One hundred consecutive hepatic resections. Blood loss, transfusion, and operative technique.
Hepatic resection is prone to significant blood loss. Adverse effects of blood loss and transfusion mandate improvements in surgical techniques to reduce blood loss and transfusion requirements. One hundred hepatic resections were carried out using a standard surgical technique that includes control of the hilar structures, extrahepatic control of the hepatic veins, and use of the Pringle maneuver. Low central venous pressure and Trendelenburg positioning were used during parenchymal transection. Data were collected retrospectively in the first 36 patients, whereas data were collected prospectively in the remaining 64 patients. Hospital mortality was 3%. Median blood loss was 450, 700, 1000, 1100, and 1500 mL for segmental, nonanatomic, lobar, extended right, and extended left resections, respectively. Major resections were more likely than minor resections to be transfused with albumin (P = .008), fresh frozen plasma (P = .009), and packed red blood cells or whole blood (P = .04). Overall transfusion of packed red blood cells or whole blood occurred in 59 of 100 patients. In the 64 patients who were followed up prospectively, the predeposit of autologous blood decreased the need for homologous transfusions from 56% to 38%. A further reduction in the transfusion rate of 25% could have been possible if all patients in the prospective group had donated 2 U of autologous blood. Patients who predeposited blood were more likely to receive transfusions and to have had a transfusion more than 24 hours after surgery than were patients who did not donate blood. The surgical techniques used results in acceptable blood loss and transfusion requirements for hepatic resection. This approach is safe, cost-effective, reproducible, and applicable for widespread use.


A prognostic model to personalize monitoring regimes for patients with incidental asymptomatic meningiomas.
Asymptomatic meningioma is a common incidental finding with no consensus on the optimal management strategy. We aimed to develop a prognostic model to guide personalized monitoring of incidental meningioma patients. A prognostic model of disease progression was developed in a retrospective cohort (2007-2015), defined as: symptom development, meningioma-specific mortality, meningioma growth or loss of window of curability. Secondary endpoints included non-meningioma-specific mortality and intervention. Included were 441 patients (459 meningiomas). Over a median of 55 months (interquartile range, 37-80), 44 patients had meningioma progression and 57 died (non-meningioma-specific). Forty-four had intervention (at presentation, n = 6; progression, n = 20; nonprogression, n = 18). Model parameters were based on statistical and clinical considerations and included: increasing meningioma volume (hazard ratio [HR] 2.17; 95% CI: 1.53-3.09), meningioma hyperintensity (HR 10.6; 95% CI: 5.39-21.0), peritumoral signal change (HR 1.58; 95% CI: 0.65-3.85), and proximity to critical neurovascular structures (HR 1.38; 95% CI: 0.74-2.56). Patients were stratified based on these imaging parameters into low-, medium- and high-risk groups and 5-year disease progression rates were 3%, 28%, and 75%, respectively. After 5 years of follow-up, the risk of disease progression plateaued in all groups. Patients with an age-adjusted Charlson comorbidity index ≥6 (eg, an 80-year-old with chronic kidney disease) were 15 times more likely to die of other causes than to receive intervention at 5 years following diagnosis, regardless of risk group. The model shows that there is little benefit to rigorous monitoring in low-risk and older patients with comorbidities. Risk-stratified follow-up has the potential to reduce patient anxiety and associated health care costs.


Norepinephrine and thyrotropin effects on the thyroid in vitro: simultaneous stimulation of iodide organification and antagonism of thyroxine release.
Norepinephrine (NE), which has previously been shown to inhibit TSH-induced T4 release by mouse thyroids in vitro, was found to stimulate iodide organification. The concentration of NE (6 X 10(-7) M) necessary to stimulate organification of iodide was 10 times less than the concentration (6 X 10(-6) M) required for inhibition of TSH-induced T4 release. Both actions of NE were exerted through an alpha-adrenergic receptor, since they were inhibited by phentolamine but not by l-propranolol. One milliunit of TSH maximally stimulated T4 release only, but larger amounts (100 mU) also stimulated organification. TSH stimulation of T4 release and organification was not affected by adrenergic antagonists and therefore was not mediated by adrenergic receptors. N6, O2-Dibutyryl cAMP and isobutylmethylxanthine, like TSH, stimulated T4 release. Their actions were inhibited by NE. However, both compounds, unlike TSH, failed to enhance organification in mouse thyroids. The effects of TSH and NE on the cAMP content of incubated mouse thyroids were also studied. TSH induced a prolonged increase in thyroidal cAMP during the 90-min incubation; this increase was unaffected by alpha- or beta-adrenergic antagonists. In contrast, NE (6 X 10(-5) M) produced a transient but significant increase in cAMP only within the first 5 min. Unlike the action of NE on organification, this short term stimulatory effect on cAMP production was mediated by a beta-adrenergic receptor, since it was blocked by l-propranolol but not by phentolamine. The following conclusions were reached: 1) stimulation of iodide organification and thyroid hormone release involves different sensitivity thresholds for TSH and NE; 2) TSH stimulation of iodide organification, hormone release, and cAMP formation is not exerted through adrenergic receptors; 3) NE stimulates organification and inhibits TSH-stimulated T4 release through alpha-adrenergic receptors, but stimulates cAMP production through beta-receptors; and 4) cAMP may not be the mediator of all TSH actions on the thyroid.


Preparation of electrotechnical products for reduction of hazardous substances compliance testing.
Potentially toxic substances in electrical and electronic equipment are increasingly regulated, yet there is little guidance regarding appropriate sample preparation and analysis for compliance testing. Printed circuit boards are likely to contain regulated substances. Unfortunately, they are very difficult to break into homogeneous components of a mass allowing ready analysis by X-ray fluorescence (XRF) spectrometry, instrumental neutron activation analysis or dissolution-based techniques such as inductively coupled plasma, yet they must be analyzed despite this heterogeneity. Analysis of unprepared samples of circuit board using microspot, hand-held, benchtop and polarizing XRF spectrometers results in unacceptable analytical precision. Shredding samples to < 0.5 mm improves analytical precision sufficientlyfor RoHS compliance screening. To improve analytical precision further, additional sample preparation by fine shredding or milling is required. Milling for 90 s using a puck and ring mill reduces mean particle size to < 90 microm, which is sufficiently fine for XRF analyses or acid dissolution. Sample loss during milling means that fine shredding is probably an optimal form of sample preparation.


Phosphoenolpyruvate carboxykinase and its potential role in the catabolism of organic acids in the flesh of soft fruit during ripening.
Previous studies of grapes and tomatoes have shown that the abundance of phosphoenolpyruvate carboxykinase (PEPCK) increases in their flesh at the start of ripening, and that this coincides with a decrease in its citrate and/or malate content. Thus, PEPCK might function in the catabolism of organic acid anions during the ripening of these fruits. In the present study, the abundance of PEPCK was determined in the flesh of blueberries, raspberries, red currants, and strawberries at different stages of their development. In addition, changes in the amounts of citrate, malate, soluble sugars, isocitrate lyase, NADP-malic enzyme, phosphoenolpyruvate carboxylase, and pyruvate, orthophosphate dikinase in the flesh were determined. PEPCK was not detected in strawberry flesh, in which there was no dissimilation of malate or citrate. In the flesh of the other fruits, the abundance of PEPCK increased during ripening to an amount that was similar to that in grapes and tomatoes. In the flesh of blueberries and red currants, PEPCK was most abundant when there was dissimilation of malate. In the flesh of raspberries, PEPCK was most abundant when there was dissimilation of malate and citrate. These results are consistent with PEPCK playing a role in the dissimilation of citrate and/or malate in the flesh of these fruits during ripening. However, PEPCK was also present in the flesh of blueberries, raspberries, and red currants when there was no dissimilation of malate or citrate, and this raises the possibility that PEPCK might have additional functions. Dissection of blueberries provided evidence that both PEPCK and phosphoenolpyruvate carboxylase were present in the same cells, and possible functions for this are discussed.


Sex Differences in the TyG Index and Cardiovascular Risk Factors in Metabolically Obese Normal Weight Phenotype.
The triglyceride glucose (TyG) index is a novel surrogate marker of insulin resistance and increases cardiovascular disease risk. We sought to explore sex differences in the relationship between TyG and cardiovascular (CV) risk factors in metabolically obese normal weight (MONW) phenotype. We analyzed data of 1208 healthy men and nonpregnant women enrolled in a population-based longitudinal study from January 2017-June 2020. MONW phenotype was defined by normal body mass index (BMI: 18-<25 kg/m2) with at least one of the following metabolic disorders (MONW phenotype): elevated blood pressure (BP), hypertriglyceridemia, hyperglycemia, and low HDL cholesterol. Multiple logistic regression analysis was performed to assess the association between elevated TyG index and the CV risk factors in women and men and was presented in odds ratio (OR) with 95% confidence interval (CI). Of 1208 subjects, 350 (29%) were MONW phenotype (mean age (years): male: 43.5 ± 12.4 and female: 43.1 ± 12.7) and 858 were metabolically healthy normal weight (MHNW; n = 858 (71%)). MONW women had higher mean values of the TyG index (8.03 ± 5.07) than men (7.47 ± 4.68). Multivariate analysis revealed that the elevated TyG index is significantly associated with MONW phenotype in women (adjusted OR: 8.73, 95% CI: 5.62-13.57) and men (aOR: 5.90, 95% CI: 4.23-8.23). TyG was found to be an excellent predictor of MONW status in both women (receiver operating characteristic (ROC) area under the curve (AUC): 0.979, 95% CI: 0.969-0.988) and men (ROC-AUC: 0.968, 95% CI: 0.952-0.983). Our study revealed that the TyG index may represent a cost-effective and informative screening tool for the high-risk MONW phenotype.


Ocular neovascularization.
Retinal and choroidal vascular diseases constitute the most common causes of moderate and severe vision loss in developed countries. They can be divided into retinal vascular diseases, in which there is leakage and/or neovascularization (NV) from retinal vessels, and subretinal NV, in which new vessels grow into the normally avascular outer retina and subretinal space. The first category of diseases includes diabetic retinopathy, retinal vein occlusions, and retinopathy of prematurity, and the second category includes neovascular age-related macular degeneration (AMD), ocular histoplasmosis, pathologic myopia, and other related diseases. Retinal hypoxia is a key feature of the first category of diseases resulting in elevated levels of hypoxia-inducible factor-1 (HIF-1) which stimulates expression of vascular endothelial growth factor (VEGF), platelet-derived growth factor-B (PDGF-B), placental growth factor, stromal-derived growth factor-1 and their receptors, as well as other hypoxia-regulated gene products such as angiopoietin-2. Although hypoxia has not been demonstrated as part of the second category of diseases, HIF-1 is elevated and thus the same group of hypoxia-regulated gene products plays a role. Clinical trials have shown that VEGF antagonists provide major benefits for patients with subretinal NV due to AMD and even greater benefits are seen by combining antagonists of VEGF and PDGF-B. It is likely that addition of antagonists of other agents listed above will be tested in the future. Other appealing strategies are to directly target HIF-1 or to use gene transfer to express endogenous or engineered anti-angiogenic proteins. While substantial progress has been made, the future looks even brighter for patients with retinal and choroidal vascular diseases.


Reinforcing effects of nicotine and non-nicotine components of cigarette smoke.
Nicotine and non-nicotine components of cigarette smoke contribute to its reinforcing effects; however, the specific role of each component in maintaining behavior has not yet been elucidated. To assess the reinforcing effects of nicotine and non-nicotine components of cigarette smoke by presenting a concurrent choice paradigm in which participants had access to intravenous (IV) nicotine infusions vs. saline (placebo) infusions and puffs from denicotinized ("denic") cigarettes vs. air (sham puffs). We also measured the effects on self-administration of prior satiation with each component. Sixteen smokers participated in seven sessions: 1) a baseline smoking assessment, used to tailor the nicotine dose per infusion; 2) two sessions for training discrimination of IV nicotine vs. saline infusions and denic smoke vs. sham puffs; and 3) four sessions assessing choice behavior after different satiation conditions. Denic smoke was self-administered more than any other alternative, including IV nicotine. IV nicotine, however, was preferred over IV saline and sham puffs. Preference for denic smoke vs. IV nicotine was inversely correlated with subjective ratings of "comfort" associated with nicotine. Smoke satiation reduced the number of denic puffs taken during choice periods, while prior nicotine administration did not affect puffing behavior. Smoking withdrawal symptoms were alleviated both by nicotine administration and by denic smoke. In established smokers, non-nicotine aspects of cigarette smoking have potent reinforcing effects. While current smoking cessation pharmacotherapies primarily address the nicotine component of cigarette addiction, future cessation strategies should also be designed to target non-nicotine factors.


Deficiency of G protein-coupled receptor Gpr111/Adgrf2 causes enamel hypomineralization in mice by alteration of the expression of kallikrein-related peptidase 4 (Klk4) during pH cycling process.
Enamel is formed by the repetitive secretion of a tooth-specific extracellular matrix and its decomposition. Calcification of the enamel matrix via hydroxyapatite (HAP) maturation requires pH cycling to be tightly regulated through the neutralization of protons released during HAP synthesis. We found that Gpr115, which responds to changes in extracellular pH, plays an important role in enamel formation. Gpr115-deficient mice show partial enamel hypomineralization, suggesting that other pH-responsive molecules may be involved. In this study, we focused on the role of Gpr111/Adgrf2, a duplicate gene of Gpr115, in tooth development. Gpr111 was highly expressed in mature ameloblasts. Gpr111-KO mice showed enamel hypomineralization. Dysplasia of enamel rods and high carbon content seen in Gpr111-deficient mice suggested the presence of residual enamel matrices in enamel. Depletion of Gpr111 in dental epithelial cells induced the expression of ameloblast-specific protease, kallikrein-related peptidase 4 (Klk4), suggesting that Gpr111 may act as a suppressor of Klk4 expression. Moreover, reduction of extracellular pH to 6.8 suppressed the expression of Gpr111, while the converse increased Klk4 expression. Such induction of Klk4 was synergistically enhanced by Gpr111 knockdown, suggesting that proper enamel mineralization may be linked to the modulation of Klk4 expression by Gpr111. Furthermore, our in vitro suppression of Gpr111 and Gpr115 expression indicated that their suppressive effect on calcification was additive. These results suggest that both Gpr111 and Gpr115 respond to extracellular pH, contribute to the expression of proteolytic enzymes, and regulate the pH cycle, thereby playing important roles in enamel formation.


Neuromuscular blockers--a means of palliation?
As we die, our respiratory pattern is altered and we seem to gasp and struggle for each breath. Such gasping is commonly seen as a clear sign of dyspnoea and suffering by families and loved ones, however, it is unclear whether it is perceived at all by the dying person. Narcotics and sedatives do not seem to affect these gasping respirations. In this issue of the Journal of Medical Ethics, we are asked to consider whether the last gasp of a dying patient could be or, perhaps, even should be avoided by administering neuromuscular blockers to palliate dying patients. For many reasons, such as our current failure to alleviate pain and distress, stories of inadequate analgesia and sedation in critically ill paralysed patients and the inability to know the intent-whether to palliate or to euthanise-it would seem that administering neuromuscular blockers should not be ethically permissible.


Characteristics of Late-Onset Amyotrophic Lateral Sclerosis in a Chinese Cohort.
This retrospective study analyzed the clinical characteristics and prognosis of the elderly amyotrophic lateral sclerosis (ALS) population in a large sample. The study included 1,005 patients with sporadic ALS admitted to Chinese PLA General Hospital between March 2011 and March 2021. We stratified the ALS patients into young and old groups using 2 cutoffs for the age at disease onset (≥65 or ≥70 years old) and compared their demographic, clinical, and survival data. The mean onset age of all patients was 52.79 ± 10.55 years, with 123 (12.24%) having a disease onset ≥65 years and 44 (4.38%) having an onset ≥70 years. There were 624 (62.1%) male patients. More bulbar-onset cases were in the late-onset group (p = 0.001). The sex distribution, time from onset to diagnosis, and the time of symptom spread from spinal or bulbar localization to a generalized localization did not differ between groups. Late-onset patients progressed more rapidly and had a significantly shorter survival. Chinese ALS patients have an earlier age at onset and a relatively smaller proportion of old onset than European and Japanese patients. Elderly patients are more likely to have bulbar onset, which is related to rapid progression and a shorter survival.


Comparison of ventriculoperitoneal shunt to lumboperitoneal shunt in the treatment of idiopathic: A monocentric, assessor-blinded, randomized controlled trial.
Ventriculoperitoneal shunt (VPS) and lumboperitoneal shunt (LPS) remain the mainstay of idiopathic normal pressure hydrocephalus (INPH). There are no randomized controlled trials completed to compare the efficacy of these 2 shunt techniques. We will conduct a monocentric, assessor-blinded, and randomized controlled trial titled "Comparison of Ventriculoperitoneal Shunt to Lumboperitoneal Shunt for the treatment of Idiopathic Normal Pressure Hydrocephalus: Phase I (COVLINPH-1)" trial and recruit patients at West China Hospital of Sichuan University since June 2021. And this trial is expected to end in December 2030. Eligible participants will be randomly assigned into LPS group and VPS group at ratio of 1:1 followed by evaluation before surgery, 1 month, 12 months, and 5 years after surgery. The primary outcome is the rate of shunt failure within 5 years. The secondary outcomes include modified Rankin Scale (mRS), INPH grading scale (INPHGS), mini-mental state examination (MMSE), and Evans index. We will calculate the rate of favorable outcome, which is defined as shunt success and an improvement of more than 1 point in the mRS at evaluation point. We will also analyze the complications throughout the study within 5 years after shunt insertion. The results of this trial will provide state-of-the-art evidence on the treatment option for patients with INPH, and will also generate the discussion regarding this subject. ChiCTR2000031555; Pre-results.


In vitro lymphoproliferative response and cytokine production in mice with experimental disseminated candidiasis.
Systemic candidiasis is an infection of Candida albicans (C. albicans) causing disseminated disease and sepsis, invariably when host defenses are compromised. We investigated the histopathological changes as well as the lymphoproliferative responses and cytokine production of splenic cells after stimulation with Concanavalin A (Con A) and Pokeweed mitogen (PWM) in mice with disseminated candidiasis. Lymphoproliferative responses were stimulated in vitro with Con A (1 µg/ml) and PWM (1 µg/ml) mitogens in Roswell Park Memorial Institute (RPMI) 1640 media, and the production of interferon (IFN)-γ and interleukin-4 (IL-4) in the supernatants was measured by enzyme-linked immunosorbent assay (ELISA). The results revealed that C. albicans organisms multiplied to a greater extent in the kidneys than in the liver and spleen of infected mice. The most predominant forms of C. albicans in different parts of the kidneys were yeast mixed with hyphal forms. Infected mice had a significantly increased proliferative response when splenocytes were stimulated with PWM (2.0±0.16) and Con A (1.9±0.19) (P<0.05). PWM and Con A-stimulated production of IFN-γ significantly tended to be higher in infected mice (PWM: 68.4±14.0 pg/ml; Con A: 53.7±17.3 pg/ml) when compared to controls (P<0.05). Stimulation with PWM and Con A showed no differences in IL-4 production between infected mice and controls. These findings demonstrated a significant increase in both cell proliferation and IFN-γ secretion in supernatants of PWM and Con A- stimulated splenocyte cultures obtained from mice with disseminated candidiasis.


Isosteric replacement of A3 Val of porcine insulin by allo-Thr.
A3 Val is important for insulin activity. It is invariant in insulins from different species studied thus far. Based on the three dimensional structure of insulin, it was thought to be involved in receptor binding. Its replacement by Leu resulted in remarkable lowering of insulin activity, indicating the crucial requirement of the side chain geometry at this position. When A3 Val was replaced by Thr, which is hydrophilic but isosteric with Val, substantial insulin activity was retained. Therefore, the isosteric requirement for receptor binding at this site is more stringent than the hydrophobic requirement. Here we report the replacement of A3 Val of porcine insulin by the unnatural allo-Thr. The in vivo biological activity of A3 allo-Thr insulin is similar to that of A3 Thr insulin or native insulin, but its receptor binding activity is 7.6% instead of 50% for A3 Thr insulin, indicating that at the A3 position the hydrophilic OH group of Thr could be more tolerated in receptor binding than the OH group of allo-Thr. The retention of insulin activity by substituting A3 Val with the unnatural isosteric allo-Thr demonstrates again the importance of isosteric interaction in the binding of insulin with its receptor.


Immunoenhanced enteral nutrition, effect on inflammatory markers in head and neck cancer patients.
The aim of our study was to evaluate the effect of enteral nutrition supplemented with arginine in inflammatory markers in surgical head and neck cancer patients. A population of 29 patients with oral and laryngeal cancer were enrolled in a randomized trial. At surgery patients were randomly allocated to two groups: (a) patients receiving an enteral diet supplements with arginine (group I, n=14); (b) patients receiving an isocaloric, isonitrogenous enteral formula (group II, n=15). The mean age was 61.1+/-10.8 y (five females/24 males). Characteristics of the patients on enrollment were similar for the two groups. Prealbumin and transferrin improved in both groups. c-reactive protein (CRP) levels decreased in both groups, (group I: 134.5+/-62.5 vs 75.3+/-51 mg/dl:P<0.05) and (group II: 103.6+/-62 vs 43.8+/-34.4 mg/dl:P<0.05). Interleukin-6 (IL-6) improved in both groups (group I: 20.35+/-11.2 vs 6.7+/-3.1 pg/ml:P<0.05) and (group II:22.8+/-40 vs 9.9+/-17.7 pg/ml:ns). Tumoral necrosis factor alpha and lymphocytes did not change. In conclusion, both formulas improved IL-6 and CRP levels. Further studies are needed to determine whether type of formula is the key in these patients or genetic background play a main role in inflammatory response.


Do physicians discuss end-of-life decisions with family members? A mortality follow-back study.
Deaths from chronic illness are often preceded by a potentially life-shortening end-of-life decision (ELD). Involving family in these ELDs may have psychosocial benefits for them and the dying person. This study aims to examine how often ELDs are discussed with relatives of the dying person and which characteristics determine their involvement in those ELDs. A questionnaire survey was conducted in 2013 among physicians attending a large, stratified and representative sample of deaths (n = 6188) in Flanders. In 72.3% of ELDs preceding death, family of the dying person were involved. Discussion of an ELD with family members was more likely when the decision was also discussed with the dying person, the ELD was made with the explicit intention to shorten life, specialized palliative care was provided or death occurred in an ICU. Involving family in end-of-life decision making appears to be related to the type of formal care services involved, communication with the dying person and the motives behind the decision. Our findings suggest a need to further expand a palliative care approach with a focus on both the dying person and their family within and across a variety of health care services.


BmK CT and 125I-BmK CT suppress the invasion of glioma cells in vitro via matrix metalloproteinase-2.
Chlorotoxin (CTX) is an established blocker of small‑conductance Cl‑ channels and has previously been demonstrated to inhibit the invasion of glioma cells. Buthus martensii Karsch chlorotoxin‑like toxin (BmK CT) is the first chlorotoxin-like peptide. The present study aimed to determine the inhibitory effect of BmK CT on the invasive ability of glioma cells, using a Transwell assay. BmK CT was subsequently radiolabeled with radionuclide 125I and its activity was compared with BmK CT. Additionally, the underlying anti‑invasive mechanism of BmK CT and 125I‑BmK CT on glioma cells was investigated by ELISA and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). It was revealed that BmK CT and 125I‑BmK CT were able to inhibit the invasion of glioma cells and that 125I‑BmK CT was superior to BmK CT. Consistent with the results of the Transwell assay, matrix metalloproteinase‑2 (MMP‑2) secretion by glioma cells was significantly reduced following treatment with BmK CT or 125I‑BmK CT (P<0.05). However, no significant differences in MMP-2 mRNA expression levels were identified by RT‑qPCR (P>0.05). In conclusion, the present study demonstrated that BmK CT and 125I‑BmK CT reduced the invasion of glioma cells via downregulation of MMP-2 expression. However, inhibition of the invasion of glioma cells was not demonstrated at the mRNA level.


Enzymatic modifications of human fibroblast and leukocyte interferons.
The sensitivity of highly purified human fibroblast interferon and partially purified human leukocyte interferon to several proteolytic and glycolytic enzymes was determined with respect to antiviral activity, isoelectric point, molecular weight, and thermal stability. Leucine aminopeptidase altered the distribution of isoelectric points for both interferons but produced little change in molecular weights; this enzyme somewhat reduced the activity of only leukocyte interferon. Treatment of fibroblast interferon with carboxypeptidases A and B did not greatly decrease antiviral activity, but it did slightly reduce the molecular weight of the interferon and substantially altered the distribution of isoelectric point values; similar treatment of leukocyte interferon caused some loss in activity, especially of the 17,000-molecular-weight species. Both interferons were inactivated rapidly by treatment with the endoproteases trypsin, pepsin, bromelain, and subtilisin. Chymotrypsin shifted the isoelectric points of both interferons, but only leukocyte interferon was significantly inactivated. Treatment with neuraminidase and beta-galactosidase changed the isoelectric point distribution but did not affect the activity or thermal stability of either interferon; such a treatment reduced the molecular weight of fibroblast interferon and the size heterogeneity of leukocyte interferon. Treatment with neuraminidase and then leucine aminopeptidase greatly reduced the activity of both interferons, especially leukocyte interferon. The data indicate that biologically active forms of fibroblast and leukocyte interferons can be distinguished by their relative sensitivity to certain proteases.


Prevention of postoperative recurrence of Crohn's disease: what does the evidence support?
Patients who have undergone intestinal resection for Crohn's disease frequently develop endoscopic and clinical recurrence of their disease. Active smokers and those with perforating disease appear to be at a higher risk of this development. A number of agents have been shown in randomized controlled trials to reduce the risk of endoscopic and clinical recurrence, but international guidelines are currently ambiguous on which agent should be used. Because there are wide-ranging efficacy estimates, safety profiles, and costs for the various proven prophylactic agents, physicians should personalize the management of individual patients based on their estimated probability of clinical recurrence (low/high). Such risk stratification could be based on the patient's clinical profile with or without early postoperative endoscopic surveillance. Ongoing trials using this "tailored" strategy should provide answers on whether this approach allows optimal selection of proven therapies.


Erythrocytes with T-state-stabilized hemoglobin as a therapeutic tool for postischemic liver dysfunction.
This study aimed to examine if T-state stabilization of hemoglobin in erythrocytes could protect against postischemic organ injury. Human erythrocytes containing three different states of Hb allostery were prepared: control Hb (hRBC), CO-Hb that is stabilized under R-state with the 6-coordinated prosthetic heme (CO-hRBC), and alpha-NO-deoxyHb stabilized under T-state (alpha-NO-hRBC). To prepare alpha-NO-RBC, deoxygenated RBC was treated with FK409, a thiol-free NO donor, at its half molar concentration to that of Hb; this procedure resulted in the 5-coordinated NO binding on the alpha-subunit heme, as judged by electron spin resonance spectrometry. Rats were subject to 20 min systemic hemorrhage to maintain mean arterial pressure at 40 mm Hg, and reperfused with one of hRBCs. This protocol for ischemia, followed by 60 min reperfusion with physiological saline, caused modest metabolic acidosis and cholestasis. Administration of hRBC or COhRBC significantly attenuated cholestasis and improved acidosis. Rats treated with alpha-NO-hRBC exhibited greater recovery of metabolic acidosis and bile excretion than those treated with hRBC or CO-hRBC, displaying the best outcome of local oxygen utilization in hepatic lobules. Half-life time of alpha-NO-RBC administered in vivo was approximately 60 min. These results suggest that T-state Hb stabilization by NO serves as a stratagem to treat postischemic organ dysfunction.


Investigation of the GnRH antagonist degarelix isomerization in biological matrices.
One of the main objectives of peptide drug design is the improvement of peptide pharmacokinetics with maintaining biological activity, which can be achieved by the complex modifications of the primary structure of the peptides. However, these changes often lead to the formation of peculiar impurities in the peptide drugs and their metabolites, which require the development of advanced analytical methods to properly assess their content. Here, we investigated the degradation of the potent long-acting GnRH antagonist degarelix in various biologic media by the tailor-made HPLC method, which allows precise determination of 5-Aph(Hyd)-degarelix isomer, an impurity found in the degarelix active pharmaceutical ingredient (API) during its manufacturing. Unexpectedly, we discovered a rapid and irreversible conversion of degarelix API into the corresponding hydantoin isomer in serum, suggesting that this impurity can be also a potential drug metabolite in vivo. This finding underlines the importance of the development of more accurate and performing analytical techniques to correctly characterize the chemical composition of the manufactured drugs and their behavior under physiological conditions.


Measurement and regulation of the culture reduction state in Clostridium acetobutylicum.
With a constant glucose feed concentration, the change in the continuous culture dillution rate resulted in an altered fermentation profile and the cellular NADH content. The cultures growing at high dillution rates demonstrated an oxidative metabolism low NADH and butanol concentrations. The low specific NADH flourescence (F/X) at high butanol production rates suggested that a rapid regeneration of NADH to NAD is essential for a high solventogenic culture activity. The culture florescence and butanol concentration remained constant in the solventogenic dilution rate range of D = 0.05-0.2 h(-1) with an inverse relationship between the specific flourescence (F/X) and the specific butanol production rate, q(B). Flourometric NADH observations were confirmed by enzymatic NADH determination. The stiochiometric "Fermentation Equation" was used to check the experimental data consistency and to investigate the role of the available biosynthetic and reduction energy on the culture metabolic activities under different growth conditions. The butanol concentration in the broth was stabilized in a fed-batch process when the culture NADH fluorescence was being controlled through the addition of fresh medium.


Computer system for definition of the quantitative geometry of musculature from CT images.
The computer system for quantitative determination of musculoskeletal geometry from computer tomography (CT) images has been developed. The computer system processes series of CT images to obtain three-dimensional (3D) model of bony structures where the effective muscle fibres can be interactively defined. Presented computer system has flexible modular structure and is suitable also for educational purposes.


Tailoring relaxation dynamics and mechanical memory of crumpled materials by friction and ductility.
Crumpled sheets show slow mechanical relaxation and long lasting memory of previous mechanical states. By using uniaxial compression tests, the role of friction and ductility on the stress relaxation dynamics of crumpled systems is investigated. We find a material dependent relaxation constant that can be tuned by changing ductility and adhesive properties of the sheet. After a two-step compression protocol, nonmonotonic aging is reported for polymeric, elastomeric and metal sheets, with relaxation dynamics that are dependent on the material's properties. These findings can contribute to tailoring and programming of crumpled materials to get desirable mechanical properties.


p62 Deficiency Enhances α-Synuclein Pathology in Mice.
In Lewy body disease (LBD) such as dementia with LBs and Parkinson's disease, several lines of evidence show that disrupted proteolysis occurs. p62/SQSTM1 (p62) is highly involved with intracellular proteolysis and is a component of ubiquitin-positive inclusions in various neurodegenerative disorders. However, it is not clear whether p62 deficiency affects inclusion formation and abnormal protein accumulation. To answer this question, we used a mouse model of LBD that lacks p62, and found that LB-like inclusions were observed in transgenic mice that overexpressed α-synuclein (Tg mice) with or without the p62 protein. p62 deficiency enhanced α-synuclein pathology with regard to the number of inclusions and staining intensity compared with Tg mice that expressed p62. To further investigate the molecular mechanisms associated with the loss of p62 in Tg mice, we assessed the mRNA and protein levels of several molecules, and found that the neighbor of the brca1 gene (NBr1), which is functionally and structurally similar to p62, is increased in Tg mice without p62 compared with control Tg mice. These findings suggest that p62 and NBR1 affect the pathogenesis of neurodegenerative diseases through the cooperative modulation of α-synuclein aggregation.


Nerve conduction studies in infants and children.
The electrophysiologic evaluation of peripheral nerves may provide critically important information, both with respect to diagnosis and prognosis, in the child with a suspected neuromuscular disorder. However, special attention to various technical considerations is necessary to avoid misleading results. Utilizing these techniques, both hereditary and acquired neuropathies may be identified and characterized. The latter has become especially important in view of recent advances in the treatment of acquired demyelinating neuropathies.


Prediction of gestational age based on foetal ultrasonographic biometric measurements in light breed horses.
A table was generated, based on foetal ultrasonographic measurements in light breed mares, for each day of gestation beginning with day 100, to provide the predicted value of four biometric parameters: biparietal diameter (BPD), eye approximated volume (EyV), foetal aortic diameter (AortD) and femur length (FL). Using this table, day of gestation was successfully predicted in 23 Quarter Horses (QH) with known mating or ovulation dates. BPD, EyV and FL were the best foetal age predictors between 100- and 200-days gestation predicting within 2 weeks of the actual day of gestation, while BPD and EyV were best between 200 and 300 days (within 3 weeks), and EyV was best after 300 days (within 3 weeks).


Piecemeal Mechanism Combining Sprouting and Intussusceptive Angiogenesis in Intravenous Papillary Formation Induced by PGE2 and Glycerol.
Recently, we demonstrated that in human intravascular papillary endothelial hyperplasia (IPEH), vein wall vascularization occurs in association with myriad papillae, a large part of which formed in the vascularized vein wall. Previously, using an animal model, we observed that PGE2 and glycerol administration around the femoral vein originates intense vascularization of the vein wall from its intimal endothelial cells (ECs). This vascularization is similar to that in IPEH. The aim of this study is to assess the mechanism of papillary formation, using this model after demonstrating papillary development in neo-vascularized femoral vein walls. In semithin and ultrathin sections, the sequential vascular and papillary development was as follows: (a) activation of vein intimal ECs, (b) sprouting of intimal ECs towards the vein media layer and microvessel development, (c) interconnection between neighboring microvessels originated elementary loops, which encircled vein wall components and formed papillae. The encircling ECs formed the papillary cover, and the encircled component formed the core. The papillae showed a similar structure to that of folds and pillars in intussusceptive angiogenesis, and (d) origin of secondary and complex loop systems by interconnection of neighboring elementary loops and by splitting of papillae by new loops, with abundant papillary development. In conclusion, the results support a piecemeal angiogenic mechanism in papillary formation, with association of sprouting and intussusceptive types of angiogenesis. Further studies are needed to assess whether the intravascular papillae described in several pathologic processes, including vessel tumors, such as Dabska's tumor, retiform hemangioendothelioma, and angiosarcoma, follow a similar mechanism. Anat Rec, 2017. © 2017 Wiley Periodicals, Inc. Anat Rec, 300:1781-1792, 2017. © 2017 Wiley Periodicals, Inc.


A comparison of partner notification effectiveness in African-, Caribbean-, and United States-born HIV-infected Blacks in New York City.
HIV disproportionately affects blacks more than other groups in New York City (NYC) as well as nationally. Partner Services (PS) are a proven way to interrupt HIV transmission. In NYC, PS outcomes are worse among blacks compared to other race/ethnic groups. We compared PS outcomes by country of birth to identify opportunities for improved PS effectiveness. We assessed number of sex and needle-sharing partners elicited, notified, and HIV tested, as well as number of newly identified HIV cases and number of interviews needed to identify a new HIV case. Between July 2006 and December 2008, 1049 African Americans, 285 Caribbean-born blacks, and 168 African-born blacks were interviewed for HIV Partner Services. African Americans, Caribbean-born blacks, and African-born blacks had average age of 38, 37, and 43 years and 10.0%, 0.4%, and 0.6% reported intravenous drug use, respectively. All groups had a high proportion of cases with an AIDS-defining CD4 count at HIV diagnosis (30.8%, 34.5%, and 47.9% for African Americans, Africans, and Caribbeans, respectively). The number needed to interview to find one newly diagnosed HIV case was high for all groups (66, 57, and 56 among African Americans, Africans, and Caribbeans, respectively), indicating all had similarly low rates of new HIV case identification. NYC blacks and their partners are benefiting from current Partner Services outreach as PS did identify new HIV cases among partners from all 3 groups. However, further work is needed to improve HIV case finding from Partner Services in these groups, and additional measures to promote early diagnosis are needed.


Nursing our most valuable natural resource: school age children.
Delivering nursing services in the school setting is both a challenge and an opportunity. Challenges include meeting the multiple health needs of students within communities experiencing dwindling healthcare resources, and working within an academic rather than a healthcare system. The author reviews the variety of service models--direct care of illnesses and emergencies, consultation and supervision of school personnel for delegated tasks, and health promotion and collaboration with other community service providers--and illustrates the complexity of the school nurse's role with a case study.


The Medicinal Rod: Slave Health and Redhibition Law in George Washington Cable's The Grandissimes.
The economic transactions and litigation necessary for slavery to function, coupled with the South's honor culture, meant skepticism and posturing frequently attended the buying and selling of enslaved people. This atmosphere provided opportunities for enslaved individuals familiar with the symbiotic ways their health and value intertwined to manipulate owners by feigning illness or adopting behaviors contrary to those of a "sound and sane" captive under Louisiana's redhibitory (slave warranty) law. Such actions offered a chance at preserving that which slavery denied its victims: proximity to family, a reduced chance of being sold, and an opportunity to exert agency within a strictly oppressive system. In dramatizing these paradoxes, George Washington Cable's The Grandissimes illustrates the vile hollowness of owners' paternalistic attitudes towards the enslaved, acknowledges the subjectivity and will of enslaved individuals, and castigates the return of slavery-like conditions in the form of the convict lease system.


In situ measurements of chemical distributions in a deep-sea hydrothermal vent field.
Large changes in the concentration of sulfide around a hydrothermal vent in the Galápagos Rift provide direct evidence for the consumption of sulfide by the organisms of the vent community. These changes were detected with a new chemical analyzer capable of measuring silicate, sulfide, oxygen, and temperature on the sea floor at depths of 2500 meters. More than 10,000 measurements showed systematic variations in the sulfide and oxygen concentrations due to biogenic oxidation of sulfide in the hydrothermal solutions. Silicate concentration was highly correlated with temperature, but different trends were observed at different locations.


Experimental investigation of mass analysis using an island of stability with a quadrupole with 2.0% added octopole field.
We describe experimental investigations of mass analysis using an island of stability with a linear quadrupole with a 2.0% added octopole field. The island is formed by auxiliary quadrupole excitation. The experiments confirm the results of previous computer simulations (Konenkov et al., J. Am. Soc. Mass Spectrom. 2007; 18: 826-834). With the resolving direct current (dc) applied to the quadrupole so that the Mathieu parameter a > 0, conventional mass analysis with applied radio-frequency (rf) and dc and no auxiliary excitation is possible. In this case, use of an island of stability yields similar peak shape and resolution. However, with the polarity of the resolving dc reversed, so that a < 0, only very low resolution can be obtained; the added octopole prevents conventional mass analysis. By using a stability island when a < 0, the resolution is substantially improved. In this case the use of an island allows mass analysis under conditions where the added octopole field severely degrades conventional analysis. The experiments also confirm the position of the island predicted by the simulations.


[On the assessment of spontaneous respiratory changes in respiration-olfactometry].
Alterations of breathing pattern evoked by odors are used to proof the integrity of the olfactory system in the sense of respiratory olfactometry. Spontaneous breathing changes normally cannot be distinguished from evoked changes. It is therefore necessary to repeatedly apply stimuli with a pure odorant during tidal breathing, randomized with neutral air, to detect a majority of olfactory-evoked respiratory changes. In 26 adult normosmics, 15 H2S and 15 neutral air stimuli (duration: 2 sec) were randomly applied with an interstimulus interval of at least 1 minute, using a flow olfactometer. Before stimulation, the respiration was not allowed to scatter significantly (variation coefficient of the duration of inspiration (DIN) and expiration (DEX) ≤0.1). A respiratory response was fulfilled when respectively DIN and DEX of the stimulus breath exceeded the double standard deviation of the same parameters in the five preceding respiratory cycles. The number of reactions was normalized to the number of stimuli and to the off-line as regularly calculated respiratory complexes and thus reaction indices were formed. H2S stimuli clearly evoked more respiratory changes than neutral air stimuli. DIN and DEX decreased with olfactory stimulation more frequently than with an application of neutral air. Extensions of DIN and DEX were not different between the two stimulus qualities. Only the randomized nasal stimulation with an odor and a blank makes the respiration-olfactometry meaningful.


Polymorphism, duplication, and IS1-mediated rearrangement in the chromosomal his-rfb-gnd region of Escherichia coli strains with group IA and capsular K antigens.
Individual Escherichia coli strains produce several cell surface polysaccharides. In E. coli E69, the his region of the chromosome contains the rfb (serotype O9 lipopolysaccharide O-antigen biosynthesis) and cps (serotype K30 group IA capsular polysaccharide biosynthesis) loci. Polymorphisms in this region of the Escherichia coli chromosome reflect extensive antigenic diversity in the species. Previously, we reported a duplication of the manC-manB genes, encoding enzymes involved in GDP-mannose formation, upstream of rfb in strain E69 (P. Jayaratne et al., J. Bacteriol. 176:3126-3139, 1994). Here we show that one of the manC-manB copies is flanked by IS1 elements, providing a potential mechanism for the gene duplication. Adjacent to manB1 on the IS1-flanked segment is a further open reading frame (ugd), encoding uridine-5'-diphosphoglucose dehydrogenase. The Ugd enzyme is responsible for the production of UDP-glucuronic acid, a precursor required for K30 antigen synthesis. Construction of a chromosomal ugd::Gm(r) insertion mutation demonstrated the essential role for Ugd in the biosynthesis of the K30 antigen and confirmed that there is no additional functional ugd copy in strain E69. PCR amplification and Southern hybridization were used to examine the distribution of IS1 elements and ugd genes in the vicinity of rfb in other E. coli strains, producing different group IA K antigens. The relative order of genes and, where present, IS1 elements was established in these strains. The regions adjacent to rfb in these strains are highly variable in both size and gene order, but in all cases where a ugd homolog was present, it was found near rfb. The presence of IS1 elements in the rfb regions of several of these strains provides a potential mechanism for recombination and deletion events which could contribute to the antigenic diversity seen in surface polysaccharides.


A compendium of cytogenetic abnormalities in myelofibrosis: molecular and phenotypic correlates in 826 patients.
Among 826 patients with primary myelofibrosis (PMF) and analysable metaphases on cytogenetic studies, 352 (42·6%) had abnormal karyotype, of which 240 (68·2%) were sole aberrations and 48 (13·6%) were complex; the most frequent abnormalities were 20q- (23·3%), 13q- (18·2%), +8 (11·1%), +9 (9·9%), chromosome 1q+ (9·7%) and -7/7q- (7·1%). Phenotypic correlates included: abnormal karyotype with anaemia (P = 0·02), leucopenia (P < 0·01) and thrombocytopenia (P < 0·01); complex karyotype with younger age (P = 0·04) and thrombocytopenia (P < 0·01); leucopenia with 20q-, +8 and -7/7q- and thrombocytopenia with 20q- and -7/7q-. Cytopenias were less likely to occur with 13q-. 476 patients were annotated for JAK2/CALR/MPL mutations; abnormal karyotype frequencies were 43% in JAK2, 42% CALR, 33% MPL mutated and 34% triple-negative cases (P = 0·3). A proportion of patients were also screened for ASXL1, EZH2, IDH1, IDH2, SRSF2, U2AF1 and SF3B1 mutations; in all instances, mutational frequencies were higher in patients with normal karyotype, reaching significance for ASXL1 (P = 0·02) and U2AF1 (P = 0·01). 13q- was associated with mutant CALR (P = 0·03), +9 with mutant JAK2 (P = 0·02) and 20q- with mutant SRSF2 (P = 0·02). The current PMF study provides detailed cytogenetic information and correlations with mutations and clinical phenotype.


Early Systemic Anti-neoplastic Treatment Post SARS-CoV-2 Infection in Patients with Breast Cancer.
It is unclear how soon after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection it is safe to resume systemic anti-neoplastic treatment in patients with cancer. We assessed the risk of admissions or postponed treatment cycle in vaccinated patients with breast cancer receiving early systemic anti-neoplastic treatment following SARS-CoV-2 infection. This was a retrospective cohort study conducted during Omicron SARS-CoV-2 outbreak in Israel, January-July 2022. SARS-CoV-2 cohort included 30 vaccinated patients with breast cancer with SARS-CoV-2 infection 7-14 days prior to systemic treatment. All patients had resolved symptoms and a negative antigen detection test on the day of treatment. The pre-coronavirus disease 2019 (COVID-19) pandemic cohort consisted of 49 matched patients with breast cancer treated with systemic anti-neoplastic agents during 2019. In 30 vaccinated patients with breast cancer who received systemic anti-neoplastic treatment 7-14 days following SARS-CoV-2 infection, compared with 49 matched patients treated in 2019, the rates of emergency department (ED) visits (13% versus 6%, respectively), hospitalizations (3% versus 4%), next cycle of treatment given per protocol (90% versus 88%), and death (0% versus 0%) were similar. In a cohort of vaccinated patients with breast cancer who received systemic anti-neoplastic treatment 7-14 days after SARS-CoV-2 infection, we did not observe substantially higher rates of ED visits, hospitalizations, or deaths compared with a similar cohort of pre-COVID-19 patients with breast cancer. Most patients received the next planned cycle on time. Early resumption of systemic anti-neoplastic treatment following SARS-CoV-2 infection in vaccinated patients with breast cancer with a negative antigen test at the day of treatment appeared to be safe. Additional data on larger cohorts and other malignancies are needed to support clinical guidelines.


Efficient antibody production in the methylotrophic yeast Ogataea minuta by overexpression of chaperones.
A production system for a therapeutic monoclonal antibody was developed using the methylotrophic yeast Ogataea minuta IFO10746. The genetically engineered O. minuta secreted a detectable amount of anti-TRAIL receptor antibody into the culture supernatant, and the secreted antibody was purified by multiple column chromatography steps. In the purification process, both fully and partially assembled antibodies were detected and isolated. The fully assembled antibody from O. minuta showed almost the same biological activity as that derived from mammalian cells despite the distinct glycosylation profile, whereas the partially assembled antibody showed no cytotoxic activity. To increase the production of active antibody in O. minuta, we overexpressed selected chaperone proteins (included protein disulfide isomerase (OmPDI1), thiol oxidase (OmERO1), and immunoglobulin heavy chain binding protein (OmKAR2)) known to assist in the proper folding (in the endoplasmic reticulum) of proteins destined for secretion. Each of these chaperones enhanced antibody secretion, and together these three factors yielded 16-fold higher antibody accumulation while increasing the ratio of the fully assembled antibody compared to that from the parental strain. Supplementation of a rhodanine-3-acetic acid derivative (R3AD_1c), an inhibitor of O-mannosylation, further increased the secretion of the correctly assembled antibody. These results indicated that the co-overexpression of chaperones is an effective way to produce the correctly assembled antibody in O. minuta.


Therapeutic effects of KRM-II-81, positive allosteric modulator for α2/3 subunit containing GABAA receptors, in a mouse model of Dravet syndrome.
Introduction: Dravet syndrome (DS) is an intractable epilepsy syndrome concomitant with neurodevelopmental disorder that begins in infancy. DS is dominantly caused by mutations in the SCN1A gene, which encodes the α subunit of a voltage-gated Na channel. Pre-synaptic inhibitory dysfunction is regarded as the pathophysiological mechanism, but an effective strategy for ameliorating seizures and behavioral problems is still under development. Here, we evaluated the effects of KRM-II-81, a newly developed positive allosteric modulator for α 2/3 subunit containing GABAA receptors (α2/3-GABAAR) in a mice model of DS both in vivo and at the neuronal level. Methods: We used knock-in mice carrying a heterozygous, clinically relevant SCN1A mutation (background strain: C57BL/6 J) as a model of the DS (Scn1a WT/A1783V mice), knock-in mouse strain carrying a heterozygous, clinically relevant SCN1A mutation (A1783V). Seizure threshold and locomotor activity was evaluated by using the hyperthermia-induced seizure paradigm and open filed test, respectively. Anxiety-like behavior was assessed by avoidance of the center region in locomotor activity. We estimated a sedative effect by the total distance traveled in locomotor activity and grip strength. Inhibitory post synaptic currents (IPSCs) were recorded from a hippocampal CA1 pyramidal neuron in an acutely prepared brain slice. Results: KRM-II-81 significantly increased the seizure threshold of Scn1a WT/A1783V mice in a dose-dependent manner. A low dose of KRM-II-81 specifically improved anxiety-like behavior of Scn1a WT/A1783V mice. A sedative effect was induced by relatively high dose of KRM-II-81 in Scn1a WT/A1783V mice, the dose of which was not sedative for WT mice. KRM-II-81 potentiated IPSCs by increasing its decay time kinetics. This effect was more prominent in Scn1a WT/A1783V mice. Discussion: Higher activation of α2/3-GABAAR by KRM-II-81 suggests a compensatory modification of post synaptic inhibitory function against presynaptic inhibitory dysfunction in Scn1a WT/A1783V. The increased sensitivity for KRM-II-81 may be relevant to the distinct dose-dependent effect in each paradigm of Scn1a WT/A1783V mice. Conclusion: Selective activation for α2/3-GABAAR by KRM-II-81 could be potential therapeutic strategy for treating seizures and behavioral problems in DS.


Nanofibrous Scaffolds Containing Hydroxyapatite and Microfluidic-Prepared Polyamidoamin/BMP-2 Plasmid Dendriplexes for Bone Tissue Engineering Applications.
The aim of this study is to fabricate functional scaffolds to gene delivery bone morphogenetic protein-2 (BMP-2) plasmid for bone formation in bone tissue engineering. Dendriplexes (DPs) of generation 4 polyamidoamin (G4-PAMAM)/BMP-2 plasmid were prepared through microfluidic (MF) platform. The physiochemical properties and toxicity of DPs were evaluated by DLS, AFM, FESEM and MTT assay. In order to create a suitable environment for stem cell growth and differentiation, poly-l-lactic acid (PLLA) and poly-l-lactic acid/poly (ethylene oxide) (PLLA/PEO) scaffolds containing hydroxyapatite nanoparticles (HA) and DPs were fabricated by the electrospinning method. The osteogenic potency of the scaffolds on human adipose tissue-derived mesenchymal stem cells (hASCs) was investigated. The results revealed that tuning the physical properties of DPs by adjusting flow parameters in microfluidic platform can easily improve the cell viability compared to conventional bulk mixing method. Also, the result showed that the presence of HA and DPs in PLLA/PEO scaffold enhanced alkaline phosphatase (ALP) activity and increased the amount of deposited Ca, as well as, related to osteogenesis gen markers. This study indicated that on using the MF platform in preparation of DPs and loading them along with HA in PLLA/PEO scaffold, the osteogenic differentiation of hASCs could be tuned.


Incorporating principles and practical wisdom in research ethics education: a preliminary study.
Researchers are faced with daily ethical decisions that are subtle and nuanced. However, research ethics training has primarily focused on formal guidelines, general ethical principles, and historically noteworthy cases of research abuse, which may not prepare researchers to respond to everyday dilemmas in research. This study characterized researchers' responses to ethical dilemmas with the goal of aligning research ethics education programs with the demands of practice. As a preliminary study, the authors conducted 23 semistructured interviews with senior researchers and research administrators engaged in research with human subjects at the University of Washington and affiliated institutions in 2004. Transcripts were reviewed for research conflicts and strategies used to resolve conflicts identified by participants. Participants referenced two distinct methods of reasoning when faced with conflicts: formal guidelines and practical wisdom. Formal guidelines include established goals, boundaries, and absolutes. Practical wisdom, an Aristotelian concept involving intuitions developed through experience, facilitates responses to everyday dilemmas and new situations. Developing practical wisdom requires researchers to calibrate their own intuitions about right and wrong. Three practices were reported to contribute to this development: self-reflection, sincere skepticism, and open dialogue with colleagues. These reflections from the senior researchers suggest a need to expand the scope of ethics education programs to include a focus on the development of researchers' pragmatic decision making in addition to the formal rules that govern research. Further research should explore effective educational and institutional strategies that can foster researchers' development in ethical decision making and conduct.


Synchronization of pathogenic protozoans.
Protozoans are single-cell eukaryotes and many of the best studied protozoans are parasitic to humans (e.g., Plasmodium falciparum causing malaria and Trypanosoma brucei causing sleeping sickness). These organisms are distantly related to humans but with retained eukaryotic type of cellular processes, making them good model systems for studies of the evolution of basic processes like the cell cycle. Giardia intestinalis causes 250 million cases of diarrhea yearly and is one of the earliest diverging protozoans. It has recently been possible to synchronize its cell cycle using compounds that inhibit different steps of the cell cycle and the detailed protocol is described here.


Discovery of a Series of Indane-Containing NBTIs with Activity against Multidrug-Resistant Gram-Negative Pathogens.
The rise of multidrug-resistant (MDR) Gram-negative bacteria is a major global health problem necessitating the discovery of new classes of antibiotics. Novel bacterial topoisomerase inhibitors (NBTIs) target the clinically validated bacterial type II topoisomerases with a distinct binding site and mechanism of action to fluoroquinolone antibiotics, thus avoiding cross-resistance to this drug class. Here we report the discovery of a series of NBTIs incorporating a novel indane DNA binding moiety. X-ray cocrystal structures of compounds 2 and 17a bound to Staphylococcus aureus DNA gyrase-DNA were determined, revealing specific interactions with the enzyme binding pocket at the GyrA dimer interface and a long-range electrostatic interaction between the basic amine in the linker and the carboxylate of Asp83. Exploration of the structure-activity relationship within the series led to the identification of lead compound 18c, which showed potent broad-spectrum activity against a panel of MDR Gram-negative bacteria.


Effect of vaccines against pancreas disease in farmed Atlantic salmon.
Pancreas disease (PD) caused by salmonid alphavirus (SAV) continues to negatively impact salmon farming. To assess the effect on growth and mortality of three vaccines against PD, two controlled field designs were employed: one controlled field study with individual marked fish (PIT tag) assessing three PD vaccines and three controls groups, and a second controlled field study with group marked fish (Maxilla) comparing two PD vaccines against controls. In addition, a descriptive study using whole cages compared fish immunized with two different PD vaccines against controls. The target populations experienced a natural PD outbreak where both SAV 2 and SAV 3 were identified. Only one of the PD vaccines provided statistically significant improvements in harvest weight of 0.43 kg (CI: 0.29-0.57) and 0.51 kg (CI: 0.36-0.65) compared with the control in the PIT tag and the Maxilla study, respectively. In the latter, a significant reduction in mortality of 1.31 (CI:0.8-1.8) per cent points was registered for the same vaccine compared with controls. These results aligned with the growth and PD-specific mortality registered in the descriptive Cage study. The data in this study show a difference in the efficacy of PD vaccines in farmed Atlantic salmon.


Comparison between pulsatile GnRH therapy and gonadotropins for ovulation induction in women with both functional hypothalamic amenorrhea and polycystic ovarian morphology.
Ovulation induction in patients having both functional hypothalamic amenorrhea (FHA) and polycystic ovarian morphology (PCOM) has been less studied in the literature. As results remain contradictory, no recommendations have yet been established. To compare pulsatile GnRH therapy versus gonadotropins for ovulation induction in "FHA-PCOM" patients and to determine if one treatment strikes as superior to the other. A 12-year retrospective study, comparing 55 "FHA-PCOM" patients, treated either with GnRH therapy (38 patients, 93 cycles) or with gonadotropins (17 patients, 53 cycles). Both groups were similar, defined by low serum LH and E2 levels, low BMI, excessive follicle number per ovary and/or high serum AMH level. Ovulation rates were significantly lower with gonadotropins (56.6% versus 78.6%, p = 0.005), with more cancellation and ovarian hyper-responses (14% versus 34% per initiated cycle, p < 0.005). Pregnancy rates were significantly higher with GnRH therapy, whether per initiated cycle (26.9% versus 7.6%, p = 0.005) or per patient (65.8% versus 23.5%, p = 0.007). In our study, GnRH therapy was more successful and safer than gonadotropins, for ovulation induction in "FHA-PCOM" patients. If results were confirmed by prospective studies, it could become a first-line treatment for this population, just as it is for FHA women without PCOM.


Development of Fluorescent FRET Probes for "Off-On" Detection of L-Cysteine Based on Gold Nanoparticles and Porous Silicon Nanoparticles in Ethanol Solution.
A new type of fluorescence "off-on" probe was designed for L-Cysteine (L-Cys) based on the fluorescence resonance energy transfer (FRET) between negatively charged amino-capped porous silicon nanoparticles (SiNPs) and positively charged citrate-stabilized Au nanoparticles (AuNPs). In this proposed FRET immunosensor, novel water-soluble amino-conjugated porous SiNPs in ethanol with excellent photoluminescence properties act as the energy donor. Excellent quenching efficiency between SiNPs-ethanol and citrate-stabilized AuNPs by electrostatic interaction via FRET provides an ideal "off-state" (turn-off). The addition of L-Cys leads to releasing the adsorbed AuNPs from the surface of SiNPs and hence the fluorescence emission of SiNPs-ethanol is restored (turn-on), which means the coordination ability of the thiols with AuNPs is stronger than that of the electrostatic interaction. The fluorescence intensity of SiNPs-AuNPs in ethanol is sensitive to L-Cys, and such a restored fluorescence is proportional to the concentration of L-Cys. The method will broadly benefit the development of a new thiol biosensor based on nanostructured porous materials, and the proposed procedure is also expected to develop a variety of functional nanoparticles to form other novel kinds of nanosensors.


Living Bioelectrochemical Composites.
Composites, in which two or more material elements are combined to provide properties unattainable by single components, have a historical record dating to ancient times. Few include a living microbial community as a key design element. A logical basis for enabling bioelectronic composites stems from the phenomenon that certain microorganisms transfer electrons to external surfaces, such as an electrode. A bioelectronic composite that allows cells to be addressed beyond the confines of an electrode surface can impact bioelectrochemical technologies, including microbial fuel cells for power production and bioelectrosynthesis platforms where microbes produce desired chemicals. It is shown that the conjugated polyelectrolyte CPE-K functions as a conductive matrix to electronically connect a three-dimensional network of Shewanella oneidensis MR-1 to a gold electrode, thereby increasing biocurrent ≈150-fold over control biofilms. These biocomposites spontaneously assemble from solution into an intricate arrangement of cells within a conductive polymer matrix. While increased biocurrent is due to more cells in communication with the electrode, the current extracted per cell is also enhanced, indicating efficient long-range electron transport. Further, the biocomposites show almost an order-of-magnitude lower charge transfer resistance than CPE-K alone, supporting the idea that the electroactive bacteria and the conjugated polyelectrolyte work synergistically toward an effective bioelectronic composite.


Clinical indications for the investigation of porphyria: case examples and evolving laboratory approaches to its diagnosis in New Zealand.
Patients with porphyria present in a diverse and unusual variety of ways and most clinicians will see only a few cases, if any, during their professional lives. Porphyria may present (1) with acute symptoms, which may be abdominal pain, neurological or psychiatric; (2) with skin rash or photosensitivity; or (3) with a putative family history. Screening for latent porphyria has been greatly facilitated by fluorescence emission scanning of plasma and by mutational analysis. Our reference laboratory has recently diagnosed several cases of the less common types of porphyria, which we postulate is due to the availability of these methods and to the changing population of New Zealand. Accurate screening and diagnosis of porphyria is important, as an acute porphyric attack is life-threatening and preventable. Retrospective diagnosis may be difficult.


Comparative evaluation of iodoacids removal by UV/persulfate and UV/H2O2 processes.
To develop a cost-effective method for post-formation mitigation of iodinated disinfection by-products, degradation of iodoacids by UV, UV/PS (persulfate), and UV/H2O2 was extensively investigated in this study. UV direct photolysis of 4 iodoacids followed first-order kinetics with rate constants in the range of 2.43 × 10(-4)-3.02 × 10(-3) cm(2) kJ(-1). The derived quantum yields (Ф254) of the 4 iodoacids range from 0.13 to 0.34, respectively. A quantitative structure-activity relationship (QSAR) model was subsequently established and applied to predict the direct photolysis rates of 6 other structurally similar iodoacids whose standards are commercially unavailable. At a UV dose of 140 mJ cm(-2) which is typically applied for disinfection of drinking water, the removal percentages of 4 iodoacids were only between 3.35% and 34.7%. Thus, ICH2CO2H (IAA), the most photo-recalcitrant species, was selected as the target compound for removal in the UV/PS and UV/H2O2 processes. The IAA degradation rates decreased with increasing pH from 3 to 11 in both processes. Humic acid (HA) and HCO3(-) had inhibitory effects on IAA degradation in both processes. Cl(-) adversely affected the IAA degradation in the UV/PS process but had no effect in the UV/H2O2 process. Generally, in the deionized (DI) water, surface water, treated drinking water, and secondary effluent, UV/PS process is more effective than UV/H2O2 process for IAA removal, based on the same molar ratio of oxidant: IAA. SO4(-) generated in the UV/PS process yields a greater mineralization of IAA than HO in the UV/H2O2 process. IO3(-) was the predominant end-product in the UV/PS process, while I(-) was the major end-product in the UV/H2O2 process. The respective contributions of UV, HO, and SO4(-) for IAA removal in the UV/PS process were 7.8%, 14.7%, and 77.5%, respectively, at a specific condition (1.5 μM IAA, 60 μM oxidant, and pH 7). Compared to UV/H2O2 process, UV/PS was also observed as more cost-effective process based on the electrical energy per order (EE/O) and chemical cost.


Nano-porous architecture of N-doped carbon nanorods grown on graphene to enable synergetic effects of supercapacitance.
A novel nano-porous 3D architecture of N-doped carbon nanorods arrays grown on the surface of graphene has been prepared by carbonizing polyaniline/graphene oxide (PANI-GO) composite with PANI nanorod arrays on both sides of GO nanosheets. The obtained carbon materials are entirely composed of regularly grown carbon nanorods on graphene with height of about 100 nm and width about 30 nm, showing porous property due to the decomposition of PANI chains. The morphology of PANI grown on GO at the different growth stages was investigated to demonstrate the mechanism of the finally hierarchical architecture formation. Due to its large specific surface area and incorporation of the nitrogen groups into the carbon matrix, the obtained 3D carbon material enhances the ionic transport and the super-capacitance by synergetic effect of both double-layer and faradaic capacitances. This study provides a controllable approach to fabricate hierarchical carbon material based on conducting polymers and graphene oxide with promising applications in the high-rate electrode material of supercapacitors.


Purification and Properties of Acid Stable Xylanases from Aspergillus kawachii.
Five extracellular endo-xylanases were recognized in the culture broth of shochu koji mold (Aspergillus kawachii, IFO 4308), and three major xylanases (XylA, XylB, and XylC) were purified and characterized. The molecular masses of XylA, XylB, and XylC were 35,000, 26,000, and 29,000, and isoelectric points were pH 6.7, 4.4, and 3.5, respectively. Amino acid compositions and other properties were studied and these three xylanases were found to be greatly different in their properties. These three xylanases, XylA, XylB, and XylC, were stable between pH 3-10, 3-10, and 1-9 and the optimum pHs were 5.5, 4.5, and 2.0, respectively. Consequently, these xylanases were acid stable xylanases, especially XylC was an acidophilic xylanase (acid xylanase). These xylanases produced various xylooligosaccharides including xylose from xylan and the main product was xylobiose in all xylanases. The production of acid xylanase (XylC) was enhanced with a low initial pH of the medium.


Regional differences in oxalate absorption by rat intestine: evidence for excessive absorption by the colon in steatorrhoea.
Clinical studies suggest that steatorrhoea can be associated with excessive absorption of dietary oxalate. We examined the influence of bile salts, Ca++, and long-chain fatty acid on the absorption of oxalate and water by rat intestine in vivo. Absorption was measured under steady-state conditions during single-pass infusions. Each intestinal segment served as its own control. In jejunum, 10 mM taurocholate, the principal salt in rat bile, depressed absorption of oxalate and water. Absorption was not depressed further by Ca++ or linoleic acid. In ileum, 10 mM taurocholate did not inhibit absorption. Linoleic acid, 2 mM, depressed absorption of both oxalate and water. In colon 10 mM taurocholate decreased absorption. Net water transport was depressed further when linoleic acid was added to the infusion, but oxalate absorption was enhanced. Ca++ negated these effects of linoleic acid. It is concluded that long-chain fatty acids may enhance the absorption of oxalate from the rat colon. This observation may be relevant to understanding hyperoxaluria in patients with steatorrhoea.


[Dr Alexander M. Kellas and the first Mount Everest expedition].
In 1921 the government of Tibet gave permission for a British party to attempt Mount Everest from the northern Tibetan side. Little was known about the physiological and medical problems associated with ascents to extreme altitudes. The person who knew most about these topics was Dr. Alexander Kellas, lecturer in medical chemistry at the Middlesex Hospital Medical School. He had made a number of expeditions to the Sikkim Himal and the Tibetan border before the first world war, and had become increasingly interested in the problems caused by altitude. He was invited to join the Everest expedition but died on the approach march at Kampa Dzong on the Tibetan plateau, within sight of the mountain. Before he went on the expedition Kellas wrote an article entitled A consideration of the possibility of ascending Mt. Everest. This paper was never published, but the manuscript exists in the archives of the Royal Geographical Society and the Alpine Club in London. As Kellas saw it, the main issue was whether sufficient adaptation could occur to allow a climber to ascend from a camp at about 7,700 m to the summit (8,848 m) in one day without supplementary oxygen. His conclusion was that this was possible and, in fact, the first such ascent by Habeler and Messner in 1978 started from a camp at 7,900 m. Kellas calculated the pressure on the summit to be 251 mmHg, a more accurate figure than estimates based on the "Standard Atmosphere" Kellas estimated maximum oxygen uptake at the summit to be 970 ml/min, and the current value is thought to be about 1,070 ml/min. His estimates of the climbing rate near the summit closely parallels the rate of Habeler and Messner. Kellas had a talent for asking the right questions. He applied his considerable knowledge of physiology to the topic of high altitude, and his suggestions and recommendations were of consistently high quality. He deserves to be better known, both for his geographical surveys and for his pioneer work on high altitude medicine and acclimatisation. The 1921 expedition, after many failed attempts, discovered a possible route to the top of Mount Everest, which was used on all the summit attempts between the two world wars. The route went from Kharta, over the pass Lhakpa La, across East Rongbuk glacier and up via the north col.


Recent Approaches to Design and Analysis of Electrical Impedance Systems for Single Cells Using Machine Learning.
Individual cells have many unique properties that can be quantified to develop a holistic understanding of a population. This can include understanding population characteristics, identifying subpopulations, or elucidating outlier characteristics that may be indicators of disease. Electrical impedance measurements are rapid and label-free for the monitoring of single cells and generate large datasets of many cells at single or multiple frequencies. To increase the accuracy and sensitivity of measurements and define the relationships between impedance and biological features, many electrical measurement systems have incorporated machine learning (ML) paradigms for control and analysis. Considering the difficulty capturing complex relationships using traditional modelling and statistical methods due to population heterogeneity, ML offers an exciting approach to the systemic collection and analysis of electrical properties in a data-driven way. In this work, we discuss incorporation of ML to improve the field of electrical single cell analysis by addressing the design challenges to manipulate single cells and sophisticated analysis of electrical properties that distinguish cellular changes. Looking forward, we emphasize the opportunity to build on integrated systems to address common challenges in data quality and generalizability to save time and resources at every step in electrical measurement of single cells.


Regulation of CYP1A2 by histone deacetylase inhibitors in mouse hepatocytes.
Cytochrome P450 1A2 (CYP1A2) is constitutively expressed in the mouse liver, but the constitutive expression progressively declines to an undetectable level in isolated hepatocytes. In this study, CYP1A2 was induced in hepatocytes exposed to the histone deacetylase inhibitors trichostatin A (TSA) and sodium butyrate (SB), but only well after constitutive CYP1A2 expression was silenced. However, cotreatment with the arylhydrocarbon receptor (AhR) ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and either TSA or SB reduced the induction of CYP1A2 with the same time course as TSA or SB increased its induction. These results suggest that histone modification is involved in CYP1A2 regulation in hepatocytes through pathways that are independent of AhR.


Ferritin synthesis is controlled by iron-dependent translational derepression and by changes in synthesis/transport of nuclear ferritin RNAs.
Ferritin synthesis, maintained at a very low basal rate when extracellular iron levels are low, is elevated up to 50-fold when iron levels are increased. Previous examinations of this iron-dependent activation have concluded that changes in ferritin synthesis results from selective translational activation conferred by an "iron response element" that lies near the 5' terminus of all known ferritin mRNAs. By placing an iron response element in an optimal position in other mRNAs, we find the iron response element to be a potent translational repressor whose influence can only partially be abrogated under optimal inducing conditions. Further, we show that the 25- to 50-fold iron-mediated increase in ferritin synthesis results from coupling a 5- to 6-fold iron-dependent translational derepression with a similar 5- to 6-fold nuclear-dependent increase in mRNA level.


End-of-life caregiver interactions with health care providers: learning from the bad.
This qualitative study examined caregivers' perceptions of and stories about their interactions with nonhospice health care providers. The sample included 27 family caregivers who provided the majority of in-home physical care and emotional support to the elders for at least 6 months. Caregivers described some startling interactions that suggest the need to reevaluate health care professionals' abilities and attitudes concerning their treatment and communication with family caregivers and their elders dying from life-limiting illnesses.


Association of the AURKA and AURKC gene polymorphisms with an increased risk of gastric cancer.
Single nucleotide polymorphisms (SNPs) in mitotic checkpoint genes can contribute to susceptibility of human cancer, including gastric cancer (GC). We aimed to investigate the effects of Aurora kinase A (AURKA), Aurora kinase B (AURKB), and Aurora kinase C (AURKC) gene polymorphisms on GC risk in Slovenian population. We genotyped four SNPs in AURKA (rs2273535 and rs1047972), AURKB (rs2241909), and AURKC (rs758099) in a total of 128 GC patients and 372 healthy controls using TaqMan allelic discrimination assays to evaluate their effects on GC risk. Our results showed that genotype frequencies between cases and controls were significantly different for rs1047972 and rs758099 (P < 0.05). Our study demonstrated that AURKA rs1047972 TT and (CC + CT) genotypes were significantly associated with an increased risk of gastric cancer. Our results additionally revealed that AURKC rs758099 TT and (CC + CT) genotypes were also associated with increased GC risk. In stratified analysis, genotypes TT and (CC + CT) of AURKA rs1047972 SNP were associated with increased risk of both, intestinal and diffuse, types of GC. In addition, AURKC rs758099 TT and (CC + CT) genotypes were positively associated with increased intestinal type GC risk, but not with an increased diffuse type GC risk. Based on these results, we can conclude that AURKA rs1047972 and AURKC rs758099 polymorphisms could affect the risk of GC development. Further larger studies are needed to confirm these findings. © 2016 IUBMB Life, 68(8):634-644, 2016.


[Interferon production in suspensions of swine and human leukocytes on media containing aminopeptide, hemohydrolysate and bovine amniotic fluid].
Production of different kinds of interferon in imported media (medium 199, Eagle's medium) greatly impedes the organization of large-scale interferon production in this country. The search for national substitutes of these media is important therefore. This paper deals with a comparative study of the use of national aminopeptide and hemohydrolysate preparations for interferon production. It was shown that substitution of serum with BAF in preparations of human and swine interferon increased their activity 2- and 4-fold, respectively, and the amount of protein was reduced 10-fold. Specimens of interferons produced with the addition of AP were less effective because the amount of protein increased 4-20-fold while the antiviral activity increased insignificantly. The most effective were interferon preparations obtained with hemohydrolysate medium. Their activity being similar, the amount of protein in interferon samples increased 11/2-2-fold only. Thus, aminopeptide and hemohydrolysate can effectively substitute medium 199 for production of leukocyte human and swine interferon.


Giving Meaning to Alzheimer's Disease: An Experimental Study Using a Framing Approach.
The prejudices and stereotypes associated with Alzheimer's disease (AD) are based on contents that highlight the most negative and dramatic aspects of the symptoms of this disease. This is reinforced by the preponderance of negative representations related to AD. In this paper, these representations have been analyzed as frames. The most dominant frame used to represent dementia is "body-mind dualism," and its alternative is the "body-mind unity" counter-frame. This study aims to define the impact of different messages inspired by the Dualism frame and the Unity counter-frame in the collective representation and meaning given to AD. Posters of simulated health campaigns about AD were designed, combining contents related to the Dualism frame and the Unity counter-frame, and were shown to 85 participants. To analyze the impact of both types of frameworks, a pre-post record of the attitudes of the participants toward AD was conducted using the Attitudes toward Dementia Scale. The assessment of the campaign and the self-report emotions were also registered. Participants' eye movements were measured during exposure to the posters. Attitudes toward dementia were found to be significantly more positive following exposure to discourses characteristic of the Unity counter-frame. Likewise, the dualism discourse evoked more sadness and anger. Moreover, the posters that used the discourse of the Unity counter-frame were considered more useful. We concluded that attitudes toward dementia are influenced by framing. Designing suitable health campaigns using these frames could help mitigate the attitudes and stigma associated with AD.


The Association between Postgraduate Studies, Gender and Qualifying Dental School for Graduates Qualifying from UK Dental Schools between 2000 and 2009.
Various factors will influence a dental graduate's decision to undertake postgraduate education and training, including encouragement from family, partners and staff at individual dental schools, although there is currently little information available regarding the number and distribution (by dental school) of recent dental graduates undertaking postgraduate studies. The aim of this study was to analyse data on postgraduate qualifications achieved by dentists who graduated from UK dental schools between 2000 and 2009 and relate this to graduate gender. Data were collected from the General Dental Council (GDC) in an anonymous electronic format, analysed and ordered by year of graduation, dental school, gender and type of postgraduate qualification. Nearly one-quarter (24%) of the dentists that graduated between 2000 and 2004 completed postgraduate studies, with more females (26%) than males (23%) obtaining further postgraduate qualifications. Overall, Bristol produced the largest proportion of graduates completing postgraduate study (39%) and of these the largest proportion of female graduates (45%). Glasgow produced the largest proportion of male graduates completing postgraduate study (37%). Membership of the Faculty of Dental Surgery (MFDS), one of the Royal Colleges, was the most popular postgraduate qualification obtained followed by Membership of the Faculty of General Dental Practitioners UK (MFGDP). This study provides insight into postgraduate studies undertaken by UK dental graduates. An increasing proportion of females are gaining Bachelor of Dental Surgery (BDS) qualifications and therefore the number of female dental graduates obtaining postgraduate qualifications is likely to increase further. This also suggests the male domination of the dental profession is likely to decrease.


Protein adsorption onto nanoparticles induces conformational changes: Particle size dependency, kinetics, and mechanisms.
The use of nanomaterials in bioapplications demands a detailed understanding of protein-nanoparticle interactions. Proteins can undergo conformational changes while adsorbing onto nanoparticles, but studies on the impact of particle size on conformational changes are scarce. We have shown that conformational changes happening upon adsorption of myoglobin and BSA are dependent on the size of the nanoparticle they are adsorbing to. Out of eight initially investigated model proteins, two (BSA and myoglobin) showed conformational changes, and in both cases this conformational change was dependent on the size of the nanoparticle. Nanoparticle sizes ranged from 30 to 1000 nm and, in contrast to previous studies, we attempted to use a continuous progression of sizes in the range found in live viruses, which is an interesting size of nanoparticles for the potential use as drug delivery vehicles. Conformational changes were only visible for particles of 200 nm and bigger. Using an optimized circular dichroism protocol allowed us to follow this conformational change with regard to the nanoparticle size and, thanks to the excellent temporal resolution also in time. We uncovered significant differences between the unfolding kinetics of myoglobin and BSA. In this study, we also evaluated the plausibility of commonly used explanations for the phenomenon of nanoparticle size-dependent conformational change. Currently proposed mechanisms are mostly based on studies done with relatively small particles, and fall short in explaining the behavior seen in our studies.


Exosomes: A Friend or Foe for Osteoporotic Fracture?
The clinical need for effective osteoporotic fracture therapy and prevention remains urgent. The occurrence and healing of osteoporotic fracture are closely associated with the continuous processes of bone modeling, remodeling, and regeneration. Accumulating evidence has indicated a prominent role of exosomes in mediating multiple pathophysiological processes, which are essential for information and materials exchange and exerting pleiotropic effects on neighboring or distant bone-related cells. Therefore, the exosomes are considered as important candidates both in the occurrence and healing of osteoporotic fracture by accelerating or suppressing related processes. In this review, we collectively focused on recent findings on the diagnostic and therapeutic applications of exosomes in osteoporotic fracture by regulating osteoblastogenesis, osteoclastogenesis, and angiogenesis, providing us with novel therapeutic strategies for osteoporotic fracture in clinical practice.


Endogenous endophthalmitis due to Salmonella typhimurium.
Endogenous endophthalmitis due to Salmonella typhimurium is reported in a 1-year-old child. Despite vigorous antibiotic therapy, the child's vision deteriorated, and loss of light perception occurred in the infected eye. Endophthalmitis is a very rare complication of salmonellosis, and it should alert physicians because of its severe damage to the eye.


Three cases of oculo-facio-cardio-dental (OFCD) syndrome.
Oculo-facio-cardio-dental (OFCD) syndrome is a rare condition with ocular, facial, cardiac, and dental disorders. The purpose of this report is to describe three Japanese female patients by using facial measurements, tooth measurements, and cephalometric analysis. Facial measurements, tooth measurements, and the results of cephalometric analyses were compared with Japanese standards using panoramic roentgenograms, dental casts, and lateral cephalograms. A long face and thick upper and lower lips were noted for all cases from facial measurements, and a long crown length and long dental root were noted for all cases from tooth measurements. Relative mandibular protrusion and a large anterior lower facial height, accompanied by a steep mandible and large gonial angle, were seen from the cephalometric analyses. The results of a long face and long dental root were in accordance with previous reports. In addition, thick upper and lower lips, relative mandibular protrusion, large anterior lower facial height accompanied by a steep mandible, and a large gonial angle may be characterized as specific features of this syndrome.


Following up patients after treatment for anaphylaxis.
Anaphylaxis is defined as a severe, life-threatening, generalised or systemic hypersensitivity reaction. Diagnosis is based on the presenting symptoms and signs which classically develop rapidly, typically evolving over minutes but in some cases hours. Various combinations of airway and/or breathing and/or circulatory problems are possible, as well as urticaria, and hypotension. Skin and/or mucosal changes (typically urticaria and/or angioedema) are seen in around 75% of cases, but importantly these features alone are insufficient for a diagnosis of anaphylaxis. As soon as possible after successful emergency treatment, timed blood samples should be taken for the mast cell tryptase (MCT) test. Serum samples need to be taken within 1-2 hours but no later than 4 hours from the onset of symptoms. It is important to document the acute clinical features (record BP, respiratory rate etc) and the time course of the onset of symptoms/signs and their resolution. Because of the risk of relapse patients should be observed for 6-12 hours after the onset of symptoms. Children under 16 years should be admitted and supervised by a paediatrician. An adrenaline injector device for intramuscular use only, should be prescribed as an interim measure before referral to a specialist allergy clinic. Referral to a specialist allergy service (or specialist paediatric service), is strongly recommended. Diagnosis can be confirmed, and further investigations organised.


Identification of the phosphorylation targets of symbiotic receptor-like kinases using a high-throughput multiplexed assay for kinase specificity.
Detecting the phosphorylation substrates of multiple kinases in a single experiment is a challenge, and new techniques are being developed to overcome this challenge. Here, we used a multiplexed assay for kinase specificity (MAKS) to identify the substrates directly and to map the phosphorylation site(s) of plant symbiotic receptor-like kinases. The symbiotic receptor-like kinases nodulation receptor-like kinase (NORK) and lysin motif domain-containing receptor-like kinase 3 (LYK3) are indispensable for the establishment of root nodule symbiosis. Although some interacting proteins have been identified for these symbiotic receptor-like kinases, very little is known about their phosphorylation substrates. Using this high-throughput approach, we identified several other potential phosphorylation targets for both these symbiotic receptor-like kinases. In particular, we also discovered the phosphorylation of LYK3 by NORK itself, which was also confirmed by pairwise kinase assays. Motif analysis of potential targets for these kinases revealed that the acidic motif xxxsDxxx was common to both of them. In summary, this high-throughput technique catalogs the potential phosphorylation substrates of multiple kinases in a single efficient experiment, the biological characterization of which should provide a better understanding of phosphorylation signaling cascade in symbiosis.


Determination of 7-piperidino-1,2,3,5-tetrahydroimidazo[2,1-b]quinazolin-2-one (DN-9693) in human plasma and urine by high-performance liquid chromatography with electrochemical detection: application to studies on disposition of DN-9693 in healthy human volunteers.
A procedure for the determination of a newly developed antiadhesive agent, 7-piperidino-1,2,3,5-tetrahydroimidazo[2,1-b]quinazolin-2-on e (DN-9693; 1), in human plasma and urine at low nanogram levels by high-performance liquid chromatography with electrochemical detection is described. Plasma and urine samples were made alkaline with 1 M KOH solution and extracted with chloroform. After removing organic solvent, the extracts reconstituted in methanol were applied to a reversed-phase octadesyl-packed column and eluted with a methanol: phosphate buffer (pH 8.0) mobile phase. The applied potentials were set at 0.80 and 0.95 V versus a silver-silver chloride reference electrode for the analyses of plasma and urine specimens, respectively. The method provides a recovery of greater than 90% and a within-day precision of less than 5%. The minimum quantifiable levels for 1 in plasma and urine using 1 mL of sample are 1 and 4 ng/mL, respectively. The method has been successfully used for the determination of 1 in plasma and urine samples from healthy human volunteers after intravenous infusion of the drug.


Retention or loss of v-mil sequences after propagation of MH2 virus in vivo or in vitro.
During propagation of the defective avian retrovirus MH2 in the presence of replication-competent helper virus, deletion of portions of the viral genome occurred frequently. After transformation of quail cells in vitro, v-mil sequences were lost, leading to populations of MH2 viruses which were highly deficient for mil gene expression but which could transform macrophage and fibroblast cells in vitro with high efficiency. In contrast, after induction of tumors in quail with mil-deficient MH2 viral stocks, a majority of the tumor DNAs contained mil+ proviruses, suggesting that there is selection for retention of the v-mil gene in vivo and that the mil protein may play a role in the oncogenicity of MH2 virus. We also isolated MH2-transformed cell lines which contained deleted proviruses arising from packaging and subsequent integration of the subgenomic v-myc-encoding mRNA. Some of these cell lines produced viruses which encoded abnormal v-myc proteins and had altered in vitro transforming properties. These altered phenotypes may be caused by mutations within the v-myc gene.