Phosphorylation of Vif and its role in HIV-1 replication.
Vif is a 23-kDa protein encoded by human immunodeficiency virus, type 1 (HIV-1) which is important for virion infectivity. Here, we describe the phosphorylation of HIV-1 Vif and its role in HIV-1 replication. In vivo studies demonstrated that Vif is highly phosphorylated on serine and threonine residues. To identify phosphorylation sites and characterize the Vif kinase(s), Vif was expressed in Escherichia coli and purified for use as a substrate in in vitro kinase assays. The purified Vif protein was phosphorylated in vitro on serine and threonine residues by a kinase(s) present in both cytosol and membrane fractions. Phosphorylation of Vif was stimulated by phorbol 12-myristate 13-acetate and inhibited by staurosporine and hypericin, a drug with potent anti-HIV activity. The Vif kinase(s) was resistant to inhibitors of protein kinase C, cAMP-dependent kinase, and cGMP-dependent kinase, suggesting that it is distinct from these enzymes. To identify the phosphorylation sites, 32P-labeled Vif was digested by V8 protease and the peptides were resolved by reverse-phase high performance liquid chromatography. Radioactive peptide sequencing identified three phosphorylation sites within the C terminus, Ser144, Thr155, and Thr188. Two-dimensional tryptic phosphopeptide mapping indicated that these sites are also phosphorylated in vivo. Both Ser144 and Thr188 are contained in the recognition motifs (R/KXXS*/T* and R/KXXXS*/T*) used by serine/threonine protein kinases such as cGMP-dependent kinase and PKC. Ser144 is present in the motif SLQXLA, which is the most highly conserved sequence among all lentivirus Vif proteins. Mutation of Ser144 to alanine resulted in loss of Vif activity and >90% inhibition of HIV-1 replication. These studies suggest that phosphorylation of Vif by a serine/threonine protein kinase(s) plays an important role in regulating HIV-1 replication and infectivity.


Arachidonic acid closes innexin/pannexin channels and thereby inhibits microglia cell movement to a nerve injury.
Pannexons are membrane channels formed by pannexins and are permeable to ATP. They have been implicated in various physiological and pathophysiological processes. Innexins, the invertebrate homologues of the pannexins, form innexons. Nerve injury induces calcium waves in glial cells, releasing ATP through glial pannexon/innexon channels. The ATP then activates microglia. More slowly, injury releases arachidonic acid (ArA). The present experiments show that ArA itself reduced the macroscopic membrane currents of innexin- and of pannexin-injected oocytes; ArA also blocked K(+) -induced release of ATP. In leeches, whose large glial cells have been favorable for studying control of microglia migration, ArA blocked glial dye-release and, evidently, ATP-release. A physiological consequence in the leech was block of microglial migration to nerve injuries. Exogenous ATP (100 µM) reversed the effect, for ATP causes activation and movement of microglia after nerve injury, but nitric oxide directs microglia to the lesion. It was not excluded that metabolites of ArA may also inhibit the channels. But for all these effects, ArA and its non-metabolizable analog eicosatetraynoic acid (ETYA) were indistinguishable. Therefore, ArA itself is an endogenous regulator of pannexons and innexons. ArA thus blocks release of ATP from glia after nerve injury and thereby, at least in leeches, stops microglia at lesions.


Disease specificity and dynamics of changes in IgA class anti-endomysial antibodies in celiac disease.
We evaluated the sensitivity and kinetics of serum IgA class anti-endomysial antibodies in the diagnosis of celiac disease (CD) as established by the criteria of the European Society for Pediatric Gastroenterology and Nutrition (ESPGAN). Eighty-four cases that satisfied the ESPGAN criteria for CD were evaluated for IgA-EmA titers during various phases of establishing the diagnosis. Thirty-three cases were infants and children less than 5 years of age undergoing intestinal biopsies for symptoms of CD and 51 were previously diagnosed adults. Of the 33 children, 11 were untreated and symptomatic and were IgA-EmA positive at initial presentation. Twenty-two children previously controlled on a gluten-free diet (GFD) exhibited IgA-EmA titers during gluten challenge. Furthermore, the antibody levels declined in all cases (usually to negative) when the patients were again placed on a GFD for 6-12 months. Changes in intestinal histopathology paralleled the changes in antibody titers in six cases undergoing serial biopsies. Of the 51 adult patients with proven CD who were prescribed a GFD for at least 12 months, IgA-EmA were detected in 10 cases who were noncompliant to their GFD, whereas the antibodies was found in only 1 of the remaining 41 patients strictly adhering to their diet. The sera of 140 aged-matched children with various intestinal problems, 87 healthy adults, and 67 patients with dermatological diseases served as controls and were also IgA-EmA negative. On the basis of these findings, we suggest a role for the IgA-EmA as a serological screening test for active CD. It further offers the potential for monitoring compliance to diet in established cases of CD and also indicates the proper timing for biopsy in patients undergoing evaluation of CD.


Stem cell factor and c-kit gene expression and protein localization in the sheep ovary during fetal development.
The aim of this study was to investigate stem cell factor and c-kit gene expression and protein localization in the mesonephros and ovary of sheep fetuses at different days of gestation, using RNA in situ hybridization and immunohistochemical procedures. At days 24 and 26 of gestation, stem cell factor mRNA and protein were present in cells throughout the developing gonad and mesonephros. From day 28 to day 40 of gestation, stem cell factor mRNA and protein became increasingly localized to the cortical region of the ovary, where most germ cells were present as actively proliferating oogonia. From day 40 to day 90 of gestation, stem cell factor mRNA and protein localization were confined mainly to the ovarian cortex. Moreover, within the cortical region, stem cell factor mRNA was low or absent where follicles were first forming and highest in the outer ovarian cortex, where germ cells were undergoing mitosis or the early stages of meiosis. In contrast, stem cell factor protein was present in newly forming follicles, as well as in mitotic and meiotic germ cells, which is consistent with the presence of both membrane-bound and soluble forms of this ligand. However, by day 90 of gestation, both stem cell factor mRNA and protein were observed in the granulosa cells of most (> 90%) primordial follicles. C-kit mRNA and protein were observed from day 24 of gestation in both germ cells and somatic cells but, with increasing gestational age, preferentially in germ cells (for example, pre-meiotic germ cells and both isolated oocytes and follicle-enclosed oocytes). C-kit protein, but not mRNA, was also observed in germ cells that were undergoing meiosis. The results indicate that the cells containing stem cell factor mRNA within the ovary up to day 90 of gestation originated from the gonadal blastema and from cells that migrated from the mesonephros before day 28 of gestation. Since stem cell factor mRNA was absent in both mesonephric cells migrating after day 28 of gestation and in regions where follicles were first forming, it is suggested that these later migrating mesonephric cells are the progenitors of the granulosa cells in the first forming follicles. In conclusion, during follicle formation, c-kit mRNA is localized to germ cells whereas c-kit, together with stem cell factor protein, is localized to both germ cells and somatic cells, consistent with the hypothesis that the presence of this receptor-ligand pair is essential to prevent apoptosis.


Carcinoid tumor of the middle ear.
Although carcinoid tumors are labeled as neuroendocrine tumors they can also originate in tissue lacking neuroendocrine cells, such as that in the middle ear. Symptoms of a carcinoid tumor in the middle ear are common ear symptoms such as fullness, pain and hearing loss. Carcinoid tumors have also been considered to be slow-growing. Both these aspects can easily lead to a relatively late diagnosis of carcinoid tumor of the middle ear. The diagnosis is made histologically, and the tumor is primarily treated surgically. In the follow-up of patients, octreotide scanning has proved to be a sensitive method in cases of both recurrence and metastasis. Our patient was a 34-year-old, otherwise healthy female with left-sided acute otitis media and facial palsy in her left ear. She had also suffered from the same symptoms 4 years earlier. She was treated with an operation, and the histologic diagnosis was a carcinoid tumor. In the follow-up of the patient we used octreotide scanning.


In vitro Effects of Methylprednisolone Acetate on Equine Deep Digital Flexor Tendon-Derived Cells.
Primary deep digital flexor tendon (DDFT) pathologies and those accompanying degenerative changes of navicular bone fibrocartilage are major causes of lameness associated with navicular disease. Intrasynovial corticosteroids are mainstay in the treatment due to the anti-inflammatory effects, but their effect on DDFT cell biosynthesis are unknown. The objective of this in-vitro study was to investigate the effects of methylprednisolone acetate (MPA) on cells isolated from the dorsal fibrocartilaginous region of forelimb DDFTs (DDFT-derived cells) of 5 horses (aged 11-17 years). Non-adherent aggregate cultures were established from third passage cells over a 72 to 96-h duration prior to treating with medium containing 0 (control), 0.05 and 0.5 mg/mL MPA for 24 h. Tendon and cartilage extracellular matrix (ECM) related gene expression, cell aggregate and culture medium GAG contents, culture medium collagen and MMP-3 and-13 concentrations were measured. After 24 h of treatment, only the higher MPA concentration (0.5 mg/mL) significantly down-regulated tendon ECM related genes; whereas, both MPA doses significantly down-regulated cartilage ECM related genes. MPA treatment did not affect the total GAG content of DDFT-derived cells or total GAG, soluble collagen and MMP-3 and-13 contents in culture medium compared to untreated controls. Future studies to determine the response of DDFT-derived cells with longer exposure times to corticosteroids and in the presence of inflammatory cytokines are necessary. These results are a first step in assessing the effects of intrasynovial medications on equine DDFT, for which currently no information exists.


Two-stent telescoping technique for securing an atrially embolized venous stent.
Coronary sinus stenosis or atresia is a rare problem in congenital heart disease. It is sometimes associated with ventricular dysfunction. In this setting, relief of the obstruction may improve myocardial perfusion and contractility. During percutaneous stent deployment within the coronary sinus, distal embolization of the stent into the atrial chambers occurred. The patient had double inlet left ventricle, severe ventricular dysfunction, interrupted IVC, mechanical aortic valve, and a Fontan circulation. The intervention was performed as a rescue procedure prior to transplantation referral. In this report we describe use of telescoping technique, i.e., deploying the distal end of a long stent within the embolized stent while traversing its proximal end across the stenosed coronary sinus ostium. This secured the embolized stent and at the same time relieved the coronary sinus stenosis.


Neutrophils increase paracellular permeability of restituted ischemic-injured porcine ileum.
We have previously shown minimal evidence of neutrophil infiltration during early reperfusion of porcine ischemic ileum. However, we noted marked neutrophil infiltration 6 to 18 hours after ischemia during mucosal repair. We postulated such neutrophil infiltration would disrupt restituting epithelium. Pigs were pretreated with anti-CD11/CD18 monoclonal antibody, superoxide dismutase-polyethylene glycol, or saline solution before inducing 1 hour of ischemia. Pigs recovered for up to 18 hours, after which mucosal repair was assessed. One hour of ischemia induced loss of 19 +/- 7% of the villous epithelial surface area. Epithelial restitution covered the mucosal defect within 2 hours, although full recovery of mucosal barrier function required 6 hours. By 18 hours, a significant decrease in transepithelial electrical resistance and increase in transmucosal mannitol flux was noted despite the continued presence of complete epithelial coverage. Accumulation of neutrophils within restituting epithelium was noted on histologic examination, associated with electron-microscopic evidence of widened paracellular spaces. Pretreatment with anti-CD11/CD18 monoclonal antibody and superoxide dismutase-polyethylene glycol significantly reduced neutrophil infiltration and normalized transepithelial electrical resistance and mannitol fluxes. Mucosal inflammation during epithelial repair resulted in increased paracellular permeability as neutrophils traversed restituted epithelium. Blocking neutrophil adhesion or scavenging superoxide prevented mucosal dysfunction in recovering tissue.


[Application of acellular bovine pericardium patch in implant based immediate breast reconstruction].
Objective: To examine the clinical effect of acellular bovine pericardium patch in implant based immediate breast reconstruction. Methods: The clinicopathological information of 141 breast cancer patients, who admitted to Department of Breast Reconstruction and Oncoplastic Surgery, Tianjin Medical University Cancer Hospital, underwent immediate mammoplasty with implants combined with acellular bovine pericardium patches were analyzed from June 2016 to October 2019. All patients were female, with the age of (38.8±8.5) years (range: 13 to 60 years). The body mass index was (21.9±2.5) kg/m2 (range: 16.0 to 32.3 kg/m2). There were 39 cases of duct carcinoma in situ, 46 cases of stage Ⅰ, 40 cases of stage Ⅱ and 16 cases of stage Ⅲ. All patients received nipple-areola-sparing mastectomy or skin-sparing mastectomy with sentinel lymph node biopsy or axillary lymph node dissection, and prosthesis implantation with sub-pectoralis combined with breast patch. The correlation of clinicopathological characters and complications was assessed by t test, χ2 test, Fisher's exact probability method and Logistic regression. Pre-and post-operative aesthetic, quality of life scores were recorded. Results: The operation time (M(IQR)) was 3.6(1.5) hours (range: 3.0 to 6.5 hours). The early postoperative complication rate was 22.0% (31/141), prosthesis removal was the main postoperative complication, accounting for 64.5% (20/31) of the total complications, of which 15 cases occurred in the first 30 patients. The follow-up time was 28(8) months (range: 20 to 53 months), The most frequent long-term complications were capsular contracture and implant displacement, with the incidence of 11.2% (14/125) and 10.4% (13/125), respectively. Multivariate analysis showed that prosthesis volume ≥300 ml (OR=8.173, 95%CI: 1.302 to 51.315, P=0.021) and peri-areolar incision (OR=7.809, 95%CI: 2.162 to 28.211, P<0.01) were independent relative factors for the occurrence of short-term postoperative local complications. After 2 years of operation, the score of breast appearance satisfaction was 71.7±15.5, postoperative effect satisfaction was 90.4±9.5, psychological satisfaction was 90.7±17.1, sexual satisfaction was 70.1±25.1. The immediate postoperative satisfaction rate at discharge was 95.4% (134/141), and 17.6% (22/125) of patients had the intention to received revision surgery. Conclusions: Prosthesis volume ≥300 ml and peri-areolar incision were independent realtive factors for short-term local complications after bovine pericardium patch combined with prosthesis implantation in the immediate breast reconstruction. After completing the learning curve, the postoperative complications of the procedure could be decreased.


Impact of the nature of the capillary wall on plasma refilling during hemodialysis.
Our aim was to clarify the impact of the nature of the capillary wall, defined by the contribution of large (LP), small (SP), and ultrasmall (UP) pores, on plasma refilling in a hemodialysis session. This study included data from 78 patients. The relative blood volume change (ΔBV%) was monitored using a Crit-Line monitor. A bioimpedance device was used to measure extracellular and intracellular fluid volumes, and the excess fluid mass (MExF) was calculated. We simulated blood volume change (sΔBV%) based on a three-pore model. Hydraulic permeability of the capillary wall (LpS) and fractional contribution of LP to LpS (αLP) were determined by fitting sΔBV to ΔBV. The total refilling volume (TVref) was calculated from the total ultrafiltration volume and total blood volume change. Values were standardized to a body surface area of 1.73 m2 and are denoted by the subscript BSA. LpS and αLP were 3.09 (2.32, 4.68) mL/mmHg/min and 0.069 (0.023, 0.109), respectively. The standardized regression coefficient (β) of the ultrafiltration rate (UFRBSA) and initial excess fluid mass (MExF,BSA,0) by multiple linear regression analysis of TVref,BSA without (Model 1) and with (Model 2) αLP were as follows: UFRBSA, 0.714/<0.001 (β/p); MExF,BSA,0, 0.247/<0.001 (Model 1); UFRBSA, 0.799/<0.001; MExF,BSA,0, 0.066/0.237; and αLP, -0.327/<0.001 (Model 2). The impact of volume overload (MExF,BSA,0) on plasma refilling became insignificant with the addition of αLP in the model, suggesting that the nature of the capillary wall described by inter-endothelial gaps (LP) may have a greater impact on plasma refilling than volume overload.


Recognition of complex patterned substrates by heteropolymer chains consisting of multiple monomer types.
We propose a statistical mechanical model of surface pattern recognition by heteropolymers with quenched monomer sequence distribution. The chemically heterogeneous pattern consists of different adsorption sites specifically distributed on a surface. The heteropolymer sequence is complementary with respect to the pattern. The concepts of recognition probability and recognition temperature are introduced. The algorithm for calculating the recognition probability is based on efficient recurrence procedures for evaluating the single-chain partition function of a chain macromolecule consisting of multiple monomer types, which interact with multiple types of adsorption sites. The temperature dependencies of the recognition probability are discussed. We address the critical role of the commensurability between the heteropolymer sequence and the distribution of the surface adsorbing sites on the polymer adsorption. Also, we address the question of how many types of monomer units in the heteropolymer are required for unambiguous recognition of compact target patterns. It is shown that perfect pattern recognition can be achieved for the strong-adsorption regime in the case of specifically structured compact patterns with multifunctional adsorption sites and heteropolymers with multiple monomer types when the degeneracy of the ground state is suppressed. The pattern recognition ability increases with the number of different types of monomer units and complementary adsorption sites. For random heteropolymers and patterns, the free energy change associated with the recognition process decreases linearly with increasing this number. Correlated random heteropolymers are capable of recognizing related patterns on a random background.


[Imputation of missing data].
In medical research missing data are sometimes inevitable. Different missingness mechanisms can be distinguished: (a) missing completely at random; (b) missing by design; (c) missing at random, and (d) missing not at random. If participants with missing data are excluded from statistical analyses, this can lead to biased study results and loss of statistical power. Imputation methods can be applied to estimate missing values; multiple imputation gives a good idea of the inaccuracy of the reconstructed measurements. The most common imputation methods assume that missing data are missing at random. Multiple imputation contributes greatly to the efficiency and reliability of estimates because maximum use is made of the data collected. Imputation is not meant to obviate low-quality data.


Retrospective evaluation of palliative care in Romania: the perspective of the involved professionals.
Palliative care at home is a new service provision in Romania. This study evaluated retrospectively, after the patient's death, the provision of care at home from the perspective of professionals directly involved in the care process. The evaluation included assessment of the role of the professional, the functioning of the team, communication with patients and family, and overall judgment. The study is part of the development of palliative care at home, by multidisciplinary teams in Romania. Over a period of eighteen months five teams with a total of 19 professionals, provided palliative care for 103 patients with cancer in its terminal stage. During this period 80 patients died. A questionnaire was sent to the professionals involved. All responded resulting in 181 evaluations. These evaluations are the basis for analysis. The data were analyzed using SPSS. Of the 181 cases evaluated, 63 indicated an emotional burden on the professionals. GP's and nurses reported such a burden more frequently. A lack of knowledge of how to treat a special case was reported 56 times. This was especially so when a case was judged to be different from other cases, which often included (unexpected) complications. GP's reported cases with complications more frequently than oncologists and nurses. The multidisciplinary teams functioned very satisfactorily in treating patients. Although communication with patients/families was generally judged positively, communication with patients was sometimes viewed as problematic. The overall verdict was that the care delivered to the 80 patients was positive. Most professionals reported that they would deal with the cases in the same way again. The patients who received palliative care and died, were treated well according to the professionals who evaluated the care process. Improvement of palliative care services at home may be achieved by provision of additional information on (acute) complications. Since palliative care at home is a new phenomenon in Romania, professionals working closely with the patient have to learn to cope with the emotional burden certain cases may include.


Sclerostin Inhibition: A Novel Target for the Treatment of Postmenopausal Osteoporosis.
Osteoporosis, a widespread skeletal disorder with a substantial economic load, is characterized by increased porosity of the bones resulting in vulnerability to fractures. When activated, the canonical Wnt signaling pathway results in osteoblastogenesis and bone formation. A Wnt ligand forms a complex with low-density lipoprotein receptor-related proteins 5 and 6 (Lrp5/6) and stimulates intracellular signaling cascades, leading to nuclear translocation of β-catenin and transcription of downstream molecules involved in osteoblast differentiation, maturation, and survival. Sclerostin (SOST), a glycoprotein produced by osteocytes, is an extracellular Wnt antagonist that blocks the binding of Wnt ligands to Lrp5/6, preventing the activation of the pathway and osteoblast-mediated bone formation subsequently. Inhibition of SOST represents a new therapeutic paradigm for the treatment of osteoporosis. Monoclonal antibodies to SOST include romosozumab, blosozumab, and setrusumab. With its unique dual effect of increasing bone formation (anabolic action) and decreasing bone resorption, the Food and Drug Administration approved romosozumab, a promising new treatment for postmenopausal osteoporosis. Its efficacy and safety have been established in trials. However, patients at high risk of cardiovascular or cerebrovascular events should not be prescribed romosozumab.


Pediatric pulmonology year in review 2015: Part 3.
Our journal covers a broad range of research and scholarly topics related to children's respiratory disorders. For updated perspectives on the rapidly expanding knowledge in our field, we will summarize the past year's publications in our major topic areas, as well as selected publications in these areas from the core clinical journal literature outside our own pages. The current review (Part 3) covers articles on asthma, diagnostic testing/endoscopy, respiratory complications of neuromuscular disorders, and rare lung diseases. Pediatr Pulmonol. 2016;51:747-753. © 2016 Wiley Periodicals, Inc.


Elevated levels of S-nitrosoalbumin in preeclampsia plasma.
The availability of nitric oxide (NO), which is required for the normal regulation of vascular tone, may be decreased in preeclampsia, thus contributing to the vascular pathogenesis of this pregnancy disorder. Because ascorbate is essential for the decomposition of S-nitrothiols and the release of NO, we speculated that the ascorbate deficiency typical of preeclampsia plasma might result in decreased rates of decomposition of S-nitrosothiols. We tested the hypothesis that total S-nitrosothiol and S-nitrosoalbumin concentrations are increased in preeclampsia plasma, reflecting a decreased release of NO from these major reservoirs of NO. Gestationally matched plasma samples were obtained (before labor or intravenous MgSO(4)) from 21 women with preeclampsia and 21 women with normal pregnancy, and plasma samples were also obtained from 12 nonpregnant women of similar age and body mass index during the follicular phase of the menstrual cycle. All were nonsmokers. The assay included ultraviolet-induced decomposition of S-nitrosothiols to liberate NO captured by a florigenic reagent, 4,5-diaminofluoresceine, to produce diaminofluoresceine-Triazole. Preeclampsia plasma contained significantly higher concentrations of total S-nitrosothiols (11.1+/-2.9 nmol/mL) than normal pregnancy samples (9.4+/-1.5 nmol/mL). Even greater differences were found between preeclampsia plasma and plasma samples from normal pregnancies and nonpregnant women (294+/-110, 186+/-25, and 151+/-25 pmol/mg protein, respectively) when S-nitrosothiol content was expressed per milligram protein. The albumin fraction contained 49.4% of total plasma S-nitrosothiols in the control samples and 53.7% and 56.8% of plasma S-nitrosothiols in normal pregnancy and preeclampsia, respectively. The level of S-nitrosoalbumin was significantly higher in preeclampsia than in normal pregnancy or nonpregnancy plasma (6.3+/-1.4, 5.1+/-0.7, and 4.2+/-1.0 nmol/mL, respectively). The increased concentration of S-nitrosoalbumin in preeclampsia almost completely accounted for the increased levels of S-nitrosothiols in total plasma. Due to combined increases in nitrosothiols and decreases in protein, the preeclampsia plasma concentration of S-nitrosoalbumin was greatly increased on a per milligram of protein basis (271% and 186% compared with normal nonpregnancy and normal pregnancy plasma, respectively). We conclude that S-nitrosoalbumin and total S-nitrosothiol concentrations are significantly increased in preeclampsia plasma and may reflect insufficient release of NO groups in this condition.


Laboratory diagnosis of congenital syphilis by immunoglobulin M (IgM) and IgA immunoblotting.
We screened cord blood or serum samples from 101 infants at risk for congenital syphilis and serum samples from their mothers for immunoglobulin G (IgG), IgM, and IgA antibodies to Treponema pallidum by western blotting (immunoblotting). Clinical evaluation showed that six infants had signs and/or symptoms consistent with congenital syphilis. The sera from five of these infants were IgM blot positive, and four were IgA blot positive. Four asymptomatic infants had serologic evidence of congenital syphilis. The sera from three of these infants were IgM blot positive, and two were IgA blot positive. However, the IgM reactivity of the serum from one asymptomatic infant, which was also IgA positive, was abolished by protein G treatment. An IgM capture enzyme-linked immunosorbent assay corroborated the presence of IgM antibodies in six of seven IgM blot-reactive sera. Overall, for detection of symptomatic congenital syphilis, a sensitivity of 83% for IgM blotting and 67% for IgA blotting was obtained. The significance of positive IgM or IgA Western blots for asymptomatic infants requires further study to confirm infection in these infants.


Epimedium flavonoids counteract the side effects of glucocorticoids on hypothalamic-pituitary-adrenal axis.
Our previous studies demonstrated that the epimedium herb, when simultaneously used with GCs, counteracted suppressive effects of GCs on the HPA axis without adverse influence on the therapeutic action of GCs. Here, total flavones were extracted from the epimedium flavonoids (EFs) and then used to investigate whether EFs provide protective effects on the HPA axis. We found that GCs induced a significant decrease in body weight gain, adrenal gland weight gain, and plasma adrenocorticotropin (ACTH) and corticosterone levels. After treatment with EFs, body weight gain, adrenal gland weight gain, and plasma corticosterone level were significantly restored, whilst plasma ACTH level was partially elevated. EFs were also shown to promote cell proliferation in the outer layer of adrenal cortex and to enhance the migration of newly divided cells toward the inner layer. To elucidate the underlying mechanisms, the mRNA expression of insulin-like growth factor II (IGF-II) was measured, and EFs significantly upregulated IGF-II expression. Our results indicated that EFs counteract the suppression of the HPA axis induced by GCs. This may involve both the ACTH and IGF-II pathways and thereby promote regeneration of the adrenal cortex suggesting a potential clinical application of EFs against the suppressive effects of GCs on the HPA axis.


Mini Monoka silicone monocanalicular lacrimal stents: Subjective and objective outcomes.
BACKGROUND. Canalicular injuries and anomalies are relatively common. Despite this, controversy still exists regarding the indications for surgery and best surgical methods. The most favourable intervention would be one that is simple, associated with a high level of subjective and objective outcome, and does not threaten the uninjured or unaffected part of the lacrimal drainage system. This study assesses outcomes of intervention with the Mini Monoka silicone monocanalicular lacrimal stent. METHOD. A single cohort, hospital-based study with retrospective and prospective components. Participants were all identifiable patients at Bristol Eye Hospital in whom the Mini Monoka silicone monocanalicular stent was used for any indication. Retrospective analysis of patient records for all related pre-operative, operative and postoperative data was performed. Prospective analysis of subjective outcome via confidential patient questionnaire and objective outcome via clinical examination was performed. RESULTS. Complete data were obtainable in 13 patients (14 canaliculi) of the identified 22 patients (23 canaliculi). Follow-up was from 12 to 70 months (mean 39 months). A high level of subjective outcome was noted with no patients experiencing significant or disabling symptoms. In terms of objective outcome, 79% achieved canalicular patency. The main complications with this technique were premature stent loss (29%) and stent migration (14%). CONCLUSIONS. The Mini Monoka monocanalicular stent is a safe, effective and simple surgical technique that, unlike bicanalicular procedures, does not threaten the uninjured / unaffected part of the lacrimal drainage system.


Lack of association of VEGF promoter polymorphisms with sporadic ALS.
The authors tested the association of three vascular endothelial growth factor (VEGF) promoter polymorphisms with sporadic ALS (SALS) to verify the results of a previous study and to investigate their modifier effects on the subphenotypes of SALS in a large family-based and case-control cohort of North American white subjects (N = 1,603). They did not find any association of the VEGF promoter polymorphisms with SALS or its subphenotypes, suggesting that they do not have a direct causal role in ALS.


"Physician, heal thyself": how teaching holistic medicine differs from teaching CAM.
The term complementary and alternative medicine (CAM) has been adopted to describe a system of health care not generally recognized as part of mainstream medical practice. It is often conflated with an older term, holistic medicine, which can briefly be defined as the art and science of healing the whole person-body, mind, and spirit-in relation to that person's community and environment. Coursework in CAM is now offered in at least two thirds of U.S. medical schools. There is also a growing number of courses in the medical humanities and in spirituality and health. However, courses explicitly designed to introduce students to the principles and practices of holistic medicine are unusual. The author describes the fundamental differences between CAM and holistic medicine, highlighting holistic medicine's emphasis on the promotion of healthy lifestyles for practitioners and patients alike. He argues that offering physicians-to-be more coursework in holistic medicine could lay the groundwork for future physicians' adopting and modeling healthy lifestyles.


Residential arrangements for adults with cerebral palsy--California, 1988.
In the United States, an estimated 274,000 persons have cerebral palsy (CP), a neurologic condition defined as a group of nonprogressive disorders in which an abnormality of the central nervous system can result in motor dysfunction (e.g., paresis, involuntary movement, and incoordination). CP is the third leading cause of the need for assistance with basic life activities and the fifth leading cause of activity limitation. An adult with CP may require adaptive housing to improve accessibility (e.g., to entrances and toilet facilities), attendant care to assist with activities of daily living, and/or nursing care to meet specific health-care needs. These needs can be met through a variety of residential accommodations. However, because the residential environment has a considerable impact on well-being and quality of life, the accommodations should be in the least restrictive environment and, when possible, community-based. This report summarizes an assessment of characteristics of adults with CP to determine which factors are associated with placement in a more restrictive environment.


Role of Autoantibodies to N-Methyl-d-Aspartate (NMDA) Receptor in Relapsing Herpes Simplex Encephalitis: A Retrospective, One-Center Experience.
Post-herpes simplex virus encephalitis relapses have been recently associated with autoimmunity driven by autoantibodies against N-methyl-d-aspartate (NMDA) receptors. Because it offers different treatment options, determination of this condition is important. Between 2011 and 2014, 7 children with proven diagnosis of herpes simplex virus encephalitis were identified in a university hospital of Istanbul. Two patients had neurologic relapse characterized mainly by movement disorders 2 to 3 weeks after initial encephalitis. The first patient received a second 14 days of acyclovir treatment together with antiepileptic drugs and left with severe neurologic sequelae. The second patient was found to be NMDA receptors antibody positive in the cerebrospinal fluid. She was treated with intravenous immunoglobulin and prednisolone. She showed substantial improvement, gradually regaining lost neurologic abilities. Post-herpes simplex virus encephalitis relapses may frequently be immune-mediated rather than a viral reactivation, particularly in children displaying movement disorders like choreoathetosis. Immunotherapy may provide benefit for this potentially devastating condition, like the case described in this report.


Effects of Methyl, Ester, and Amine Surface Groups on Microbial Activity and Communities in Nitrifying Biofilms.
The objective of this study was to determine how different attachment surface chemistries affected the initial and long-term performance and microbial populations of nitrifying biofilms under well-controlled hydrodynamic mixing conditions. While much previous research has focused on the effects of surface properties such as hydrophobicity on bacterial attachment in pure cultures, this study evaluated the effects of specific functional groups on mixed culture composition and functional behavior. Three surfaces with varying hydrophobicity and charge were evaluated for biofilm community development and performance: unmodified poly(dimethylsiloxane) (PDMS), which included terminal methyl groups and was relatively hydrophobic (P-Methyl), PDMS silanized with ester groups (P-Ester), which was uncharged and relatively hydrophilic, and PDMS modified with amine groups (P-Amine), which possessed a positive charge and was the most hydrophilic. The surface chemistries of the three attachment surfaces were characterized by contact angle goniometry, Fourier transform infrared (FTIR) spectroscopy, and X-ray photoelectron spectroscopy (XPS). These surfaces were inoculated with dilute activated sludge, and biofilms were grown in rotating annular bioreactors for 80 days, with experimental triplicates. Nitrification rates increased most rapidly in P-Amine biofilm reactors, and their biofilm communities contained significantly more Nitrosomonas (p < 0.05) than those on the other surfaces in early growth stages (days 40-50). From days 50-60, the P-Amine surface biofilm had significantly higher nitrate production rates than the P-Methyl and P-Ester biofilms. The biofilms grown on the P-Amine and P-Methyl surfaces were significantly (p < 0.05) more diverse than the P-Ester biofilms, containing higher relative abundances of the order Rhizobiales, including a significantly higher abundance of the nitrifying genus Nitrobacter (p < 0.05), which coincided with higher rates of nitrate generation. Conversely, biofilms grown on the uncharged hydrophilic P-Ester surface were consistently less productive and had lower diversity than biofilms on the other surfaces. These results indicate that surface chemistry may be a useful design parameter to improve the performance of nitrifying biofilm systems for wastewater treatment and that surface chemistry affects mixed biofilm community composition.


Detection of DNA damage by space radiation in human fibroblasts flown on the International Space Station.
Although charged particles in space have been detected with radiation detectors on board spacecraft since the discovery of the Van Allen Belts, reports on the effects of direct exposure to space radiation in biological systems have been limited. Measurement of biological effects of space radiation is challenging due to the low dose and low dose rate nature of the radiation environment, and due to the difficulty in distinguishing the radiation effects from microgravity and other space environmental factors. In astronauts, only a few changes, such as increased chromosome aberrations in their lymphocytes and early onset of cataracts, are attributed primarily to their exposure to space radiation. In this study, cultured human fibroblasts were flown on the International Space Station (ISS). Cells were kept at 37°C in space for 14 days before being fixed for analysis of DNA damage with the γ-H2AX assay. The 3-dimensional γ-H2AX foci were captured with a laser confocal microscope. Quantitative analysis revealed several foci that were larger and displayed a track pattern only in the Day 14 flight samples. To confirm that the foci data from the flight study was actually induced from space radiation exposure, cultured human fibroblasts were exposed to low dose rate γ rays at 37°C. Cells exposed to chronic γ rays showed similar foci size distribution in comparison to the non-exposed controls. The cells were also exposed to low- and high-LET protons, and high-LET Fe ions on the ground. Our results suggest that in G1 human fibroblasts under the normal culture condition, only a small fraction of large size foci can be attributed to high-LET radiation in space.


[Specific aspects of mobility and changes in the shape of the dural sac in functional lumbosacral myelography].
The purpose of the work was an analysis of lateral myelograms of the lumbosacral spine in the neutral position, in anteflexion and retroflexion from the aspect of the mobility of the dural sac and its relationship to the spinal canal. The group comprised 50 patients with clinically obvious discopathies and other vertebrogenic syndromes. The areas of the dural sac were measured planimetrically and evaluated by means of a computer. It was revealed: 1. The Soinal canal and dural sac are not static variables but dynamic ones. By movements of the lumbosacral spine the size of the dural sac changes and so does its shape. These phenomena can be evaluated not only qualitatively but also quantitatively. 2. The anteroposterior diameter of the dural sac on the myelograms in anteflexion and in retroflexion changes significantly, as compared with the neutral position, it increases during anteflexion and diminishes during retroflexion. 3. The area of the dural sac increases during anteflexion and diminishes in retroflexion, as compared with the neutral position. 4. Changes in the shape of the dorsal portion of the dural sac revealed in the segment of the discopathy are considered to be due to hypertrophy of the yellow ligaments.


The evolution of the psychology of the self: toward a mature narcissism.
Since the publication of Heinz Kohut's last book in 1984, "self psychology" has become much more diverse in its perspectives and even fragmented in its applications. Perhaps due in large part to Kohut's own inconsistencies and lack of clarity on certain major points of theory and technique, self psychology currently lacks a clear definition and has become increasingly marginalized in the larger psychoanalytic community, even as some concepts have been absorbed into more general psychoanalytic usage. Major theoretical and clinical concepts in self psychology remain foggy in their definition and in their clinical application. This article will outline an essential paradigm for a psychoanalytic self psychology, given the author's background in social science, and its connection to particular psychoanalytic values. Focal changes in post-Kohutian self psychology that conflict with these views are briefly reviewed; changes that I saw at odds with not only the basic orientation of self psychology but also with psychoanalysis itself. Changes in the selfobject concept and in how the role of early trauma is understood in the context of Kohut's concept of optimal frustration are discussed. A clinical illustration demonstrates my concern that the introduction of the relational perspective in self psychology appears to interfere with psychoanalytic goals per se; that heavy reliance on present reality and "optimal gratification" is at the expense of access to the unconscious and interferes with achievement of specific analytic goals regarding the acquisition of maturity in the narcissistic realm. These conclusions relocate self psychology within the fabric of psychoanalysis and its emphasis on the self as internal.


Lower Critical Solution Temperature Behavior in Polyelectrolyte Complex Coacervates.
In light of recent experimental observations of lower critical solution temperature (LCST) in polyelectrolyte complex coacervates (Ali, S. et al. ACS Macro Lett. 2019, 8, 289-293), we explore its possible mechanisms on the basis of a slight modification of our theory (Adhikari, S. et al. J. Chem. Phys. 2018, 149, 163308). We explore the consequences of the temperature dependence of the solvent dielectric constant (ε) and the solvent-polymer interaction parameter (χ) on the complex coacervates' phase behavior. The results show that the temperature dependence of the solvent dielectric constant and solvent-polymer interaction parameter can result in a complex phase behavior involving two disjoint unstable regions on the temperature (T)-polyelectrolyte concentration (ϕ p) plane. Comparison of phase diagrams constructed for different possible temperature dependencies of ε and χ shows that the experimentally observed LCST behavior is obtained only if the solvent dielectric constant decreases and the solvent-polymer interaction parameter increases with increasing temperature. Preferential partitioning of salt into the polyelectrolyte poor phase is predicted for all possible combinations of temperature dependencies of χ and ε considered in this work.


Vitamins C and E alone and in combination partly protect against chronic ethanol-induced toxicity in rat erythrocytes.
Oxidative stress is part of the mechanisms involved in ethanol toxicity. We investigated effects of vitamins C (VC), E (VE) and the combination of VC+VE on chronic ethanol-induced toxicity in rat erythrocytes. The following groups were treated for 30 days: control (C), VC (200 mg/kg), VE (200 mg/kg), VC (200 mg/kg) + VE (200 mg/kg), ethanol 4 g/kg, ethanol + VC, ethanol + VE and ethanol + VC + VE. The doses of vitamins and ethanol were selected for per kilogram of animal's body weight. Blood samples collected at the end of treatments were analyzed for erythrocyte osmotic fragility and plasma scavenging activity. The washed erythrocytes were used to determine superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and malondialdeyde (MDA). Ethanol induced erythrocyte fragility (p < 0.001) and increased lipid peroxidation (p < 0.001) in rat erythrocytes. Furthermore, there were significant decreases in plasma scavenging (p < 0.001), SOD (p < 0.001), CAT (p < 0.01) and GPx (p < 0.001) activities in erythrocytes of ethanol-treated animals. Although VC or VE alone restored all of ethanol-induced disturbances to near normal (p > 0.05) but there were still significant differences compared to control animals. Combination therapy completely corrected ethanol-induced erythrocyte fragility, lipid peroxidation and prooxidant/antioxidant imbalance. We showed the beneficial effects of VC and VE combination in decreasing erythrocyte fragility and lipid peroxidation in both ethanol and control groups. Therefore this combination treatment may provide a new potential alternative for prevention of ethanol toxicity which deserves consideration and further examination.


[Personality, achievement and health in social change].
All countries with modern industrial societies are undergoing epochal changes. These processes include globalization, internationalization and structural alterations in the work environment which have social and psychophysical consequences for the individual. These problems are rarely discussed concerning their pathogenic or health-supporting effects. Transformations of society primarily change the relationship between the demands, expectations and requirements placed on the individual and his or her abilities and competencies necessary to find solutions. This challenge simulataneously offers chances and risks for an individual's personality development, namely new demands on an individual's regulation of potential and achievement. Not only is competence in the workplace required, but social skills, emotional and self-reflective qualities are vital as well. Developing these abilities requires two tasks. One promotes and develops such skills in the individual. The second would promote sound political decisions enabling the skills in the individual to be actualized.


Ultrahigh-Strength Porous Ceramic Composites via a Simple Directional Solidification Process.
Porous ceramics possess great application potential in various fields. However, the contradiction between their pores and their strength have significantly hampered their applications. In this study, we present a simple directional solidification process that relies on its in situ pore forming mechanism to fabricate Al2O3/Y3Al5O12/ZrO2 porous eutectic ceramic composites with a highly dense and nanostructured eutectic skeleton matrix and a lotus-type porous structure. The flexural strength of this porous ceramic composite with a porosity of 34% is 497 MPa at ambient temperature, which is a new record of the strength of all current porous ceramics. This strength can remain at 324 MPa when the temperature increases up to 1773 K because of its refined lamellar structure and strong bonding interface. We demonstrate an interesting application of the directional solidification in efficiently preparing the ultrahigh-strength porous ceramic with high purity. The findings will open a window to the strength of porous ceramics.


Characterization of a glucuronide metabolite of 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and its dose-dependent excretion in the urine of mice and rats.
Following analysis by reversed-phase HPLC, a previously uncharacterized metabolite of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) was found in the urine of A/J mice treated with NNK. Treatment with beta-glucuronidase converted the metabolite to a peak that co-eluted with 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL). Treatment with sulfatase or beta-glucuronidase plus saccharic acid 1,4-lactone did not change the retention time of the metabolite. These data suggested that the unknown metabolite was a glucuronic acid conjugate of NNAL. Upon isolation and purification of larger quantities of the metabolite from the urine of A/J mice, CD-1 mice and F344 rats, 1H and 13C NMR and MS confirmed that the unknown metabolite was 4-(methylnitrosamino)-1-(3-pyridyl)-1-butyl beta-D-glucopyranosiduronic acid (NNAL Glu). To determine the quantitative relationship between NNK dose and NNAL Glu production and to compare the importance of glucuronidation relative to other metabolic pathways, [5-3H]NNK was administered to F344 rats and A/J mice at doses of 500-0.005 mumol/kg. At 500 mumol/kg, NNAL Glu accounted for 22% of the total urinary excretion of NNK in A/J mice, and for 8% in F344 rats 48 h after dosing. The proportions of excreted glucuronide and NNAL decreased with diminishing doses of NNK, yielding undetectable levels of each metabolite in both mice and rats at a dose of 0.005 mumol/kg NNK. Since substantial amounts of metabolites formed via alpha-hydroxylation and N-oxidation pathways were observed at the lower doses of NNK, these data demonstrate that NNAL glucuronidation is a quantitatively unimportant metabolic pathway at low doses of NNK.


Delusional misidentification syndromes and dementia: a border zone between neurology and psychiatry.
The delusional misidentification syndromes (DMSs) are psychopathologic phenomena in which a patient consistently misidentifies persons, places, objects, or events. Although often described in relation to psychotic states including schzofrenia, it is, nevertheless, widely considered that these syndromes have an anatomical basis because of their frequent association with organic brain disease; studies have pointed to the presence of identifiable lesions, especially in the right frontal lobe and adjacent regions, in a considerable proportion of patients. The purpose of this article is to examine the phenomenon in people with dementia. We searched the electronic databases for original research and review articles on DMS in patients with dementia using the search terms "Delusional Misidentification Syndrome, Capgras syndrome, Fregoli syndrome, reduplicative paramnesia, and dementia." The DMSs are a frequent problem in dementia. The violence and dangerousness in patients with dementia having these syndromes are well documented, and forensic aspects are highlighted. Pathogenetic viewpoint and management are considered.


Group A streptococcus (GAS) carbohydrate as an immunogen for protection against GAS infection.
Previous studies have shown that human serum containing anti-group A streptococcus carbohydrate (GAS CHO) antibodies were opsonic for different M protein-carrying serotypes. To investigate the role that anti-GAS CHO antibodies play in passive and active protection, mice were immunized subcutaneously or intranasally with GAS CHO conjugated to tetanus toxoid, and mortality and oral colonization were monitored after challenge with live GAS. Compared with control mice, immunized mice were significantly protected against systemic or nasal challenge with GAS. Furthermore, studies of serum samples and throat cultures from Mexican children revealed an inverse relationship between high serum titers of anti-GAS CHO antibodies and the presence of GAS in the throat. Anti-GAS CHO antibodies were also tested for cross-reactivity with human tissues and cytoskeletal proteins. No cross-reactivity was observed in either assay. The present study demonstrates that GAS CHO is both immunogenic and protective against GAS infections.


Chemical Derivatization in Flow Analysis.
Chemical derivatization for improving selectivity and/or sensitivity is a common practice in analytical chemistry. It is particularly attractive in flow analysis in view of its highly reproducible reagent addition(s) and controlled timing. Then, measurements without attaining the steady state, kinetic discrimination, exploitation of unstable reagents and/or products, as well as strategies compliant with Green Analytical Chemistry, have been efficiently exploited. Flow-based chemical derivatization has been accomplished by different approaches, most involving flow and manifold programming. Solid-phase reagents, novel strategies for sample insertion and reagent addition, as well as to increase sample residence time have been also exploited. However, the required alterations in flow rates and/or manifold geometry may lead to spurious signals (e.g., Schlieren effect) resulting in distorted peaks and a noisy/drifty baseline. These anomalies can be circumvented by a proper flow system design. In this review, these aspects are critically discussed mostly in relation to spectrophotometric and luminometric detection.


Geographical Information Systems and canine faecal contamination: the experience in the city of Naples (southern Italy).
Geographical Information Systems (GIS) can be used to make transect sampling when non-moving objects are to be counted, involving choosing a line or series of lines along which the counts are to take place. This approach has been used by us in order to study canine faecal contamination in the city of Naples (southern Italy), and to evaluate the consequent presence of canine parasitic elements. A GIS was constructed utilizing the geo-referenced digital photographs and the cadastral maps of Naples. In order to uniformly evaluate the canine faecal contamination throughout the city, a grid representing sub-areas of 1 km x 700 m was overlaid on the city map within the GIS. The territory of Naples was divided in 218 equal, rectangular sub-areas. In each sub-area a 1 km transect was drawn, and digitalized on-screen in the GIS. Canine faeces (copros in Greek) were counted along the transects in 143 sub-areas. In these sub-areas, 415 copros were collected and examined. Out of the 143 sub-areas, 141 contained canine copros. The results of the negative binomial regression model showed a positive association between the number of copros and the human population density. Out of the total of 415 copros, 70 (16.9%) were positive for parasitic elements; eggs of Toxocara canis, Toxascaris leonina, Ancylostoma caninum and Trichuris vulpis were found, as well as oocysts of Isospora canis.


Bacterial endosymbiont of the slender pigeon louse, Columbicola columbae, allied to endosymbionts of grain weevils and tsetse flies.
The current study focuses on a symbiotic bacterium found in the slender pigeon louse, Columbicola columbae (Insecta: Phthiraptera). Molecular phylogenetic analyses indicated that the symbiont belongs to the gamma subdivision of the class Proteobacteria and is allied to Sodalis glossinidius, the secondary symbiont of tsetse flies (Glossina spp.) and also to the primary symbiont of grain weevils (Sitophilus spp.). Relative-rate tests revealed that the symbiont of C. columbae exhibits accelerated molecular evolution in comparison with the tsetse fly symbiont and the weevil symbiont. Whole-mount in situ hybridization was used to localize the symbiont and determine infection dynamics during host development. In first- and second-instar nymphs, the symbionts were localized in the cytoplasm of oval bacteriocytes that formed small aggregates on both sides of the body cavity. In third-instar nymphs, the bacteriocytes migrated to the central body and were finally located in the anterior region of the lateral oviducts, forming conspicuous tissue formations called ovarial ampullae. In adult females, the symbionts were transmitted from the ovarial ampullae to developing oocytes in the ovarioles. In adult males, the bacteriocytes often disappeared without migration. A diagnostic PCR survey of insects collected from Japan, the United States, Australia, and Argentina detected 96.5% (109/113) infection, with a few uninfected male insects. This study provides the first microbial characterization of a bacteriocyte-associated symbiont from a chewing louse. Possible biological roles of the symbiont are discussed in relation to the host nutritional physiology associated with the feather-feeding lifestyle.


Linear Size Contraction of Ligand Protected Ag29 Clusters by Substituting Ag with Cu.
There are only a few examples of atomically precise, ligand protected, bimetallic coinage metal clusters in which molecular structure remains essentially unchanged over a wide composition range starting from the corresponding homometallic species. Such model systems are particularly useful to study the dynamics of alloy formation on the nanoscale. Here we demonstrate the unusual reactivity of solvated metalloid-superatom Ag29(BDT)12(PPh3)4 (BDT = 1,3 benzenedithiol) clusters toward semiconducting Cu12S6(DPPPT)4 (DPPPT = bis(diphenylphosphino)pentane) clusters as an efficient way to exchange multiple copper atoms into the atomically precise silver clusters without changing overall the structure type. Concentration-dependent UV-vis absorption and online mass spectrometry shows that 14 Cu atoms can be exchanged into the silver cluster. Beyond the 14 Cu atom exchange, the cluster degrades to smaller thiolates. Information on cluster structures is obtained from high-resolution ion mobility mass spectrometry, which shows a linear decrease in collision cross section (CCS) with each Ag/Cu exchanged. Several isomeric structures are calculated by density functional theory (DFT), and their calculated collision cross sections are used to identify the most stable isomers for each Ag/Cu exchange product. Ag/Cu exchange is essentially limited to the cluster surface/shell. The core appears not to be involved.


Effects of flunixin meglumine on pyrexia and bioenergetic variables in postparturient dairy cows.
Study objectives were to determine whether a nonsteroidal antiinflammatory drug would reduce parturition-induced inflammation and fever and consequently improve appetite, bioenergetic parameters, and production variables in transitioning dairy cows. Multiparous cows (n = 26) were randomly assigned to 1 of 2 treatments beginning at parturition: 1) flunixin meglumine (FM; 2.2 mg/kg of BW; Banamine, 50 mg/mL, Schering-Plough Animal Health, Kenilworth, NJ), or 2) saline (control) at 2.0 mL/45.5 kg of BW. All treatments were administrated i.v. daily for the first 3 d in milk (DIM). Individual milk yield and dry matter intake (DMI) were recorded daily for the first 35 DIM. Rectal temperature was measured daily at 0700 and 1600 h for the first 7 DIM. Milk composition was determined on 2, 7, 14, 21, 28, and 35 DIM and blood plasma was collected on 1, 2, 3, 4, 7, 14, 21, 28, and 35 DIM. Body weight and body condition score were determined on -7, 1, 7, 14, 21, 28, and 35 DIM. Flunixin meglumine treatment slightly increased rectal temperature (38.99 vs. 38.76 degrees C) during the first 7 DIM and reduced overall DMI (22.04 vs. 19.48 kg/d), but there were no treatment differences in overall milk yield (35.2 kg/d), 3.5% fat-corrected milk (37.6 kg/d), energy-corrected milk (37.7 kg/d), DMI (2.97% of BW), or overall energy balance (-2.32 Mcal/d). There were no treatment differences in milk fat (3.91%), protein (3.32%), or lactose (4.57%). Treatment had no effect on plasma glucose (66.5 mg/dL) or nonesterified fatty acids (553 microEq/L), but plasma urea nitrogen tended to be less in FM-treated cows (16.4 vs. 14.5 mg/dL). Daily FM administration to cows for the first 3 d after parturition slightly increased rectal temperatures by 0.23 degrees C, reduced feed intake, and did not improve production or energetic variables during the first 35 DIM in transition dairy cows.


Diagnostic issues in second opinion consultations in prostate pathology.
The increase in early detection of prostate cancer in the Asian population has bolstered second opinion consultations in prostate pathology in this region. In this review, we aimed to identify the spectrum of lesions and queries submitted to a pathologist with uropathology interest at the Singapore General Hospital. Request forms for second opinion prostate consultations from pathologists and clinicians were retrieved from central laboratory records and a specialist's correspondence files within 2004-2007. Histomorphological queries raised in the referrals, and comparison of original diagnosis and Gleason scoring with the review diagnoses were collated. Discordant diagnoses (183/323, 57%) were more common than concordant diagnoses (143/326, 44%) between original diagnosis and subspecialist review. The majority of discordances comprised initial undergrading of Gleason scores (132/183, 72%; p < 0.01) especially in needle core biopsies. Among the significantly altered pathological diagnoses, 24 cases (13%; p < 0.01) were changed from benign to malignant (n = 9, 5%; p < 0.01), and malignant to benign (n = 11, 6%; p < 0.01) or high grade prostatic intraepithelial neoplasia (n = 4, 2%; p < 0.01). Benign mimics of malignancy such as atypical adenomatous hyperplasia and atrophy, and small foci of adenocarcinoma, were some examples of morphological pitfalls. Prostate biopsy review is important because of management and prognostic implications that vary among Gleason scores, and predictive parameters that are detailed in prostate pathology reports.


Characterization of a saccharide-binding protein from Talisia esculenta seeds with trypsin inhibitory activity.
Some proteins exhibit characteristics that suggest they have a primary, if not an exclusive role in nutrient reserve storage. The best studied examples are the storage proteins that accumulate specifically in developing seeds. Some of these protein demonstrate biological activities that could contribute to resistance to pest, pathogens or abiotic stresses. In this study we present the biochemical characterization and cloning of the major protein from seeds of T. esculenta (Talisin), a member of the Sapindaceae family. The N-terminal sequence of the protein isolated was used to produce a degenerated primer. This primer allowed the amplification of the Talisin cDNA by RTPCR from mRNA of the T. esculenta seeds protein. The sequence analysis of the cloned cDNA, demonstrated a 756 bp sequence encoding a peptide of 198 amino acids. The deduced peptide presented high similarity to a typical VSP, the 22-kDa protein in lychee (73 %) and 50.0 % identity to Theobroma bicolor reserve protein. Identities of 52.0 % and 44.0 % to trypsin inhibitors from Treobroma mammosum and Populus tremula respectively. In conclusion, we may suggest that Talisin could be a seed storage protein with affinity properties, i.e. interacts with carbohydrates and trypsin enzyme.


Recurrent Admissions for Acute Decompensated Heart Failure Among Patients With and Without Peripheral Artery Disease: The ARIC Study.
Background Peripheral artery disease (PAD) is both a common comorbidity and a contributing factor to heart failure. Whether PAD is associated with hospitalization for recurrent decompensation among patients with established heart failure is uncertain. Methods and Results Since 2005, the ARIC (Atherosclerosis Risk in Communities) study has conducted active surveillance of hospitalized acute decompensated heart failure (ADHF), with events verified by physician review. From 2005 to 2016, 1481 patients were hospitalized with ADHF and discharged alive (mean age, 78 years; 69% White). Of these, 207 (14%) had diagnosis of PAD. Those with PAD were more often men (55% versus 44%) and smokers (17% versus 8%), with a greater prevalence of coronary artery disease (72% versus 52%). Patients with PAD had an increased risk of at least 1 ADHF readmission, both within 30 days (11% versus 7%) and 1 year (39% versus 28%) of discharge from the index hospitalization. After adjustments, PAD was associated with twice the hazard of ADHF readmission within 30 days (HR, 2.02; 95% CI, 1.14-3.60) and a 60% higher hazard of ADHF readmission within 1 year (HR, 1.60; 95% CI, 1.25-2.05). The 1-year hazard of ADHF readmission associated with PAD was stronger with heart failure with reduced ejection fraction (HR, 2.01; 95% CI, 1.29-3.13) than preserved ejection fraction (HR, 1.04; 95% CI, 0.69-1.56); P for interaction=0.05. Conclusions Patients with ADHF and concomitant PAD have a higher likelihood of ADHF readmission. Strategies to prevent ADHF readmissions in this high-risk group are warranted.


[Changes in Xenopus oocyte nucleus and cytoplasm organization after actin filaments depolymerization by latrunculin].
Actin-containing filaments have been visualized inside the Xenopus oocyte nuclei due to combination of fluorescence and transmission electron microscopy. It has been shown that these filaments contact with nucleoli, spherical bodies and nuclear pore complexes. The incubation of oocytes with actin-depolymerizing latrunculin causes membrane vesiculation in the cytoplasm, and disruption of the nucleoplasm and nuclear envelope integrity. We suppose that actin-containing filaments belong to crucial cell components which are involved in coordination of nuclear-cytoplasmic interactions as well as distribution and transport of intranuclear components in growing Xenopus oocytes.


Structural analysis of proteolytic products of MHC class II-invariant chain complexes generated in vivo.
The MHC class II alpha beta heterodimer associates with invariant (I) chain in the endoplasmic reticulum and remains associated until the complex reaches a post-Golgi compartment. During early stages of transport, I chain blocks peptide binding to alpha beta dimers. I chain is proteolytically cleaved in a post-Golgi compartment releasing alpha beta dimers that can bind antigenic peptides and transport them to the cell surface. Human B lymphoblastoid cell lines grown in leupeptin, a sulfhydryl protease inhibitor, accumulate a partial proteolytic product of the I chain called leupeptin-induced protein (LIP). LIP remains associated with alpha beta dimers. We find, using chemical cross-linking, sucrose gradient sedimentation, and size exclusion chromatography, that the alpha beta LIP complex retains the nine-subunit structure described for alpha beta I complexes. Unlike the alpha beta I complex, in certain detergents the alpha beta LIP nonamer is unstable and dissociates into trimers containing one alpha, beta, and LIP molecule. This finding emphasizes the reported stoichiometry of the alpha beta I complex as a nine-subunit structure comprised of three alpha beta I trimers. Also, these data indicate that the region(s) of I chain necessary for retaining the nonameric structure lie within the LIP fragment, but that domains to the C-terminus of the LIP cleavage site act to further stabilize the nine chain structure. In addition, alpha beta I complexes containing forms of human I chain encoding the p35/p43 N-terminal cytoplasmic extension responsible for endoplasmic reticulum retention can transport to post-Golgi proteolytic compartments where LIP is formed.


Primary excision compared with re-excision of extremity soft tissue sarcomas--is anything new?
Soft tissue sarcomas (STS) are rare and are commonly excised outside of a sarcoma center without appropriate preoperative planning. Studies have shown varying results in survival and outcome when comparing patients undergoing re-excision to patients undergoing a single, planned excision. This retrospective study evaluated 278 patients treated for STS of the extremities between January 2000 and July 2006. One hundred seventy-two patients had a primary excision while 106 patients had a sarcoma re-excised. Survival curves for disease-free survival, metastasis-free survival, and local recurrence-free survival were calculated using competing risk analysis for both groups. After adjusting for high-risk variables, our results indicate that re-excision is a proxy for smaller, low-grade tumors which tend to have a better survival profile. Death due to sarcoma and distant metastases were correlated with high-grade and large tumors. The presence of positive microscopic margins was the strongest predictor of local recurrence (P < 0.05). There were no differences in death, metastases, or local recurrence between the two groups after adjusting for high-risk variables. Survival advantages previously reported with STS re-excision serve as proxy for tumors that have a better survival profile.


Phosphole modified pentathienoacene: synthesis, electronic properties and self-assembly.
A series of new ladder π-conjugated materials, phosphole modified pentathienoacene (PO-PTA), are synthesized and characterized. Single-crystal X-ray results demonstrate that methyl-disubstituted PO-PTA forms a face-to-face dimer structure driven by π-π interactions. The investigations of optical properties showed that the oxidized phosphole moiety in this ladder system can effectively narrow the band gap. PO-PTA is a promising building block in π-conjugated polymers and oligomers for optoelectronic applications. The derivative of PO-PTA, obtained by introducing four long alkyl chains, can self-assemble into one-dimensional (1D) fibers based on intermolecular π-π interactions, dipole-dipole interactions and van der Waals interactions. Interestingly, the uniform and well-ordered monolayers were also obtained for PO-PTA derivative on a HOPG (highly oriented pyrolytic graphite) surface.


Diagnostic usefulness of endorectal magnetic resonance imaging with dynamic contrast-enhancement in patients with localized prostate cancer: mapping studies with biopsy specimens.
New diagnostic criteria for dynamic magnetic resonance (MR) imaging in prostate cancer are presented. The diagnostic usefulness of endorectal MR imaging with dynamic contrast-enhancement in localized prostate cancer and the validity of these criteria were evaluated. Eighteen untreated patients who were suspected of localized prostate cancer were included in the study. They received endorectal dynamic MR imaging before systematic sextant needle biopsy. First. a mapping study with the findings of MR images and histopathology of biopsy specimens was performed in eight patients out of 18 to compare the difference in T2-weighted images with the endorectal coil and the body coil in the same individuals. Second, another mapping study was performed in all 18 patients by analyzing the findings of endorectal dynamic MR images. For the diagnosis of prostate cancer in MR imaging, we offered diagnostic criteria from our experience in addition to those in plain T2-weighted images from the literature. The overall diagnostic rates of endorectal dynamic MR imaging were 88.9% in accuracy, 100% in sensitivity, and 81.8% in specificity. In the comparison of the endorectal and body coils in T2-weighted images in eight patients, there was no difference in the diagnostic rates except for one more histopathologic false positive portion in endorectal MR imaging. In the second mapping study in 18 patients, the diagnostic rates were 92.6% in accuracy, 88.9% in sensitivity and 93.3% in specificity. Endorectal dynamic imaging raised the diagnostic sensitivity from 77.8 to 88.9%. The data demonstrated the validity of this diagnostic criteria and the diagnostic usefulness of endorectal dynamic MR imaging in localized prostate cancer.


Chebulagic acid, a hydrolyzable tannin, exhibited antiviral activity in vitro and in vivo against human enterovirus 71.
Human enterovirus 71 is one of the major causative agents of hand, foot and mouth disease in children under six years of age. Presently, no vaccines or antiviral drugs have been clinically available to employ against EV71. In this study, we demonstrate that treatment with chebulagic acid reduced the viral cytopathic effect on rhabdomyosarcoma cells with an IC50 of 12.5 μg/mL. The utilization of the chebulagic acid treatment on mice challenged with a lethal dose of enterovirus 71 was able to efficiently reduce mortality and relieve clinical symptoms through the inhibition of viral replication. Chebulagic acid may represent a potential therapeutic agent to control infections to enterovirus 71.


Energy-based de novo protein folding by conformational space annealing and an off-lattice united-residue force field: application to the 10-55 fragment of staphylococcal protein A and to apo calbindin D9K.
The conformational space annealing (CSA) method for global optimization has been applied to the 10-55 fragment of the B-domain of staphylococcal protein A (protein A) and to a 75-residue protein, apo calbindin D9K (PDB ID code), by using the UNRES off-lattice united-residue force field. Although the potential was not calibrated with these two proteins, the native-like structures were found among the low-energy conformations, without the use of threading or secondary-structure predictions. This is because the CSA method can find many distinct families of low-energy conformations. Starting from random conformations, the CSA method found that there are two families of low-energy conformations for each of the two proteins, the native-like fold and its mirror image. The CSA method converged to the same low-energy folds in all cases studied, as opposed to other optimization methods. It appears that the CSA method with the UNRES force field, which is based on the thermodynamic hypothesis, can be used in prediction of protein structures in real time.


Creating and evaluating a data-driven curriculum for central venous catheter placement.
Central venous catheter placement is a common procedure with a high incidence of error. Other fields requiring high reliability have used Failure Mode and Effects Analysis (FMEA) to prioritize quality and safety improvement efforts. To use FMEA in the development of a formal, standardized curriculum for central venous catheter training. We surveyed interns regarding their prior experience with central venous catheter placement. A multidisciplinary team used FMEA to identify high-priority failure modes and to develop online and hands-on training modules to decrease the frequency, diminish the severity, and improve the early detection of these failure modes. We required new interns to complete the modules and tracked their progress using multiple assessments. Survey results showed new interns had little prior experience with central venous catheter placement. Using FMEA, we created a curriculum that focused on planning and execution skills and identified 3 priority topics: (1) retained guidewires, which led to training on handling catheters and guidewires; (2) improved needle access, which prompted the development of an ultrasound training module; and (3) catheter-associated bloodstream infections, which were addressed through training on maximum sterile barriers. Each module included assessments that measured progress toward recognition and avoidance of common failure modes. Since introducing this curriculum, the number of retained guidewires has fallen more than 4-fold. Rates of catheter-associated infections have not yet declined, and it will take time before ultrasound training will have a measurable effect. The FMEA provided a process for curriculum development. Precise definitions of failure modes for retained guidewires facilitated development of a curriculum that contributed to a dramatic decrease in the frequency of this complication. Although infections and access complications have not yet declined, failure mode identification, curriculum development, and monitored implementation show substantial promise for improving patient safety during placement of central venous catheters.


Interactions and reactions of transition metal clusters with the interior of single-walled carbon nanotubes imaged at the atomic scale.
Clusters of transition metals, W, Re, and Os, upon encapsulation within a single-walled carbon nanotube (SWNT) exhibit marked differences in their affinity and reactivity with the SWNT, as revealed by low-voltage aberration-corrected high-resolution transmission electron microscopy (AC-HRTEM). Activated by an 80 keV electron beam, W reacts only weakly with the SWNT, Re creates localized defects on the sidewall, and Os reacts readily causing extensive defect formation and constriction of the SWNT sidewall followed by total rupture of the tubular structure. AC-HRTEM imaging at the atomic level of structural transformations caused by metal-carbon bonding of π- and σ-character demonstrates what a crucial role these types of bonds have in governing the interactions between the transition metal clusters and the SWNT. The observed order of reactivity W < Re < Os is independent of the metal cluster size, shape, or orientation, and is related to the metal to nanotube bonding energy and the amount of electronic density transferred between metal and SWNT, both of which increase along the triad W, Re, Os, as predicted by first-principles density functional theory calculations. By selecting the appropriate energy of the electron beam, the metal-nanotube interactions can be controlled (activated or precluded). At an electron energy as low as 20 keV, no visible transformations in the nanotube in the vicinity of Os-clusters are observed.


Genotypic identification of presumptive Streptococcus pneumoniae by PCR using four genes highly specific for S. pneumoniae.
It was previously reported that two oligonucleotide primer sets (spn9802 and spn9828) for discriminating Streptococcus pneumoniae from pneumococcus-like oral streptococcal isolates using PCR had been developed. In this study, PCR amplification of the lytA, ply, spn9802 and spn9828 genes was used to identify presumptive S. pneumoniae. Two genetic groups were identified by analysing sputum samples from 28 patients with community-acquired pneumonia: the lytA-positive, ply-positive, spn9802-positive and spn9828-negative group, and the lytA-positive, ply-positive, spn9802-positive and spn9828-positive group. Isolates of the former group were resistant to optochin, while those of the latter group showed susceptibility to optochin. The lytA-positive, ply-positive, spn9802-negative and spn9828-negative isolates, and lytA-positive, ply-positive, spn9802-negative and spn9828-positive isolates, were not detected in sputum from patients with pneumonia. Subsequently, a total of 92 saliva samples from healthy individuals was screened by PCR using these primer sets. The lytA-positive, ply-positive, spn9802-positive and spn9828-negative group was identified more frequently in saliva from healthy children than in saliva from older healthy individuals and patients with pneumonia. The lytA-positive, ply-positive, spn9802-positive and spn9828-positive group was found frequently in saliva from healthy children, and in saliva and sputum from patients with pneumonia. This study demonstrates a rapid, optimal screening method for the genotypic identification of presumptive S. pneumoniae by PCR using four genes highly specific for S. pneumoniae.


Rh(III) -Catalyzed C-H Olefination of Benzoic Acids under Mild Conditions using Oxygen as the Sole Oxidant.
Phthalide skeletons have been synthesized for the first time through a Rh(III) -catalyzed C-H olefination of benzoic acids under mild conditions using oxygen as the sole oxidant. Aromatic acids bearing a variety of functional groups could react with diverse alkenes to afford the desired cyclized lactones or uncyclized alkenylarenes in moderate-to-excellent yields.


[Current developments in systemic therapy of breast carcinoma].
New developments in hormone- and chemotherapy of metastasizing breast cancer comprise replacement of ovariectomy by LH-RH antagonists in premenopausal women, new antiestrogens with less residual estrogenic activity for use mainly in postmenopausal patients as well as new potent inhibitors of aromatase with fewer adverse effects also for postmenopausal women. These new drugs will possibly provide better efficacy with fewer side effects, but at higher cost. In chemotherapy the search for more potent regimes continues. Experiments are under way with high-dose chemotherapy plans in conjunction with autologous bone marrow transplantation and hemopoietic growth factors; however, the result of these latter studies are so far disappointing. It has become clear that in nonaggressive metastasizing cancer of the breast high rates of remission seem not to be the ultimate goal. At this stage of disease the natural course is very heterogeneous; therefore, chemotherapy has to be adapted to risk. The goal must be improvement of quality of life for the remaining life-span by palliation of symptomatic disease while inducing as few side effects of treatment as possible. New; more potent chemotherapeutic agents are not at sight. Research has recently produced derivatives of already known agents producing fewer adverse effects at equal efficacy. Decisive for the choice of primary chemotherapy are prognostic indicators of the disease. For most cases, tumor specific therapeutic modalities have to be optimized.


Effect of hyperthermia on intracellular pH in human U-87 MG glioblastoma cells.
The effect of hyperthermia at 43 degrees C on intracellular pH (pHi) in human U-87 MG glioblastoma cells was studied by using the fluorescent probe 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein-pentaacetoxymethyl ester. The presence of Na+/H+ antiporter activity in the cells were demonstrated by the Na(+)-dependent increase in intracellular pH (pHi) after cellular acidification in the absence of HCO3-. Hyperthermia at 43 degrees C caused significant decrease in pHi. The acidification was readily reversible by cooling the cells back down to 37 degrees C. The pHi change was inhibited by the addition of 1 mM amiloride in the incubation medium. Amiloride and hyperthermia exhibited a synergistic effect in suppressing thymidine incorporation into the cells.


Exploration of shared gene signature with development of pre-eclampsia and cervical cancer.
Background: The relationship between pre-eclampsia (PE) and cervical cancer (CC) has drawn more attention recently, while little is known about the shared pathogenesis of CC and PE. In the present research, we aimed to generate the shared gene network as well as the prognostic model to reveal the development of CC and PE. Methods: The transcription data of CC and PE patients were obtained and enrolled into weighted gene co-expression network (WGCNA) analysis. Disease-specific modules in CC and PE were determined to discover the shared genes. The expression patterns of genes at protein level were examined by HPA database. Further, LASSO penalty regression and Cox analysis were applied to create a prognostic signature based on the shared genes, with survival curves and ROC plots employed to confirm the predictive capacity. To uncover the function roles and pathways involved in signature, gene set enrichment analysis (GSEA) was conducted. Finally, the immune infiltration status in CC was depicted using CIBERSORT algorithms. Results: WGCNA determined three hub modules between CC and PE. A total of 117 shared genes were obtained for CC and PE and mainly enriched in cell proliferation, regulation of cell development and neuron differentiation. Then, we created a robust prognostic model based on the 10 shared genes by performing stepwise Cox analyses. Our proposed model presented a favorable ability in prognosis forecast and was correlated with the infiltration of immunocytes including B cells, macrophages and T cells. GSEA disclosed that high-risk group was involved in cancer-related pathways. Conclusion: The present project identified the shared genes to uncover the pathogenesis of CC and PE and further proposed and validated a prognostic signature to accurately forecast the clinical outcomes of CC patients.


Hyperoside ameliorates the progression of osteoarthritis: An in vitro and in vivo study.
Osteoarthritis (OA) is a common degenerative joint disease. The pathogenesis of OA is closely related to inflammatory responses and apoptosis of chondrocytes. Hyperoside (Hyp), a natural flavonoid compound, exerts multiple bioactivities in various diseases. Our study aims to investigate the anti-arthritic effects of Hyp and delineate the potential mechanism at the cellular level. Murine chondrocytes were stimulated with interleukin-1β (IL-1β) with or without Hyp treatment. CCK-8 assay was used to evaluate the cytotoxic effect of Hyp. DCFH-DA was used to detect intracellular ROS. Annexin V-FITC/PI method was applied to examine apoptosis of chondrocytes. The anti-arthritic effects of Hyp and related mechanisms were investigated by examining and analyzing relative markers through quantitative PCR, western blot analysis and immunofluorescent staining. C57BL/6 mice were performed the destabilized medial meniscus (DMM) surgery to establish OA model and then injected intraperitoneally with Hyp (20 mg/kg)) for 4 or 8 weeks. Finally, mice were sacrificed and knee joints were collected for histological observation and analysis. Hyp inhibited IL-1β-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Additionally, Hyp attenuated IL-1β-induced destruction of the extracellular matrix (ECM) by downregulating the expression of MMPs and ADAMTS5, and meanwhile upregulating the expression of collagen II, aggrecan, and SOX9. Also, Hyp pretreatment reduced IL-1β-induced overproduction of ROS and apoptosis of chondrocytes. Mechanistically, Hypexerted anti-inflammatory effects by partly suppressing the PI3K/AKT/NF-κB and the MAPK signaling pathways, enhancing the Nrf2/HO-1 to limit the activation of NF-κB. Moreover, Hyp played an anti-apoptotic effect via the Nrf2/ROS/BAX/Bcl-xlaxis. In vivo, cartilage degradation was attenuated with a lower OARSI score in Hyp-treated group compared to the DMM group. Our study demonstrated that anti-arthritic effects of Hyp in vitro and in vivo, indicating Hyp might serve as a potential agent for the treatment of OA.


Inhibitory effect of sulphated polysaccharide porphyran on nitric oxide production in lipopolysaccharide-stimulated RAW264.7 macrophages.
Porphyran, extracted from an edible red alga (Porphyra yezoensis), is a sulphated polysaccharide with a wide variety of biological activities including anti-tumour, antioxidant and immuno-modulating activities. In this study, we examined the effect of porphyran on nitric oxide (NO) production in mouse macrophage cell line RAW264.7 cells. Although no significant activity of porphyran to induce NO or tumour necrosis factor-α (TNF-α) production in RAW264.7 cells was observed at the concentration range tested (10-500 µg/ml), it was found for the first time that porphyran inhibited NO production and expression of inducible nitric oxide synthase (iNOS) in RAW264.7 cells stimulated with lipopolysaccharide (LPS). In the presence of 500 µg/ml porphyran, NO production and expression of iNOS in LPS-treated RAW264.7 cells were completely suppressed. On the other hand, porphyran showed only a marginal effect on the secretion of TNF-α from LPS-stimulated RAW264.7 cells. Electrophoretic mobility shift assay (EMSA) using infrared dye labelled oligonucleotide with nuclear factor-κB (NF-κB) consensus sequence suggested that porphyran inhibited the LPS-induced NF-κB activation. The LPS-inducible nuclear translocation of p65, and the phosphorylation and degradation of IκB-α were also inhibited by the pre-treatment with porphyran. Our results obtained in in vitro analysis suggest that porphyran suppresses NO production in LPS-stimulated macrophages by the blocking of NF-κB activation.


SUV39H2 methylates and stabilizes LSD1 by inhibiting polyubiquitination in human cancer cells.
LSD1 is a histone lysine demethylase, which is highly expressed in multiple types of human cancer. Although its roles in transcriptional regulation have been well-studied, functional regulation of LSD1 by post-translational modifications still remains unknown. Here, we demonstrate that the histone lysine methyltransferase SUV39H2 trimethylated LSD1 on lysine 322. Knockdown of SUV39H2 resulted in a decrease of LSD1 protein even though the mRNA levels were unchanged. SUV39H2-induced LSD1 methylation suppresses LSD1 polyubiquitination and subsequent degradation. In addition, we also observed indirect effect of SUV39H2 overexpression on LSD1-target genes. Our results reveal the regulatory mechanism of LSD1 protein through its lysine methylation by SUV39H2 in human cancer cells.


Factors influencing emergency intubation in the pre-hospital setting--a multicentre study in the German Helicopter Emergency Medical Service.
Definitive airway control by endotracheal intubation (ETI) is standard of care in pre-hospital airway management. However, there are specific factors that may influence and complicate ETI. Prospective, descriptive study at three German Helicopter Emergency Medical Services (HEMS) over a 1-yr period. We examined the success and complication rate for field intubation performed by trauma anaesthetists. In 342 patients (9.3%) ETI was performed. The overall success rate was 100%; in 87.4% the first attempt was successful, whereas in 11.1% a second and in 1.5% a third ETI attempt was necessary. No patient required a surgical intervention. Limited access to the patient was found upon arrival at the scene in 20.2% of the patients and in 9.6% of the patients at the time of ETI attempt. An orotracheal ETI technique was used in all patients. In the patients in whom only one ETI attempt was necessary for successful intubation, the assessment of ETI conditions was rated 'very good' or 'good' in 94.7%, but in those requiring a second or third ETI attempt this was reduced to 68.6 and 20.0%, respectively. Difficulties encountered during ETI included blood (19.9%), vomit/debris (15.8%) and secretions (13.8%) in the upper airway; anatomical reasons (11.7%), patient position (9.6%) and surrounding conditions (9.1%), making laryngoscopy more difficult. Despite various factors increasing the difficulties in managing the airway in the field, definitive airway control by ETI seems to be safe practice.


The Hidden Strengths of Weak Theories.
There has been a strong tradition of assuming that concepts, and their patterns of formation might be best understood in terms of how they are embedded in theory-like sets of beliefs. Although such views of concepts as embedded in theories have been criticized on five distinct grounds, there are reasonable responses to each of these usual objections. There is, however, a newly emerging concern that is much more challenging to address - people's intuitive theories seem to be remarkably impoverished. In fact, they are so impoverished it is difficult to see how they could provide the necessary structure to explain differences between concepts and how they might form in development. One response to this recent challenge is to abandon all views of concept structure as being related to people's intuitive theories and see concepts as essentially structure-free atoms. The alternative proposed here argues that our very weak theories might in fact do a great deal of work in explaining how we form concepts and are able to use them to successfully refer.


Nucleophilicity Prediction via Multivariate Linear Regression Analysis.
The concept of nucleophilicity is at the basis of most transformations in chemistry. Understanding and predicting the relative reactivity of different nucleophiles is therefore of paramount importance. Mayr's nucleophilicity scale likely represents the most complete collection of reactivity data, which currently includes over 1200 nucleophiles. Several attempts have been made to theoretically predict Mayr's nucleophilicity parameters N based on calculation of molecular properties, but a general model accounting for different classes of nucleophiles could not be obtained so far. We herein show that multivariate linear regression analysis is a suitable tool for obtaining a simple model predicting N for virtually any class of nucleophiles in different solvents for a set of 341 data points. The key descriptors of the model were found to account for the proton affinity, solvation energies, and sterics.


Hepatitis a outbreak in a public school in Rio de Janeiro, Brazil.
From June 1 to July 1 1999, an outbreak involving 25 cases of hepatitis A occurred in a public school in Rio de Janeiro, Brazil. Since these cases were notified to the State Health Department, the National Reference Center for Hepatitis Viruses (CNRHV) was required to investigate the extent of hepatitis A virus (HAV) dissemination. Blood samples from all students were tested for IgM and total anti-HAV antibodies using a commercial enzyme-linked immunoassay (ELISA). At the same time, a questionnaire was completed in order to identify possible risk factors for HAV infection. The environmental investigation showed that there was no fecal contamination of the water supply. The epidemiological investigation demonstrated that almost 50% of this population was susceptible to HAV infection and probably person-to-person transmission was the principal mode of virus dissemination. In this situation, a massive vaccination campaign could control the HAV infection.


Exploring security and privacy issues in hospital information system: an Information Boundary Theory perspective.
A small community hospital (67 beds) in Central New York was undergoing a major technological change within the organization, as they move from the use of several legacy information systems to a hospital-wide information system. The focus of the present research is to explore the privacy and security information issues using a framework called Information Boundary Theory [Stanton, 2002]. IBT explains the motivational factors that lead to the revelation or disclosing of information.


Concept mapping: promoting science learning in BN learners in Australia.
We reviewed the process of using concept mapping as a methodology to teach science to RNs in a bachelor degree of nursing program in Australia. The technique was used for two classes of students in the science unit during 1995 with work packages and independent student learning being the focus of the unit. Both instructors and students found the experience mainly positive. It helped the students become more independent learners, have a greater confidence in their knowledge of science, and allowed their understanding of science in their practice to become more meaningful and personalized. For the instructors it allowed them to concentrate on students with learning difficulties because they had less face-to-face teaching hours. Major issues arising from the process related to how to balance science content and the techniques of concept mapping and how to more effectively assess the learning outcomes of the students. Concept mapping is a useful technique to link science and nursing practice. Not only can it be used in academic programs to teach the science underlying practice, it can be used in the clinical area to teach patients. As a technique it works well in staff development and continuing education where the focus is on elucidating the rationale for inuring practice and providing evidence for the value of this practice.


Investigation of the effect of nocodazole on the PI3K/Akt signaling pathway in COS7 cells.
A number of studies have demonstrated that nocodazole suppresses Akt phosphorylation; however, the underlying molecular mechanism remains unclear. In the current study, the mechanism of nocodazole‑induced suppression of Akt phosphorylation was investigated. The cell cycle was analyzed using flow cytometry and EGF receptor dimerization was evaluated through a cross-linking assay. Immunoprecipitation experiments were performed to investigate the interaction between P85 and EGFR and cell migration was evaluated through a wound healing assay. COS7 cells were observed to be rounded following a 24‑h treatment with nocodazole, and the results revealed that ~45% of COS7 cells were arrested at the G2/M phase and that the cyclin B1 expression level was greatly increased. EGF‑mediated Akt phosphorylation was markedly inhibited in nocodazole‑treated cells. In addition, the levels of internalized EGF‑EGFR complexes in nocodazole‑treated cells were reduced, and EGF‑EGFR dimerization was found to be affected by nocodazole. Akt phosphorylation in COS7 cells was demonstrated to be overridden by AG1478 and wortmannin. The results also showed that p85 did not bind to activated EGFR in nocodazole‑treated cells, and that nocodazole and protein inhibitors reduced cell migration. In summary, these results indicate that nocodazole inhibits the PI3K/Akt pathway by interfering with the binding of p85 binding to activated EGFR and further affects the growth of cells.