Analysis of an immunodominant epitope of topoisomerase I in patients with systemic sclerosis.
In this paper an immunodominant epitope of Topoisomerase I is described. An epitope expression sublibrary was constructed from Topoisomerase I cDNA. The subclones were screened with an antiserum from a patient with systemic sclerosis (SSc). The positive clones defined one immunodominant B cell epitope (epitope III), which was located at the carboxyterminal part of the protein. The epitope, 52 amino acids in length, neither contains the p30gag sequence nor the suggested active site Tyr-723, both presumed antibody recognition sites. More than 70% of our anti-TopoI sera recognize this epitope III, indicating that it is a major recognition site of the anti-TopoI autoantibodies in SSc sera. DNA relaxation experiments show that all sera that recognize epitope III and most sera with antibodies to other epitopes inhibit Topoisomerase I activity.


Gestational and smoking effects on peptidase activity in the placenta.
Angiotensin converting enzyme (ACE) and leucine aminopeptidase (LAP) regulate fetally and maternally generated peptides in the placenta. In this study, ACE-like activity was found to be decreased and LAP-like activity increased with increasing days of gestation in rat placental tissues forming the fetal:maternal interface. Membrane-associated ACE-like and LAP-like activities in the placenta of smokers were also found to be significantly higher than their respective activities in placenta of nonsmokers. Our collective findings suggest that gestational and environmentally-induced changes in placental peptidase activities may account for variable peptide hormone and/or therapeutic peptide metabolism in the placenta.


Increased synthesis of nitric oxide in rat brain cortex due to halogenated volatile anesthetics confirmed by EPR spectroscopy.
Halogenated volatile anesthetics (HVAs) are considered to be inhibitors of nitric oxide synthase (NOS). On other hand, NO mediates the vasodilation produced by HVAs. Thus, both increase and decrease of NO concentration in brain tissues are possible during anesthesia. Previously, we have observed an increase of NO content in rat brain cortex under halothane anesthesia. The goal of this study was to determine whether the observed phenomenon was general for this anesthetic group, if it was specific for brain cortex, and if the NO increase was due changes in NOS activity. NO scavengers were injected to adult rats 30 min prior to anesthesia. Rats were anesthetized by inhalation of an O2 mixture with volatile anesthetics (1.5% for halothane; 1% for isoflurane, 2% for sevoflurane). After 30 min of anesthesia, rats were decapitated and brain cortex, cerebellum, liver, heart, kidneys and testes were dissected, frozen in liquid nitrogen and subjected to EPR spectroscopy. Nitric oxide content was determined quantitatively based on the intensity of the NO-Fe-DETC complex spectrum and its comparison with the calibration curve. In rats anesthetized with HVAs, we observed a greater than twofold increase of NO content in brain cortex as compared to the nonanesthetized animals. No significant changes were detected in other organs. The NOS inhibitor N(omega)-nitro-L-arginine abolished the increase of NO content in brain produced by volatile anesthetics. The action of volatile anesthetics is coupled with an increase of NO content in the cortex dependent on NOS activity.


[Macro-CK type 1: a methodological study approach and its clinical significance].
Description of an analytical procedure in order to show the atypical-CK carriers that simulate an abnormal raising of CK-MB circulating levels in dosages for immune inhibition. This procedure was applied to two samples taken from patients of such a type, immune form neoplastic and cardiac pathology. A systematic dosage of a "real" MB isoenzyme is desirable in all the cases of very raised CK-MB activity.


Effect of ammodytin L from Vipera ammodytes on L-6 cells from rat skeletal muscle.
Ammodytin L (AMDL) is a myotoxic phospholipase-like protein from the venom of Vipera ammodytes with a serine in position 49 instead of an aspartate, therefore this toxin is devoid of phospholipase activity, and the membrane-damaging effect does not involve any step of phospholipase activity. The aim of the present study was to analyze the effect of AMDL on L-6 cells from rat skeletal muscle to investigate its mechanism of action and the role of calcium ions in its muscle-damaging activity. Our data indicate that the effect of ammodytin L is strongly dependent on the degree of cell differentiation. Low doses of myotoxin gave rise to a marked release of creatine kinase in myotubes differentiated from L-6 myoblasts and the presence of calcium ions plays a role in the cytotoxic effect. The presence of EGTA in the incubation buffer reduced by 50% the release of creatine kinase. No membrane damage was observed in myoblasts, but there was a significant increase of intracellular calcium concentration measured with Fura-2. A non-specific membrane effect of AMDL was ruled out using platelets as reference cells: no platelet aggregation pattern and no increase in intracellular calcium were observed.


Direct isolation, culture and transplant of mouse skeletal muscle derived endothelial cells with angiogenic potential.
Although diseases associated with microvascular endothelial dysfunction are among the most prevalent illnesses to date, currently no method exists to isolate pure endothelial cells (EC) from skeletal muscle for in vivo or in vitro study. By utilizing multicolor fluorescent-activated cell sorting (FACS), we have isolated a distinct population of Sca-1(+), CD31(+), CD34(dim) and CD45(- )cells from skeletal muscles of C57BL6 mice. Characterization of this population revealed these cells are functional EC that can be expanded several times in culture without losing their phenotype or capabilities to uptake acetylated low-density lipoprotein (ac-LDL), produce nitric oxide (NO) and form vascular tubes. When transplanted subcutaneously or intramuscularly into the tibialis anterior muscle, EC formed microvessels and integrated with existing vasculature. This method, which is highly reproducible, can be used to study the biology and role of EC in diseases such as peripheral vascular disease. In addition this method allows us to isolate large quantities of skeletal muscle derived EC with potential for therapeutic angiogenic applications.


Synaptic mitochondria are critical for mobilization of reserve pool vesicles at Drosophila neuromuscular junctions.
In a forward screen for genes affecting neurotransmission in Drosophila, we identified mutations in dynamin-related protein (drp1). DRP1 is required for proper cellular distribution of mitochondria, and in mutant neurons, mitochondria are largely absent from synapses, thus providing a genetic tool to assess the role of mitochondria at synapses. Although resting Ca2+ is elevated at drp1 NMJs, basal synaptic properties are barely affected. However, during intense stimulation, mutants fail to maintain normal neurotransmission. Surprisingly, FM1-43 labeling indicates normal exo- and endocytosis, but a specific inability to mobilize reserve pool vesicles, which is partially rescued by exogenous ATP. Using a variety of drugs, we provide evidence that reserve pool recruitment depends on mitochondrial ATP production downstream of PKA signaling and that mitochondrial ATP limits myosin-propelled mobilization of reserve pool vesicles. Our data suggest a specific role for mitochondria in regulating synaptic strength.


Importance of the geographic barriers to promote gene drift and avoid pre- and post-Columbian gene flow in Mexican native groups: Evidence from forensic STR Loci.
To analyze the origin, structure, relationships, and recent admixture in Mexican Native groups based on 15 STRs commonly used in human identification. We analyzed 39 Mexican Native population samples using STR databases based on the AmpFlSTR® Identifiler kit (n = 3,135), including Mexican-Mestizos (admixed), European and African populations, as reference. Based upon effective population size (Ne) differences, Native groups were clustered into three regions: i) Center-Southeast groups, characterized by larger Ne, migration rate (Nm), genetic diversity (He), and relative homogeneity principally in the Yucatan Peninsula; ii) Isolated southern groups from Chiapas and Oaxaca, characterized by lower Ne, Nm, and He (i.e. higher isolation and genetic differentiation); iii) North-Northwest groups, which are similar to the previous group but are characterized by generating the widest gene flow barrier in the Pre-Hispanic Mexican territory, and currently by elevated admixture in some northern Native groups. Despite the relative congruence between genetic relationships with cultural, linguistic, geographic criteria, these factors do not explain the present-day population structure of Native groups, excepting in those linguistically related to the Mayan that show higher homogeneity. The Isolation by distance model was demonstrated at long distances (>1,500 km), whereas geographic isolation stands as a determining factor to avoid both non-indigenous admixture and bottleneck processes. Different dynamics of gene flow and drift were observed among Mexican Native groups, highlighting the geographic barriers (mountains, canyons and jungle regions) as the main factor differentiating Pre-Hispanic populations, and eventually helping to avoid Post-European contact admixture and population bottleneck. Am J Phys Anthropol 160:298-316, 2016. © 2016 Wiley Periodicals, Inc.


Analysis of environmental Escherichia coli isolates for virulence genes using the TaqMan PCR system.
To assess the presence of virulence genes in environmental and foodborne Escherichia coli isolates using the TaqMan PCR system. Three TaqMan pathogen detection kits called O157:H7, StxI and StxII were used to investigate the presence of virulence genes in Escherichia coli isolates. All 54 foodborne E. coli O157:H7 isolates showed expected results using these kits. Ninety (15%) of 604 environmental isolates gave positive amplification with an O157:H7-specific kit. TaqMan PCR amplification products from these 90 isolates were analysed by agarose gel electrophoresis, and 90% (81 of 90) of the environmental samples contained the expected PCR product. Sixty-six of these 90 were chosen for serotyping tests and only 35% (23 of 66) showed agglutination with both anti-O157 and anti-H7 antibodies. Further ribotyping of 16 sero-positive isolates in an automated Riboprinter did not identify these to be O157:H7. Multiplex PCR with primers for eaeA, stxI and stxII genes was used to confirm the TaqMan results in 10 selected environmental isolates. All three TaqMan pathogen detection kits were useful for virulence gene analysis of prescreened foodborne O157:H7 isolates, while the O157:H7-specific kit may not be suitable for virulence gene analysis of environmental E. coli isolates, because of high false positive identification. The ability to rapidly identify the presence of pathogenic E. coli in food or environmental samples is essential to avert outbreaks. These results are of importance to microbiologists seeking to use TaqMan PCR to rapidly identify pathogenic E. coli in environmental samples. Furthermore, serotyping may not be a reliable method for identification of O157:H7 strains.


Viral clearance in HCV viraemic patients with normal alanine aminotransferase after combination therapy: a controlled, open-labelled study.
In patients with chronic hepatitis C virus infection and persistently normal alanine aminotransferase levels, liver fibrosis has been reported in 0-22% of cases and advanced liver disease in 5-10% of cases. To determine whether patients with persistently normal alanine aminotransferase levels clear infection after anti-viral therapy at equal or different rates from infected patients with raised alanine aminotransferase levels. Thirty-five hepatitis C virus RNA-positive patients with fibrosis at liver histology (Group 1) were matched for genotype, sex, age and histology with patients with raised alanine aminotransferase levels (Group 2). Both groups were treated with 3 MU interferon-alpha2b plus ribavirin (1000-1200 mg) for 12 months. End-of-therapy response was achieved in 71.4%[95% confidence interval (CI), 56.4-86.3] of patients in Group 1 and in 52.3% (95% CI, 42.8-61.9) of those in Group 2 (P = 0.04). At week 72, 22 patients (62.8%; 95% CI, 46.8-78.1) in Group 1 and 50 patients (47.5%; 95% CI, 38.0-57.1) in Group 2 showed a sustained virological response (P = 0.11). Non-1 genotype was the only independent predictor of sustained response (P = 0.002), with an odds ratio of 3.45 (95% CI, 1.58-7.50). At month 3 of therapy, the positive predictive values for non-response were 100% and 96% in Groups 1 and 2, respectively. Interferon and ribavirin induce comparable sustained virological response in patients with persistently normal or raised alanine aminotransferase levels. Stage 1 fibrosis, rather than alanine aminotransferase levels, may be the criterion on which to decide whether or not to treat patients with persistently normal alanine aminotransferase levels.


Differential effects of methylphenidate on the growth of neonatal and adolescent rats.
Methylphenidate (MPH), the drug of choice in the treatment of Attention Deficit Disorders with Hyperactivity (ADD/H), has raised concern regarding its suspected potential for reducing body stature in growing patients. In a previous study we demonstrated that neonatal rats treated with MPH (35 mg/kg, SC, twice daily) showed an acute growth impairment followed by a rapid growth-rebound phenomenon. This report confirms our earlier findings in neonatal rats and extends the investigation of the growth suppressing effects of MPH to the periadolescent period of development in rats. Specifically, neonatal groups of male and female rats treated with higher and lower doses of MPH than in the original study confirmed the growth impairment and growth rebound phenomena reported earlier. Unlike neonatal rats, rats treated during the periadolescent period of development failed to show any growth impairment. These data suggest that the growth suppressing effects of MPH are the result of an acute toxicity which is readily reversible on discontinuation of the drug. Further, it is concluded that there is a low probability of long term effects on human body stature when the minimal therapeutic dose is used in clinical practice.


Angiotensin-converting enzyme insertion/deletion polymorphism is associated with cerebral white matter changes in Alzheimer's disease.
The presence of cerebral white matter changes (WMC) has been reported as an important predictor of the rapidity of cognitive decline in Alzheimer's disease (AD). The association between the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism and WMC in AD is yet to be elucidated. The present study aimed to examine the association between the ACE I/D polymorphism and WMC among AD patients in Taiwan. A total of 403 patients clinically diagnosed with AD were recruited in a cross-sectional study carried out in an area hospital in Kaohsiung, Taiwan. The ACE I/D polymorphism was genotyped, and cerebral white matter rating was carried out using the visual rating scale for age-related white matter changes. The I allele was associated with a significantly lower total age-related white matter changes scale score compared with the D allele (4.83 vs 5.93, P = 0.013). The total age-related white matter changes scale score was significantly lower for the I/I genotype than for the I/D (4.37 vs 5.87, P = 0.009) and I/D + D/D genotypes (4.37 vs 5.91, P = 0.006), with no differences observed between the I/I + I/D and the D/D genotypes (5.08 vs 6.09, P = 0.373), after adjustment for age and hypertension. A stratified analysis by sex demonstrated that the I/I genotype was associated with significant lower WMC than other genotypes in women, but not in men. The present study supports the hypothesis that the ACE I/D polymorphism is associated with the severity of WMC in patients with AD. Patients with AD who are homozygous for the I allele might be less likely to develop WMC, especially women. Geriatr Gerontol Int 2017; 17: 945-950.


Capsule Commentary on Verma et al., Prevalence and Costs of Discharge Diagnoses in Inpatient General Internal Medicine: a Multi-center Cross-sectional Study.


Embryonic stem cell/fibroblast hybrid cells with near-tetraploid karyotype provide high yield of chimeras.
Ten primary clones of hybrid cells were produced by the fusion of diploid embryonic stem (ES) cells, viz., line E14Tg2aSc4TP6.3 marked by green fluorescent protein (GFP), with diploid embryonic or adult fibroblasts derived from DD/c mice. All the hybrid clones had many characteristics similar to those of ES cells and were positive for GFP. Five hybrid clones having ploidy close to tetraploidy (over 80% of cells had 76-80 chromosomes) were chosen for the generation of chimeras via injection into C57BL blastocysts. These hybrid clones also contained microsatellites marking all ES cell and fibroblast chromosomes judging from microsatellite analysis. Twenty chimeric embryos at 11-13 days post-conception were obtained after injection of hybrid cells derived from two of three clones. Many embryos showed a high content of GFP-positive descendents of the tested hybrid cells. Twenty one adult chimeras were generated by the injection of hybrid cells derived from three clones. The contribution of GFP-labeled hybrid cells was significant and comparable with that of diploid E14Tg2aSc4TP6.3 cells. Cytogenetic and microsatellite analyses of cell cultures derived from chimeric embryos or adults indicated that the initial karyotype of the tested hybrid cells remained stable during the development of the chimeras, i.e., the hybrid cells were mainly responsible for the generation of the chimeras. Thus, ES cell/fibroblast hybrid cells with near-tetraploid karyotype are able to generate chimeras at a high rate, and many adult chimeras contain a high percentage of descendants of the hybrid cells.


Neuroprotective effects of Ilexonin A following transient focal cerebral ischemia in rats.
Ilexonin A is a compound isolated from the root of a plant commonly used in traditional Chinese medicine. The aim of the present study was to investigate the possible protective mechanism of Ilexonin A in rats subjected to occlusion of the middle cerebral artery (MCAO). Transient focal cerebral ischemia was induced by 2 h of MCAO, followed by reperfusion. Ilexonin A at doses of 20, 40 and 80 mg/kg were administered via intraperitoneal injection immediately following ischemia/reperfusion. The expression levels of glial fibrillary acidic protein (GFAP), ionized calcium‑binding adapter molecule‑1 (Iba‑1), vascular endothelial growth factor (VEGF), fetal liver kinase‑1 (Flk‑1) and Nestin were examined using immunostaining and Western blot analysis of the peri‑infarct region following ischemia/reperfusion. Ilexonin A significantly decreased the infarct volume and improved neurological deficits in a dose‑dependent manner. The expression levels of VEGF, Flk‑1 and Nestin were significantly increased in the rats treated with Ilexonin A, compared with the rats administered with saline. Following treatment with Ilexonin A, a higher number of GFAP‑positive astrocytes were found in the Ilexonin A‑treated rats at 1, 3 and 7 days, compared with the rats exposed to ischemia only, however, there were fewer astrocytes at 14 days, compared with the ischemia group. Ilexonin A significantly decreased the protein expression of Iba‑1. The results of the present study suggested that the protective effects of Ilexonin A were associated with revascularization, neuronal regeneration, and the regulation of astrocyte and microglia cell activation.


Determinants of hospital charges for coronary artery bypass surgery: the economic consequences of postoperative complications.
This is a prospective study of 500 consecutive patients having coronary artery bypass surgery; mean hospital charge from time of surgery to discharge was +11,900 +/- 12,700. Multiple regression analysis was performed using preoperative variables and postoperative complications. No preoperative clinical feature was a significant predictor of higher average charge. Sternal wound infection (p = 0.0001), respiratory failure (p = 0.0001) and left ventricular failure (p = 0.017) were associated with higher average hospital charge. The absence of any complication predicted a lower average charge, and postoperative death (4.4 +/- 4.5 days after surgery) was also associated with lower average charge. A cost equation was developed: hospital charge equalled $11,217 + $41,559 of sternal wound infection, + $28,756 for respiratory failure, + $5,186 for left ventricular failure, - $1,798 for no complication and - $6,019 for death. Recognition of the influence of complications on charges suggests that low average charges can only be achieved by surgical programs with a low complication rate.


In vivo choroidal thickness measurement.
We developed a system that permits exceptionally accurate measurements of human choroidal thickness in vivo by means of ultrasound. With minicomputer techniques, the complementary functions of the time and spectral domains of reflected sound can be used to permit measurements accurate to less than 20 mu at 10 MHz center frequency. Even more accuracy can be obtained with higher center frequencies. Choroidal thickness has been found to exceed that previously reported, with typical values of 420 mu found at the posterior pole. Even greater values are seen in regions outside the macula.


Hertz-rate metropolitan quantum teleportation.
Quantum teleportation can transfer an unknown quantum state between distant quantum nodes, which holds great promise in enabling large-scale quantum networks. To advance the full potential of quantum teleportation, quantum states must be faithfully transferred at a high rate over long distance. Despite recent impressive advances, a high-rate quantum teleportation system across metropolitan fiber networks is extremely desired. Here, we demonstrate a quantum teleportation system which transfers quantum states carried by independent photons at a rate of 7.1 ± 0.4 Hz over 64-km-long fiber channel. An average single-photon fidelity of ≥90.6 ± 2.6% is achieved, which exceeds the maximum fidelity of 2/3 in classical regime. Our result marks an important milestone towards quantum networks and opens the door to exploring quantum entanglement based informatic applications for the future quantum internet.


Modern labor epidural analgesia: implications for labor outcomes and maternal-fetal health.
Epidural analgesia is an important means of pain control during labor throughout the world. Over its historic development, it has been implicated in several undesirable outcomes, including prolongation of labor and increased need for operative delivery. These effects have emerged in some retrospective and observational studies, but such methods of investigation are highly prone to bias and are particularly ill-suited for the study of labor analgesia. In high-quality studies, including Cochrane reviews and meta-analyses, epidural analgesia has been suggested to extend the first stage of labor by 30 minutes and the second stage by 15 minutes, when compared with alternative forms of analgesia. Although this may be a reproducible effect, it may be argued that it is clinically negligible. With respect to mode of delivery, similar high-quality studies have consistently shown no increased risk of cesarean delivery associated with epidural analgesia. Some forms of epidural analgesia were associated with higher risk of assisted vaginal delivery, but the use of newer modalities has been shown to abolish this effect. Specific advancements have centered on reducing total anesthetic consumption, given that local anesthetic-induced motor block is theorized to interfere with maternal expulsive efforts in the second stage of labor. These measures include the use of low-concentration local anesthetic solutions equivalent to ≤0.1% bupivacaine, shown in meta-analyses to lead to no higher risk of assisted vaginal delivery relative to nonepidural analgesia. Additional advancements in the maintenance of analgesia include programmed intermittent epidural bolus and patient-controlled epidural analgesia, the combination of which has been shown to reduce the risk of assisted vaginal delivery, also likely mediated by reduction in local anesthetic dose. These techniques have gained popularity in the past two decades, such that studies published since 2005 show no higher risk of assisted vaginal delivery with epidural than with opioid analgesia (as reported in a Cochrane review). Labor epidural analgesia has implications for maternal and fetal health perinatally. It is known to result in transient maternal hypotension (particularly with initiation), which may progress to the level of necessitating fluid or vasopressor therapy. This is not clearly associated with any adverse outcomes. There is also a consistently higher incidence of fever in parturients receiving neuraxial anesthesia, likely of noninfectious origin, which has similarly not been associated with adverse neonatal outcomes. Finally, neonates of parturients who receive epidural analgesia have been shown to have no worse Apgar scores and more favorable acid-base status than their counterparts. These observations should serve to reassure providers that modern labor analgesia, as currently understood, is not consistently associated with any significant adverse outcomes for the parturient or fetus. In this review, we describe variations of modern labor epidural analgesia, conduct an in-depth review of current literature on its use, and explore the most up-to-date evidence on its implications for the progression and outcomes of labor, including the pertinent maternal and fetal side effects.


[Adherence to neurologic treatment. Lessons from multiple sclerosis].
In light of increasing prevalence rates of chronic conditions and a growing elderly population, patients' nonadherence to medical regimes reflects a significant problem in modern health care. In the past, patients were primarily held responsible for problematic adherence. Over the past decades this attitude has changed dramatically, and it is now acknowledged that lack of adherence reflects a problem requiring different approaches and interventions on many different levels. Here we highlight central aspects and consequences of adherence problems in long-term therapy. Factors affecting treatment motivation in multiple sclerosis patients are summarized focusing on neurologic treatment issues, and specific intervention strategies for patients, doctors, and nurses are discussed.


Dissection of the MKK3 Functions in Human Cancer: A Double-Edged Sword?
The role played by MKK3 in human cancer is controversial. MKK3 is an evolutionarily conserved protein kinase that activates in response to a variety of stimuli. Phosphorylates, specifically the p38MAPK family proteins, contribute to the regulation of a plethora of cellular processes such as proliferation, differentiation, apoptosis, invasion, and cell migration. Genes in carcinogenesis are classified as oncogenes and tumor suppressors; however, a clear distinction is not always easily made as it depends on the cell context and tissue specificity. The aim of this study is the examination of the potential contribution of MKK3 in cancer through a systematic analysis of the recent literature. The overall results reveal a complex scenario of MKK3's involvement in cancer. The oncogenic functions of MKK3 were univocally documented in several solid tumors, such as colorectal, prostate cancer, and melanoma, while its tumor-suppressing functions were described in glioblastoma and gastric cancer. Furthermore, a dual role of MKK3 as an oncogene as well as tumor a suppressor has been described in breast, cervical, ovarian, liver, esophageal, and lung cancer. However, overall, more evidence points to its role as an oncogene in these diseases. This review indicates that the oncogenic and tumor-suppressing roles of MKK3 are strictly dependent on the tumor type and further suggests that MKK3 could represent an efficient putative molecular target that requires contextualization within a specific tumor type in order to adequately evaluate its potential effectiveness in designing novel anticancer therapies.


Stimulation of the periaqueductal gray subdues sensitized pain in morphine- and meperidine-dependent rats.
Because increased tolerance of narcotics is marked by progressive deactivation of the descending antinociceptive system, a question was raised whether stimulation of the periaqueductal gray matter (PAG) would have any electroanalgetic effect in animals adapted to increasing doses of narcotics. The daily dose of morphine (10 mg/kg) administered to rats was increased on alternate days by 10 mg/kg to 100 mg/kg/day. To another group, the daily dose of meperidine was increased from 15 mg/kg by 15 mg/kg to 150 mg/kg/day. Electrophysiological experiments were conducted under chloralose and urethane anesthesia 16 h after the last injection of morphine or meperidine. Spike potentials evoked from individual neurons of the nucleus ventralis posterolateralis by single-pulse stimulation of the sciatic nerve were accumulated in poststimulus time histograms. For nociceptive neurons the histograms were characterized by a short-latency activity peak and at least two late (270 and 420 ms) peaks. For non-nociceptive neurons the histograms had no late activity peaks. In control rats, stimulation of the PAG (400 ms at 70/s) prior to each sciatic nerve pulse reorganized the late activity peaks of the nociceptive neurons: a single late peak occurred during the 280 to 400 ms poststimulus interval, indicating suppression of pain by electroanalgesia. In rats adapted to morphine or meperidine, intracarotid infusion of naloxone lowered the nociceptive threshold. Stimulation of the PAG reorganized the late peaks but only if the sciatic nerve stimulation was not increased. At the voltage used to stimulate the sciatic nerve in control animals, two separate late peaks appeared, which were subdued by PAG stimulation after intracarotid infusion of 5-hydroxytryptophan (5-HTP). These results affirmed previous findings that electroanalgesia is induced by activity in an ascending and a descending pathway, both originating from the PAG. Since the function of the descending pathway is impaired by repeated administration of narcotics, only the pathway ascending to the somesthetic thalamus can be activated to mask pain, unless 5-HTP is injected. The latter renews the functional capacity of the descending pathway and thus reinstates the full capacity of electroanalgesia.


Cranioplasty with custom-made implants: analyzing the cases of 10 patients.
The aim of this study was to assess quantitatively whether a symmetric reconstruction of the calvaria could be achieved using 3-dimensional (3D) custom-made implants and to examine any complications caused by the cranioplasty. Custom-made cranial implants were produced using data obtained from computed tomographic scanning of the defect using computer-aided design and rapid prototyping techniques. Polymethylmethacrylate was used as the reconstruction material and the implant was cast from a silicone rubber mold. These implants were used in 10 patients (9 men and 1 woman) who previously received a craniectomy. The symmetry gained after cranioplasty was quantified by volumetric analysis using 3D reconstructed postoperative computed tomographic imaging. Any complications after cranioplasty also were recorded. The average follow-up was 42.5 months (range, 7 to 85 mo). The esthetic appearance of all patients was much improved. When the volume of the reconstructed right calvaria was compared with the left calvaria, the difference was not statistically significant (P > .05). There were 2 cases of complications. One exhibited a transient seroma collection. Another had a wrinkle formation in the forehead. No infectious episodes or signs of plate rejection were encountered. The custom-made implants for cranioplasty showed a significant improvement in morphology. The implants may be very useful for repairing large and complex-shaped cranial defects. The technique may be useful for the bone reconstruction of other sites.


Effect of sulfinpyrazone on ventricular arrhythmia, prostaglandin synthesis, and catecholamine release following coronary artery occlusion in the cat.
We examined if inhibition of endogenous prostaglandin (PG) synthesis reduced the severity of ventricular arrhythmia and the incidence of ventricular fibrillation (VF) following occlusion of the left anterior descending coronary artery (LAD) in anesthetized cats. We also determined whether the PGs were interacting in a facilitory manner with the sympathetic nervous system to produce arrhythmia and VF after LAD occlusion. Sulfinpyrazone, an inhibitor of cyclo-oxygenase enzyme, or vehicle was administered intravenously to cats 1 h before LAD occlusion. Sulfinpyrazone completely (p less than 0.001) inhibited the release of 6-keto-PGF1 alpha into the great cardiac vein following LAD occlusion. Sulfinpyrazone (100 mg/kg) significantly (p less than 0.001) reduced the amount of ventricular arrhythmia and the incidence of VF (p less than 0.05) in the 1st h after LAD occlusion. In addition to 6-keto-PGF1 alpha sulfinpyrazone also (p less than 0.001) inhibited the increase in plasma norepinephrine from the heart due to sympathetic nervous system stimulation following LAD occlusion. Since sulfinpyrazone was ineffective in increasing the dose of digoxin required to produce arrhythmia and death, sulfinpyrazone apparently did not depress cardiac excitability. Finally, the extent of infarction resulting from LAD occlusion was not different in sulfinpyrazone-treated animals compared with control. These data indicate that sulfinpyrazone, by inhibiting endogenous PG synthesis in the heart following LAD occlusion, may prevent a facilitory interaction between PGs and the sympathetic nervous system that contributes, in part, to the development of ventricular arrhythmia and VF normally associated with this event.


Solvent-free, supersoft and superelastic bottlebrush melts and networks.
Polymer gels are the only viable class of synthetic materials with a Young's modulus below 100 kPa conforming to biological applications, yet those gel properties require a solvent fraction. The presence of a solvent can lead to phase separation, evaporation and leakage on deformation, diminishing gel elasticity and eliciting inflammatory responses in any surrounding tissues. Here, we report solvent-free, supersoft and superelastic polymer melts and networks prepared from bottlebrush macromolecules. The brush-like architecture expands the diameter of the polymer chains, diluting their entanglements without markedly increasing stiffness. This adjustable interplay between chain diameter and stiffness makes it possible to tailor the network's elastic modulus and extensibility without the complications associated with a swollen gel. The bottlebrush melts and elastomers exhibit an unprecedented combination of low modulus (∼100 Pa), high strain at break (∼1,000%), and extraordinary elasticity, properties that are on par with those of designer gels.


How would I manage a case of essential thrombocythaemia presenting with an ischaemic toe.
Essential thrombocythaemia (ET) is an acquired myeloproliferative disorder. The phenotypic and biological heterogeneity of ET can make management of individual cases problematic, especially in the era of changing ideas on the molecular pathogenesis of this disease process. This case discussion will explore the evidence base and rationale that guides treatment of a 46-year-old individual with ET presenting with an ischaemic episode.


Perforated Meckel's diverticulum in an adult due to faecolith: A case report and review of literature.
Meckel's diverticulum (MD) is a persistent remnant of the vitelointestinal duct and is present in 2% of population [1]. It is the most common congenital malformation of the gastrointestinal tract. It can present clinically as haemorrhage, diverticulitis, intussusception, chronic ulceration, intestinal obstruction and perforation. Complicated presentation, especially bleeding, tends to be more common in the paediatric group, whereas intestinal obstruction is more common in adults [2]. Patients with a perforation of Meckel's diverticulum by an enterolith are rare and may present with right iliac fossa pain, which mimics acute appendicitis.


Gallbladder carcinoma update: multimodality imaging evaluation, staging, and treatment options.
The purpose of this article is to review the epidemiology, multimodality imaging findings, differential diagnosis, pathologic staging, and current treatment options of gallbladder carcinoma. Understanding the characteristic appearances of primary gallbladder carcinoma at multiple imaging modalities can facilitate diagnosis and enable more accurate staging for triage to extended resection or an alternate therapy.


Different techniques for diagnostic arthroscopy. A randomized comparative study.
A randomized comparison between the anterolateral approach and the Needlescope technique for knee joint arthroscopy in 121 consecutive patients is presented. The central approach was used a reference, and the results of the other two techniques were compared with the results of this technique in the same patient. It is shown that the Needlescope necessitates a multiple puncture technique to give reliable information. When so used the results were comparable with those obtained with Storz arthroscopes and the central approach. On the other hand, the anterolateral approach gave less accurate results, mainly because of the difficulties encountered in examining the posterior compartments.


[The PhD program "Multimorbidity in Old Age and Selected Care Problems" Center for Human and Health Sciences of the Charité-Universitätsmedizin Berlin].
The PhD Program concerns the care of multimorbid geriatric patients. It takes a multidisciplinary approach in combining different perspectives on delivering care to the elderly. Students have completed courses of studies such as nursing, social education, sociology, psychology, or public health. Likewise, doctoral projects are overseen by members of various disciplines including medicine. These disciplines are represented in the Center for Human and Health Sciences of the Charité-Universitätsmedizin Berlin, where the PhD Program is located. The doctoral projects are grouped into three main topics--Care Problems and Interventions, Quality of Life and Self-Determination, and Care Structures and Quality.


'What Brings Him Here Today?': Medical Problem Presentation Involving Children with Autism Spectrum Disorders and Typically Developing Children.
Conversation and discourse analyses were used to examine medical problem presentation in pediatric care.Healthcare visits involving children with ASD and typically developing children were analyzed. We examined how children's communicative and epistemic capabilities, and their opportunities to be socialized into a competent patient role are interactionally achieved. We found that medical problem presentation is designed to contain a 'pre-visit' account of the interactional and epistemic work that children and caregivers carry out at home to identify the child's health problems; and that the intersubjective accessibility of children's experiences that becomes disrupted by ASD presents a dilemma to all participants in the visit. The article examines interactional roots of unmet healthcare needs and foregone medical care of people with ASD.


Restricting sugar or carbohydrate intake does not impact physical activity level or energy intake over 24 h despite changes in substrate use: a randomised crossover study in healthy men and women.
To determine the effects of dietary sugar or carbohydrate restriction on physical activity energy expenditure, energy intake, and physiological outcomes across 24 h. In a randomized, open-label crossover design, twenty-five healthy men (n = 10) and women (n = 15) consumed three diets over a 24-h period: moderate carbohydrate and sugar content (MODSUG = 50% carbohydrate [20% sugars], 15% protein, 35% fat); low sugar content (LOWSUG = 50% carbohydrate [< 5% sugars], 15% protein, 35% fat); and low carbohydrate content (LOWCHO = 8% carbohydrate [< 5% sugars], 15% protein, 77% fat). Postprandial metabolic responses to a prescribed breakfast (20% EI) were monitored under laboratory conditions before an ad libitum test lunch, with subsequent diet and physical activity monitoring under free-living conditions until blood sample collection the following morning. The MODSUG, LOWSUG and LOWCHO diets resulted in similar mean [95%CI] rates of both physical activity energy expenditure (771 [624, 919] vs. 677 [565, 789] vs. 802 [614, 991] kcal·d-1; p = 0.29] and energy intake (2071 [1794, 2347] vs. 2195 [1918, 2473] vs. 2194 [1890, 2498] kcal·d-1; P = 0.34), respectively. The LOWCHO condition elicited the lowest glycaemic and insulinaemic responses to breakfast (P < 0.01) but the highest 24-h increase in LDL-cholesterol concentrations (P < 0.001), with no differences between the MODSUG and LOWSUG treatments. Leptin concentrations decreased over 24-h of consuming LOWCHO relative to LOWSUG (p < 0.01). When energy density is controlled for, restricting either sugar or total dietary carbohydrate does not modulate physical activity level or energy intake over a 24-h period (~ 19-h free-living) despite substantial metabolic changes. NCT03509610, https://clinicaltrials.gov/show/NCT03509610.


Assessment of some diterpenoids in commercial distilled gin.
In the present study the qualitative and quantitative determination of diterpenoids in commercial distilled gin was carried out. This widely consumed juniper-based spirit is aromatized using Juniper (Juniperus communis) berries. Although juniper reportedly contains several diterpenic compounds, no studies have addressed the diterpenic composition of juniper-based spirits or beverages. With this objective, here we followed a multilevel factorial experimental design to optimize a direct immersion-solid phase microextraction (DI-SPME) method coupled to gas chromatography/mass spectrometry and analyzed eight commercial brands of gin. With total concentrations ranging from 10 to 190 microg L(-1), manool, manoyl oxide and trans-totarol were the most abundant diterpenoids of the 10 identified or tentatively identified at variable but not negligible concentrations in the distilled gin samples. The diterpenic composition allowed the brands to be differentiated. This indicates that these compounds contribute to the sensory characteristics of the distinct commercial brands, thus guaranteeing the authenticity and consequently the quality of the product.


Variability in HER2 expression between primary colorectal cancer and corresponding metastases.
This study aimed to investigate the HER2 status of primary colorectal cancer (CRC) and corresponding metastases, and determine the correlation between HER2 and clinicopathological characteristics. We collected the clinicopathological features of 98 CRC patients and 66 patients which were previously evaluated for the KRAS status. The tissue samples of primary CRC tumors (n = 98), noninvolved colorectal mucosa (n = 98), paired lymph nodes (n = 98, 69 patients had positive metastatic nodes), and liver metastases (n = 22) were evaluated using immunohistochemistry for HER2. The kappa test was used to evaluate the concordance rate of HER2 status. Survival analysis was established according to the Kaplan-Meier method. HER2 overexpression was more common in primary tumors among the younger patients (P < 0.05). No survival significance was revealed in the HER2 status. The HER2-positive rate was 11.2% for the primary CRC tumors, 0% for the normal adjacent mucosa tissues, 10.1% for the matched positive lymph nodes, and 31.8% for the corresponding metastasis. Seven of sixty-nine cases (10.1%) exhibited biomarker discordance in nodal metastases compared with primary tumors (κ = 0.48, P < 0.05); 6 out of 22 cases (27.3%) exhibited biomarker discrepancy in liver metastases compared with primary tumors (κ = 0.32, P > 0.05). Compared with lymph nodes, HER2 overexpression in the primary tissue was more common in KRAS wild-type patients (P < 0.05). The present study showed a high rate of discordance in matched pairs of primary tumors and metastases, suggesting that the accurate evaluation of HER2 status is essential before any therapeutic decision.


Intra-lymphatic administration of interleukin-2 (IL-2) in cancer patients: a pilot study.
Seven patients suffering from advanced metastatic tumours, unresponsive to standard therapies, were treated with 3 to 5 intra-lymphatic injections of interleukin-2 (IL-2) and IL-2-activated-peripheral blood lymphocytes (PBL). A partial (50-70%) regression was obtained in three of the patients, and complete regression in the other four. It thus seems that intra-lymphatic injections of IL-2 and PBL can be used for the treatment of certain solid tumours (e.g., hypernephroma, adenocarcinoma, seminoma, epidermoid carcinoma) without noticeable side effects. This method could advantageously replace intravenous IL-2 administration and warrants further investigation.


Mutational analysis of the Klebsiella pneumoniae nitrogenase promoter: sequences essential for positive control by nifA and ntrC (glnG) products.
ntr (nitrogen regulated) and nif (nitrogen fixation) promoters are structurally similar to each other but bear no resemblance to canonic Escherichia coli promoters. ntr promoters are normally activated by the ntrC (glnG) product, but they can also be activated by the ntrC-related Klebsiella pneumoniae nifA product. In contrast, nif promoters of K. pneumoniae such as the nitrogenase (nifH) promoter can only be nifA activated. In this paper, we report the isolation and characterization of 28 mutants of the K. pneumoniae nifH promoter. Class A mutants no longer respond to nifA-mediated transcription, and class B mutants can now respond to ntrC-mediated activation. These two classes of mutants define sequences important to nifA- and ntrC-mediated transcription. Most surprising is that a single base change is sufficient to convert a nifA-activated promoter into an ntrC-activated one.


Isoquercitrin from Argemone platyceras inhibits carbachol and leukotriene D4-induced contraction in guinea-pig airways.
Argemone platyceras is used in Mexico as a remedy for cough, bronchitis and pneumonia. The present study was performed to investigate the pharmacological anti-asthmatic properties of Argemone platyceras on airways and to identify its active principles. Methanol extracts of leaves and flowers, subsequent organic and aqueous extraction phases, and silica gel chromatography fractions were assayed on the carbachol-induced response, and/or on ovalbumin antigenic challenge, and on leukotriene D(4)-induced response of tracheae from sensitized and non-sensitized guinea-pigs. Methanol extracts, ethyl-acetate phase, and its fractions 6 and 7 inhibited the carbachol-induced contractile response. Isoquercitrin and rutin were the main compounds found in fractions 6 and 7 respectively. Isoquercitrin (fraction 6) abolished the response to ovalbumin, and decreased the contractile response to leukotriene D(4). Because of its effect on carbachol-induced contractile response, on the late-phase response to ovalbumin, and on leukotriene D(4)-induced contractile response, isoquercitrin might be highly useful in treatment of asthma.


Renal cortical mercury levels associated with experimental amalgam tattoos: effects of particle size and amount of implanted material.
Powdered dental amalgam that had passed through either a 106 microns or a 45 microns sieve was implanted subcutaneously in guinea pigs for periods of up to 2 yr. The renal cortical mercury levels associated with the 106 microns material were on average 16% of those produced by the 45 microns material. A reduction in the amount of 45 microns powder implanted, by a factor of 75%, resulted in a fall of only 27% in renal mercury concentrations. The marked effect of particle size on mercury release may be explained by the large increase in the proportion of implanted material that was degraded within phagocytic cells in the local lesions.


Development of a radioimmunoassay for idazoxan hydrochloride.
To facilitate the measurement of idazoxan in clinical trials an immunoassay capable of detecting low ng ml-1 concentrations was required. A stable derivative was prepared which, after linking to keyhole limpet hemocyanin (KLH), was subsequently immunized into sheep. Early assay development was carried out with both fluorescent (1-FITC) and iodinated (I-125) labels. The assay methodology was the same in both cases, using magnetizable solid-phase particles to which the antibody was linked. The sensitivity of the assay was such that a large sample volume was required, which in turn, led to problems of increased protein interference. The use of pepsin to digest the protein was used effectively after several blocking agents were unsuccessfully used. The limit of detection was in the region of 3 ng ml-1. Cross-reactivity studies showed that the antibody was specific for idazoxan. Intra- and inter-assay precision was 7 and 12%, respectively. Correlation with the analytical GC-MSD method was in the order of 0.90.


Exploring the acute effects of running on cerebral blood flow and food cue reactivity in healthy young men using functional magnetic resonance imaging.
Acute exercise suppresses appetite and alters food-cue reactivity, but the extent exercise-induced changes in cerebral blood flow (CBF) influences the blood-oxygen-level-dependent (BOLD) signal during appetite-related paradigms is not known. This study examined the impact of acute running on visual food-cue reactivity and explored whether such responses are influenced by CBF variability. In a randomised crossover design, 23 men (mean ± SD: 24 ± 4 years, 22.9 ± 2.1 kg/m2 ) completed fMRI scans before and after 60 min of running (68% ± 3% peak oxygen uptake) or rest (control). Five-minute pseudo-continuous arterial spin labelling fMRI scans were conducted for CBF assessment before and at four consecutive repeat acquisitions after exercise/rest. BOLD-fMRI was acquired during a food-cue reactivity task before and 28 min after exercise/rest. Food-cue reactivity analysis was performed with and without CBF adjustment. Subjective appetite ratings were assessed before, during and after exercise/rest. Exercise CBF was higher in grey matter, the posterior insula and in the region of the amygdala/hippocampus, and lower in the medial orbitofrontal cortex and dorsal striatum than control (main effect trial p ≤ .018). No time-by-trial interactions for CBF were identified (p ≥ .087). Exercise induced moderate-to-large reductions in subjective appetite ratings (Cohen's d = 0.53-0.84; p ≤ .024) and increased food-cue reactivity in the paracingulate gyrus, hippocampus, precuneous cortex, frontal pole and posterior cingulate gyrus. Accounting for CBF variability did not markedly alter detection of exercise-induced BOLD signal changes. Acute running evoked overall changes in CBF that were not time dependent and increased food-cue reactivity in regions implicated in attention, anticipation of reward, and episodic memory independent of CBF.


Did the late spring frost in 2007 and 2011 affect tree-ring width and earlywood vessel size in Pedunculate oak (Quercus robur) in northern Poland?
Trees are sensitive to extreme weather and environmental conditions. This sensitivity is visible in tree-ring widths and cell structure. In our study, we hypothesized that the sudden frost noted at the beginning of May in both 2007 and 2011 affected cambial activity and, consequently, the number and size of vessels in the tree rings. It was decided to test this hypothesis after damage to leaves was observed. The applied response function model did not show any significant relationships between spring temperature and growth. However, this method uses average values for long periods and sometimes misses the short-term effects. This is why we decided to study each ring separately, comparing them with rings unaffected by the late frost. Our study showed that the short-term effect of sudden frost in late spring did not affect tree rings and selected cell parameters. The most likely reasons for this are (i) cambial activity producing the earlywood vessels before the occurrence of the observed leaf damage, (ii) the forest micro-climate protecting the trees from the harsh frost and (iii) the temperature decline being too short-lived an event to affect the oaks. On the other hand, the visible damage may be occasional and not affect cambium activity and tree vitality at all. We conclude that oak is well-adapted to this phenomenon.


The Multi-Omics Analysis Revealed a Metabolic Regulatory System of Cecum in Rabbit with Diarrhea.
With the comprehensive prohibition of antibiotics in the feed industry in China, the incidence of diarrhea in rabbits increased, such as loss of appetite, vomiting, and excretion of atheromatous feces. In order to explore the pathological and the molecular mechanisms of the diarrhea in the rabbitry fed with antibiotic-free diet, we used microbial metagenomics, transcriptome, and non-targeted metabolomics sequencing. The results showed that the Firmicutes level was significantly decreased (p < 0.001) and the Proteobacteria level was significantly increased (p < 0.05). The functional enrichment of cecum revealed that most differentially expressed genes (DEGs) were expressed in immune, inflammatory, and metabolic processes. The enrichment of the cecal fecal metabolites focused on the bile secretion, antifolate resistance, and tryptophan metabolism pathways, which are mainly associated with inflammation. The results of correlation analysis showed that Fournierella was positively correlated with myricetin, ursolic acid, and furtherly might cause bile secretion and tryptophan metabolism disorder, aggravate intestinal inflammation, change intestinal permeability, and reduce host immunity, leading to diarrhea in rabbits. This study provides a theoretical basis for illustrating the reason for diarrhea and developing new feeds for the health of rabbits.


Combined chemoassay and mass spectrometric approach to study the reactive potential of electrophiles towards deoxynucleosides as model for DNA.
The modification of DNA by adduct formation is a potential molecular initiating event of genotoxicity. A chemoassay was established to study adduct formation of electrophiles with deoxynucleosides. Liquid chromatography-mass spectrometry was used to determine the reactivity of the model electrophiles para-benzoquinone, hydroquinone, and 1,4-naphthoquinone with deoxynucleoside (deoxyadenosine (dA), deoxyguanosine (dG), deoxycytidine (dC) and thymidine (dT)) to detect formation of adducts via constant neutral loss scan of deoxyribose (116 Da), and to elucidate adduct structures using high resolution mass spectrometry. Of the four deoxynucleosides dG was most susceptible, followed by dC and para-benzoquinone was the most reactive electrophile. With this approach five dG and four dC adducts were detected, formed by Michael addition and subsequent condensation. Also oxidation occurred with reactive oxygen species (ROS). Three of the adducts formed by benzoquinone have not been reported before. This chemoassay combined with mass spectrometry offers a way (a) to screen a large number of chemicals for their genotoxic potential, (b) to determine novel adducts that may be searched for in in vitro and in vivo studies and thus (c) to better understand the reaction of electrophiles with nucleobases.


Cytotoxic effect of nanosilver particles on testicular tissue: Evidence for biochemical stress and Hsp70-2 protein expression.
Lastly, there are growing evidences that nanosilver (NS) particles highly induce cytotoxic impacts in vitro and in vivo. Here, we analyzed the dose dependent effect of NS on histological changes, biochemical alterations and endocrine statuses, sperm parameters as well as chaperone Hsp70-2 expression. NS particles (50-60nm) were administrated in 3 doses of 0.5, 1 and 5mg/kg, intraperitoneally, for 35 days. The 0.3mL normal saline was administrated in control-sham group. Histological alterations, sperm parameters, serum levels of LH, FSH and testosterone were evaluated. Germinal and Leydig cells RNA damage, Leydig cells steroidogenic foci, the testicular and sperm total antioxidant capacity (TAC), malondialdehyde (MDA), nitric oxide (NO) levels, immunohistochemical (IHC) expression and mRNA level of Hsp70-2 were analyzed. The NS, dose dependently, resulted in enhanced germinal cells degeneration, necrosis, seminiferous tubules atrophy and decreased serum levels of LH, FSH and testosterone. Elevated germinal and Leydig cells RNA damage associated with increased sperm abnormalities were observed in NS-treated groups. Expression of Hsp70-2 was up-regulated in 0.5mg/kg, while its expression was decreased in 1 and 5mg/kg NS-treated groups. Testicular and sperm TAC levels reduced. However, the MDA and NO levels significantly (P<0.05) increased in all NS-treated groups. No histological and biochemical changes were detected in control-sham group. In conclusion, the NS particles exert their pathological impact via affecting testicular antioxidant and endocrine statuses, which in turn lead to diminished expression of Hsp70-2. Ultimately, by this mechanism NS particles adversely impact the cellular RNA, DNA and protein contents.


Double-labeling study of axonal branching within the lateral reticulocerebellar projection in the rat.
Collateral axonal branching to the cerebellum from the lateral reticular nucleus (LRN) was studied in the rat by using the fluorescent double-labeling technique. Following injection of Fast Blue (FB) into the cerebellar cortex, followed 3 days later by injection of Nuclear Yellow (NY) into a different region of the cortex, single- and double-labeled cells were found within the LRN. Most LRN-cerebellar projections were bilateral with ipsilateral preponderance, except for the projection to the paramedian lobule, which was completely ipsilateral. The dorsolateral area of the magnocellular division of the LRN contained cells whose axons branch to terminate in the rostral anterior lobe and the caudal part of the ipsilateral paramedian lobule (hindlimb areas of the cerebellar cortex), while the medial area of the LRN contained cells that supply, via collateral axonal branching, the caudal area of the contralateral anterior lobe and the rostral part of the ipsilateral paramedian lobule (forelimb areas of the cerebellum). Branched LRN-cerebellar axons projected to both hemispheres and to both sides of the caudal anterior lobe. No axonal branching was evident in the LRN-cerebellar projection to the rostral anterior lobe. The projection to the anterior and posterior lobe vermis also contained collateral axonal branching.


MRI in eye tumors.
Thirty patients with intraocular tumors were examined with MRI. A clear demarcation was seen between the melanomas and other intraocular structures such as sclera, vitreous and ciliary body. Retinal detachment, hemorrhage or choroidal-retinal layer could be seen separated from the tumor. In all our patients investigated with MRI the absence of extraocular melanoma tissue was confirmed by histopathology or by the findings during surgery. The demarcation between tumor and sclera seemed more sensitive in MRI than in ultrasonography in establishing extrascleral extension. Our study showed that MRI can be a valuable addition to ultrasonography in detecting intraocular tumors or tumor simulating lesions.


Global information network on chemicals (GINC) and its Asian component.
The Global Information Network on Chemicals (GINC) is an effort to build a global information network that links international, national, and other organizations working for the safe management of chemicals in order to exchange information and improve communications. The project was originally proposed in 1993 by one of the authors then at the National Institute of Health Sciences (NIHS) of Japan to the International Program on Chemical Safety (IPCS), which is a joint project of World Health Organization (WHO), International Labor Organization (ILO), and United Nations Environment Program (UNEP). The base support system was first implemented at NIHS using the Internet/World Wide Web (WWW) technology in 1995. The project was then endorsed by the Intergovernmental Forum on Chemical Safety (IFCS) and was adopted by the Inter-Organization Program for the Sound Management of Chemicals (IOMC). However, the base system (http://www.nihs.go.jp/GINC/index.html) has been developed and maintained solely by the NIHS group under the support of the Ministry of Health and Welfare (MHW), Japan. Asia, particularly East Asia and the Pacific region, was chosen as the feasibility study region for this project. During the period from December 1994 to July 2002, NIHS hosted eight meetings on this project held in Tokyo.


Association between levator ani avulsion and urinary incontinence in women: A systematic review and meta-analysis.
Urinary incontinence is a bothersome symptom. Although the relationship between stress urinary incontinence (SUI) and vaginal delivery is established, the pathology underlying SUI after vaginal birth remains to be elucidated. To determine whether levator ani muscle avulsion predisposes for SUI in women. Pubmed and Embase were searched for terms and their variations "levator ani muscle avulsion" and "urinary incontinence", from inception until 5 November 2019. Inclusion criterion: studies describing the relationship between urinary incontinence and levator ani muscle avulsion in women at least 1 year after delivery. Exclusion criterion: studies only analyzing the urethral sphincter or hiatus dimensions. Odds ratios were used and if not available, were calculated as means of data synthesis, adjusted odds ratios if presented by the study, random-effects model to compute a pooled estimate. Seven studies were included, accounting for 2388 women. Comparing women with and without levator ani muscle avulsion, the overall odds ratio for SUI is 0.87 (95% confidence interval 0.56-1.34), and after adjustment for possible confounders was 0.72 (95% confidence interval 0.40-1.30). There is no relationship between levator ani muscle avulsion and SUI in women.


Assessment of larval deformities and selenium accumulation in northern pike (Esox lucius) and white sucker (Catostomus commersoni) exposed to metal mining effluent.
Uranium mining and milling operations in northern Saskatchewan (Canada) release effluents with elevated levels of certain trace metals and metalloids, including selenium. The goal of the present study was to evaluate the presence of selenium-induced deformities in northern pike (Esox lucius) and white sucker (Catostomus commersoni) larvae originating from adults collected downstream of a uranium mine. Eggs were fertilized in the field and incubated in the laboratory following a two-way (crossover) analysis-of-variance experimental design to discriminate effects from maternal transfer versus those from exposure to site water in the developing embryos. Selenium concentrations in northern pike and white sucker eggs (8.02 and 4.89 microg/g dry wt, respectively; mean +/- standard error throughout) from the exposure site were approximately two- to threefold higher than reference (2.35 +/- 0.20 and 1.94 +/- 0.25 microg/g dry wt, respectively). Among all evaluated deformities (skeletal curvatures, craniofacial deformities, fin deformities, and edema), only edema in white sucker fry from the exposure site was slightly elevated ( approximately 3%) compared to reference. The occurrence of edema, however, can be associated with factors other than selenium (e.g., other metals and organic compounds). Both fish species displayed strong linear relationships between the selenium concentrations in eggs and other tissues (muscle, liver, kidney, and bone), suggesting that selenium concentrations in eggs could be predicted from selenium concentrations in adult tissues. The lack of a clear, toxic response in the present study is in agreement with selenium thresholds for early life-stage deformities reported in other studies, with egg selenium concentrations in northern pike and white sucker collected at the exposure site being less than the 10 microg/g (dry wt) threshold associated with the presence of deformities.


A 3D QSAR study of betulinic acid derivatives as anti-tumor agents using topomer CoMFA: model building studies and experimental verification.
Betulinic acid (BA) is a natural product that exerts its cytotoxicity against various malignant carcinomas without side effects by triggering the mitochondrial pathway to apoptosis. Betulin (BE), the 28-hydroxyl analog of BA, is present in large amounts (up to 30% dry weight) in the outer bark of birch trees, and shares the same pentacyclic triterpenoid core as BA, yet exhibits no significant cytotoxicity. Topomer CoMFA studies were performed on 37 BA and BE derivatives and their in vitro anti-cancer activity results (reported as IC₅₀ values) against HT29 human colon cancer cells in the present study. All derivatives share a common pentacyclic triterpenoid core and the molecules were split into three pieces by cutting at the C-3 and C-28 sites with a consideration toward structural diversity. The analysis gave a leave-one-out cross-validation q² value of 0.722 and a non-cross-validation r² value of 0.974, which suggested that the model has good predictive ability (q² > 0.2). The contour maps illustrated that bulky and electron-donating groups would be favorable for activity at the C-28 site, and a moderately bulky and electron-withdrawing group near the C-3 site would improve this activity. BE derivatives were designed and synthesized according to the modeling result, whereby bulky electronegative groups (maleyl, phthalyl, and hexahydrophthalyl groups) were directly introduced at the C-28 position of BE. The in vitro cytotoxicity values of the given analogs against HT29 cells were consistent with the predicted values, proving that the present topomer CoMFA model is successful and that it could potentially guide the synthesis of new betulinic acid derivatives with high anti-cancer activity. The IC₅₀ values of these three new compounds were also assayed in five other tumor cell lines. 28-O-hexahydrophthalyl BE exhibited the greatest anti-cancer activities and its IC₅₀ values were lower than those of BA in all cell lines, excluding DU145 cells.


Solution structure of a cysteine rich domain of rat protein kinase C.
Intracellular protein phosphorylation by protein kinase C (PKC) plays a major role in the translation of extracellular signals into cellular events. Speculations on the structural basis for PKC activation are based on sequence homology between their cysteine-rich domains (CRD) and the DNA-binding 'zinc-fingers'. We produced a fragment comprising the second CRD (CRD2) of rat PKC-alpha and determined its three-dimensional structure in solution by NMR spectroscopy. This revealed that CRD2 adopts a globular fold allowing two non-consecutive sets of zinc-binding residues to form two separate metal-binding sites. The fold is different to those previously proposed and allows insight into the molecular topology of a family of homologous proteins.


Effect of water coagulation by seeds of Moringa oleifera on bacterial concentrations.
The effects of a Sudanese water purification method traditionally used in Sudan to treat turbid waters were studied with respect to turbidity reduction and removal of faecal indicator bacteria as well as selected enteric bacterial pathogens. Water treatment was performed at 30 degrees C with Moringa oleifera seed material as a coagulant, and the technique employed corresponded closely to that used to clarify turbid water in Sudanese villages. A turbidity reduction of 80.0-99.5% paralleled by a primary bacterial reduction of 1-4 log units (90.00-99.99%) was obtained within the first 1 to 2 h of treatment, the bacteria being concentrated in the coagulated sediment. During the 24 h observation period a secondary bacterial increase due to regrowth in the supernatant water was consistently observed for Salmonella typhimurium and Shigella sonnei, in some cases for Escherichia coli, but not for Vibrio cholerae, Streptococcus faecalis and Clostridium perfringens. The potential of the method when compared with some alternative for the improvement of rural drinking water supplies is discussed.


Chemotoxicity recovery of mitochondria in non-Hodgkin lymphoma resulting in minimal residual disease.
The mechanisms responsible for resistant disease or recurrence of non-Hodgkin lymphoma (NHL) in children cover a wide spectrum from drug resistance to genetic mutations. A unique mechanism suggesting the role of mitochondria as the key energy source is studied following a clinical observation where pediatric Burkitt lymphoma (BL) specimens from patients on therapy were found to have increased copies of mitochondria DNA (mtDNA) in specimens which were shown to be positive for minimal residual disease and/or persistent disease (MRD/PD). This study hypothesized that the mitochondria play an important role in a cell's recovery from toxicity via a compensatory increase in mtDNA. BL specimens with MRD/PD were assayed for mtDNA. An in vitro model was then designed using Ramos cell lines by exposing the lymphoma cells to varying concentrations of doxorubicin and vincristine for 1 hr; and allowing for recovery in culture over 7 days. DNA was extracted from aliquots over several days to determine mtDNA copy numbers by real-time polymerase chain reaction (PCR). Increased mtDNA copy numbers were found in clinical specimens with MRD/PD as well as in recovering Ramos cells from chemotoxicity. The recovering lymphoma cells from the chemotoxic effects appeared to compensate by increasing mtDNA content, which may contribute to the clinical residual or resistant disease in some cases of childhood BL.


Application and pharmacological mechanism of methotrexate in rheumatoid arthritis.
Methotrexate (MTX) has been used for the treatment of rheumatoid arthritis (RA) for about forty years and to date MTX remains the part of global standard of treatment for RA. The efficacy of MTX in RA is the result of multiple mechanisms of action. In order to summarize the possible pharmacological mechanisms of MTX in the treatment of RA, this review will elaborate on folate antagonism, promotion of adenosine accumulation, regulation of inflammatory signaling pathways, bone protection and maintenance of immune system function.


Fenestrated capillaries in the rat paw dermis adjacent to epidermis and skin appendages.
The fine structure of dermal blood vessels of male Wistar rats was studied in paw regions. Electron microscopy revealed a well-developed plexus of fenestrated capillaries in papillary dermis and adjacent to skin appendages. Intravenously injected ferritin escaped from some of these capillaries. It is suggested that these blood vessels play a role in the exchange of substances at sites of high epithelical turnover.


Transcranial laser doppler mapping of activation flow coupling of the rat somatosensory cortex.
Signal averaged laser Doppler (LD) through a thinned skull over the rat somatosensory cortex was used to map the spatial and temporal characteristics of activation-flow coupling, the change in regional cerebral blood flow (CBF) due to neuronal activation, in response to electrical forepaw stimulation. The location of maximal changes in amplitude of the LD response was reproducibly recorded at 4-5 mm lateral to Bregma. This location is very similar but slightly posterior and lateral to results obtained from electrophysiological, magnetic resonance imaging (MRI), and optical imaging studies. The latency of the activation-flow coupling (AFC) response was inversely correlated to response amplitude, with shorter latencies at positions of maximal amplitude.


Successful reconstruction of extensive laryngotracheal strictures after inhalation burn injury: report of a case.
We report a rare case of long segmental laryngotracheal stenosis following inhalation burn injury. The patient presented 2 months after his injury with progressive stridor and dyspnea necessitating tracheostomy. A computed tomographic scan of the neck revealed stenosis extending from the vocal cords to the top of the sternum. Repair was successfully carried out with multiple surgical procedures employing hinge-flap closure tented with autogenous tissue.


Kinetics of cellular cytotoxicity mediated by cloned cytotoxic T lymphocytes.
Kinetic methods can provide significant information concerning the mechanism of cellular cytotoxicity reactions. Previous kinetic studies of cytotoxic T lymphocytes (Tc) have been hampered by the heterogeneity of the effector cell population tested. We therefore examined the kinetics of lysis mediated by cloned, IL 2 and antigen-dependent murine Tc cells with strong cytotoxic activity that is restricted to distinct tumor-associated antigens on P815 mastocytoma target cells. Initial velocity measurements for cytotoxicity mediated by these clones fit a simple Michaelian kinetic model. Specific activity values obtained from these initial rate measurements are compared to those obtained for polyclonal Tc preparations, NK cells, and activated killer cells. Whereas the initial rate of lytic programming mediated by these cloned cells was very rapid, the rate of cytolysis mediated by the cloned cells decreased significantly within one hour. Since this decrease was observed over a wide range of E:T ratios, it is unlikely to result from product inhibition or a significant decrease in the concentration of unlysed target cells, but may be due to a decrease in the rate of programming and/or effector cell recycling. These results indicate that a simple Michaelian kinetic model is not adequate for tumor cell cytolysis by Tc cells in vitro.


Epilepsy and aging.
The intersection of epilepsy and aging has broad, significant implications. Substantial increases in seizures occur both in the elderly population, who are at a higher risk of developing new-onset epilepsy, and in those with chronic epilepsy who become aged. There are notable gaps in our understanding of aging and epilepsy at the basic and practical levels, which have important consequences. We are in the early stages of understanding the complex relationships between epilepsy and other age-related brain diseases such as stroke, dementia, traumatic brain injury (TBI), and cancer. Furthermore, the clinician must recognize that the presentation and treatment of epilepsy in the elderly are different from those of younger populations. Given the developing awareness of the problem and the capabilities of contemporary, multidisciplinary approaches to advance understanding about the biology of aging and epilepsy, it is reasonable to expect that we will unravel some of the intricacies of epilepsy in the elderly; it is also reasonable to expect that these gains will lead to further improvements in our understanding and treatment of epilepsy for all age groups.


A clinical evaluation of a new Silastic seminal fluid collection device.
A new Silastic seminal fluid collection device has been tested clinically and comparison studies have been made with both the polyethylene sheath and commercial latex condoms. In a series of 100 patients, the Silastic seminal fluid device has been found to be superior to the polyethylene sheath with regard to patient comfort and also has had greater reliability. In comparison with the latex device, the Silastic seminal fluid collection device is nearly as comfortable and has been found to be far more reliable.


Microplastics negatively impact embryogenesis and modulate the immune response of the marine medaka Oryzias melastigma.
Microplastic (MP) pollution is an emerging contaminant in aquatic environments worldwide. Nonetheless, the developmental toxicity of MPs in the early life stages of fish and the mechanisms involved are not yet fully understood. The present study investigated the effects of different concentrations of polystyrene (PS) MPs on the early development of the marine model fish the medaka Oryzias melastigma. Our results showed that waterborne exposure to PS MPs significantly delayed the hatching time, altered the heartbeat and decreased the hatching rate of embryos. Furthermore, the genes involved in cardiac development, encoding for embryo-hatching enzymes, as well as inflammatory responses were significantly upregulated. The transcriptome results showed that mainly the pathways involved in metabolism, immune response, genetic information processing and diseases were significantly enriched. These results demonstrate that PS MPs negatively impact embryogenesis and the immune response of O. melastigma.


Effects of thyroid hormones on enzymes involved in fatty acid and glycerolipid synthesis.
The influence of thyroid hormones on lipid biosynthesis was studied after administration of L-thyroxine to rats for 5 days. Their weights remained the same as those of control animals, despite an approximately 3-fold increment in plasma L-thyroxine and L-triiodothyronine concentrations. The activity of acetyl-CoA carboxylase and fatty acid synthetase as well as incorporation of tritium into fatty acids were depressed significantly in epididymal adipose tissue and enhanced significantly in livers of thyroxine-treated rats. Using antibodies specific against rat liver fatty acid synthetase, it was determined that the changes in activity of this multienzymic complex were due to alterations in amount of enzyme protein. In the presence of optimal concentrations of fatty acids, radioactive sn-glycero-3-phosphate, and co-substrates, total glycerolipid synthesis (defined in this study as the sum of newly formed radioactive mono- and diacyl-sn-glycero-3-phosphate, diglyceride, and triglyceride) was decreased significantly in adipose tissue and increased in liver and heart. Thus, administration of thyroid hormone results in tissue-specific alterations in lipid biosynthesis which, at least in the case of fatty acid synthetase, are due to changes in enzyme protein content.


Reliability and validity of DSM-IV and DSM-5 methamphetamine use disorder diagnoses using the Chinese Version of the Semi-Structured Assessment for Drug Dependence and Alcoholism (SSADDA).
The Semi-structured Assessment for Drug Dependence and Alcoholism (SSADDA) was developed to assess substance-use disorders and other psychiatric traits. We translated the SSADDA into Chinese and evaluated its inter-rater reliability and concurrent validity in diagnosing DSM-IV methamphetamine (MA) dependence and DSM-5 MA-use disorder (MUD). The sample comprised 231 participants who were interviewed using the Chinese SSADDA and the Mini-International Neuropsychiatric Interview (Chinese MINI) for concurrent validation. Of the 231 participants, 191 were interviewed by two different interviewers two weeks apart. We evaluated the inter-rater reliability and concurrent validity of the diagnoses using percent agreement and Cohen's kappa coefficient (κ). Cohen's linear weighted kappa was used to assess the reliability of DSM-5 MUD severity. It showed good inter-rater reliability and no significant differences among the DSM-5 MUD (κ = 0.71), DSM-IV MA abuse or dependence (κ = 0.72), and the DSM-IV diagnoses of MA dependence (κ = 0.66) and abuse (κ = 0.68) tested separately. The weighted kappa was 0.67 across the three DSM-5 MUD severity levels. The reliability of each individual diagnostic criterion for DSM-5 MUD ranged from fair to excellent (κ = 0.41-0.80), except for "repeated attempts to quit/control use" (κ = 0.38). The concurrent validity based on MINI-derived diagnoses ranged from good to excellent (κ = 0.65-0.78). This study shows that the Chinese version of SSADDA has good reliability and validity among Chinese MA users.


Relationship between the water-exchangeable fraction of PAH and the organic matter composition of sediments.
The sorption of PAH on 12 different sediments was investigated and was correlated to their corresponding organic matter (OM) content and quality. For this purpose, the OM was precisely characterized using thermal analysis consisting in the successive combustion and quantification of the increasingly thermostable fractions of the OM. Simultaneously, the water-exchangeable fraction of the sorbed PAH defined as the amount of PAH freely exchanged between the water and the sediment (by opposition to the PAH harshly sorbed to the sediments particles) was determined using a passive sampler methodology recently developed. The water concentrations, when the sediment-water system is equilibrated, were also assessed which allows the determination of the sediment-water distribution coefficients without artifacts introduced by the non water-exchangeable fraction of PAH. Hence, the present study provides the distribution coefficients of PAH between the water and 4 different OM fractions combusted at a specific temperature range. The calculated distribution coefficients demonstrate that the sedimentary OM combusted at the intermediate temperature range (between 300 °C and 450 °C) drives the reversible sorption of PAH while the inferred sorption to the OM combusted at a lower and higher temperature range does not dominate the partitioning process.


Distribution of chemical forms of mercury in sediments from abandoned ponds created during former gold mining operations in Colombia.
Total mercury (THg) and methylmercury (MeHg) were studied in sediments from 27 abandoned gold mining ponds (AGMPs) through small-scale artisanal gold mining in the district of San Juan in Chocó region of Colombia. The AGMPs were abandoned in the last century (1997) and were grouped into three distinct groups (2-6; 7-12; 13-20 years). Overall concentration (in ng g-1) pattern of THg in sediments varied from 39.06 to 1271.32 (avg. 209.57) with 174.81 (13-20 years), 205.56 (7-12 years) and 248.33 (2-6 years) respectively. MeHg concentrations accounted for 3.3-10.9% (avg. 6.5%) of THg and were significantly correlated with THg during all periods. Correlations between organic matter (OM) vs MeHg and THg were negative in the oldest pools, signifying a "dilution effect" or "natural burial" of THg and MeHg. Results for sequential extraction indicate that the fraction of elemental Hg (Hg-e) and organo chelated Hg (Hg-o) represent the main chemical forms of Hg in the sediments, regardless of the abandonment period, whereas the bioavailable fraction was only 0.12-1.65% of THg. The significant statistical relationship between MeHg, THg and OM suggests that these parameters control the distribution, mobility, toxicity and bioavailability of Hg in the sediments of these abandoned ponds. Evaluation of THg with sediment quality guidelines indicates that the values are on the higher side for Threshold effect concentration and Upper continental crust. Comparing of MeHg with many other regions outside Colombia is a worrying factor and needs immediate attention to protect the human health.


Aortic dissection: prompt diagnosis and emergency treatment are critical.
Diagnosing aortic dissection requires a high index of suspicion, as it may mimic other more common conditions that cause chest pain. Prompt diagnosis is key, as it requires emergency evaluation and treatment for optimal chances of survival. This paper reviews key clinical features as well as laboratory and imaging tests.


EAU-ESMO consensus statements on the management of advanced and variant bladder cancer-an international collaborative multi-stakeholder effort: under the auspices of the EAU and ESMO Guidelines Committees†.
Although guidelines exist for advanced and variant bladder cancer management, evidence is limited/conflicting in some areas and the optimal approach remains controversial. To bring together a large multidisciplinary group of experts to develop consensus statements on controversial topics in bladder cancer management. A steering committee compiled proposed statements regarding advanced and variant bladder cancer management which were assessed by 113 experts in a Delphi survey. Statements not reaching consensus were reviewed; those prioritised were revised by a panel of 45 experts before voting during a consensus conference. Online Delphi survey and consensus conference. The European Association of Urology (EAU), the European Society for Medical Oncology (ESMO), experts in bladder cancer management. Statements were ranked by experts according to their level of agreement: 1-3 (disagree), 4-6 (equivocal), 7-9 (agree). A priori (level 1) consensus was defined as ≥70% agreement and ≤15% disagreement, or vice versa. In the Delphi survey, a second analysis was restricted to stakeholder group(s) considered to have adequate expertise relating to each statement (to achieve level 2 consensus). Overall, 116 statements were included in the Delphi survey. Of these, 33 (28%) statements achieved level 1 consensus and 49 (42%) statements achieved level 1 or 2 consensus. At the consensus conference, 22 of 27 (81%) statements achieved consensus. These consensus statements provide further guidance across a broad range of topics, including the management of variant histologies, the role/limitations of prognostic biomarkers in clinical decision making, bladder preservation strategies, modern radiotherapy techniques, the management of oligometastatic disease and the evolving role of checkpoint inhibitor therapy in metastatic disease. These consensus statements provide further guidance on controversial topics in advanced and variant bladder cancer management until a time where further evidence is available to guide our approach.


Contents of cry genes and insecticidal toxicity of Bacillus thuringiensis strains from terrestrial and aquatic habitats.
Two Bacillus thuringiensis collections from terrestrial and aquatic habitats were compared in order to study the possible interrelationships between habitat and biological characteristics (serovar, cry genes content and toxicity). Bacillus thuringiensis strains were characterized by serology, PCR, and one-dose treatment against the noctuids Helicoverpa armigera and Spodoptera exigua, and the dipteran Tipula oleracea. A total of 12 and 10 different serovars were identified within terrestrial and aquatic strains, respectively. The number of non-toxic strains was greater in aquatic (41.6%) than in terrestrial habitats (5.3%). The genes cry1C, cry1D and cry1E were significantly more frequent in the terrestrial habitat. The cry1B gene was very frequent within thuringiensis strains. A high diversity was found in terms of serovars present and cry genes content in both collections. The relative frequency of individual cry genes was different in both collections, and a serovar-dependent distribution of the cry1B gene was found. Some strains sharing the same set of cry genes differed in their toxicity, suggesting important differences in gene expression. The inter-relationships between serology, cry gene content and toxicity may allow a better understanding of B. thuringiensis ecology.


Phosphorylation of a 72-kDa nucleoprotein (NP-72) in HL-60 cells is mediated by the double-stranded DNA-dependent protein kinase (DNA-PK).
Nuclear extracts prepared from human leukemic cells grown in the presence of cell differentiation-inducing agents showed a significant increase in the phosphorylation of a nuclear protein referred to as NP-72. The NP-72 phosphorylation was greatly increased by the addition of purified HeLa DNA-PK and was reduced by the inclusion of a DNA-PK-specific monoclonal antibody to the assay mixture. Phosphoamino acid analysis showed serine (60%) and threonine (40%) residues to be the phosphate acceptors. Western blot analysis of control and TPA-treated nuclear extracts detected multiple immunoreactive protein bands. The two most prominent species migrated as 300- and 210-kD proteins on SDS-PAGE, possibly representing an intact and a processed form of DNA-PK. The 300-kD form of DNA-PK was only detected in TPA-treated nuclear extracts, raising the possibility that cell differentiation is associated with the down-regulation and/or the inhibition of a protease(s) capable of regulating DNA-PK turnover.


Myomaker and Myomerger Work Independently to Control Distinct Steps of Membrane Remodeling during Myoblast Fusion.
Classic mechanisms for membrane fusion involve transmembrane proteins that assemble into complexes and dynamically alter their conformation to bend membranes, leading to mixing of membrane lipids (hemifusion) and fusion pore formation. Myomaker and Myomerger govern myoblast fusion and muscle formation but are structurally divergent from traditional fusogenic proteins. Here, we show that Myomaker and Myomerger independently mediate distinct steps in the fusion pathway, where Myomaker is involved in membrane hemifusion and Myomerger is necessary for fusion pore formation. Mechanistically, we demonstrate that Myomerger is required on the cell surface where its ectodomains stress membranes. Moreover, we show that Myomerger drives fusion completion in a heterologous system independent of Myomaker and that a Myomaker-Myomerger physical interaction is not required for function. Collectively, our data identify a stepwise cell fusion mechanism in myoblasts where different proteins are delegated to perform unique membrane functions essential for membrane coalescence.


Recovery of brain from deficiency of essential fatty acids in rats.
Rats were made visibly deficient in essential fatty acids by feeding a deficient diet and were then fed a diet containing 5.0 per cent by weight corn oil as a source of essential fatty acids. After 11-13 weeks on deficient diet the weights of the brain were 25% less than those of rats on control diets. After 5 weeks on the control diet, deficient animals had regained normal brain weight and composition and had lost the deficiency symptoms. Lipids were extracted from the brains and analysed qualitatively and quantitatively, the overall effect of essential fatty acid deficiency being a reduction in the proportion of cerebrosides and sphingomyelin, the appearance of a high proportion of eicosatrienoic acid (20:3) and the reduction of arachidonic acid (20: 4) and the other essential fatty acids. It is considered that essential fatty acid deficiency retards maturing of brains and the present data show that this effect is reversible.


Properties of an ideal PET perfusion tracer: new PET tracer cases and data.
An ideal positron emission tomography (PET) tracer should be highly extractable by the myocardium and able to provide high-resolution images, should enable quantification of absolute myocardial blood flow (MBF), should be compatible with both pharmacologically induced and exercise-induced stress imaging, and should not require an on-site cyclotron. The PET radionuclides nitrogen-13 ammonia and oxygen-15 water require an on-site cyclotron. Rubidium-82 may be available locally due to the generator source, but greater utilization is limited because of its relatively low myocardial extraction fraction, long positron range, and generator cost. Flurpiridaz F 18, a novel PET tracer in development, has a high-extraction fraction, short positron range, and relatively long half-life (as compared to currently available tracers), and may be produced at regional cyclotrons. Results of early clinical trials suggest that both pharmacologically and exercise-induced stress PET imaging protocols can be completed more rapidly and with lower patient radiation exposure than with single-photon emission computerized tomography (SPECT) tracers. As compared to SPECT images in the same patients, flurpiridaz F 18 PET images showed better defect contrast. Flurpiridaz F 18 is a potentially promising tracer for assessment of myocardial perfusion, measurement of absolute MBF, calculation of coronary flow reserves, and assessment of cardiac function at the peak of the stress response.


Severe hypoxemia in children with upper airway obstruction during sleep does not lead to significant changes in heart rate.
Life-threatening cardiac arrhythmias, including bradyarrhythmias, are well-known sequelae of obstructive sleep apnea syndrome (OSAS) in adults and are associated with apnea and severe hypoxemia. Since the clinical expression of OSAS in children is different, we questioned whether arrhythmias are a common feature of pediatric OSAS. Therefore, we analyzed R-R interval patterns from 12 subjects (age 8 months to 14 years) with OSAS in detail. The diagnosis of pediatric OSAS is based on clinical signs of loud snoring and paradoxical respiratory efforts, as well as elevated end-tidal carbon dioxide tension (PETCO2) and major phasic decreases of oxyhemoglobin saturation (SaO2). Two hundred and nine episodes of severe hypoxemia, defined as SaO2 values < or = 85% lasting > or = 30 seconds, were identified. Only 29 (14%) of these episodes were associated with obstructive apneas. The R-R intervals were analyzed before and during each desaturation. Compared to baseline, mean and minimum R-R intervals (RRmean, RRmin) decreased during the desaturation episodes (P < 0.05), while maximum R-R interval (RRmax) increased (P < 0.05). Although the absolute changes in all parameters were significantly different from baseline, the magnitude of these changes was small, with a mean of -5.3%, -12.2%, and 10.4% for RRmean, RRmin, and RRmax, respectively. No child had life-threatening arrhythmias. Although severe hypoxemia existed in these children, the magnitude of the R-R interval changes was modest, and profound bradycardia as described in adults, was rare.(ABSTRACT TRUNCATED AT 250 WORDS)


Genome-wide association study for host response to bovine leukemia virus in Holstein cows.
The mechanisms of leukemogenesis induced by bovine leukemia virus (BLV) and the processes underlying the phenomenon of differential host response to BLV infection still remain poorly understood. The aim of the study was to screen the entire cattle genome to identify markers and candidate genes that might be involved in host response to bovine leukemia virus infection. A genome-wide association study was performed using Holstein cows naturally infected by BLV. A data set included 43 cows (BLV positive) and 30 cows (BLV negative) genotyped for 54,609 SNP markers (Illumina Bovine SNP50 BeadChip). The BLV status of cows was determined by serum ELISA, nested-PCR and hematological counts. Linear Regression Analysis with a False Discovery Rate and kinship matrix (computed on the autosomal SNPs) was calculated to find out which SNP markers significantly differentiate BLV-positive and BLV-negative cows. Nine markers reached genome-wide significance. The most significant SNPs were located on chromosomes 23 (rs41583098), 3 (rs109405425, rs110785500) and 8 (rs43564499) in close vicinity of a patatin-like phospholipase domain containing 1 (PNPLA1); adaptor-related protein complex 4, beta 1 subunit (AP4B1); tripartite motif-containing 45 (TRIM45) and cell division cycle associated 2 (CDCA2) genes, respectively. Furthermore, a list of 41 candidate genes was composed based on their proximity to significant markers (within a distance of ca. 1 Mb) and functional involvement in processes potentially underlying BLV-induced pathogenesis. In conclusion, it was demonstrated that host response to BLV infection involves nine sub-regions of the cattle genome (represented by 9 SNP markers), containing many genes which, based on the literature, could be involved to enzootic bovine leukemia progression. New group of promising candidate genes associated with the host response to BLV infection were identified and could therefore be a target for future studies. The functions of candidate genes surrounding significant SNP markers imply that there is no single regulatory process that is solely targeted by BLV infection, but rather the network of interrelated pathways is deregulated, leading to the disruption of the control of B-cell proliferation and programmed cell death.


A novel technique to assess rotational deformities in lower extremities using CT-based motion analysis.
Rotational deformities following intramedullary (IM) nailing of tibia has a reported incidence of as high as 20%. Common techniques to measure deformities following IM nailing of tibia are either based on clinical assessment, plain X-rays or Computed Tomography (CT) comparing the treated leg with the uninjured contralateral side. All these techniques are based on examiners manual calculation inherently subject to bias. Following our previous rigorous motion analysis and symmetry studies on hemi pelvises, femurs and orthopaedic implants, we aimed to introduce a novel fully digital technique to measure rotational deformities in the lower legs. Following formal institutional approval from the Imperial College, CT images of 10 pairs of human lower legs were retrieved. Images were anonymized and uploaded to a research server. Three dimensional CT images of the lower legs were bilaterally reconstructed. CT-based motion analysis (CTMA) was used and the mirrored images of the left side were merged with the right side proximally as stationary and distally as moving objects. Discrepancies in translation and rotation were automatically calculated. Our study population had a mean age of 54 ± 20 years. There were six males and four females. We observed a greater variation in translation (mm) of Centre of Mass (COM) in sagittal plane (95% CI - 2.959-.292) which was also presented as rotational difference alongside the antero-posterior direction or Y axis (95% CI .370-1.035). In other word the right lower legs in our study were more likely to be in varus compared to the left side. However, there were no statistically significant differences in coronal or axial planes. Using our proposed fully digital technique we found that lower legs of the human adults were symmetrical in axial and coronal plane. We found sagittal plane differences which need further addressing in future using bigger sample size. Our novel recommended technique is fully digital and commercially available. This new technique can be useful in clinical practice addressing rotational deformities following orthopaedic surgical intervention. This new technique can substitute the previously introduced techniques.