The effects of in utero and lactational exposure to chloroform on postnatal growth and glucose tolerance in male Wistar rats.
Water chlorination results in the formation of trihalomethanes (THMs) including chloroform. In human stud-ies, fetal growth restriction has been associated with exposure to THMs during pregnancy and impaired fetal growth has been associated with an increased risk of type 2 diabetes. Therefore, the objective of this study was to determine the effect of in utero and lactational exposure to chloroform on birthweight and postnatal indicators of type 2 diabetes. Female Wistar rats were given chloroform (0 microg/L, 75 microg/L) in their drinking water for 2 wk prior to mating until parturition (in utero exposure only) or until weaning (in utero+lactational exposure). At postnatal d 1 (PND1) pups of dams exposed to chloroform had significantly higher serum glucose levels and lower insulin levels, but this effect was not due to b<FONT "Optima">-cell depletion in the neonatal pancreas. Glucose homeostasis in response to a glucose challenge was not changed by chloroform treatment. Chloroform exposure did not affect birthweight; however, offspring of dams exposed to chloroform had significantly impaired postnatal growth. Although fetal and neonatal exposure to chloroform did not elicit physiological changes associated with the onset of type 2 diabetes, there were physiological changes resulting in impaired postnatal growth.


Dapsone-Induced Granulomatous Cholestatic Hepatitis Unmasked by Steroid Taper: A Case Report.
Dapsone is known to cause drug-induced liver injury (DILI) but can rarely induce the formation of hepatic granulomas. We describe a patient with jaundice who demonstrated granulomas on liver biopsy in response to dapsone. Her symptoms were only evident once steroids, used to also treat her pyoderma gangrenosum, had been tapered. In this case, a 67-year-old female was hospitalized due to 1 day of jaundice. She had started dapsone and prednisone concurrently 7 weeks ago to treat her pyoderma gangrenosum. Steroids were discontinued 4 days prior to symptoms. Her laboratories were notable for newly elevated alkaline phosphatase (756 U/L), aspartate transaminase (199 U/L), alanine transaminase (273 U/L), and total bilirubin (12.6 mg/dL). Dapsone was held due to suspicion for DILI. A liver biopsy was performed and disclosed non-necrotizing hepatic granulomas. After infectious and autoimmune causes were excluded, dapsone was determined to be the cause of her hepatic granulomas. Her bilirubin and liver enzymes steadily normalized over the next 4 weeks following discontinuation of dapsone. Thus, dapsone-related liver injury may present following a steroid taper if dapsone and steroids had been initially started together. Hepatic granulomas, though rare, can be seen when dapsone causes DILI.


Vorinostat and sorafenib increase CD95 activation in gastrointestinal tumor cells through a Ca(2+)-de novo ceramide-PP2A-reactive oxygen species-dependent signaling pathway.
The targeted therapeutics sorafenib and vorinostat interact in a synergistic fashion to kill carcinoma cells by activating CD95, and this drug combination is entering phase I evaluation. In this study, we determined how CD95 is activated by treatment with this drug combination. Low doses of sorafenib and vorinostat, but not the individual drugs, rapidly increased reactive oxygen species (ROS), Ca(2+), and ceramide levels in gastrointestinal tumor cells. The production of ROS was reduced in Rho zero cells. Quenching ROS blocked drug-induced CD95 surface localization and apoptosis. ROS generation, CD95 activation, and cell killing was also blocked by quenching of induced Ca(2+) levels or by inhibition of PP2A. Inhibition of acidic sphingomyelinase or de novo ceramide generation blocked the induction of ROS; however, combined inhibition of both acidic sphingomyelinase and de novo ceramide generation was required to block the induction of Ca(2+). Quenching of ROS did not affect drug-induced ceramide/dihydro-ceramide levels, whereas quenching of Ca(2+) reduced the ceramide increase. Sorafenib and vorinostat treatment radiosensitized liver and pancreatic cancer cells, an effect that was suppressed by quenching ROS or knockdown of LASS6. Further, sorafenib and vorinostat treatment suppressed the growth of pancreatic tumors in vivo. Our findings show that induction of cytosolic Ca(2+) by sorafenib and vorinostat is a primary event that elevates dihydroceramide levels, each essential steps in ROS generation that promotes CD95 activation.


Sperm Release at Spermiation Is Regulated by Changes in the Organization of Actin- and Microtubule-Based Cytoskeletons at the Apical Ectoplasmic Specialization-A Study Using the Adjudin Model.
The mechanism that regulates sperm release at spermiation is unknown. Herein, we used an animal model wherein rats were treated with adjudin, 1-(2,4-dichlorobenzyl)-1H-indazole-3-carbohydrazide, via oral gavage to induce premature release of elongating/elongated spermatids, followed by round spermatids and spermatocytes. Spermatid release mimicking spermiation occurred within 6 to 12 hours following adjudin treatment and, by 96 hours, virtually all tubules were devoid of elongating/elongated spermatids. Using this model, we tracked the organization of F-actin and microtubules (MTs) by immunofluorescence microscopy, and the association of actin or MT regulatory proteins that either promote or demolish cytoskeletal integrity through changes in the organization of actin microfilaments or MTs by coimmunoprecipitation. Adjudin treatment induced an increase in the association of (1) epidermal growth factor receptor pathway substrate 8 (an actin barbed-end capping and bundling protein) or formin 1 (an actin nucleator) with actin and (2) end-binding protein 1 (an MT stabilizing protein) with MT shortly after adjudin exposure (at 6 hours), in an attempt to maintain spermatid adhesion to the Sertoli cell at the apical ectoplasmic specialization (ES). However, this was followed by a considerable decline of their steady-state protein levels, replacing with an increase in association of (1) actin-related protein 3 (a branched actin nucleator that converts actin filaments into a branched/unbundled network) with actin and (2) MT affinity-regulating kinase 4 (an MT destabilizing protein kinase) with MTs by 12 hours after adjudin treatment. These latter changes thus promoted actin and MT disorganization, leading to apical ES disruption and the release of elongating/elongated spermatids, mimicking spermiation. In summary, spermiation is a cytoskeletal-dependent event, involving regulatory proteins that modify cytoskeletal organization.


[Microionophoretic study of the cholinoceptor neurons of the visual cortex of cats].
The optic cortex unit responses to the microionophoretic acetylcholine application were studied in acute experiments on unanesthetized cats restrained with diplacine. Of total amount of registered units 59% were sensitive to acetylcholine and revealed excitative (predominantly) or inhibitory responses.


Identification of further proteolytic cleavage sites in the Southampton calicivirus polyprotein by expression of the viral protease in E. coli.
Southampton virus (SV) is a human enteric calicivirus with a positive-sense RNA genome which encodes a protease as part of a large precursor polyprotein. Expression vectors based on pRSET were constructed carrying the entire 3C-like viral protease (3Cpro) sequence together with flanking sequences from a region of the viral genome 3'-distal to the putative helicase-encoding region. Expression from these vectors in E. coli resulted in discrete protein products with smaller than expected molecular sizes. This confirmed that an active viral protease was produced in E. coli and that the protease was capable of cleaving the expressed protein at defined sites. Expressed cleavage products surrounding the protease region of the viral polyprotein were separated by SDS-PAGE, transferred to PVDF membranes and subjected to N-terminal sequence analysis. Cleavage occurred at an EG dipeptide at the N terminus of the putative VPg (961E/GKNKG) and also at the protease/polymerase boundary (1280E/GGDKG). The N terminus of the protease was released from the VPg C terminus at an EA dipeptide in the sequence 1099E/APPTL. These studies demonstrate that SV enteric calicivirus encodes a 3C-like protease with a specificity similar to the picornaviral 3C protease and that the SV polyprotein is cleaved into at least six mature products.


Immunoreactive endogenous ouabain in primary aldosteronism and essential hypertension: relationship with plasma renin, aldosterone and blood pressure levels.
To investigate the role of ouabain in human hypertension and to establish whether immunoreactive endogenous ouabain is secreted by the adrenal gland under the influence of dopaminergic regulation. We measured plasma levels of endogenous ouabain by immunoassay, together with other variables, including plasma renin activity and aldosterone levels, in 91 clinically selected hypertensives and 19 healthy volunteers. We also measured endogenous ouabain in adrenal venous blood and the effect of DA2 dopaminergic receptor blockade and stimulation. After a thorough clinical evaluation, 64 patients were diagnosed with essential hypertension and 24 with primary aldosteronism. Plasma levels of endogenous ouabain were higher in essential hypertensives than in controls. Multiple regression analysis showed a significant relationship of mean blood pressure with plasma endogenous ouabain, age and body mass index, but not with other measured parameters. The plasma levels of endogenous ouabain were more than two standard deviations above the mean value for normotensives in 45% of patients with essential hypertension in whom plasma renin activity was normal. Higher plasma levels of endogenous ouabain were found in patients with aldosterone excess, specifically affecting 56% of 17 patients with surgically confirmed adrenal cortical adenoma and one (14%) of seven patients with idiopathic causes. Removal of adenomas lowered blood pressure in half of the patients in whom plasma levels of endogenous ouabain normalized after surgery. Plasma endogenous ouabain levels were similar in venous blood from the adrenal and inferior vena cava, and plasma levels were not influenced by DA2 dopaminergic blockade and stimulation. Approximately half of Caucasian patients with essential hypertension and with hyperaldosteronism exhibit elevated circulating levels of endogenous ouabain. The latter do not appear to be secondary to hypertension, are unrelated to plasma renin activity, and may not involve adrenal type-2 dopaminergic receptors.


Intravascular volume administration: a contributing risk factor for intracranial hemorrhage during extracorporeal membrane oxygenation?
The objective of this study was to determine the relationship between the frequency and total volume of intravascular volume administration and the development of intracranial hemorrhage during venoarterial extracorporeal membrane oxygenation. In a retrospective, matched, case-control study, 24 newborns who developed an intracranial hemorrhage during venoarterial extracorporeal membrane oxygenation treatment were compared with 40 control subjects. Both groups were analyzed for gestational age, gender, race, Apgar scores at 1 and 5 minutes, birth weight, cardiopulmonary resuscitation before venoarterial extracorporeal membrane oxygenation, age at the start of treatment, duration of treatment, worst arterial blood gas sample preceding treatment, activated clotting time values, need for platelet transfusions, mean blood pressure, and the use of inotropics and steroids before the treatment. For both groups, total number and volume of intravascular infusions of normal saline, pasteurized plasma protein solution, erythrocytes, and platelets during the first 24 hours of treatment were determined. Variables were analyzed in their relationship to intracranial hemorrhage by using univariate and multivariate conditional logistic regression. The only statistically significant difference in patient characteristics between the case patients and control subjects was arterial blood gas values. Newborns who developed intracranial hemorrhage during the treatment received both a statistically significantly higher number and a statistically significantly higher total volume of intravascular volume administrations compared with control patients. After adjustment for pH, Paco(2), and Pao(2) in the multivariate analysis, we found a significant relation between the development of intracranial hemorrhage and >8 infusions or >300 mL of volume infusion in the first 8 hours and >10 infusions in the first 24 hours of treatment. The number and total volume of intravascular volume administration in the first 8 and 24 hours of venoarterial extracorporeal membrane oxygenation treatment are statistically significantly related to the development of intracranial hemorrhage.


[The reactivity of the arterial vessels of the ovary].
The responsiveness of the ovarian vascular muscle to administration of adrenaline, noradrenaline, serotonin, acetylcholine, histamine, angiotensin II and prostaglandin F2 alpha was studied in isolated ovarian artery rings of women, cow and rabbit. The data obtained show that the ovarian artery responsiveness differs in various species and that the responsiveness of the ovarian vascular muscle depends on the functional state of the ovary.


Chemoprotective effect of nanocurcumin on 5-fluorouracil-induced-toxicity toward oral cancer treatment.
Cancer of oral cavity is the uncontrolled expansion of damaged cell within the mouth cavity. 5-fluorouracil (5-FU) chemotherapy was focused to kill the cancer cell, but it would affect the surrounding normal cells during oral cancer treatment. This study included the evaluation of chemoprotective effects of curcumin (CU), as an herbal remedy, on 5-FU-induced-cytotoxicity toward oral cancer treatment, loaded within a nanocarrier system. CU was combined with 5-FU chemotherapy as a combinational drug-delivery system to evaluate synergistic effects. Nanoformulation of CU (nano-CU) and nanoformulation of 5-FU (nano-FU) were prepared by employing homogenization with high-energy sonication. The characterizations of prepared nanoformulations were evaluated on the basis of particle size, zeta potential, and polydispersity index (PDI) values. The chemopreventive effect of nano-CU on 5-FU induced-toxicity and synergistic efficacy were optimized through different in-vitro assays. The average particle size of nano-CU and nano-FU were up to 200 nm, negatively-charged, and shown up to 4th-day control release of the drug within the acceptable concentration. IC50 value for growth inhibition was calculated as 47.89 and 26.19 μg/ml, respectively, for nano-CU and nano-FU. OCC was pretreated with nano-CU and shown the protective effect by reducing 5-FU induced-cytotoxicity by preventing normal cells through reduced viability. The DPPH-indicated fluorescence-tagged cells had quantified for antioxidant effect as it reduces intracellular reactive oxygen species level in OCC. Along with alteration in cell protein expression, Blc2, and Bax, shows enhanced apoptosis rate in OCC. Nano-CU provides chemoprotective nature towards 5-FU induced-toxicity, along with synergistic effects in oral cancer treatment.


Essential role for the lectin pathway in collagen antibody-induced arthritis revealed through use of adenovirus programming complement inhibitor MAp44 expression.
Previous studies using mannose-binding lectin (MBL) and complement C4-deficient mice have suggested that the lectin pathway (LP) is not required for the development of inflammatory arthritis in the collagen Ab-induced arthritis (CAIA) model. MBL, ficolins and collectin-11 are key LP pattern recognition molecules that associate with three serine proteases-MASP-1, MASP-2, and MASP-3-and with two MBL-associated proteins designated sMAP and MBL-associated protein of 44kDA (MAp44). Recent studies have shown that MAp44, an alternatively spliced product of the MASP-1/3 gene, is a competitive inhibitor of the binding of the recognition molecules to all three MASPs. In these studies, we examined the effect of treatment of mice with adenovirus (Ad) programmed to express human MAp44 (AdhMAp44) on the development of CAIA. AdhMAp44 and Ad programming GFP (AdGFP) expression were injected i.p. in C57BL/6 wild type mice prior to the induction of CAIA. AdhMAp44 significantly reduced the clinical disease activity (CDA) score by 81% compared with mice injected with AdGFP. Similarly, histopathologic injury scores for inflammation, pannus, cartilage and bone damage, as well as C3 deposition in the cartilage and synovium, were significantly reduced by AdhMAp44 pretreatment. Mice treated with AdmMAp44, programming expression of mouse MAp44, also showed significantly decreased CDA score and histopathologic injury scores. In addition, administration of AdhMAp44 significantly diminished the severity of Ross River virus-induced arthritis, an LP-dependent model. Our study provides conclusive evidence that an intact complement LP is essential to initiate CAIA, and that MAp44 may be an appropriate treatment for inflammatory arthritis.


Perforation of dental gloves during prosthodontic treatments as assessed by the conductivity and water inflation tests.
The incidence of latex glove perforation during prosthodontic treatment was investigated on 122 occasions using two methods, a conductivity test and a water inflation test. Latex glove perforation was detected in 38.5% of the treatments by the conductivity test and in 27.9% by the water inflation test. The perforation went unrecognized in 74.7% of the occurrences (35/47 incidents) using the conductivity test and in 64.7% (22/34) when the water inflation test was used. Of the total 55 glove perforations, 21 perforations were detected only by conductivity test, 3 were detected only by the water inflation test, and 31 perforations were detected by both methods, which suggested that the conductivity test is more sensitive than the water inflation test for the detection of glove perforation. The results of this research suggested that even when latex gloves are worn, the risk to prosthodontists of exposure to body fluids remains in four of every six treatments, often without the awareness of the prosthodontist.


In vivo and in vitro correlation of pulmonary MAP kinase activation following metallic exposure.
Residual oil fly ash (ROFA) is a particulate pollutant produced in the combustion of fuel oil. Exposure to ROFA is associated with adverse respiratory effects in humans, induces lung inflammation in animals, and induces inflammatory mediator expression in cultured human airway epithelial cells (HAEC). ROFA has a high content of transition metals, including vanadium, a potent tyrosine phosphatase inhibitor that we have previously shown to disregulate phosphotyrosine metabolism and activate mitogen-activated protein kinase (MAPK) signaling cascades in HAEC. In order to study MAPK activation in response to in vivo metal exposure, we used immunohistochemical methods to detect levels of phosphorylated protein tyrosines (P-Tyr) and the MAPKs ERK1/2, JNK, and P38 in lung sections from rats intratracheally exposed to ROFA. After a 1-h exposure to 500 microg ROFA, rat lungs showed no histological changes and no significant increases in immunostaining for either P-Tyr or phospho-(P-) MAPKs compared to saline-instilled controls. At 4 h of exposure, there was mild and variable inflammation in the lung, which was accompanied by an increase in specific immunostaining for P-Tyr and P-MAPKs in airway and alveolar epithelial cells and resident macrophages. By 24 h of exposure, there was a pronounced inflammatory response to ROFA instillation and a marked increase in levels of P-Tyr and P-MAPKs present within the alveolar epithelium and in the inflammatory cells, while the airway epithelium showed a continued increase in the expression of P-ERK1/2. By comparison, HAEC cultures exposed to 100 microg/ml ROFA for 20 min resulted in marked increases in P-Tyr and P-MAPKs, which persisted after 24 h of exposure. P-Tyr levels continued to accumulate for up to 24 h in HAEC exposed to ROFA. These results demonstrate in vivo activation in cell signaling pathways in response to pulmonary exposure to particulate matter, and support the relevance of in vitro studies in the identification of mechanisms of lung injury induced by pollutant inhalation.


Short-term effects of air pollution on wheeze in asthmatic children in Fresno, California.
Although studies have demonstrated that air pollution is associated with exacerbation of asthma symptoms in children with asthma, little is known about the susceptibility of subgroups, particularly those with atopy. This study was designed to evaluate our a priori hypothesis that identifiable subgroups of asthmatic children are more likely to wheeze with exposure to ambient air pollution. A cohort of 315 children with asthma, 6-11 years of age, was recruited for longitudinal follow-up in Fresno, California (USA). During the baseline visit, children were administered a respiratory symptom questionnaire and allergen skin-prick test. Three times a year, participants completed 14-day panels during which they answered symptom questions twice daily. Ambient air quality data from a central monitoring station were used to assign exposures to the following pollutants: particulate matter ≤ 2.5 μm in aerodynamic diameter, particulate matter between 2.5 and 10 μm in aerodynamic diameter (PM10-2.5), elemental carbon, nitrogen dioxide (NO2), nitrate, and O3. For the group as a whole, wheeze was significantly associated with short-term exposures to NO2 [odds ratio (OR) = 1.10 for 8.7-ppb increase; 95% confidence interval (CI), 1.02-1.20] and PM10-2.5 (OR = 1.11 for 14.7-μg/m3 increase; 95% CI, 1.01-1.22). The association with wheeze was stronger for these two pollutants in children who were skin-test positive to cat or common fungi and in boys with mild intermittent asthma. A pollutant associated with traffic emissions, NO2, and a pollutant with bioactive constituents, PM10-2.5, were associated with increased risk of wheeze in asthmatic children living in Fresno, California. Children with atopy to cat or common fungi and boys with mild intermittent asthma were the subgroups for which we observed the largest associations.


Neutrophil accumulation promotes intimal hyperplasia after photochemically induced arterial injury in mice.
The role of leukocytes in the pathogenesis of coronary arterial disease has become a focus for clinical research. The aim of this study was to determine whether neutrophil accumulation would participate in the development of intimal hyperplasia after endothelial injury in mice, and whether d-myo-inositol hexakisphosphate (phytic acid) which inhibits the binding of L- and P-selectin to sialyl Lewis(X) could inhibit the development of intimal hyperplasia. Endothelial injury was inflicted in one femoral artery via the photochemical reaction between systemically injected rose bengal and transillumination with green light (wavelength: 540 nm). Scanning electron microscopic observation at 3 days after the injury showed an increase in the number of leukocytes adhering to the injury site. Histological observation at 21 days showed that in the neutropenia group administered anti-neutrophil antibody and in the phytic acid-treated group the progression of intimal hyperplasia was significantly attenuated by comparison with the corresponding control groups. These results suggest that neutrophil accumulation contributes to the initiation and/or development of intimal hyperplasia and L- and/or P-selectin may participate in their mechanisms.


Binding and endocytosis of glycoproteins and neoglycoproteins by isolated rabbit hepatocytes.
Rabbit hepatocytes were isolated by a collagenase perfusion technique, and used to study the binding and endocytosis of the glycoprotein, asialo-orosomucoid, and the neoglycoprotein, Gal39-bovine serum albumin. Both of these proteins contain exposed galactosyl residues, and were avidly bound by the lectin on the hepatic parenchymal cell surface. Steady state and kinetic experiments performed at 2 degrees C and at 37 degrees C revealed the presence of two apparent classes of binding sites totalling 4.7 X 10(5) sites/cell at 2 degrees C, and 6.3 X 10(5) sites/cell at 37 degrees C. At 37 degrees C, both classes of sites participated in internalization of bound ligand. The cells were capable of internalizing about 60 000 molecules/min per cell. The process appeared to be first-order, with a rate constant k = 0.098 min-1 and t1/2 = 7.1 +/- 0.6 min. Binding could be inhibited by galactose-containing compounds, EGTA, and by anti-(hepatic lectin) immunoglobulin G. The inhibition by antibody appeared to be reversible upon removal of antibody-containing medium.


Successful Ultrasound-Guided Superficial Cervical Plexus Block Treatment for Head and Neck Pain with an Unusually Delayed Onset Following Ventriculoperitoneal Shunt: A Case Report.
Background and Objectives: Ventriculoperitoneal (VP) shunt placement is the most common treatment for cerebrospinal fluid diversion. Head and neck pain occurring after a long period following VP shunt insertion is rarely reported. Here, we present a rare case of head and neck pain occurring 2 years after surgery due to irritation of the superficial cervical plexus by the VP shunt. Case Description: A 46-year-old female patient received VP shunt placement surgery. Two years after the surgery, she experienced a left temporal headache with neck pain on the left side, which extended to the left para-auricular and fascial region. Ultrasound (US) scanning revealed that the VP shunt passed within the superficial cervical fascia and through the left sternocleidomastoid muscle (SCM). Additionally, friction of the branches of the superficial cervical plexus and of the greater auricular and lesser occipital nerves caused by the VP shunt was found underneath the lateral border of the SCM. Subsequently, the blocking and hydro-release of the left superficial cervical plexus were performed. After four series of treatments, the patient's head and neck pain vanished, and the frequency of the headaches was substantially reduced. The patient was regularly followed-up in the outpatient department of neurosurgery. Conclusions: Head and neck pain caused by the malpositioning of a VP shunt catheter with an unusually delayed onset is a rarely reported complication and could be easily neglected. Patients with head and neck pain following VP shunt insertion should be checked using US scanning to identify the potential origin of the pain and receive adequate treatments. Intraoperative US-guided tunnelling is suggested to avoid the malpositioning of the VP shunt catheter.


Cleavage of atrophin-1 at caspase site aspartic acid 109 modulates cytotoxicity.
Dentatorubropallidoluysian atrophy (DRPLA) is one of eight autosomal dominant neurodegenerative disorders characterized by an abnormal CAG repeat expansion which results in the expression of a protein with a polyglutamine stretch of excessive length. We have reported recently that four of the gene products (huntingtin, atrophin-1 (DRPLA), ataxin-3, and androgen receptor) associated with these open reading frame triplet repeat expansions are substrates for the cysteine protease cell death executioners, the caspases. This led us to hypothesize that caspase cleavage of these proteins may represent a common step in the pathogenesis of each of these four neurodegenerative diseases. Here we present evidence that caspase cleavage of atrophin-1 modulates cytotoxicity and aggregate formation. Cleavage of atrophin-1 at Asp109 by caspases is critical for cytotoxicity because a mutant atrophin-1 that is resistant to caspase cleavage is associated with significantly decreased toxicity. Further, the altered cellular localization within the nucleus and aggregate formation associated with the expanded form of atrophin-1 are completely suppressed by mutation of the caspase cleavage site at Asp109. These results provide support for the toxic fragment hypothesis whereby cleavage of atrophin-1 by caspases may be an important step in the pathogenesis of DRPLA. Therefore, inhibiting caspase cleavage of the polyglutamine-containing proteins may be a feasible therapeutic strategy to prevent cell death.


Validity of the Expressed Emotion Adjective Checklist (EEAC) in Caregivers of Children with Mood Disorders.
Expressed Emotion (EE; criticism/hostility and emotional overinvolvement) displayed in family interactions has been associated with the presence and poorer course of multiple disorders in adults and children. As such, validating appropriate tools for measuring EE could have important implications for research and clinical practice. Child EE measures are limited though there are some established methods of assessing EE in adults. The Expressed Emotion Adjective Checklist (EEAC), a self-report measure of EE, has demonstrated validity with adults but has not been evaluated in child samples. The present study examined reliability, stability, and validity of the EEAC in measuring EE in caregivers of children with mood disorders. EEAC scores were associated with the criticism component of the Five Minute Speech Sample (FMSS), a commonly used EE measure in children. EEAC scores were also stable and predicted manic symptom severity and global impairment one year later. These data suggest the EEAC may be a useful self-report measure of EE in children.


Ultrastructure and life cycle of the lipid-containing bacteriophage phi 6.
An electron microscopic study of the lipid-containing bacteriophage phi 6 revealed an electron dense compact inner core of 30 nm in diam., which apparently contains the nucleic acid of the virus. This inner particle is surrounded by a complex polyhedral capsid with an outer diam. of 50 nm. Outside this is the envelope, which gives the virus a total diam. of 65 to 75 nm. The envelope, which has a thickness of a unit membrane, could be removed by treating the phage with Triton X-100. A definite structure is seen inside the envelope of the phage tail. In infection, phages are attached by their tails to the host cell pili. Occasional pili with a few attached phages were seen in a phage resistant mutant. In the course of the infection phages were also seen attached to the outer membrane of the cell. In a phage-tolerant mutant many normal-looking pili with adsorbed phages were visible, but we could never see phage-cell membrane associations. The membrane of the phage appears to fuse with the bacterial outer membrane and 50 nm virus particles could be seen in the periplasmic space of the bacterium, probably attached to the cytoplasmic membrane. Newly formed 50 nm particles appeared 45 min post infection (p.i.) cnetrally in the host cell. Assembly of the envelope also began at this time and by 80 minutes p.i. all the 50 nm particles were covered by the virus membrane. At no stage were phages seen in the periphery of the bacterium. Mature phages were finally released by a rupture of the host cell without spheroplast formation.


Sjögren's syndrome: autoantibodies to cellular antigens. Clinical and molecular aspects.
Autoantibodies to cellular autoantigens are usually found in sera of patients with systemic autoimmune rheumatic diseases. Patients with Sjögren's syndrome (SS) frequently present autoantibodies to both organ and non-organ-specific autoantigens. The most commonly detected autoantibodies are those directed against the ribonucleoproteins Ro/SSA and La/SSB. The presence of the antibodies in SS is associated with early disease onset, longer disease duration, parotid gland enlargement, higher frequency of extraglandular manifestations and more intense lymphocytic infiltration of the minor salivary glands. Over the past several years, the structure and function of these autoantigens have been extensively studied. Several centers, using different techniques, have investigated the B cell epitopes on the protein components Ro 60 kD, Ro 52kD, and La 48 kD. Finally, increased evidence of direct involvement of anti-Ro/SSA and anti-La/SSB autoantibodies in the pathogenesis of tissue injury has been contributed by several studies.


Targeted antigen presentation using crosslinked antibody heteroaggregates.
We have targeted protein antigens to antigen-presenting cells in vitro by using antibody heteroaggregates containing an antibody against a protein antigen covalently crosslinked to an antibody against a target structure on the surface of the antigen-presenting cells. Antigen presentation was assessed by measurement of lymphokine released by antigen-specific T cell hybridomas. Depending on the experimental conditions, the crosslinked antibodies decreased the amount of antigen required to give a response by the hybridomas by factors of 10(2) to 10(3). Enhanced presentation occurred when antigen was targeted to MHC class I and class II molecules, surface immunoglobulin, or Fc gamma receptors on the surface of the murine B cell lymphoma-hybridoma, TA3. An enhancement of antigen presentation also occurred when antigen was targeted to surface IgD, or class I and class II MHC molecules on murine splenic B cells, and when antigen was targeted to class I and class II molecules on irradiated adherent spleen cells. No response was seen when antigen was targeted to Fc gamma R on B cells or adherent spleen cells. The ability of each crosslinked antibody to enhance presentation paralleled the total amount of each that bound to the surface of the antigen-presenting cells. Antigen presentation, mediated by crosslinked antibody, was antigen-specific and I-A restricted. The presentation of one antigen by using crosslinked antibody did not result in enhanced presentation of a second, bystander antigen. These results suggest that a novel means of stimulating immune responses may be possible in vivo, by targeting antigen to surface structures on antigen-presenting cells.


Advances in the agent-based modeling of economic and social behavior.
In this review we discuss advances in the agent-based modeling of economic and social systems. We show the state of the art of the heuristic design of agents and how behavioral economics and laboratory experiments have improved the modeling of agent behavior. We further discuss how economic networks and social systems can be modeled and we discuss novel methodology and data sources. Lastly, we present an overview of estimation techniques to calibrate and validate agent-based models and show avenues for future research.


Impaired responsiveness of platelets to epinephrine due to α2A adrenoreceptor deficiency in Male Chinese.
Epinephrine is known as a weak, but important, agonist for platelet activation. It has been reported that the responsiveness of platelets to epinephrine was markedly impaired in 6% of Caucasians and in 16% of Japanese. The purpose of this study was to screen and characterize this abnormality in healthy Taiwanese Chinese volunteers. We used aggregometry, flow cytometry and platelet function analyzer (PFA)-100 system to assess in 50 healthy male volunteers the responsiveness of platelets to epinephrine stimulation. Using α2A adrenoceptor antagonist BRL44408 maleate competition and a [(3)H]yohimbin binding assay, we evaluated α2A adrenoceptors on platelets. The aggregation of platelets after stimulation with 10 μM of epinephrine indicated two distinct groups of study participants: 24 (48.0%) good- and 26 (52.0%) impaired-responders to epinephrine. Flow cytometric analysis of platelets after stimulated with 1 μM epinephrine showed that glycoprotein (GP) IIb/IIIa and P-selectin expression of epinephrine good- and impaired-responders were 27.1 ± 11.0% vs. 9.9 ± 5.4% (p = 0.003) and 12.2 ± 6.2% vs. 3.6 ± 3.5% (p < 0.001), respectively. The PFA-100 system showed that epinephrine-impaired-responders had a longer collagen-epinephrine induced closure time. Good-responder platelets incubated with BRL44408 maleate had an impaired response to epinephrine stimulation. [(3)H]yohimbine binding studies showed fewer α2A adrenoreceptors on the platelets of epinephrine-impaired-responders than on those of good-responders. The prevalence of impaired responsiveness to epinephrine was high and probably due to α2A adrenoreceptor deficiency in male Taiwanese Chinese.


Load and release of gambogic acid via dual-target ellipsoidal-Fe3O4@SiO2@mSiO2-C18@dopamine hydrochloride -graphene quantum dots-folic acid and its inhibition to VX2 tumor cells.
Ellipsoidal-Fe3O4@SiO2@mSiO2-C18@dopamine hydrochloride-graphene quantum dots-folic acid (ellipsoidal-HMNPs@PDA-GQDs-FA), a dual-functional drug carrier, was stepwise constructed. Theα-Fe2O3ellipsoidal nanoparticles were prepared by a hydrothermal method, and then coated with SiO2by Stöber method. The resulting core-shell structure, Fe3O4@SiO2@mSiO2-C18magnetic nano hollow spheres, abbreviated as HMNPs, was finally grafted with graphene quantum dots (GQDs), dopamine hydrochloride (PDA) and folic acid (FA) by amide reaction to obtain HMNPs@PDA-GQDs-FA. Transmission electron microscopy, Fourier transform infrared spectroscopy, fluorescence spectroscopy and element analysis proved the successful construction of the HMNPs@PDA-GQDs-FA nanoscale carrier-cargo composite. The carrier HMNPs@PDA-GQDs-FA has higher load (51.63 ± 1.53%) and release (38.56 ± 1.95%) capacity for gambogic acid (GA). Cytotoxicity test showed that the cell survival rate was above 95%, suggesting the cytotoxicity of the carrier-cargo was very low. The cell lethality (74.91 ± 1.2%) is greatly improved after loading GA because of the magnetic targeting of HMNPs, the targeting performance of FA to tumor cells, and the pH response to the surrounding environment of tumor cells of PDA. All results showed that HMNPs@PDA-GQDs-FA had good biocompatibility and could be used in the treatment of VX2 tumor cells after loading GA.


[The estimation of weight-loss programmes and using of slimming preparations among young women].
The aim of this study was to evaluate the range of weight-loss programmes and behaviours associated with the use of slimming supplements observed among young women. The study in the form of questionnaire survey was carried out among 300 female students of the West Pomeranian University of Technology in Szczecin during late autumn 2009. Analysis of the responses revealed that most of the women had tried to lose weight ever during their lives, many of them within the past six months. The respondents undertook weight-loss programmes regardless of their BMI, which may imply a disturbed perception of one's own body. In order to reduce body weight, the women applied various methods, usually modified diet and use slimming preparations. Among the preparations, the women most frequently mentioned appetite inhibitors or fat burning and thermogenesis enhancers. During the use of slimming enhancers, many women also changed their dietary patterns; such alterations were not necessarily nutritionally correct, due to either lack of knowledge on the principles of proper nutrition (which apply also during a weight-loss action) or due to abandoning these rules when applying the slimming aids. The observed dietary behaviours of the women may underlie serious nutrition-related health problems when they reach their middle age, but can also result in nutritional disorders in the near future.


A clinical histologic study of bovine hydroxyapatite in combination with autogenous bone and fibrin glue for maxillary sinus floor augmentation. Results after 6 to 8 months of healing.
Biopsies were taken from 16 out of 20 consecutive referral patients 6 to 8 months after maxillary sinus floor augmentation with a mixture of bovine hydroxyapatite (BH), autogenous bone particles and fibrin glue. Four days prior to biopsy retrieval the patients were given a single dose of tetracycline to label bone forming sites. Fluorescence microscopy of 100 microm thick sections revealed active bone formation in conjunction with the BH particles in 14 of 15 specimens analysed. Light microscopy and morphometry of ground sections from 16 patients showed various amounts of mineralised bone tissue in all except one specimen. In the latter case, the BH particles were encapsulated by a dense fibrous connective tissue. Sections from the augmented areas were occupied by non-mineralized tissue (54.1+12.6%), lamellar bone (21.2+24.5%), BH particles (14.5+10.3%) and woven bone (10.2+13.4%). The non-mineralized tissue seen in bone forming areas consisted of a loose connective tissue, rich of vessels and cells, and in the periphery of a more dense fibrous connective tissue. Woven bone with large and scattered osteocyte lacunae was bridging between the BH particles and the lamellar trabecular bone. There were no signs of resorption of the BH particles. The lamellar bone appeared to have originated from the recipient site and was seldom in contact with the BH particles. It is concluded that the tested implant material has bone conducting properties. The bone associated with the BH particles after 6 to 8 months of healing was mainly woven.


Levodopa improves motor deficits but can further disrupt cognition in a macaque Parkinson model.
Levodopa effectively relieves motor symptoms in Parkinson's disease (PD), but has had inconsistent effects on cognition, even worsening some aspects of cognitive functioning. Therefore, remediation of PD cognitive deficits is a major unmet need. However, drug development efforts have been hampered by lack of an animal model in which motor and cognitive deficits can be examined simultaneously. Cynomolgus monkeys were trained to perform cognitive tasks and then chronically exposed to MPTP to slowly produce cognitive and motor deficits of parkinsonism. Administration of L-dopa to these animals dose dependently improved motor functioning, but did not significantly improve cognitive performance. At doses that maximally improved motor function, additional cognitive deficits were observed. The present model of MPTP-induced parkinsonism recapitulates important motor and cognitive aspects of PD. Results with L-dopa mirror data derived from PD patients. This model should allow more efficient testing of potential PD therapeutics to evaluate motor and cognitive functions simultaneously. © 2012 Movement Disorder Society.


Transferrin receptors of Neisseria meningitidis: promising candidates for a broadly cross-protective vaccine.
Production of a meningococcal vaccine capable of generating long-lasting immunity in all age groups is still a high priority worldwide. Iron-regulated outer-membrane proteins have attracted considerable attention in recent years and it has become increasingly evident that the meningococcal transferrin-binding proteins, TBP1 and TBP2, have characteristics compatible with a safe and broadly cross-reactive vaccine candidate. Both TBPs are surface-exposed and immunogenic in man and animals, and antibodies to their native structure are bactericidal to homologous and many heterologous strains. These include strains from various serogroups, serotypes and serosubtypes, with no obvious correlation between bactericidal activity and the identity of the strains or the molecular mass of the heterogeneous TBP2 molecule. A meningococcal vaccine based on, or enriched with, undenatured TBPs from one or more strains, in combination with conventional polysaccharide-based vaccines, might increase the spectrum of strains against which protection can be achieved to include serogroup B strains. In this review, the structure-function and immunological properties of TBP1 and TBP2 are discussed.


Excipient foods: designing food matrices that improve the oral bioavailability of pharmaceuticals and nutraceuticals.
The oral bioavailability of many lipophilic bioactive agents (pharmaceuticals and nutraceuticals) is limited due to various physicochemical and physiological processes: poor release from food or drug matrices; low solubility in gastrointestinal fluids; metabolism or chemical transformation within the gastrointestinal tract; low epithelium cell permeability. The bioavailability of these agents can be improved by specifically designing food matrices that control their release, solubilization, transport, metabolism, and absorption within the gastrointestinal tract. This article discusses the impact of food composition and structure on oral bioavailability, and how this knowledge can be used to design excipient foods for improving the oral bioavailability of lipophilic bioactives. Excipient foods contain ingredients or structures that may have no bioactivity themselves, but that are able to promote the bioactivity of co-ingested bioactives. These bioactives may be lipophilic drugs in pharmaceutical preparations (such as capsules, pills, or syrups) or nutraceuticals present within food matrices (such as natural or processed foods and beverages).


Acidity rate and fluoride content of consumed beverages in kerman/iran.
High consumption of nonalcoholic beverages is one of the causes of dental erosion. Annual consumption of beverages in Iran is 48 LPCY (Lit per capita per year) that is equal to 144 bottles per capita in year and is relatively a high rate. Taking this into consideration, the present study was planned to determine the pH and fluoride content of beverages consumed in Kerman, south east of Iran. In a descriptive cross-sectional study during 2006-2007, samples of any type of beverages in Kerman were taken once every 4 months and each sample was examined for its acidity and fluoride content in ambient and refrigerator temperatures. A total of 744 experiments were conducted on 62 brands of beverages with different colors to determine the acidity and on 31 brands of beverages to determine their fluoride content. Acidity was determined by pH-meter in the ambient temperature and refrigerator temperature (+4◦C). Fluoride content was determined using specific electrode method. Data analysis was done by SPSS software package. Mean acidity of the consumed beverages in Kerman was 2.55 and 2.78 mg/L in the ambient temperature and refrigerator temperature, respectively. Mean fluoride concentration in the temperature of the ambient and refrigerator were 1.13 and 1.27 mg/L respectively. Therefore beverages consumption in Kerman had a higher acidity than national standard. Since some of the consumed beverages in Kerman are distributed and consumed in other parts of Iran as well, necessary precautions for controlling dental erosion especially in Kerman residents should be taken.


The 2D and 3D MR arthrographic description of aponeurotic expansion of supraspinatus tendon and biceps tendon anomaly in a large patient cohort.
To describe the aponeurotic expansion of supraspinatus tendon (AEST) and biceps tendon abnormalities with magnetic resonance (MR) arthrographic examinations and determine their prevalence in patients, we performed a high-resolution 3D direct MR arthrography. This was a retrospective study of 700 shoulder MR arthrograms performed between May 2010 and January 2022. Extension in the coronal plane of an AEST on 3D fat-suppressed T1-weighted volumetric interpolated breath-hold examination (VIBE) MR arthrography was identified. Based on its morphology, the AEST on MR arthrography was divided into four subtypes: absence of tendinous thickness in the bicipital synovial surface or intra-synovial tendon-like structure in the bicipital groove, thin and flat tendinous thickness ≥1 mm of bicipital synovium, oval tendinous structure less than half the size of the adjacent biceps tendon, oval tendinous structure more than half the size of the adjacent biceps tendon, and oval tendinous structure larger than the adjacent biceps tendon. Based on its origin and termination, aponeurotic expansions can be divided into three subtypes: proximal pulley zone, middle humeral neck zone, and distal myotendinous junction zone. Association with the biceps synovium of the AEST was categorized into three types: intra-synovial, extra-synovial, and trans-synovial. An AEST in the anterior shoulder joint in 3D VIBE MR arthrography images was identified in 63 (9%) of 700 arthrograms. The most common arthrographic type of AEST was type 1-this was detected in 39 of 63 patients. The most common course type of the AEST was anteriorly midline. The most common distal insertion type was at the tenosynovial sheath of the long head of the biceps tendon (LHBT) in the middle humeral neck zone-this was detected in 31 of 63 patients. There were only 10 MR arthrograms biceps tendon abnormality, including 4 biceps agenesis and 6 split ruptures. A 2D and high-resolution 3D MR arthrography can demonstrate the anatomical detail around the bicipital groove and facilitate the differentiation between a biceps tendon anomaly and an AEST. On high-resolution 3D MR arthrographic images, the AEST tends to be in the anterior midline and anteromedial portions of the biceps synovium with intra-synovial, extra-synovial, and trans-synovial courses and its three different insertion types.


Preimplantation genetic diagnosis: technology and clinical applications.
Preimplantation genetic diagnosis (PGD) is a method by which embryos formed through in vitro fertilization (IVF) can be tested for single-gene disorders or chromosome abnormalities prior to embryo transfer. This enables couples to significantly improve their chances of having a healthy child. PGD is an important addition to conventional prenatal diagnosis for genetic disorders. PGD is a complex combination of various technologies that requires close collaboration of a team of specialists for optimal patient care. This review article will cover patient management, assisted reproductive technologies including IVF and PGD as well as indications for PGD. Clinical vignettes from The Froedtert Hospital and Medical College of Wisconsin Reproductive Medicine Clinic PGD Program will be presented, including the first single-gene disorder PGD performed in Wisconsin. These vignettes highlight the importance of a detailed family history, use of PGD for cases of recurrent miscarriage, and the use of PGD for spinal muscular atrophy.


Genetic and voltage-clamp analysis of a Drosophila potassium channel.
These genetic and voltage-clamp studies have produced evidence that strongly supports the hypothesis that the Shaker locus codes for at least one molecular component of the IA channel. Mutations at the Shaker locus have effects that are specific to IA, and thus a general membrane component is not produced by this locus. Regarding the function of the Shaker product itself, there is evidence that it is at least involved in the inactivation and recovery process of the channel. The evidence so far is also consistent with there being a single Shaker product per channel. Regarding its genetic and developmental properties, IA is apparently completely independent of the other major voltage-activated K+ channel in the membrane, IK. However, only after molecular information becomes available will it become clear whether or not these experiments have revealed clues about the molecular nature of these channels. Ultimately, voltage-clamp experiments, even combined with resourceful genetic tools, can only reveal circumstantial information about the molecular structure of channels. It is hoped, however, that these studies have stimulated interest in the Shaker locus and that the way has been well prepared for molecular studies of the IA channel. The actual molecular characterization of the Shaker locus is already under way (cf. Jan et al., this volume; M. Tanouye , pers. comm .).


[Effect of biliary decompression on the results of the surgical treatment of gunshot wounds of the liver].
The purpose of this study consisted in providing experimental grounds to prove the efficiency of the biliferous tract decompression at surgical treatment of gunshot wounds of the liver. The experiments were carried out on 39 dogs. It was ascertained that within 3 to 5 days after gunshot injury to the liver the conditions for normal bile discharge from the liver into the duodenum get hampered. The decompression of the biliferous system in the postoperative period resulted in the decrease of the severity and duration of posttraumatic bile decompression and exerted an appreciable effect upon the lowering of the lethality.


Dietary lysophospholipids reduce lymphatic cholesterol transport compared with dietary phospholipids in thoracic lymph-duct cannulated rats.
Dietary phospholipids have been traditionally known to affect micelle formation. Egg yolk-derived lysophospholipids (LysoPL) are commercially available. We investigated the effects of dietary LysoPL on lymphatic lipid transport. We also compared sn-1 LysoPL and sn-2 LysoPL, which have different fatty acyl esterification positions. Thoracic lymph duct-cannulated rats were fed a diet supplemented with egg yolk-derived sn-1 LysoPL, sn-2 LysoPL, or phospholipids (PL). The amount of lymphatic lipid transport was also evaluated. Time courses of transport were applied to the one-compartment model as one of the pharmacokinetic analyses. The solubility of cholesterol in bile acid micelles was measured. Compared to the PL diet, the sn-1 and sn-2 LysoPL diets significantly reduced the lymphatic transport of cholesterol. There were no differences in the lymphatic PL and TAG transport. There was no difference in cholesterol transport between the sn-1 LysoPL group and the sn-2 LysoPL group; however, the transport rate constant at a decrease in lymphatic cholesterol was lower in the sn-1 LysoPL group than in the sn-2 LysoPL group. Cholesterol solubility in bile acid micelles was significantly decreased in the sn-1 LysoPL and sn-2 LysoPL groups compared to that in the PL group. Dietary LysoPL affects the behavior of intestinal cholesterol and suppresses lymphatic cholesterol transport.


Aphthous Ulcer of the Vocal Cord Leading to Hoarseness.
Acute or chronic laryngitis, laryngeal trauma, vocal cord paralysis, cysts of the vocal cord, and benign or malignant tumors of the vocal folds are frequent causes of hoarseness. It is important to identify the cause of hoarseness to rule out potential malignancies. In this case report, we describe a patient with a bilateral aphthous ulcer in the vocal cords who was admitted with hoarseness and had been previously diagnosed with Behcet disease. Behcet disease is characterized by mucosal aphthous lesions, which are found primarily in oral and genital regions. To the best of our knowledge, this is the first instance of a patient with bilateral aphthous ulcer in the vocal cords.


Brain growth of the domestic pig (Sus scrofa) from 2 to 24 weeks of age: a longitudinal MRI study.
An animal model with brain growth similar to humans, that can be used in MRI studies to investigate brain development, would be valuable. Our laboratory has developed and validated MRI methods for regional brain volume quantification in the neonatal piglet. The aim of this study was to utilize the MRI-based volume quantification technique in a longitudinal study to determine brain growth in domestic pigs from 2 to 24 weeks of age. MRI data were acquired from pigs 2-24 weeks of age using a 3-dimensional magnetization-prepared gradient echo sequence on a Magnetom Trio 3-tesla imager. Manual segmentation was performed for volume estimates of total brain, cortical, diencephalon, brainstem, cerebellar and hippocampal regions. Logistic modeling procedures were used to characterize brain growth. Total brain volume increased 130% (±12%) and 121% (±7%) from 2 to 24 weeks in males and females, respectively. The maximum increase in total brain volume occurred about the age of 4 weeks and 95% of whole brain growth occurred by the age of 21-23 weeks. Logistical modeling suggests there are sexually dimorphic effects on brain growth. For example, in females, the cortex was smaller (p = 0.04). Furthermore, the maximum growth of the hippocampus occurred about 5 weeks earlier in females than males, and the window for hippocampal growth was significantly shorter in females than males (p = 0.02, p = 0.002 respectively). These sexual dimorphisms are similar to what is seen in humans. In addition to providing important data on brain growth for pigs, this study shows pigs can be used to obtain longitudinal MRI data. The large increase in brain volume in the postnatal period is similar to that of human neonates and suggests pigs can be used to investigate brain development.


Effects of maintaining normal plasma colloid osmotic pressure on renal function and excretion of sodium and water after major surgery. A randomized study.
A prospective, randomized study of patients undergoing abdominal aortic surgery was undertaken to determine the effects of maintaining normal plasma colloid osmotic pressure (COPp) on postoperative renal function and excretion of water and electrolytes. Two groups of 13 patients were given whole blood transfusions to replace blood loss. One group (ALB) received 80 g albumin on the day of operation and 20 g albumin the following three days. The other group (NON-ALB) received no extra albumin. The glomerular filtration rate (GFR) did not change significantly from preoperative values on the first or fourth postoperative day in any of the groups. The differences between the groups were non-significant. COPp could not be correlated to either GFR or sodium or fluid balances. Postoperative sodium and water retention was found to depend on the amount of infused sodium. Unexplained fluid shifts to a "third space" did not occur. Administration of albumin in addition to quantitative blood loss replacement is unnecessary and expensive.


A novel contact model for AFM indentation experiments on soft spherical cell-like particles.
The use of the simple Hertz model for the analysis of Atomic Force Microscopy (AFM) force-distance curves measured on soft spherical cell-like particles leads to significant underestimations of the objects Young's modulus E. To correct this error, a mixed double contact model (based on the simple Hertz model and the Johnson-Kendall-Roberts (JKR) model) was derived. The model considers two independent particle deformation sites: (i) the upper part of the particle is deformed by the AFM indenter, (ii) the bottom part is deformed by the substrate, which is usually unnoticed. It becomes apparent that for soft particles even small forces between substrate and particle can influence the resulting force-distance curves. For instance we show, that a gravity-induced compression on the particle bottom side can have significant influence on the measurements. To highlight these observations, the deviation of the particle Young's modulus E between the simple Hertz model and our model is calculated. This error strongly depends on the ratio of the three involved radii: (i) the radius of the AFM indenter, (ii) the radius of the particle and (iii) the radius of the substrate as well as on the acting gravity force. Overall, the analysis suggests that for nanoscopic indenters the deviation is negligible, whereas the use of microscopic indenters results in significant errors that can be corrected via the presented model. This is important especially for very soft particles, since larger indenters can achieve higher signal to noise ratios. Furthermore, the applicability of the model was confirmed by indentation experiments on hydrogel microbeads. The mixed double contact model is applicable to a large range of indenter geometries and can be adapted for other contact models.


Selective peripheral nerve recording using simulated human median nerve activity and convolutional neural networks.
It is difficult to create intuitive methods of controlling prosthetic limbs, often resulting in abandonment. Peripheral nerve interfaces can be used to convert motor intent into commands to a prosthesis. The Extraneural Spatiotemporal Compound Action Potentials Extraction Network (ESCAPE-NET) is a convolutional neural network (CNN) that has previously been demonstrated to be effective at discriminating neural sources in rat sciatic nerves. ESCAPE-NET was designed to operate using data from multi-channel nerve cuff arrays, and use the resulting spatiotemporal signatures to classify individual naturally evoked compound action potentials (nCAPs) based on differing source fascicles. The applicability of this approach to larger and more complex nerves is not well understood. To support future translation to humans, the objective of this study was to characterize the performance of this approach in a computational model of the human median nerve. Using a cross-sectional immunohistochemistry image of a human median nerve, a finite-element model was generated and used to simulate extraneural recordings. ESCAPE-NET was used to classify nCAPs based on source location, for varying numbers of sources and noise levels. The performance of ESCAPE-NET was also compared to ResNet-50 and MobileNet-V2 in the context of classifying human nerve cuff data. Classification accuracy was found to be inversely related to the number of nCAP sources in ESCAPE-NET (3-class: 97.8% ± 0.1%; 10-class: 89.3% ± 5.4% in low-noise conditions, 3-class: 70.3% ± 0.1%; 10-class: 52.5% ± 0.3% in high-noise conditions). ESCAPE-NET overall outperformed both MobileNet-V2 (3-class: 96.5% ± 1.1%; 10-class: 84.9% ± 1.7% in low-noise conditions, 3-class: 86.0% ± 0.6%; 10-class: 41.4% ± 0.9% in high-noise conditions) and ResNet-50 (3-class: 71.2% ± 18.6%; 10-class: 40.1% ± 22.5% in low-noise conditions, 3-class: 81.3% ± 4.4%; 10-class: 31.9% ± 4.4% in high-noise conditions). All three networks were found to learn to differentiate nCAPs from different sources, as evidenced by performance levels well above chance in all cases. ESCAPE-NET was found to have the most robust performance, despite decreasing performance as the number of classes increased, and as noise was varied. These results provide valuable translational guidelines for designing neural interfaces for human use.


Simultaneous oxidation of Hg0 and NH3-SCR of NO by nanophase Ce x Zr y Mn z O2 at low temperature: the interaction and mechanism.
Simultaneous oxidation of Hg0 and NH3-SCR of NO by catalyst is one of the key methods for co-purification of coal-fired flue gas. Till now, the interaction between the oxidation of Hg0 and NH3-SCR of NO and its mechanism have not clarified. In this study, a series of nanophase Ce x Zr y Mn z O2 was prepared for the simultaneous oxidation of Hg0 and NH3-SCR of NO at low temperature. The catalysts were characterized using surface area analysis, X-ray diffraction, temperature-programmed techniques, and several types of microscopy and spectroscopy. The experimental results indicated that the Ce0.47Zr0.22Mn0.31O2 exhibited superior Hg0 removal efficiency (> 99%) and NO conversion efficiency (> 90%) even at 150 °C, and it also exhibited a good durability in the presence of SO2 and H2O. The excellent performance of Ce0.47Zr0.22Mn0.31O2 on co-purifying Hg0 and NO was due to the stronger synergistic effects of Ce-Zr-Mn in Ce0.47Zr0.22Mn0.31O2 than that of the others, which was illustrated by the characterization results of XPS, XRD, and FT-IR. Moreover, it was found that the NO conversion of Ce0.47Zr0.22Mn0.31O2 could be slightly influenced by Hg0 and was decreased about 4% to the max, while that of Hg0 could rarely be affected by the selected catalytic reduction process of NO. It might be due to the co-purification mechanism of NO and Hg0. The mechanism of the simultaneous oxidation of Hg0 and NH3-SCR of NO was mainly due to the synergetic effect on the mobility of surface oxygen and the activation of lattice oxygen of Ce0.47Zr0.22Mn0.31O2. The effect of the oxidation of Hg0 on the NH3-SCR of NO was mainly due to the absorbed Hg0/Hg2+ on the surface of Ce0.47Zr0.22Mn0.31O2, which attenuated the formation of NH3(ad), -NH2(ad), and NH4+ on its acid sites. Similarly, the NH3-SCR of NO process could hardly influence the oxidation of Hg0 when NO and Hg0 were co-purified.


Suppression of potyvirus infection by coexpressed closterovirus protein.
A tobacco etch virus (TEV)-based expression vector has been used for insertion of several ORFs derived from the unrelated beet yellows virus (BYV). Hybrid TEV variants expressing the BYV capsid protein, 20-kDa protein, or HSP70 homolog systemically infected Nicotiana tabacum and stably retained BYV sequences. In contrast, insertion of the ORF encoding BYV leader proteinase (L-Pro) resulted in severely impaired systemic transport and accumulation of recombinant TEV. Progeny of this virus underwent various deletions affecting the L-Pro sequence and mitigating the defects in virus spread. Model experiments involving several spontaneous and engineered mutants indicated that the central domain of BYV L-Pro was responsible for the defect in hybrid virus accumulation, whereas full-size L-Pro was required for maximal debilitation of systemic transport. Strikingly, BYV L-Pro expression did not debilitate systemic infection of hybrid TEV in Nicotiana benthamiana plants. No major defects in replication or encapsidation of recombinant RNA were revealed in N. tabacum protoplasts. These results indicated that BYV L-Pro specifically interfered with TEV systemic transport and accumulation in a host-dependent manner and suggested a potential utility of closterovirus L-Pro as an inhibitor of potyvirus infection. In addition, it was demonstrated that the 107-amino-acid-residues-long N-terminal part of the TEV helper component proteinase is not essential for systemic infection.


Obesity, diabetes, and hyperlipidemia in a central Australian aboriginal community with a long history of acculturation.
To determine the age- and sex-specific prevalence of diabetes and to examine associations between related anthropometric and metabolic abnormalities in an Aboriginal community in central Australia with a long history of acculturation. We used a cross-sectional survey of 353 adults > 15 yr of age (87% response rate) and measured the following parameters: weight, height, circumferences of waist and hips; glucose, insulin, cholesterol, triglyceride, and high-density lipoprotein cholesterol in fasting plasma; and plasma glucose and insulin 2 h after 75 g oral glucose. The prevalence of diabetes was 29.6% in survey participants > 35 yr of age and 5.3% in those < 35 yr of age. Impaired glucose tolerance also occurred with higher frequency in those > 35 yr of age (14.8 vs. 4.7%). Of those > 35 yr of age, 75% of the women and 51% of the men were overweight or obese, with a body mass index > or = 25 kg/m2. A large insulin response to oral glucose was evident, with the upper tertile of the 2-h insulin response six times higher than the lower tertile (113 +/- 43 vs. 19 +/- 8 mU/L). Hyperinsulinemia showed a strong, positive association with impaired glucose tolerance, body mass index, waist-to-hip ratio, cholesterol, and triglyceride levels and a negative association with high-density lipoprotein cholesterol levels. Cholesterol levels were on average 0.5 mM higher in men than in women. Deteriorations in carbohydrate and lipid metabolism occurred before 40 yr of age: diabetes, body mass index, waist-to-hip ratio, and fasting triglycerides and cholesterol concentrations peaked and high-density lipoprotein cholesterol concentrations reached their nadir at the end of the fourth decade. These data suggest that any intervention programs developed to prevent or reduce diabetes prevalence in this population should be targeted at adolescents and young adults.


Involvement of Mfn2, Bcl2/Bax signaling and mitochondrial viability in the potential protective effect of Royal jelly against mitochondria-mediated ovarian apoptosis by cisplatin in rats.
The current study aimed to assess cisplatin-mediated ovarian apoptosis in a rat model by Royal jelly (RJ). Thirty female adult albino rats (180-200 g) were divided into three groups (n=10): saline (0.9% NaCl, IP) was given to the control group, the cisplatin group: received (5 mg/kg/once a week IP) for 5 successive weeks, the RJ+Cis. group: received RJ (100 mg/kg/ day PO daily), and Cisplatin (5 mg/kg/once per week IP) for 5 successive weeks. At the end of the experiment, rats were sacrificed and their ovaries were isolated and used for biochemical analysis, molecular investigations and morphometric assessment as well as histological study. Moreover, blood samples were collected for determination of follicle-stimulating hormone (FSH), luteinizing hormone (LH), Estradiol, progesterone and anti-mullerian hormone (AMH). The current study clarified that RJ given to rats prior to cisplatin significantly increased the ovarian and uterine weights, in addition to follicular count at P˂0.05 compared to rats injected only with cisplatin. Moreover, it restored normal ovarian histological structure with a concurrent reduction in FSH, and LH levels, and increased AMH and ovarian hormone concentrations at P˂0.05 compared to cisplatin group. Also, RJ decreased the ovarian antioxidant/oxidative imbalance harmonized with significant suppression of inducible nitric oxide synthase and increase of quinone oxidoreductase 1 mRNA expression at P˂0.05 compared to cisplatin group. We concluded that RJ could alleviate mitochondrial-induced ovarian apoptosis caused by cisplatin via increasing anti-apoptotic Bcl2, and diminishing pro-apoptotic Bax with a concomitant increase of Mfn2 mRNA and protein expressions.


Reproductive isolation and morphogenetic evolution in Drosophila analyzed by breakage of ethological barriers.
This article reports the breaking of ethological barriers through the constitution of soma-germ line chimeras between species of the melanogaster subgroup of Drosophila, which are ethologically isolated. Female Drosophila yakuba and D. teissieri germ cells in a D. melanogaster ovary produced functional oocytes that, when fertilized by D. melanogaster sperm, gave rise to sterile yakuba-melanogaster and teissieri-melanogaster male and female hybrids. However, the erecta-melanogaster and orena-melanogaster hybrids were lethal, since female D. erecta and D. orena germ cells in a D. melanogaster ovary failed to form oocytes with the capacity to develop normally. This failure appears to be caused by an altered interaction between the melanogaster soma and the erecta and orena germ lines. Germ cells of D. teissieri and D. orena in a D. melanogaster testis produced motile sperm that was not stored in D. melanogaster females. This might be due to incompatibility between the teissieri and orena sperm and the melanogaster seminal fluid. A morphological analysis of the terminalia of yakuba-melanogaster and teissieri-melanogaster hybrids was performed. The effect on the terminalia of teissieri-melanogaster hybrids of a mutation in doublesex, a regulatory gene that controls the development of the terminalia, was also investigated.


Resection Prolongs Overall Survival for Nonmetastatic Midgut Small Bowel Neuroendocrine Tumors: A National Cancer Data Base Study.
This study aimed to understand if resection (RS) for nonmetastatic small bowel neuroendocrine tumors (SBNETs) prolongs 5-year overall survival. Patients from National Cancer Data Base with primary histologically confirmed SBNETs from 2007 to 2016 were included. Patients younger than 18 years, with the disease in the duodenum/Meckel diverticulum or metastatic disease were excluded. We assessed 5-year survival rates using Kaplan-Meier curves and multivariate Cox proportional hazards regression after RS, nonresection surgical management (NRS), or no resection (NR). Multivariate models were adjusted with age, sex, race, insurance, Charlson-Deyo comorbidity score, academic facility, primary tumor location, clinical T, clinical N, stage, and grade. We identified 4180 patients. On average, patients were 64 years old (standard deviation, 12 years), male (53%), and White (84%). The majority received RS (91.8%) as opposed to NRS (4.0%) or NR (4.2%). Patients who received RS versus NR had increased survival rates (84.2% vs 73.9%; univariate log-rank, P < 0.0001; multivariate hazard ratio, 0.73; 95% confidence interval, 0.53-0.99; P = 0.04). No statistical difference in survival was observed for NRS versus NR. To our knowledge, this is the first national study to evaluate survival after RS for nonmetastatic SBNETs. Results suggest that RS of SBNETs may prolong 5-year survival.


Psilocybin-assisted therapy for reducing alcohol intake in patients with alcohol use disorder: protocol for a randomised, double-blinded, placebo-controlled 12-week clinical trial (The QUANTUM Trip Trial).
Alcohol use disorder is a difficult-to-treat psychiatric disorder and a major burden on public health. Existing treatment efficacy is moderate, and relapse rates are high. Preliminary findings suggest that psilocybin, a psychedelic compound, can safely and reliably occasion highly meaningful experiences that may spur a positive change in drinking behaviour when administered in a therapeutic context. However, the efficacy of a single psilocybin administration and its potential neurobiological underpinnings still remain unknown. To establish efficacy, we will investigate the effects of psilocybin-assisted therapy versus placebo in a randomised, double-blinded, placebo-controlled 12-week clinical trial. Ninety treatment-seeking patients, aged 20-70 years, diagnosed with alcohol use disorder will be recruited from the community via advertisement and referrals from general practitioners or specialised treatment units. The psilocybin or placebo will be administered in accordance with a protocol for psychological support before, during and after the dosing. Outcome assessments will be carried out 1, 4, 8 and 12 weeks postdosing. The primary outcome is reduction in the percentage of heavy drinking days from baseline to follow-up at 12 weeks. Key secondary outcomes are as follows: (1) total alcohol consumption, (2) phosphatidyl-ethanol, an objective biomarker for alcohol, (3) plasma psilocin, the active metabolite, to establish a possible therapeutic range, (4) the acute subjective drug experience as a possible predictor of treatment outcome and (5) neuronal response to alcohol cues and cognitive flexibility within corticostriatal pathways by use of functional MR brain imaging 1-week postdosing. Ethical approval has been obtained from the Committee on Health Research Ethics of the Capital Region of Denmark (H-20043832). All patients will be provided oral and written information about the trial before screening. The study results will be disseminated by peer-review publications and conference presentations. EudraCT 2020-000829-55 and NCT05416229.


Thrips tabaci population genetic structure and polyploidy in relation to competency as a vector of tomato spotted wilt virus.
Knowledge of population-level genetic differences can help explain variation among populations of insect vectors in their role in the epidemiology of specific viruses. Variation in competency to transmit Tomato spotted wilt virus (TSWV) that exists among populations of Thrips tabaci has been associated with the presence of cryptic species that exhibit different modes of reproduction and host ranges. However, recent findings suggest that vector competency of T. tabaci at any given location depends on the thrips and virus populations that are present. This study characterizes the population genetic structure of T. tabaci collected from four locations in North Carolina and examines the relationship between population genetic structure and variation in TSWV transmission by T. tabaci. Mitochondrial COI sequence analysis revealed the presence of two genetically distinct groups with one characterized by thelytokous, parthenogenetic reproduction and the other by arrhenotokous, sexual reproduction. Using a set of 11 microsatellite markers that we developed to investigate T. tabaci population genetic structure, we identified 17 clonal groups and found significant genetic structuring among the four NC populations that corresponded to the geographic locations where the populations were collected. Application of microsatellite markers also led to the discovery of polyploidy in this species. All four populations contained tetraploid individuals, and three contained both diploid and tetraploid individuals. Analysis of variation in transmission ofTSWV among isofemale lines initiated with individuals used in this study revealed that 'clone assignment,' 'virus isolate' and their interaction significantly influenced vector competency. These results highlight the importance of interactions between specific T. tabaci clonal types and specific TSWV isolates underlying transmission of TSWV by T. tabaci.


A Comparative Analysis of Selection Pressures Suffered by Mitochondrial Genomes in Two Planthopper Species with Divergent Climate Distributions.
Mitochondrial DNA (mtDNA) has been widely used as a valuable tool in studies related to evolution and population genetics, under the implicit assumption of neutral evolution. However, recent studies suggest that natural selection also plays a significant role in shaping mitochondrial genome evolution, although the specific driving forces remain elusive. In this study, we aimed to investigate whether and how climate influences mitochondrial genome evolution by comparing the selection pressures acting on mitochondrial genomes between two rice planthoppers, Sogatella furcifera (Horváth) and Laodelphax striatellus (Fallén), which have different climate distributions. We employed the dN/dS method, MK test and Tajima's D tests for our analysis. Our results showed that the mitochondrial genomes of the two species appear to undergo predominantly purifying selection, consistent with the nearly neutral evolution model. However, we observed varied degrees of purifying selection among the 13 protein-coding genes. Notably, ND1, ND2, ND6, COIII, and ATP8 exhibited significantly stronger purifying selection and greater divergence between the two species compared to the other genes. Additionally, we observed relatively stronger purifying selection in the mitochondrial genomes of S. furcifera compared to L. striatellus. This difference could be attributed to varying metabolic requirements arising from distinct habitats or other factors that are unclear here. Furthermore, we speculate that mito-nuclear epistatic interactions may play a role in maintaining nonsynonymous polymorphisms, particularly for COI and COII. Overall, our results shed some light on the influence of climate on mitochondrial genome evolution.


Ammonia decomposition catalysis using lithium-calcium imide.
Lithium-calcium imide is explored as a catalyst for the decomposition of ammonia. It shows the highest ammonia decomposition activity yet reported for a pure light metal amide or imide, comparable to lithium imide-amide at high temperature, with superior conversion observed at lower temperatures. Importantly, the post-reaction mass recovery of lithium-calcium imide is almost complete, indicating that it may be easier to contain than the other amide-imide catalysts reported to date. The basis of this improved recovery is that the catalyst is, at least partially, solid across the temperature range studied under ammonia flow. However, lithium-calcium imide itself is only stable at low and high temperatures under ammonia, with in situ powder diffraction showing the decomposition of the catalyst to lithium amide-imide and calcium imide at intermediate temperatures of 200-460 °C.


Immunoidentification of type XII collagen in embryonic tissues.
We have generated a monoclonal antibody against a synthetic peptide whose sequence was derived from the nucleotide sequence of a cDNA encoding alpha 1(XII) collagen. The antibody, 75d7, has been used to identify the alpha 1(XII) chain on immunoblots of SDS-PAGE tendon extracts as a 220-kD polypeptide, under reducing conditions. Amino-terminal amino acid sequence analysis of an immunopurified cyanogen bromide fragment of type XII collagen from embryonic chick tendons gave a single sequence identical to that predicted from the cDNA, thus confirming that the antibody recognizes the type XII protein. Immunofluorescence studies with the antibody demonstrate that type XII collagen is localized in type I-containing dense connective tissue structures such as tendons, ligaments, perichondrium, and periosteum. With these data, taken together with previous results showing that a portion of the sequence domains of type XII collagen is similar to domains of type IX, a nonfibrillar collagen associated with cross-striated fibrils in cartilage, we suggest that types IX and XII collagens are members of a distinct class of extracellular matrix proteins found in association with quarter-staggered collagen fibrils.


The impact of COVID-19 on oncology professionals-one year on: lessons learned from the ESMO Resilience Task Force survey series.
COVID-19 has had a significant impact on the well-being and job performance of oncology professionals globally. The European Society for Medical Oncology (ESMO) Resilience Task Force collaboration set out to investigate and monitor well-being since COVID-19 in relation to work, lifestyle and support factors in oncology professionals 1 year on since the start of the pandemic. An online, anonymous survey was conducted in February/March 2021 (Survey III). Key outcome variables included risk of poor well-being or distress (expanded Well-Being Index), feeling burnout (single item from expanded Well-Being Index), and job performance since COVID-19. Longitudinal analysis of responses to the series of three surveys since COVID-19 was carried out, and responses to job demands and resources questions were interrogated. SPSS V.26.0/V.27.0 and GraphPad Prism V9.0 were used for statistical analyses. Responses from 1269 participants from 104 countries were analysed in Survey III: 55% (n = 699/1269) female, 54% (n = 686/1269) >40 years, and 69% (n = 852/1230) of white ethnicity. There continues to be an increased risk of poor well-being or distress (n = 464/1169, 40%) and feeling burnout (n = 660/1169, 57%) compared with Survey I (25% and 38% respectively, P < 0.0001), despite improved job performance. Compared with the initial period of the pandemic, more participants report feeling overwhelmed with workload (45% versus 29%, P < 0.0001). There remain concerns about the negative impact of the pandemic on career development/training (43%), job security (37%). and international fellowship opportunities (76%). Alarmingly, 25% (n = 266/1086) are considering changing their future career with 38% (n = 100/266) contemplating leaving the profession. Oncology professionals continue to face increased job demands. There is now significant concern regarding potential attrition in the oncology workforce. National and international stakeholders must act immediately and work closely with oncology professionals to draw up future-proof recovery plans.


Genomic relationships and speciation times of human, chimpanzee, and gorilla inferred from a coalescent hidden Markov model.
The genealogical relationship of human, chimpanzee, and gorilla varies along the genome. We develop a hidden Markov model (HMM) that incorporates this variation and relate the model parameters to population genetics quantities such as speciation times and ancestral population sizes. Our HMM is an analytically tractable approximation to the coalescent process with recombination, and in simulations we see no apparent bias in the HMM estimates. We apply the HMM to four autosomal contiguous human-chimp-gorilla-orangutan alignments comprising a total of 1.9 million base pairs. We find a very recent speciation time of human-chimp (4.1 +/- 0.4 million years), and fairly large ancestral effective population sizes (65,000 +/- 30,000 for the human-chimp ancestor and 45,000 +/- 10,000 for the human-chimp-gorilla ancestor). Furthermore, around 50% of the human genome coalesces with chimpanzee after speciation with gorilla. We also consider 250,000 base pairs of X-chromosome alignments and find an effective population size much smaller than 75% of the autosomal effective population sizes. Finally, we find that the rate of transitions between different genealogies correlates well with the region-wide present-day human recombination rate, but does not correlate with the fine-scale recombination rates and recombination hot spots, suggesting that the latter are evolutionarily transient.


Precision Medicine in Neonates: Future Perspectives for the Lung.
Bronchopulmonary dysplasia (BPD) is the most common complication of pre-term birth with long lasting sequelae. Since its first description more than 50 years ago, many large randomized controlled trials have been conducted, aiming to improve evidence-based knowledge on the optimal strategies to prevent and treat BPD. However, most of these intervention studies have been performed on a population level without regard for the variation in clinical and biological diversity (e.g., gestational age, ethnicity, gender, or disease progression) between patients that is driven by the complex interaction of genetic pre-disposition and environmental exposures. Nevertheless, clinicians provide daily care such as lung protective interventions on an individual basis every day despite the fact that research supporting individualized or precision medicine for monitoring or treating pre-term lungs is immature. This narrative review summarizes four potential developments in pulmonary research that might facilitate the process of individualizing lung protective interventions to prevent development of BPD. Electrical impedance tomography and electromyography of the diaphragm are bedside monitoring tools to assess regional changes in lung volume and ventilation and spontaneous breathing effort, respectively. These non-invasive tools allow a more individualized optimization of invasive and non-invasive respiratory support. Investigation of the genomic variation in caffeine metabolism in pre-term infants can be used to optimize and individualize caffeine dosing regimens. Finally, volatile organic compound analysis in exhaled breath might accurately predict BPD at an early stage of the disease, enabling clinicians to initiate preventive strategies for BPD on an individual basis. Before these suggested diagnostic or monitoring tools can be implemented in daily practice and improve individualized patient care, future research should address and overcome their technical difficulties, perform extensive external validation and show their additional value in preventing BPD.


Two dimensional NMR spectroscopic approaches for exploring plant metabolome: A review.
Today, most investigations of the plant metabolome tend to be based on either nuclear magnetic resonance (NMR) spectroscopy or mass spectrometry (MS), with or without hyphenation with chromatography. Although less sensitive than MS, NMR provides a powerful complementary technique for the identification and quantification of metabolites in plant extracts. NMR spectroscopy, well appreciated by phytochemists as a particularly information-rich method, showed recent paradigm shift for the improving of metabolome(s) structural and functional characterization and for advancing the understanding of many biological processes. Furthermore, two dimensional NMR (2D NMR) experiments and the use of chemometric data analysis of NMR spectra have proven highly effective at identifying novel and known metabolites that correlate with changes in genotype or phenotype. In this review, we provide an overview of the development of NMR in the field of metabolomics with special focus on 2D NMR spectroscopic techniques and their applications in phytomedicines quality control analysis and drug discovery from natural sources, raising more attention at its potential to reduce the gap between the pace of natural products research and modern drug discovery demand.


Stable STIM1 Knockdown in Self-Renewing Human Neural Precursors Promotes Premature Neural Differentiation.
Ca2+ signaling plays a significant role in the development of the vertebrate nervous system where it regulates neurite growth as well as synapse and neurotransmitter specification. Elucidating the role of Ca2+ signaling in mammalian neuronal development has been largely restricted to either small animal models or primary cultures. Here we derived human neural precursor cells (NPCs) from human embryonic stem cells to understand the functional significance of a less understood arm of calcium signaling, Store-operated Ca2+ entry or SOCE, in neuronal development. Human NPCs exhibited robust SOCE, which was significantly attenuated by expression of a stable shRNA-miR targeted toward the SOCE molecule, STIM1. Along with the plasma membrane channel Orai, STIM is an essential component of SOCE in many cell types, where it regulates gene expression. Therefore, we measured global gene expression in human NPCs with and without STIM1 knockdown. Interestingly, pathways down-regulated through STIM1 knockdown were related to cell proliferation and DNA replication processes, whereas post-synaptic signaling was identified as an up-regulated process. To understand the functional significance of these gene expression changes we measured the self-renewal capacity of NPCs with STIM1 knockdown. The STIM1 knockdown NPCs demonstrated significantly reduced neurosphere size and number as well as precocious spontaneous differentiation toward the neuronal lineage, as compared to control cells. These findings demonstrate that STIM1 mediated SOCE in human NPCs regulates gene expression changes, that in vivo are likely to physiologically modulate the self-renewal and differentiation of NPCs.


Response properties of visual interneurons to motion stimuli in the praying mantis, Tenodera aridifolia.
Intracellular responses of motion-sensitive visual interneurons were recorded from the lobula complex of the mantis, Tenodera aridifolia. The interneurons were divided into four classes according to the response polarity, spatial tuning, and directional selectivity. Neurons of the first class had small, medium, or large receptive fields and showed a strong excitation in response to a small-field motion such as a small square moving in any direction (SF neurons). The second class neurons showed non-directionally selective responses: an excitation to a large-field motion of gratings in any direction (ND neurons). Most ND neurons had small or medium-size receptive fields. Neurons of the third class had large receptive fields and exhibited directionally selective responses: an excitation to a large-field motion of gratings in preferred direction and an inhibition to a motion in opposite, null direction (DS neurons). The last class neurons had small receptive fields and showed inhibitory responses to a moving square and gratings (I neurons). The functional roles of these neurons in prey recognition and optomotor response were discussed.


Pharmacoscreening, molecular dynamics, and quantum mechanics of inermin from Panax ginseng: a crucial molecule inhibiting exosomal protein target associated with coronary artery disease progression.
Exosomes, microvesicles, carry and release several vital molecules across cells, tissues, and organs. Epicardial adipose tissue exosomes are critical in the development and progression of coronary artery disease (CAD). It is hypothesized that exosomes may transport causative molecules from inflamed tissue and deliver to the target tissue and progress CAD. Thus, identifying and inhibiting the CAD-associated proteins that are being transported to other cells via exosomes will help slow the progression of CAD. This study uses a systems biological approach that integrates differential gene expression in the CAD, exosomal cargo assessment, protein network construction, and functional enrichment to identify the crucial exosomal cargo protein target. Meanwhile, absorption, distribution, metabolism, and excretion (ADME) screening of Panax ginseng-derived compounds was conducted and then docked against the protein target to identify potential inhibitors and then subjected to molecular dynamics simulation (MDS) to understand the behavior of the protein-ligand complex till 100 nanoseconds. Finally, density functional theory (DFT) calculation was performed on the ligand with the highest affinity with the target. Through the systems biological approach, Mothers against decapentaplegic homolog 2 protein (SMAD2) was determined as a potential target that linked with PI3K-Akt signaling, Ubiquitin mediated proteolysis, and the focal adhesion pathway. Further, screening of 190 Panax ginseng compounds, 27 showed drug-likeness properties. Inermin, a phytochemical showed good docking with -5.02 kcal/mol and achieved stability confirmation with SMAD2 based on MDS when compared to the known CAD drugs. Additionally, DFT analysis of inermin showed high chemical activity that significantly contributes to effective target binding. Overall, our computational study suggests that inermin could act against SMAD2 and may aid in the management of CAD.


Production of infectious hepatitis C virus of various genotypes in cell cultures.
A unique hepatitis C virus (HCV) strain JFH-1 has been shown to replicate efficiently in cell culture with production of infectious HCV. We previously developed a DNA expression system containing HCV cDNA flanked by two self-cleaving ribozymes to generate HCV particles in cell culture. In this study, we produced HCV particles of various genotypes, including 1a (H77), 1b (CG1b), and 2a (J6 and JFH-1), in the HCV-ribozyme system. The constructs also contain the secreted alkaline phosphatase gene to control for transfection efficiency and the effects of culture conditions. After transfection into the Huh7-derived cell line Huh7.5.1, continuous HCV replication and secretion were confirmed by the detection of HCV RNA and core antigen in the culture medium. HCV replication levels of strains H77, CG1b, and J6 were comparable, whereas the JFH-1 strain replicates at a substantially higher level than the other strains. To evaluate the infectivity in vitro, the culture medium of JFH-1-transfected cells was inoculated into naive Huh7.5.1 cells. HCV proteins were detected by immunofluorescence 3 days after inoculation. To evaluate the infectivity in vivo, the culture medium from HCV genotype 1b-transfected cells was inoculated into a chimpanzee and caused a typical course of HCV infection. The HCV 1b propagated in vitro and in vivo had sequences identical to those of the HCV genomic cDNA used for cell culture transfection. The development of culture systems for production of various HCV genotypes provides a valuable tool not only to study the replication and pathogenesis of HCV but also to screen for antivirals.


Spatial spectral modeling for robust MRSI.
We propose a novel spatial spectral model for the reconstruction of magnetic resonance spectroscopic imaging (MRSI) signal. We penalize the compartmentalized spatial total variation norm of the signal to exploit the spatial properties of the metabolite peaks. The spectral signal is modeled as a sparse linear combination of spikes and polynomials to capture the peaks and baseline induced by unsuppressed water and lipids. We also use the high-resolution map of the magnetic field distribution within the slice to model the image acquisition, thus correcting for intra-voxel line shape distortions. The spectral model enables the stable recovery of the signal even in challenging spatial regions, while the spatial model suppresses the spectral leakage from extra-cranial fat and inter-voxel crosstalk. We acquire the MRSI signal using EPSI, while the high-resolution 3-D MRI information is derived using Dixon scans. The reconstruction of phantom and in vivo MRSI data demonstrate a significant improvement in spectral quality and accuracy over classical MRSI schemes.


Vibrio parahaemolyticus FlaJ, a homologue of FliS, is required for production of a flagellin.
The flaA locus of Vibrio parahaemolyticus encodes one of the four polar flagellin genes, the flagellum-capping protein HAP2, and three additional flagellar genes. Sequence analysis downstream of the gene encoding HAP2 revealed the region to be similar to the fliD (HAP2) locus of Pseudomonas aeruginosa. The deduced protein sequences for the newly identified genes suggest that one protein belongs to the family of transcriptional regulatory proteins known to interact with sigma (54), one may be a rod component of the flagellum, and one resembles the FliS protein. fliS is an essential flagellar gene in many bacteria; however its function is not clear. The V. parahaemolyticus polar flaC flagellin gene was poorly expressed in Escherichia coli Production of FlaC was stimulated by provision of the flaA locus in trans. Dissection of this locus revealed that the fliS-like gene, flaJ, was required for increased expression of flaC. Stimulation by FlaJ occurred in E.coli mutants defective in either the master flagellar-controlling operon or the gene encoding the flagellar sigma (28). Therefore the effect of FlaJ was not mediated through flagellar proteins. Nor was it mediated through sigma (54) for enhanced FlaC production was observed in mutants with defects in the gene encoding sigma (54).


An observation on combined use of chemotherapy and traditional Chinese medicine to relieve cancer pain.
We have treated 50 patients with stage III, VI malignant tumors confirmed by pathology. The patients were divided into two groups. One group was treated by combination of chemotherapy and traditional Chinese medicine (treatment group); the other only by chemotherapy (control group). The effect of cancer treatment was evaluated according to the criteria of WHO. The results showed that the effective rate was 80% in treatment group and 52% in control group. The pain relieving rate was 68% in treatment group and 40% in control group (P < 0.01). This fact demonstrates that the application of traditional Chinese medicine can invigorate blood circulation, eliminate blood stasis, soften hardness and dissolve the mass, nourish blood and increase vigor. This kind of application can not only enhance the effect of cancer treatment but also increase the cancer pain relieving rate.