus.
Voltage-dependent K(+) channels (Kv) control repolarization and membrane potential in electrically excitable cells. In addition, Kv channels are involved in the maintenance of vascular smooth muscle tone, insulin release, epithelial K(+) transport, cell proliferation and leukocyte activation. Kv1.3 and Kv1.5 are widely distributed throughout the body and are involved in a variety of physiological processes taking place in the immune system, brain and muscle. Since the developmental pattern of Kv channels has an essential role in the maturing human, we aimed to study Kv1.3 and Kv1.5 channels in 8-10 week human fetal tissues. We chose that gestational age because all organs are in place and the nervous system, although not fully developed. However, the human embryo is undergoing major changes, which will lead to a defined adult pattern. Our results indicated that numerous tissues expressed Kv1.3 and Kv1.5. While Kv1.3 overlapped with the central and peripheral nervous systems, Kv1.5 was mostly localized in the central nervous system. In addition, both channels were abundantly expressed in the hematopoietic fetal liver. Finally, Kv1.5 heavily stained skeletal muscle and heart, whereas Kv1.3 was slightly present. This is the first study to analyze Kv1.3 and Kv1.5 in human during the beginning of fetal development.


Oropharyngeal carriage of macrolide-resistant viridans group streptococci: a prevalence study among healthy adults in Belgium.
Viridans group streptococci (VGS) are gaining significance as reservoirs of resistance determinants for respiratory tract pathogens. Our aim was to investigate healthy adults for oropharyngeal carriage of VGS that are resistant to macrolides, as well as to other common antibiotics. Macrolide-resistant VGS were isolated from throat samples of 154 healthy Belgian adults, and phenotyped and genotyped for erm(A), erm(B) and mef(A). In vitro susceptibilities to 10 antimicrobials and the presence of tetracycline resistance genes were also determined. Carriage was detected in 71% of the population screened, from whom 157 unique, macrolide-resistant VGS were isolated. A constitutive (cMLS) phenotype was present in 105 isolates, of which 102 isolates carried either erm(B) or erm(B) + mef(A). The remaining three isolates did not present with any of the genes studied. All 45 isolates showing the M phenotype carried mef(A). The least abundant inducible (iMLS) isolates (n = 7) carried either erm(B) or erm(B) + mef(A). The most abundant macrolide-resistant VGS species was Streptococcus mitis (51%). Co-resistance to tetracycline was identified in 114 isolates, of which tet(M) was present in 105, tet(O) in two and both tet(M) and tet(O) in one, while the remaining six isolates did not present with either gene. tet(M) was also present in four tetracycline susceptible and two intermediately resistant isolates. Fluoroquinolone resistance (ciprofloxacin MIC > or = 4 mg/L) was detected in 16 isolates. Resistance to telithromycin, penicillin and chloramphenicol was appreciably low. This study highlights a high oropharyngeal carriage of macrolide-resistant VGS and its co-resistance to tetracycline and fluoroquinolones among healthy Belgian adults.


Hypoglycemia: a factor associated with low survival rate of neonatal piglets infected with transmissible gastroenteritis virus.
The main purpose of this work was to study changes in the balance of fluids, electrolytes and blood metabolites in neonatal piglets with severe transmissible gastroenteritis. Six two day old conventional piglets were infected with transmissible gastroenteritis virus while six others were used as normal controls. Blood samples were collected in heparin when the infected piglets were moribund. The following variables were measured: packed red cell volume, total plasma protein and bicarbonate, blood pH, blood urea nitrogen and plasma glucose, creatinine, chloride, inorganic phosphorus, sodium, potassium, magnesium and calcium. Vomiting and diarrhea appeared 12 to 24 hours postinoculation in the infected piglets and they were moribund one or two days later. Before becoming moribund, most of the piglets fell rapidly into a lethargic and comatose state. The most evident changes in their blood variables were an increase in packed cell volume, total protein, blood urea nitrogen, phosphorus and magnesium levels and a decrease in pH and bicarbonate concentration as well as a severe hypoglycemia. The results suggest that severe hypoglycemia coupled with metabolic acidosis and dehydration might be an important factor contributing to the high mortality rates caused by transmissible gastroenteritis in neonatal piglets. The hypoglycemia results from a combination of the inadequate glucose metabolism inherent to neonatal piglets and the acute maldigestion and malabsorption resulting from the diffuse and severe villous atrophy induced by the virus.


Persistent hiccups and vomiting with multiple cranial nerve palsy in a case of zoster sine herpete.
A 76-year-old man came to our hospital complaining of hiccups and vomiting lasting for five days. A neurological examination showed dysfunction of cranial nerves V, VII, VIII, IX and X on the left side. Cerebrospinal fluid polymerase chain reaction for varicella zoster virus-DNA was positive. The patient responded well to treatment with intravenous acyclovir and steroids. To the best of our knowledge, this is the first case report of zoster sine herpete presenting with persistent hiccups and vomiting. It is important to keep in mind that herpes zoster can present with symptoms that closely resemble those of intractable hiccups and nausea of neuromyelitis optica. Early detection of the virus is critical for making appropriate treatment decisions.


Pharmacokinetics of high-dose simvastatin in refractory and relapsed chronic lymphocytic leukemia patients.
To evaluate the pharmacokinetics of simvastatin at the maximum tolerated dose (MTD) of 7.5 mg/kg, twice daily, in the context of a pilot trial enrolling patients with recurrent and refractory chronic lymphocytic leukemia. Patients received simvastatin orally at MTD for 7 days during a 21-day cycle for 6 cycles. Blood samples were collected during cycle 1. Simvastatin lactone and carboxylate concentrations were measured in plasma and peripheral blood mononuclear cells (PBMCs) using a validated HPLC-MS/MS assay. Patients accrued to this study showed high variability in their exposure to simvastatin. Exposure was dose proportional (AUC and C max) as compared to those receiving standard hyperlipidemia therapy. Peak plasma concentrations ranged from 0.08 to 2.2 and from 0.03 to 0.6 μM for simvastatin lactone and carboxylate, respectively. Our study shows that when simvastatin is administered at its MTD, only low micro-molar concentrations are achieved in plasma and PBMCs, which is consistent with the results observed in previous studies with lovastatin, but far lower than the concentrations required for anticancer effects in vitro. However, whether simvastatin at its MTD can confer therapeutic benefits to patients still remains to be determined.


High-throughput single-molecule mapping links subtelomeric variants and long-range haplotypes with specific telomeres.
Accurate maps and DNA sequences for human subtelomere regions, along with detailed knowledge of subtelomere variation and long-range telomere-terminal haplotypes in individuals, are critical for understanding telomere function and its roles in human biology. Here, we use a highly automated whole genome mapping technology in nano-channel arrays to analyze large terminal human chromosome segments extending from chromosome-specific subtelomere sequences through subtelomeric repeat regions to terminal (TTAGGG)n repeat tracts. We establish detailed maps for subtelomere gap regions in the human reference sequence, detect many new large subtelomeric variants and demonstrate the feasibility of long-range haplotyping through segmentally duplicated subtelomere regions. These features make the method a uniquely valuable new tool for improving the quality of genome assemblies in complex DNA regions. Based on single molecule mapping of telomere-terminal DNA fragments, we provide proof of principle for a novel method to estimate telomere lengths linked to distinguishable telomeric haplotypes; this single-telomere genotyping method may ultimately enable delineation of human cis elements involved in telomere length regulation.


Prevalence and determinants of inadequate use of folic acid supplementation in Japanese pregnant women: the Japan Environment and Children's Study (JECS).
The aim of the study was to clarify the prevalence and determinants of inadequate use of folic acid supplementation in pregnant Japanese women. This study was part of the Japan Environment and Children's Study, a nationwide and government-funded birth cohort study. We collected information on the use of folic acid supplementation before and during pregnancy and characteristics of participants using self-administered questionnaire. Among 9849 women who completed the data collection for this study, the prevalence of inadequate users was 92.6% of the total population and varied from 84.5% to 96.2% among regions. On the basis of multivariate logistic regression analysis, younger age, not married, lower family income, multipara, natural conception and no history of spontaneous abortion were found to be determinants for inadequate users of folic acid supplementation. Most Japanese pregnant women show inadequate folic acid supplementation use. Japanese women of child-bearing age need to be specifically informed about the need for periconceptional intake of folic acid to prevent neural tube defects.


Adult intussusception induced by the stump of the jejunal loop after liver transplantation.
Intussusception occurs rarely in adults compared with children, and in the case of adults, some organic disease, such as a benign or malignant tumor, can be found at the leading edge of the intussusception in about 90% of adult cases. This study reports a case of adult intussusception in which the stump of the jejunal loop in a Roux-en-Y bilioenteric reconstruction acted as a leading tip of the intussusception.


Survival and chromate reducing ability of Pseudomonas aeruginosa in industrial effluents.
The ability of a chromate-reducing Pseudomonas aeruginosa strain, isolated from tannery effluent, to survive and reduce chromate in the effluent of a tannery and an electroplating unit was evaluated. The test strain survived in the native tannery effluent but numbers fell sharply in the native electroplating effluent. Supplementation with a carbon (C), nitrogen (N) and phosphorus (P) source supported bacterial multiplication and chromate reduction in both types of effluents with almost equal efficiency. Chromate reduction, however, was not observed in the absence of C, N or P supplement, or in the chromate-reducing strain.


Potential fluoride exposure from selected food crops grown in high fluoride soils in the Makueni County, south-eastern Kenya.
Makueni County, located in south-eastern Kenya, faces challenges such as limited potable water and restricted food supplies as the result of semi-aridity. High fluoride (F) concentrations have been reported in drinking water with resultant dental fluorosis affecting the local population. To determine the potential F exposure through the consumption of food crops grown in the area, F concentration was assessed in the main five locally grown and consumed crops. Additionally, the water-soluble F fraction was determined from 30 soil samples with mineralogical determination of 20 samples. Mean F concentration in the food crops was in the order; 700, 288, 71.2, 36.6, and 29 mg/kg in kale, cowpeas leaves, green grams, cowpeas (legume portion), and maize, respectively. The F concentration in farm soils ranged from 0 to 3.47 mg/kg (mean of 0.87 mg/kg) and showed a significant strong positive correlation (p = 0.03, r = 0.89) with F values in the crops. Apatite, muscovite, and biotite were identified as the F-rich minerals present. While considering two hypothetical F absorption fractions (75 and 100%), the estimated average daily dose (EADD) of F from consuming the crops ranged between 0.004 and 65.17 mg/kg/day where the highest values were from the vegetables. Most of these values were higher than the F reference dose (RfD) of 0.06 mg/kg. The estimated EADD values of several hypothetical meals prepared from the analyzed crops revealed that steamed kale and maize porridge pose the highest health risk of F associated diseases to the local population, whereas boiled cowpeas pose no health risk. Children, due to their higher daily energy requirement and low body weight, were the most vulnerable group at risk of high daily F intake relative to the RfD. These results suggest that consumption of the analyzed food crops in Makueni County may significantly contribute to F related diseases in the local population. This creates a food security issue for the area because of the potential health risks associated with these crops which are highly relied upon in the semi-arid area with a limited selection of food crops available and viable to grow.


Low prevalence of rheumatoid arthritis among patients with pre-existing type 2 diabetes mellitus.
Type 2 diabetes mellitus (T2DM) is a non-autoimmune disease characterized by chronic hyperglycemia and increased non-enzymatic glycation of amino groups. Glycation occurs through a series of events eventually leading to the formation of irreversible "advanced glycation end-products" (AGEs). AGEs may affect the function of long-lived proteins, including cytokines, immunoglobulins and their receptors, resulting in a "less active" immune system. We aimed to test the hypothesis that a common inflammatory chronic disease, such as rheumatoid arthritis (RA), in which the earliest event is an inflammatory response to unknown stimulus, has a lower prevalence in these patients than in normoglycemic, non-diabetic subjects. In this study, we compared the prevalence of RA in a prospectively followed outpatient cohort of patients with T2DM patients (n=1,630) with a control, matched, non-diabetic population (n=1,630). Among non-diabetic controls, 13 patients (prevalence 0.80%) with RA were identified. An almost 3-fold lower prevalence of RA (0.25%) was found in consecutive patients with T2DM (P=0.029). Most of the RA cases among participants with T2DM were diagnosed early after diabetes onset. The onset of RA in patients with T2DM occurred at significantly older age (64±15 years) as compared to the non-diabetes group (48±18 years; P=0.004). The prevalence of RA is lower and occurs in an older age in patients with pre-existing T2DM in comparison with people without T2DM.


Pharmacists' opinions about and compliance with recommendations for intravenous admixture practices.
Hospital pharmacy departments nationwide were surveyed to determine compliance with and opinions on the importance of recommended practices for preparation of i.v. admixtures. Questionnaires that included 84 recommendations by the National Coordinating Committee on Large-Volume Parenterals, the American Society of Hospital Pharmacists, and the Joint Commission on Accreditation of Hospitals were mailed to 625 randomly selected acute-care hospitals. Questions covered the compounding area and recommended equipment; selection, education, and training of personnel; compounding procedures; quality assurance; labeling and record keeping; and reference materials and professional services. The response rate was 43% (198 usable replies). Responses indicated that 42 (50%) of the recommended practices were followed in fewer than 75% of the hospitals, 25 (30%) in fewer than 50% of the hospitals, and 17 (20%) in fewer than 25% of the hospitals. Twenty of the recommendations were considered marginal in importance. Adherence to published guidelines for i.v. admixture services varies among hospitals in the United States; compilation of a single set of guidelines should be considered.


Tissue injury and the inflammatory response to pediatric cardiac surgery with cardiopulmonary bypass: a descriptive study.
There are few detailed descriptions of the inflammatory response to cardiac surgery with cardiopulmonary bypass (CPB) in children beyond 24 h postoperatively. This is especially true for the antiinflammatory cytokines and the extent of tissue injury. The aim of the current study was to describe the inflammatory and injury responses in uncomplicated pediatric cardiac surgery with CPB, where methylprednisolone and modified ultrafiltration (MUF) were used. Blood samples were collected up to 48 h postoperatively. Cytokines (tumor necrosis factor-alpha and interleukin-6, -1beta, -10, and -1ra), complement (C3d and C4d) and coagulation system (prothrombin activation fragments 1 and 2 and antithrombin III) activation, neutrophil elastase, and the resulting tissue injury (creatine kinase, lactate dehydrogenase, alanine transaminase, amylase, and gamma-glutamyl transferase) were measured. The proinflammatory cytokine release varied widely, in contrast to a clear-cut antiinflammatory response. Cytokine concentrations did not decrease immediately after MUF, and no rebound increases later in the postoperative period were observed. The coagulation system, but not complement, was activated. There was a late release of C-reactive protein. Tissue injury could be quantified biochemically without evidence of hepatic or pancreatic dysfunction. In this group of uncomplicated subjects, the antiinflammatory cytokine and tissue injury responses were well defined, in contrast to a variable proinflammatory cytokine release. This was accompanied by activation of the coagulation system but not of complement. Concentrations of inflammatory mediators did not decrease immediately after MUF, and there was no evidence for rebound release later in the postoperative period.


Frizzled-4 C-terminus Distal to KTXXXW Motif is Essential for Normal Dishevelled Recruitment and Norrin-stimulated Activation of Lef/Tcf-dependent Transcriptional Activation.
The carboxy (C)-termini of G protein coupled receptors (GPCR) dictate essential functions. The KTXXXW motif C-terminus of Frizzleds (FZD) has been implicated in recruitment of Dishevelled (DVL). Through study of FZD4 and its associated ligand Norrin, we report that a minimum of three residues distal to the KTXXXW motif in the C-terminal tail of Frizzled-4 are essential for DVL recruitment and robust Lef/Tcf-dependent transcriptional activation in response to Norrin.


A neurocognitive psychometrics account of individual differences in attentional control.
Attention control processes play an important role in many substantial psychological theories but are hard to reliably and validly measure on the subject-level. Therefore, associations between individual differences in attentional control and other variables are often inconsistent. Here we propose a novel neurocognitive psychometrics account of attentional control that integrates model parameters from the dual-stage two-phase model (Hübner et al., 2010), a mathematical model of selective attention, with neural correlates of conflict processing (i.e., latencies of the stimulus-locked lateralized readiness potential) in a multilayer structural equation model framework. We analyzed data from 150 participants who completed the Eriksen Flanker task while their EEG was recorded and used the neurocognitive psychometric approach to distinguish between two sequential stages of information-processing-target selection and response selection. Model parameters and neural correlates showed convergent validity and could be meaningfully related to each other. Together, these neurocognitive process parameters jointly explained 37 % of the variance in individual differences in higher-order cognitive abilities that were assessed with a battery of intelligence tests and working memory tasks. Individuals with greater cognitive abilities were not only better at focusing their attention on the target stimulus but also at subsequent response-selection. All in all, our results support the idea that individual differences in attentional control processes are related to individual differences in cognitive abilities. Moreover, they provide hope that the measurement crisis of individual differences in attentional control can be overcome by integrating measurement approaches from related disciplines such as mathematical psychology and cognitive neuroscience. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy: an imaging mimic of multiple sclerosis. A report of two cases.
To describe the imaging findings of two cases of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) mimicking multiple sclerosis. Two cases presenting with neurological signs and symptoms were referred for magnetic resonance imaging (MRI) evaluation of the brain. Case 1 was a 36-year-old female patient presenting with recurrent headaches and recent onset numbness in the fingers of the right hand. Neurological examination showed a mild sensory deficit in the right hand. Case 2 was a 31-year-old female patient presenting with attacks of right-sided numbness of the face and body. The neurological examination revealed a sensory loss in the face and brisk deep tendon reflexes. Routine MRI sequences showed two types of lesions in both cases: 'punctate' hyperintense lesions on T(2)-weighted images (T(2)WI)/fluid-attenuated inversion recovery (FLAIR) images, hypointense on T(1)-weighted images (T(1)WI) and 'diffuse' white matter lesions, hyperintense on T(2)WI/FLAIR sequences and isointense to hypointense on T(1)WI. All lesions showed no contrast enhancement. Both cases were previously clinically and radiologically diagnosed as multiple sclerosis. There was a strong family history consistent with recurrent infarctions in other family members of both patients. Both cases were later diagnosed as CADASIL by skin biopsy/genetic linkage studies and follow-up. The cases showed that CADASIL causes stroke-like episodes in adults and can mimic multiple sclerosis on imaging. Clinical evaluation and MRI findings allow a differentiation of the two entities.


Variability in the labeling of asthma among pediatricians.
Few studies have examined variability among physicians in the perception and interpretation of asthma symptoms. We report the results of a pilot study to investigate the variability of symptom description and diagnostic labeling and nomenclature among a group of clinicians using standardized audiovisual presentations of asthma. Practicing pediatricians in Wisconsin recruited from an electronic mailing list were shown the International Study of Asthma and Allergies in Childhood (ISAAC) video questionnaire online, and asked to describe the symptoms and signs they observed and suggest possible diagnostic labels for each presentation. A total of 113 pediatricians (mean age = 43 years; 56% female) responded to ≥1 of the 5 video scenes. The number of practitioners who described the principal symptom(s) of asthma depicted in the 5 sequences ranged from 5.5% for Scene 5 (featuring both dyspnea and wheeze), to 100% for Scene 4 (featuring cough). The number who suggested label of 'asthma' as a possible cause of the presentations ranged from 69.7% for Scene 3 (featuring nocturnal wheeze), to 92.7% for Scene 2 (featuring exercise induced wheeze). There is important unexplained variation in the perceptions and labeling of asthma symptoms among pediatricians. These differences may influence the likelihood of diagnosis and the apparent prevalence of asthma. Many participants suggested that the ISAAC video be used in the education and training of pediatricians.


Testicular blood flow, vascular permeability, and testosterone production after stimulation of unilaterally cryptorchid adult rats with human chorionic gonadotropin.
Testicular blood flow, testosterone production, and the formation of testicular interstitial fluid (IF) were studied in unilaterally cryptorchid rats, basally, 8 h and 24 h after treatment with 200 IU human CG (hCG). Testicular blood flow was lower in the abdominal testis in both control rats and hCG-treated rats than in the scrotal testis within the same treatment group. The scrotal testicular blood flow increased significantly 24 h after hCG treatment, but not after 8 h. In the abdominal testis, there was a significant increase of blood flow 8 h after hCG, but not 24 h after. The formation of IF was subnormal in the abdominal testes of control rats, but this was corrected in hCG-treated rats, where there was a significant increase of IF in both abdominal and scrotal testes. The total endothelial surface of small blood vessels was decreased in abdominal testes. Testosterone concentration in the spermatic vein was significantly lower on the abdominal side than on the scrotal side in both control and hCG-treated rats. The concentration of testosterone was lower in IF on the abdominal side in control rats, but after hCG the testosterone concentration was similar in both scrotal and abdominal testes, indicating a trapping of testosterone in the abdominal testis. The outflow of testosterone in the spermatic vein was significantly increased at both 8 and 24 h after hCG from both the scrotal and abdominal testes, although it was always smaller from the abdominal testis. The lower secretion of testosterone from the abdominal testis after hCG was mainly due to reduced blood flow and not to any disability of the Leydig cells of abdominal testes to produce testosterone.


[A clinical comparative study on treatment of severe newly diagnosed immune thrombocytopenia by recombinant human thrombopoietin combined with glucocorticoid].
To evaluate the efficacy and safety of recombinant human thrombopoietin (rhTPO) combined with glucocorticoid in treatment of severe newly diagnosed primary immune thrombocytopenia (ITP). From June 2009 to December 2012, 24 male patients and 38 female patients with the diagnosis of severe primary ITP in our hospital were randomized into trial group (31 cases) or control group (31 cases), the median age was 50 years (range: 21-84 years). Trial group was treated with rhTPO combined with glucocorticoid, and control group was treated with glucocorticoid only. At the day 3, 7 and 14 from the beginning of treatment, the average platelet count (APC) in trial group[(35.5±24.9)×10⁹/L, (135.2±94.9)×10⁹/L and (192.0±109.1)×10⁹/L]were significantly higher than that in control group[(24.5±15.6)×10⁹/L, (78.2±121.9)×10⁹/L and (95.8±60.5)×10⁹/L, P=0.022, 0.009 and 0.001, respectively]. There was no significant difference in APC between the two groups at day 28 and 90 after treatment[(147.8±59.1)×10⁹/L vs (105.1±56.9)×10⁹/L, P=0.243; (137.4±52.3)×10⁹/L vs (104.3±59.8)×10⁹/L, P=0.568, respectively]. At the day 7, 14 and 28, the complete response rates in trial group were 61.3%, 87.1% and 80.6%, which were also significantly higher than that in control group (16.1%, 29.0% and 48.3%, P=0.000, 0.000 and 0.004, respectively). The median time to response in trial group was 3 days while in the control group was 5 days; the median duration of complete response in trial group was 76 days while in the control group was 54 days. In trial group, there were 4 cases treated with platelet transfusion, while in control group there were 11 cases, respectively. For patients with severe primary ITP, rhTPO combined with glucocorticoid could rapidly increase the platelet count, significantly improve the complete response rate and prolonged the effect with a low incidence of tolerable adverse events compared to single use of glucocorticoid. rhTPO combined with glucocorticoid could be a new therapeutic choice to those patients.


Organ-specific contribution to circulating C7 levels by the bone marrow and liver in humans.
Many cells types can produce complement component C7, although the major site of C7 synthesis is unknown. Conversion from recipient to donor allotype following organ transplantation has demonstrated the synthetic sites of several complement proteins, but in the case of C7 this was not possible until recently. A novel C7 polymorphism (C7 M/N) has been described based on the reactivity with the monoclonal antibody WU 4-15 which identifies in allotype of C7 (C7 M). Bone marrow and hepatic C7 production was quantified in bone marrow transplant and liver transplant recipients, respectively, where a mismatch for the C7 allotypes distinguished by the monoclonal antibody had occurred. In the bone marrow transplant group, one informative transplant was identified and donor-derived C7 was detected by enzyme-linked immunosorbent assay. It contributed to 18-27% of the total circulating C7 during the post-transplant phase and was increased during episodes of inflammation. In the liver transplant group, the hepatic contribution to the C7 levels were 30% and 52%, respectively, in two patients identified prospectively. A further three informative liver transplant patients were identified retrospectively and in these individuals, 56-62% of the circulating C7 was liver-derived. This study demonstrates that the majority of the circulating C7 is derived from the liver and bone marrow with a lesser contribution from other sources. These findings provide further support for the concept that locally secreted complement proteins have an important role in inflammation.


Selective determination of volatile N-nitrosamines by derivatization with diethyl chlorothiophosphate and gas chromatography with flame photometric detection.
A selective and sensitive method was developed for the determination of volatile N-nitrosamines by gas chromatography (GC). After denitrosation of N-nitrosamines with hydrobromic acid, the resulting secondary amines were converted into their N-diethylthiophosphoryl derivatives and then measured by GC using a DB-1701 capillary column with flame photometric detection. The calibration graphs for N-nitrosamines in the range 0.05-1 nmol were linear and sufficiently reproducible for quantitative determination. This method was successfully applied to cigarette smoke samples without prior clean-up. N-Nitrosamines and secondary amines were completely separated by extraction with diethyl ether containing 25% 2-propanol. Overall recoveries of N-nitrosamines added to cigarette smoke samples were 83-110%. By using this method, N-nitrosamines in these samples could be determined without any interference from coexisting substances. Analytical results for the contents of N-nitrosamines and secondary amines in mainstream and sidestream smokes of several cigarettes are presented.


200 kHz 5.5 W Yb(3+): YVO4-based chirped-pulse regenerative amplifier.
A chirped-pulse regenerative amplifier based on a Yb:YVO4 (Yb:YVO) crystal was demonstrated for the first time, to our knowledge. A novel wavelength-independent off-axis technique was used to deliver 26.6 W of pump power with a low quantum defect. It was seeded by means of a 120 fs Yb:YVO4 oscillator. It generates as much as 5.5 W of average output power with chirped pulses and 4.2 W with 200 fs compressed pulses at a 200 kHz repetition rate. A maximum pulse energy of 140 μJ was obtained at pulse repetition frequencies up to 25 kHz at the central wavelength of 1018 nm.


Illness perception and cardiovascular risk factors in patients with type 2 diabetes: cross-sectional questionnaire study.
AIM. To investigate illness perception in patients with type 2 diabetes mellitus and its association with the degree of control over relevant cardiovascular risk factors. METHODS. A cross-sectional questionnaire-based study was performed from June 2007 to March 2008. A stratified random sample of 46 Croatian general practitioners was asked to select, using systematic sampling, the first 6 patients with type 2 diabetes mellitus aged > or =18 years who visited them for consultation during the study period. Data on 250 patients included patient illness perception assessment (Brief Illness Perception Questionnaire, IPQ), cardiovascular risk factors, and socio-demographic data. RESULTS. The patients' mean age was 63.0+/-10.9 years and mean duration of diabetes was 9.3+/-7.8 years. The patients' illness perception assessment on an 11-point (0 to 10) scale showed the highest median scores (interquartile range): 10 (8 to 10) for "timeline" and 8 (7 to 9) for "treatment control," followed by 7 (5 to 8) for "personal control," 7 (5 to 9) for "understanding," 5 (3 to 7) for "consequences," 6 (4 to 7) for "concern," and 5 (2 to 7) for "emotional response." The lowest score was 3 (1 to 5) for "identity." Multivariate logistic regression showed that the Brief IPQ item "concern" (P<0.001) was a significant predictor of body mass index; "personal control" (P<0.001) and "concern" (P=0.048) were significant predictors of fasting blood glucose; "treatment control" (P=0.009) was a significant predictor of total cholesterol; and "understanding" (P=0.010) was a significant predictor of blood pressure. CONCLUSION. As patients' beliefs seem to be associated with the degree of control over cardiovascular risk factors, they should be included in routine clinical assessments.


Altered mRNA expression related to the apoptotic effect of three xanthones on human melanoma SK-MEL-28 cell line.
We previously demonstrated that α-mangostin, γ-mangostin, and 8-deoxygartanin have significant cytotoxic effects on human melanoma SK-MEL-28 cell line. The current study revealed the underlying mechanisms. α-Mangostin (7.5 μg/mL) activated caspase activity, with a 3-fold and 4-fold increased caspase 8 and 9 activity, respectively. The molecular mechanisms were investigated by qRT-PCR for mRNA related to cell cycle arrest in G1 phase (p21(WAF1) and cyclin D1), apoptosis (cytochrome C, Bcl-2, and Bax), and survival pathways (Akt1, NFκB, and IκBα). α-Mangostin significantly upregulated mRNA expression of cytochrome C and p21(WAF1) and downregulated that of cyclin D1, Akt1, and NFκB. γ-Mangostin significantly downregulated mRNA expression of Akt1 and NFκB and upregulated p21(WAF1) and IκBα. 8-Deoxygartanin significantly upregulated the mRNA expression of p21(WAF1) and downregulated that of cyclin D1 and NFκB. The three xanthones significantly inhibited the mRNA expression of the BRAF V600E mutation. Moreover, α-mangostin and γ-mangostin significantly downregulated Akt phosphorylation at Ser473. In conclusion, the three xanthones induced an inhibitory effect on SK-MEL-28 cells by modulating the molecular targets involved in the apoptotic pathways.


Second-order theories for extensional vibrations of piezoelectric crystal plates and strips.
An infinite system of two-dimensional (2-D) equations for piezoelectric plates with general symmetry and faces in contact with vacuum is derived from the 3-D equations of linear piezoelectricity in a manner similar to that of previous work, in which an infinite system of 2-D equations for plates with electroded faces was derived. By using a new truncation procedure, second-order equations for piezoelectric plates with faces in contact with either vacuums or electrodes are extracted from the aforementioned infinite systems of equations, respectively. The second-order equations for plates with or without electrodes are shown to predict accurate dispersion curves by comparing to the corresponding curves from the 3-D equations in a range up to the cut-off frequencies of the first symmetric thickness-stretch and the second symmetric thickness-shear modes without introducing any correction factors. Furthermore, a system of 1-D second-order equations for strips with rectangular cross section is deduced from the 2-D second-order equations by averaging variables across the narrow width of the plate. The present 1-D equations are used to study the extensional vibrations of barium titanate strips of finite length and narrow rectangular cross section. Predicted frequency spectra are compared with previously calculated results and experimental data.


Drug dependence as a disorder of neural plasticity: focus on dopamine and glutamate.
Drug addiction, as a disease, has grown to reach the level of a social illness. Psychostimulants, opiates, alcohol, nicotine and cannabis abuse affects millions worldwide and virtually all classes of modern society. In spite of the enormous proportions of its spread, intimate neurobiological mechanisms leading to distintictive features of this pathological status, such as craving for the abused substance and loss of control over intake, remain largely obscure and pharmacotherapies sadly unsatisfactory. In the last decade, preclinical and clinical research in this field has made great progress to improve our understanding of the brain mechanisms which form the basis of this illness. The review of recent literature, which represents the focus of the present paper, leads to the emerging consensus that an alteration of physiological mechanisms of neural plasticity within the brain dopamine and glutamate systems may underlie some of the behavioral abnormalities occurring during the dependence cycle. In particular, a reduction of dopamine neuronal activity and glutamate neurotransmission at the level of the ventrotegmental area, after withdrawal from chronic administration of drugs of abuse, may work in concert with alterations in other forebrain areas, such as the nucleus accumbens and the amygdaloid complex. In addition, following prolonged periods of abstinence, even after somatic withdrawal signs have vanished, responsiveness of these systems to drugs of abuse remains abnormal. This suggests that these two neurotransmitters may play a substantial role in the long-lasting, enduring changes typical of the addictive process and may represent ideal targets for pharmacological intervention aimed at normalizing forms of neural plasticity impaired after chronic drug intake.


A comparison of false-negative responses for full threshold and SITA standard perimetry in glaucoma patients and normal observers.
To compare false-negative (FN) rates for Full Threshold (FULL) and Swedish Interactive Threshold Algorithm (SITA) Standard automated perimetry in healthy control subjects and glaucoma patients. One eye of 19 glaucoma patients and 5 normal control subjects underwent 5 visual field examinations within 6 weeks on the Humphrey Field Analyzer II using the 24-2 FULL and SITA testing procedures. The order of presentation of test procedures was counterbalanced across participants. We obtained FN response procedures that presented stimuli 9 dB greater than a previously determined threshold value at specific locations in the visual field (FULL) or up to 20 dB greater than the threshold value (SITA), and evaluated whether the patient responded to these suprathreshold stimuli. We compared the percentage of FN responses for the FULL and SITA tests before (Peridata output) and after postprocessing of data. The percentage of FNs was almost twice as high for FULL testing compared with the 2 SITA analyses. The SD of FN percentages was half as large for SITA postprocessing compared with FULL and SITA Peridata procedures. Because the distributions were not normally distributed, a Kruskal-Wallis analysis of variance was performed and revealed a statistically significant difference (H=13.094, P<0.001) for FN percentages among the 3 evaluations. FULL test procedures yield about twice as many FNs as SITA procedures. This may be related to the SITA method of testing for FN in relatively normal areas as opposed to any location (even if it has low sensitivity) with FULL. Because FN responses can be due to either high variability in damaged areas or pathologic variations, these revised FN procedures provide greater assistance in properly interpreting visual field results.


Three cases of primary acquired melanosis of the conjunctiva as a manifestation of the atypical mole syndrome.
We report three patients with the atypical mole syndrome (AMS) [also known as dysplastic naevus or FAMMM syndrome] who presented with primary acquired melanosis (PAM). PAM is a melanocytic lesion of the conjunctiva which may progress to conjunctival melanoma. The association of this rare condition with the AMS phenotype in three individuals suggests that PAM may be a conjunctival manifestation of the AMS.


Analysis of peripheral doses for base of tongue treatment by linear accelerator and helical TomoTherapy IMRT.
The purpose of this study was to compare the peripheral doses to various organs from a typical head and neck intensity-modulated radiation therapy (IMRT) treatment delivered by linear accelerator (linac) and helical TomoTherapy. Multiple human CT data sets were used to segment critical structures and organs at risk, fused and adjusted to an anthropomorphic phantom. Eighteen contours were designated for thermoluminescent dosimeter (TLD) placement. Following the RTOG IMRT Protocol 0522, treatment of the primary tumor and involved nodes (PTV70) and subclinical disease sites (PTV56) was planned utilizing IMRT to 70Gy and 56 Gy. Clinically acceptable treatment plans were produced for linac and TomoTherapy treatments. TLDs were placed and each treatment plan was delivered to the anthropomorphic phantom four times. Within 2.5 cm (one helical TomoTherapy field width) superior and inferior to the field edges, normal tissue doses were on average 45% lower using linear accelerator. Beyond 2.5 cm, the helical TomoTherapy normal tissue dose was an average of 52% lower. The majority of points proved to be statistically different using the Student's t-test with p > 0.05. Using one method of calculation, probability of a secondary malignancy was 5.88% for the linear accelerator and 4.08% for helical TomoTherapy. Helical TomoTherapy delivers more dose than a linac immediately above and below the treatment field, contributing to the higher peripheral doses adjacent to the field. At distances beyond one field width (where leakage is dominant), helical TomoTherapy doses are lower than linear accelerator doses.


Septal q waves as an indicator of risk of mortality in elderly patients with chronic heart failure.
The absence of electrocardiographic septal q wave is a recognized marker of left ventricular disease. We aimed to investigate the prognostic significance of absent septal q waves in elderly (age >65 years) patients with chronic heart failure. A total of 110 patients (mean age 73 +/- 4 years) with New York Heart Association functional class II to IV and left ventricular ejection fraction of <45% were enrolled in the study. Standard 12-lead electrocardiograms were critically analyzed for the presence or absence of septal q waves in leads I, aVL, V5, and V6. Patient survival was determined from hospital and general practitioner records and National Statistics Registry at a mean follow-up of 4 years. Septal q waves were absent in 71 and present in 39 patients. The overall mortality rate was 47% (43 patients). The incidence of death was 49% (36 patients) in the group with no septal q waves and 18% (7 patients) in those who demonstrated septal q waves. On univariate analysis by Cox proportional hazard method, absence of septal q waves was found to be a strong marker of poor prognosis in CHF (P =.003, hazard ratio 1.40, 95% CI 1.10-1.67). Kaplan-Meier survival curves showed a significant difference in survival independent of age, New York Heart Association functional class, peak VO2, and QRS duration between these 2 groups. Absence of the normal septal q wave on 12-lead electrocardiography, which may indicate structural heart disease and myocardial fibrosis, is an independent predictor of poor prognosis in elderly patients with CHF.


Connecting artificial intelligence and primary care challenges: findings from a multi stakeholder collaborative consultation.
Despite widespread advancements in and envisioned uses for artificial intelligence (AI), few examples of successfully implemented AI innovations exist in primary care (PC) settings. To identify priority areas for AI and PC in Ontario, Canada. A collaborative consultation event engaged multiple stakeholders in a nominal group technique process to generate, discuss and rank ideas for how AI can support Ontario PC. The consultation process produced nine ranked priorities: (1) preventative care and risk profiling, (2) patient self-management of condition(s), (3) management and synthesis of information, (4) improved communication between PC and AI stakeholders, (5) data sharing and interoperability, (6-tie) clinical decision support, (6-tie) administrative staff support, (8) practitioner clerical and routine task support and (9) increased mental healthcare capacity and support. Themes emerging from small group discussions about barriers, implementation issues and resources needed to support the priorities included: equity and the digital divide; system capacity and culture; data availability and quality; legal and ethical issues; user-centred design; patient-centredness; and proper evaluation of AI-driven tool implementation. Findings provide guidance for future work on AI and PC. There are immediate opportunities to use existing resources to develop and test AI for priority areas at the patient, provider and system level. For larger scale, sustainable innovations, there is a need for longer-term projects that lay foundations around data and interdisciplinary work. Study findings can be used to inform future research and development of AI for PC, and to guide resource planning and allocation.


Evaluation of implant abutment screw head deformation in hexagonal and star designs after successive tightening and loosening.
Tightening and loosening of the prosthetic components of the implant are carried out with various screw designs. This study compared the rate of deformation of the abutment screw in two hexagonal and star screw head designs after consecutive tightening and loosening. In this study, two fixtures were mounted vertically in die stone blocks using a surveyor. Then the corresponding abutment (with a diameter of 4.5 mm and a gingival height of 2 mm; Dio-SM and Dio-UF system) was mounted on each fixture. In each fixture, six abutment screws from each manufacturer were used (12 abutment screws in total). The abutment screw head of the Dio-UF system is hexagonal, and the abutment screw head of the Dio-SM system is star-shaped. The samples were examined under a stereomicroscope (Nikon C-DS) at a magnification of×50. The image of each abutment screw head (6 abutment screws in each group) was prepared before the procedural steps and after 5, 10, and 15 times of tightening and loosening with 25-Ncm torque using a stereomicroscope. Then the differences in the surface areas of consecutive specimens (0, 5, 10, and 15 consecutive tightening and loosening rounds) between hexagonal and star-shaped abutment screws were calculated. The study results were reported via descriptive statistical methods (mean±standard deviation). Statistical analysis was performed using SPSS 24, and the significance level was defined at P<0.05. Increasing the number of tightening and loosening rounds increased the screw head surface area in both hexagonal and star shapes. At all stages, the changes in the star-shaped screw head were greater than in the hexagonal screw. These changes were statistically significant at all stages (P<0.05). In addition, there was a statistically significant difference between the area values and the number of tightening and loosening rounds separately in both screw types (P<0.001). Also, the surface areas of the head of both screws in all tightening and loosening rounds were significantly different (P<0.001). Increasing the number of tightening and loosening rounds increased the screw head surface area in both hexagonal and star-shaped geometrical forms. Also, the extent of area changes at all stages in star-shaped screw heads was greater than in hexagonal screws.


Insight into the factors influencing the backbone dynamics of three homologous proteins, dendrotoxins I and K, and BPTI: FTIR and time-resolved fluorescence investigations.
Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, combined with hydrogen/deuterium exchange technique and time-resolved fluorescence spectroscopy, has been used to investigate the changes in structure and dynamics that underlie the thermodynamic stability differences observed for three closely homologous proteins: dendrotoxins I and K, and bovine pancreatic trypsin inhibitor (BPTI). The experiments were performed on proteins under their native state and a modified form, obtained by selective reduction of a disulfide bond at the surface of the molecule, increasing slightly the backbone flexibility without changing the average structure. The data confirmed the high local as well as global rigidity of BPTI. In protein K, the exchange process was slow during the first 2 h of exchange, presumably reflecting a compact three-dimensional conformation, and then increased rapidly, the internal amide protons of the beta-strands exchanging 10-fold faster than in BPTI or protein I. The most probable destabilizing element was identified as Pro32, in the core of the beta-sheet. Protein I was found to present a 10% more expanded volume than protein K or BPTI, and there is a possible correlation between the resulting increased flexibility of the molecule and the lower thermodynamic stability observed for this protein. Interestingly, the interior amide protons of the beta-sheet structure were found to be as protected against exchange in protein I as in BPTI, suggesting that, although globally more flexible than that of Toxin K or BPTI, the structure of Toxin I could be locally quite rigid. The structural factors suspected to be responsible for the differences in internal flexibility of the two toxins could play a significant role in determining their functional properties.


Allelic losses on chromosome 4 suggest the existence of a candidate tumor suppressor gene region of about 0.6 cM in gamma-radiation-induced mouse primary thymic lymphomas.
Mouse chromosome 4 was investigated to assess the involvement of tumor suppressor genes in primary thymic lymphomas induced by gamma-irradiation. PCR analysis using microsatellite DNA polymorphic markers was performed in F1 animals generated from a cross between the strains C57BL/6J and RF/J. Microsatellite markers were selected with focus on chromosome 4 around the region containing the interferon alpha gene cluster (D4Mit17, D4Wsm1, D4Mit9, and D4Mit205 and a more distal region (D4Mit12, D4Mit54, and D4Mit13). Allelic losses were detected in 21/47 (44.7%) gamma-radiation-induced thymic lymphomas. Analysis of markers located on six other chromosomes as well as on the proximal region of chromosome 4 illustrates the specificity of the occurrences of LOH appearing on the former markers. This analysis clearly suggests the existence of a candidate tumor suppressor gene region on mouse chromosome 4 of about 0.6 cM between the markers D4Wsm1 and D4Mit9 (TLSR1, Thymic Lymphoma Suppressor Region 1). In addition, another more distal region centered at the marker D4Mit54 could be also exist (TLSR2). In most tumors, allelic losses on these chromosome regions involved the paternal RF/J allele (19 of 20).


Glucocorticoids Oppose Thymocyte Negative Selection by Inhibiting Helios and Nur77.
Glucocorticoid (GC) signaling in thymocytes shapes the TCR repertoire by antagonizing thymocyte negative selection. The transcription factors Nur77 and Helios, which are upregulated in TCR-signaled thymocytes, have been implicated in negative selection. In this study, we found that GCs inhibited Helios and, to a lesser extent, Nur77 upregulation in TCR-stimulated mouse thymocytes. Inhibition was increased by GC preincubation, and reductions in mRNA were prevented by a protein synthesis inhibitor, suggesting that GCs suppress indirectly via an intermediary factor. Upregulation of Helios in TCR-stimulated thymocytes was unaffected by deletion of Nur77, indicating Nur77 and Helios are regulated independently. Whereas CD4+ thymocytes are positively selected in wild-type AND TCR-transgenic B6 mice, loss of GC receptor expression resulted in increased negative selection. Correspondingly, Helios and Nur77 levels were elevated in TCRhiCD4+CD8+ (TCR-signaled) thymocytes. Notably, deletion of Helios fully reversed this negative selection, whereas deletion of Nur77 had no effect on CD4+CD8+ cell numbers but reversed the loss of mature CD4+ thymocytes. Thus, Nur77 and Helios are GC targets that play nonredundant roles in setting the signaling threshold for thymocyte negative selection.


Incessant ectopic atrial tachycardia and sudden death.
A patient with refractory and incessant ectopic atrial tachycardia (IEAT) is reported in whom it was possible to document, during ECG (Holter) the occurrence of aborted sudden death by spontaneous ventricular fibrillation (VF). Following the second of two attempts at surgical ablation of the origin of the IEAT, the patient has been asymptomatic without antiarrhythmic drugs and in sustained sinus rhythm for 24 months. Although we cannot exclude the residual action of amiodarone and flecainide (proarrhythmia) or the residual peripartum cardiomyopathy it is probable that the observed VF was a true complication of a cardiomyopathy induced by a chronically increased heart rate (HR). Although unclear, this VF might be considered as a form of adrenergic-dependent long QT syndrome due to early afterdepolarization in the presence of predisposing myocardial conditions.


Toxoplasma gondii inhibits the in vitro induced apoptosis of HL-60 cells.
The integrity of the host cell may represent an important prerequisite for the intracellular survival and development of obligate intracellular pathogens. In the present study, we investigated the influence of infections with the protozoan parasite Toxoplasma gondii on the rate of apoptosis in the human leukemia cell line HL-60. After infection with T. gondii tachyzoites of the strain NTE and in uninfected controls, less than 2% of the host cells showed typical signs of apoptosis, i.e. condensation of chromatin after staining with Hoechst 33258 or internucleosomal DNA fragmentation after agarose gel electrophoresis of genomic DNA. After treatment with 0.1 to 0.5 microg/ml actinomycin D for up to 16 hours, HL-60 cells considerably underwent apoptosis. However, this actinomycin D-induced apoptosis was clearly reduced after concomitant infection with T. gondii as shown by staining with Hoechst 33258 and by DNA fragmentation assay. Inhibition of apoptosis by the intracellular pathogen T. gondii might be recognized as an evasion mechanism that enables intracellular survival and establishes long-lasting persistence.


Synthesis of multisubstituted pyrroles from doubly activated cyclopropanes using an iron-mediated oxidation domino reaction.
An alternative route has been developed for the construction of multisubstituted pyrrole derivatives from readily available, doubly activated cyclopropanes and anilines using an iron-mediated oxidation domino reaction (i.e., sequential ring-opening, cyclization, and dehydrogenation reactions). This reaction uses readily available reactants and is tolerant of a broad range of substrates, with the desired products being formed in good to excellent yields.


Meta-analysis of factors affecting ontogenetic development rate in the Culex pipiens (Diptera: Culicidae) complex.
Meta-analysis of 33 studies of developmental timing of Culex pipiens s.l. Linnaeus demonstrates that development rate, or the rate of progression through immature life stadia, is primarily driven by temperature, whereas immature survival is driven by temperature, density, and variability in the environmental conditions. As expected, the linear relationship of temperature and development rate is positive for the larval period as well as development to adult emergence. However, the strength of this association varies significantly. Variation in development rate can be explained using additional environmental factors of intraspecific rearing density, sex, and study methodology. Heterogeneity in development rates even once temperature has been considered emphasizes the need for further research of multiple environmental factors and in changing environments. Immature survival is also significantly impacted by variability in environmental conditions. Development rates vary between subspecies of Cx. pipiens, but these population differences are no longer significant once an environmental factor of temperature is considered. Thus, variability in development rate of these insects appears to be primarily driven by response to certain environmental conditions rather than differences between populations. Broad patterns of phenotypic variation across latitude and 96 yr of empirical estimates were not significant once environmental rearing conditions had been considered.


Activation of NLRP3 and AIM2 inflammasomes in Kupffer cells in hepatic ischemia/reperfusion.
Inflammasome activation by danger signals in ischemia/reperfusion (I/R) injury is responsible for the sterile inflammatory response. Signals triggering formation and activation of the inflammasome involve the generation of oxidative stress. The aim of this study was to examine the molecular mechanisms of inflammasome activation and the involvement of reactive oxygen species in hepatic I/R. I/R induced the formation of nucleotide-binding domain leucine-rich repeat containing family pyrin domain containing 3 (NLRP3) and absent in melanoma 2 (AIM2) inflammasomes and the subsequent serum release of interleukin 1β. Pannexin-1 inhibitor and anti-cathepsin B antibody attenuated I/R-induced inflammasome activation and hepatic injury. The expression of the thioredoxin-interacting protein gene and the interaction between NLRP3 and the thioredoxin-interacting protein increased after I/R. Treatment with the antioxidant N-acetylcysteine significantly attenuated protein conversion of interleukin 1β after hepatic I/R. Moreover, pannexin-1 protein expression and cathepsin B release were strongly attenuated by N-acetylcysteine. The depletion of Kupffer cells with gadolinium chloride markedly decreased NLRP3 and AIM2 inflammasome expression and activation of their signaling pathways, and also reduced the level of caspase-1 protein in F4/80-positive cells. Our findings suggest that reactive-oxygen-species-mediated activation of NLRP3 and AIM2 inflammasomes leads to I/R-induced inflammatory responses in which Kupffer cells play a crucial role.


Chromatin pattern by variogram analysis.
Many cytological processes such as cell proliferation, differentiation, transformation, apoptosis, etc., are accompanied by specific chromatin changes, usually identified on the basis of the relative content of euchromatin and heterochromatin. In order to achieve a quantitative, non-subjective evaluation of the chromatin pattern, two different approaches may be undertaken, one consisting in the analysis of the several morphological features of chromatin grains (size, shape, density, arrangement, and distribution), and the second consisting in the analysis of the chromatin globally considered as a coherent texture. Although the second approach appears to be simpler and more suitable, methods of texture analysis--including those specifically designed for the analysis of the chromatin pattern--are rarely applied due mainly to the unsuitability of sampling procedures and the excessive crypticism of results. As an alternative to traditional texture analysis, we suggest a method supported by a sound mathematical theory and approximately 30 years of applications in the field of geostatistics. The method, called variogram, analyzes the intrinsic structure of data sampled at different distance intervals and directions, and outputs easily understandable results. Recently, variogram analysis has successfully been exported from geostatistics to other fields (for example, ecology and epidemiology) that make use of spatially referenced variables. Based on the fact that pixels represent a perfect array of data ordered at regular distance intervals and directions, the variogram can be adopted to explore nuclear images and recognize chromatin patterns. Variograms of different nuclei can be summarized by multivariate methods without the need of previous standardization of data. This allows comparison and discrimination of chromatin patterns from mixed cell populations. Preliminary data obtained from young neurons undergoing massive apoptosis reveal a self-consistent map of nuclear changes correlated to the degenerative process.


Psychosocial predictors of postpartum depressed mood in socioeconomically disadvantaged women.
The effects of stress, social support, and labor and delivery experiences on postpartum depressed mood were examined in an ethnically diverse sample of low-income women (N = 108). Women were interviewed on multiple occasions throughout pregnancy and then once approximately 2 months postpartum. Information on labor and delivery outcomes was abstracted from medical charts. Results indicated that women who were more satisfied with the prenatal social support they received were less likely to experience postpartum depressed mood, after controlling for prenatal depressive symptomatology. In addition, women who experienced more distressing life events during pregnancy and who reported higher levels of prenatal anxiety were also more likely to feel depressed, after controlling for all other factors in the model. Finally, women who were more satisfied with their labor and delivery experience tended to be less depressed in the early months following childbirth. Together, these variables accounted for 45% of the variance in postpartum depressed mood.


Commentary: further comments on Mycoplasma pulmonis and lymphoma in bioassays of rats.
The authors recently assessed the likelihood that lifetime cancer bioassays of aspartame, methanol, and methyl tertiary butyl ether conducted with conventional (not specific pathogen free) Sprague-Dawley rats were compromised by Mycoplasma pulmonis disease. From the tumor data and other information, the authors concluded that the rats used in these bioassays likely had M pulmonis disease and that lesions of the disease were plausibly interpreted as lymphoma. Subsequently, they analyzed the nonneoplastic lesion data from these bioassays for occurrence of inflammatory lesions and found that 2,267 of 2,960 rats (76.6%) were reported to have bronchitis, the signature lesion of M pulmonis disease, and that 633 rats (21.4%) were reported to have otitis, another common lesion of the disease. Also, documentation is now available containing serologic evidence of mycoplasma infection in the rats. In contrast, the reports of 6 National Toxicology Program bioassays based on specific pathogen-free Sprague-Dawley rats listed no instances of bronchitis or otitis. These findings provide substantial additional evidence that the bioassays in question were compromised by M pulmonis disease. Therefore, the reported induction of lymphoma in these studies should not be considered in cancer risk assessments. The authors also found that inflammatory lesions were prevalent in lymph nodes, thymus, pleura, and brain. Finally, they found that of all 328 cases of lymphoimmunoblastic lymphoma affecting the lung (the primary form of lymphoma reported), 218 (66.5%) occurred within the first 104 weeks of the studies, showing that occurrence of such lesions was not due to appearance in rats surviving beyond that interval.


The pattern of stimulation influences the amount of oscillatory work done by frog muscle.
1. A doublet is the interjection of an additional action potential at the beginning of a regular motoneuron discharge pattern. The present experiments were designed to estimate the magnitude of the effect of doublets on the capacity of isolated frog sartorius muscle to do work during oscillatory length changes. 2. For the work loop method, the muscle was subjected to sinusoidal length changes at 4 Hz. Work was calculated from the loop formed when force was plotted against length. Work done was positive when the muscle was shortening and was negative when the muscle was lengthening; net work was the difference. 3. Adding pulses at the start of the stimulus pulse train increased isometric force and markedly increased net work per cycle. Adding one pulse increased the net work by about 52%, but increased isometric force by only 24%. 4. Eighty per cent of the maximum net work was achieved by adding only three pulses at the start of a low-frequency pulse train, i.e. 80% of the work achieved with seventeen pulses could be achieved with only six pulses. 5. The maximum net work per stimulus pulse was achieved with a stimulus train that consisted of five pulses with two being additional initial pulses at 5 ms interpulse intervals, i.e. a triplet. 6. The results of the present study imply that doublet stimulation patterns may be important to reduce the fatigue that occurs during artificial neuromuscular stimulation of skeletal muscle.