Neuromyelitis optica spectrum disorders and myelin oligodendrocyte glycoprotein antibody-associated disease: current topics.
We reviewed present topics on neuromyelitis optica spectrum disorders (NMOSD) and myelin oligodendrocyte glycoprotein (MOG)-antibody-associated disease (MOGAD). The number of NMOSD-related publications have increased year by year after the discovery of aquaporin 4 (AQP4)-antibody, and those on MOGAD started to surge since 2012-2013. Recent clinic-epidemiological surveys in NMOSD suggest that some racial differences in the prevalence and the clinical course. At present, experts feel the 2015 diagnostic criteria of AQP4-antibody-seronegative NMOSD should be revised. Randomized controlled trials of monoclonal antibodies in NMOSD have demonstrated a significant risk reduction of relapse, especially in AQP4-antibody-positive cases. Meanwhile, the efficacy in seronegative NMOSD was unclear. MOGAD can show NMO and other clinical phenotypes, but the clinical manifestations and frequencies are different in children and adults. One pathological study has suggested that MOGAD is distinct from AQP4-antibody-positive NMOSD, but may share some features with multiple sclerosis and acute disseminated encephalomyelitis. Immunosuppressive therapy can reduce relapse in MOGAD, but, unlike AQP4-antibody-positive NMOSD, some MOGAD patients treated with rituximab experience relapses despite a complete B-cell depletion. Our understanding and therapy of AQP4-antibody-positive NMOSD has made a significant progress, and recent research has identified challenges in seronegative NMOSD and MOGAD.


Gender- and sex-specific sports-related injury research in emergency medicine: a consensus on future research direction and focused application.
Title IX, the commercialization of sports, the social change in sports participation, and the response to the obesity epidemic have contributed to the rapid proliferation of participation in both competitive organized sports and nontraditional athletic events. As a consequence, emergency physicians are regularly involved in the acute diagnosis, management, disposition, and counseling of a broad range of sports-related pathology. Three important and highly publicized mechanisms of injury in sports relevant to emergency medicine (EM) include concussion, heat illness, and sudden cardiac death. In conjunction with the 2014 Academic Emergency Medicine consensus conference "Gender-specific Research in Emergency Care: Investigate, Understand, and Translate How Gender Affects Patient Outcomes," a consensus group consisting of experts in EM, emergency neurology, sports medicine, and public health convened to deliberate and develop research questions that could ultimately advance the field of sports medicine and allow for meaningful application in the emergency department (ED) clinical setting. Sex differences in injury risk, diagnosis, ED treatment, and counseling are identified in each of these themes. This article presents the consensus-based priority research agenda.


Anti-PD-1 treatment-induced immediate central diabetes insipidus: a case report.
Modulating PD-1 expression can constrain tumor growth. Hodgkin's lymphoma patients commonly express PD-L1 on tumor cells. We report the case of a 60-year-old male patient with relapsed classical Hodgkin's lymphoma who suffered from immediate-onset chill, hyperthermia and polyuria following initial treatment with sintilimab, an anti-PD-1 monoclonal antibody. The results revealed central diabetes insipidus (cDI). After 3 months of treatment with glucocorticoids and desmopressin acetate, his symptoms and the results were consistent with the resolution of cDI and the treatment course was discontinued. Diabetes insipidus is a rare complication of immunotherapeutic treatment, and this is the first case report to our knowledge to have described immediate-onset cDI caused by anti-PD-1 treatment.


Joint custody: a critical analysis and appraisal.
Joint custody, now in the statutes of over half the states, is becoming the legally preferred custody arrangement. It eases the burden on the courts, and is thought to benefit all family members. Research has indicated that children generally gain from maintaining a relationship with each parent; however, the assumption that joint custody promotes cooperation between divorced parents is a research question that has yet to be answered. There is a disparity between the strength of the joint custody movement and the sufficiency of evidence that this is the optimum approach for all children.


Hormone, serum, and methylxanthine interactions in the regulation of DNA synthesis in organ-cultured rat mammary tumors.
Previous studies have shown that the growth of MTW9, a hormone-dependent rat mammary tumor, is stimulated by the presence of a mammosomatotropic tumor coimplant (MtTW10) but is inhibited by the administration of dibutyryl cyclic AMP (DBcAMP). This study examines the direct effect of serum taken from MtTW10-bearing WF rats (MtT serum) and of substances that increase cellular levels of cyclic AMP (cAMP) on DNA synthesis in organ-cultured MTW9. Explant DNA synthesis was significantly greater in the presence of MtT serum than in either normal female or male rat serum. Preparations of hormones known to be present in MtT serum failed to simulate the activity of MtT serum. Whereas 1 mM cAMP 1 mM did not alter DNA synthesis in MTW9 explants, inhibitors of cyclic nucleotide phosphodiesterases substantially suppressed the rate of [3H]thymidine incorporation into DNA. However, addition of MtT serum to cultures completely prevented the effects of these substances. This activity of MtT serum could not be duplicated by any of the following additions to the culture medium: normal female or male rat serum, National Institutes of Health (NIH) rat prolactin (2.5 micrograms/ml), NIH rat growth hormone (2.5 micrograms/ml), insulin (5 micrograms/ml), 17 beta-estradiol (5 micrograms/ml), or a hormone combination simulating the known endocrine constituents of MtT serum. The results suggested that the serum of the MtT-bearing animal contained an activity, unidentified at present, that could antagonize the growth-inhibitory action of the cAMP system and that may participate in the regulation of mammary tumor growth.


Folic acid modifies the shape of epithelial cells during morphogenesis via a Folr1 and MLCK dependent mechanism.
Folic acid supplementation can prevent neural tube defects, but the specific molecular mechanisms by which it does have not been elucidated. During neural plate morphogenesis, epithelial cell apical constriction cooperates with other events to drive tissue-bending, and when defective, can result in neural tube defects. A Rho-kinase deficient binding mutant of the apical constriction regulating protein, Shroom3 (Shroom3R1838C), is one of only a handful of mouse mutant lines with neural tube defects that can be rescued by folic acid supplementation. This provided a unique opportunity to probe the functional rescue of a protein linked to neural tube development by folic acid. Utilizing an epithelial cell culture model of apical constriction, it was observed that treatment with exogenous folic acid, as well as co-expression of the folic acid receptor Folr1, can rescue the function of the Rho-kinase binding deficient mutant of Shroom3 in vitro It was also determined that the rescuing ability of folic acid is RhoA and Rho-kinase independent but myosin light chain kinase (MLCK) and Src-kinase dependent. Inhibition of Rho-kinase-dependent apical constriction in chick embryo neural epithelium was also observed to be rescued by exogenous folic acid and that treatment with folic acid is accompanied by elevated activated myosin light chain and MLCK. Furthermore, doubly heterozygous mouse embryos lacking one copy each of Shroom3 and Folr1 exhibit a low rate of neural tube defects and also have lower levels of activated myosin light chain and MLCK. These studies suggest a novel mechanism by which folic acid modifies epithelial cell shape during morphogenesis, shedding light onto how folic acid may prevent neural tube defects.


Discrepancy in pathologic diagnosis of placental lesions.
Placentas are routinely examined by surgical pathologists, but peer review of placental diagnosis is rarely performed. To determine the frequency of discrepant placental diagnosis between general surgical pathologists and a pediatric pathologist. One hundred fourteen placentas from infants with intrauterine growth restriction (IUGR) and 170 placentas from infants appropriate for gestational age (AGA) were reviewed for 10 lesion types using standardized criteria. The review diagnosis was compared with original reports. The review identified 333 lesions, 168 in the IUGR group and 165 in AGA group. Discrepant diagnosis occurred in 137 lesions (41.1%). There was no significant difference in the frequency of discrepant diagnosis between the IUGR (44.7%) and AGA groups (37.6%) (P >.05). Most discrepancies (92.7%) were due to underdiagnosis (identified on review but not mentioned in original diagnosis), but a few (7.3%) were due to misdiagnosis (mentioned in original report but disagreed on review). The common underdiagnoses with their corresponding rates were as follows: hemorrhagic endovasculitis (84.6%), fetal thrombotic vasculopathy (75%), massive perivillous fibrin deposition (68.4%), maternal floor infarction (66.7%), retroplacental hemorrhage (60.6%), intervillous thrombus (57.1%), decidual angiopathy (33.3%), placental infarction (25.4%), acute chorioamnionitis (22.7%), and chronic villitis (21.7%). Misdiagnosis was found in 10 cases: 5 cases of infarction (review diagnosis was perivillous fibrin deposits in 4, intervillous thrombus in 1), 3 cases of acute chorioamnionitis, and 2 cases of decidual angiopathy. Among the 8 general surgical pathologists involved, the frequency of discrepant diagnosis ranged from 31.5% to 58.6% (P >.05). The intraobserver discrepancy rate for the reviewer was 4.8%, significantly lower than the discrepancy rate for the 8 general surgical pathologists. It is common for general surgical pathologists not to recognize placental lesions, which may have clinical significance. Awareness of this deficiency, standardization of diagnostic criteria, and increased knowledge in placental pathology may improve the quality of diagnosis in this area.


Occurrence and Antioxidant Activity of C1 Degradation Products in Cocoa.
Procyanidin C1 is by far the main flavan-3-ol trimer in cocoa. Like other flavan-3-ols, however, it suffers a lot during heat treatments such as roasting. RP-HPLCHRMS/MS(ESI(-))analysis applied to an aqueous model medium containing commercial procyanidin C1 proved that epimerization is the main reaction involved in its degradation (accounting for 62% of degradation products). In addition to depolymerization, cocoa procyanidin C1 also proved sensitive to oxidation, yielding once- and twice-oxidized dimers. No chemical oligomer involving the native trimer was found in either model medium or cocoa, while two C1 isomers were retrieved. C1 degradation products exhibited antioxidant activity (monitored by RPHPLC-Online TEAC) close to that of C1 (when expressed in µM TE/mg·kg-1).


Effect of Dietary Pyrroloquinoline Quinone Disodium Salt on Cognitive Function in Healthy Volunteers: A Randomized, Double-Blind, Placebo-Controlled, Parallel-Group Study.
Cognitive dysfunctions are increasing alarmingly around the world, and researchers are exploring preventive measures for improving brain performance. Pyrroloquinoline quinone (PQQ), a naturally occurring coenzyme in foods, exhibits potent antioxidant activity, and improves diverse functions which include mitochondrial activation, growth, repair, protection of nerve cells by increased expression of nerve growth factor (NGF) and NGF receptors; and suppression of fibril formation and aggregation of amyloid β. This randomized, double-blind, placebo-controlled, parallel-group clinical investigation (RCT) evaluated the efficacy and safety of PQQ disodium salt powder (mnemoPQQ®) for improved cognitive function after 12 weeks of supplementation in healthy Japanese male and female (age 40 to <80 Y). 64 healthy subjects were randomly assigned to receive either mnemoPQQ® (PQQ disodium salt: 21.5 mg/day) or a placebo over a period of 12 weeks. The efficacy of mnemoPQQ® on cognitive performance (memory, attention, judgment, and cognitive flexibility) was examined using Cognitrax as the primary outcome (primary endpoint), and forgetfulness questionnaire (DECO: Deterioration Cognitive Observee) and Mini-Mental State Examination-Japanese (MMSE-J) as the secondary outcome (secondary endpoint). A total of 58 subjects (placebo = 31; Age = 70.91 ± 3.06 Y; mnemoPQQ® group = 27; Age = 72.10 ± 3.77 Y) completed the study over a period of 12 weeks of supplementation. Significant improvements were observed on the Cognitrax's cognitive function domain score on "composite memory", "verbal memory", "reaction time", "complex attention", "cognitive flexibility", "executive function", and "motor speed" in the mnemoPQQ® group as compared to the placebo group. The DECO and the MMSE-J scores were also significantly improved in the mnemoPQQ® group. No adverse events were observed. Study demonstrates that supplementation of PQQ disodium salt is useful in improving memory, attention, judgment, and cognitive function, in middle-aged to elderly population, who feel they have become more forgetful because of aging.


Myiasis in patients with head and neck cancer: Seldom described but commonly seen.
Human myiasis is a parasitosis usually found in tropical and underdeveloped countries. It usually affects ulcerated lesions or devitalized tissues, developing after deposition of dipterous eggs. Patients with head and neck cancer are at risk to develop secondary myiasis. A representative percentage of those patients manifest with neglected and advanced tumors, usually in exposed areas and with necrotic tissues. Few case reports and small series constitute the available information about this parasitosis. Most studies have been conducted in patients with skin carcinomas, although myiasis has already been described in association with other head and neck malignancies. The authors present a series of 12 cases of myiasis secondary to head and neck cancer in addition to a literature review.


Inhibition effect of epigallocatechin gallate on the growth and biofilm formation of Vibrio parahaemolyticus.
Vibrio parahaemolyticus is a common marine foodborne pathogen that causes gastroenteritis. With the long-term use of antibiotics, many bacteria become resistant; therefore, developing antibiotic-free antimicrobial strategies is urgent. Epigallocatechin gallate (EGCG), a constituent of polyphenols present abundantly in tea extract, has broad-spectrum antibacterial activity and is non-toxic. Here, we take advantage of these properties of EGCG to evaluate its inhibition effect on the growth and biofilm formation of V. parahaemolyticus 17802, and explore its antibacterial mechanism. It was found that EGCG showed antibacterial activity against V. parahaemolyticus 17802, and the minimum inhibitory concentration (MIC) was estimated to be 128 μg ml-1 . Results of crystal violet staining and confocal laser scanning microscope (CLSM) evidenced that EGCG hindered its biofilm formation. Moreover, the swimming motility and extracellular polysaccharides were also notably inhibited. The antibacterial mechanism was further confirmed by several assays, such as scanning electron microscopy (SEM), transmission electron microscopy (TEM), and live/dead staining assay, together with membrane permeability assay, which all suggested that EGCG caused damage to cell membrane and made it lose integrity, eventually resulting in the death of V. parahaemolyticus 17802. The bactericidal activity of EGCG verified its potential as a promising candidate to combat foodborne pathogen.


Predictors of bleeding and user satisfaction during consecutive use of the levonorgestrel-releasing intrauterine system.
Consecutive use of the levonorgestrel-releasing intrauterine system (LNG-IUS) is increasing. However, little is known about factors that predict the bleeding during consecutive use. The objective of this study was to analyse the possible factors which may predict the bleeding pattern during the first year of use of a second LNG-IUS. Fertile-aged women (n = 204) who had used their first LNG-IUS for over 4 years and who opted for a second LNG-IUS were recruited. Bleeding data were reported using 90-day reference periods (RPs) starting from the last 90 days of the first LNG-IUS use (baseline), until the end of the first year of the second LNG-IUS (RPs 1-4). Demographic factors such as age, parity, body mass index, indication of LNG-IUS use or smoking could not be identified as predictors for bleeding and spotting (B/S). Mean (+/-SD) number of B/S days was 8.9 (+/-9.1) at baseline. This increased slightly during RP1 and fell to 6.4 (+/-8.1) during RP4. Compared with the mean, women with uterine fibroids or a bleeding pattern of >9 days of spotting or any bleeding at RP1 had more B/S days during RP1-4. Although the number of B/S days decreased progressively from RP1 to RP4 in the group with a bleeding pattern of >9 days of spotting or any bleeding at baseline, such a phenomenon was not observed for women with fibroids. The difference for the change in B/S days between women with and without fibroids was statistically significant at RP3 and RP4. A high degree (91.7%) of satisfaction with the bleeding pattern was observed, with amenorrhoeic women being most satisfied. Uterine B/S is reduced during consecutive use of the LNG-IUS. Women with uterine fibroids or any bleeding at baseline continued to have more B/S than other women.


Measurement properties of the Spinal Appearance Questionnaire in adolescents with idiopathic scoliosis: a systematic review.
Scoliosis is defined as a three-dimensional deformity of the spine characterized by lateral tilt and axial rotation of the vertebrae. Its magnitude in the frontal plane is identified by a Cobb angle greater than 10o. The aim of the study was to systematically examine the clinimetric properties of the Spinal Appearance Questionnaire (SAQ) in its cross-cultural adaptations in different languages. Medline (PubMed), CINAHL, EMBASE, Science Direct, PsycINFO and WorldWideScience.org databases were used for screening studies until July 16, 2022. In this study, records on the development, evaluation and translation of the SAQ instrument in adolescents with idiopathic scoliosis were included. In addition, two independent reviewers defined whether the studies were eligible and analyzed their psychometric properties of internal consistency, reliability, content validity, cross-cultural validity, construct validity and structural validity, according to the COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN). The modified GRADE was applied for evidence synthesis. A total of 95 articles were selected by title and abstract. After removing duplicates and reading and searching the references, a total of 13 studies were included in this review. The original version of the SAQ was described in English, and the instrument was translated into Polish, Canadian French, Simplified Chinese, Spanish (Europe), Danish, Traditional Chinese, Portuguese (Brazil), Korean, German, Turkish and Persian. The evidence was moderate for construct validity, low for internal consistency, and very low for reliability and cross-cultural validity; the content and structural validity properties did not present minimum data for classification. The quality of the evidence regarding the clinimetric properties of the SAQ instrument in adolescents with idiopathic scoliosis was low due to the absence of clinimetric properties or dubious methodological quality. However, for clinical practice and research, we recommend the use of the instrument to assess the self-perception of the spine in adolescents. For future translations and adaptations, we recommend the use of the COSMIN guidelines.


Role of aluminium hydrides in localised corrosion of aluminium revealed by operando Raman spectroscopy.
Filiform corrosion (FFC) is characteristic of metals such as aluminium and magnesium, usually takes place on coated metals, and spreads from coating defects in the form of filaments with a width on the order of 100 μm. In this work, in situ and operando Raman spectroscopy and optical microscopy were used to characterize the composition and distribution of corrosion products inside growing filaments. The filament head contains water (OH stretching modes, 3000-3600 cm-1), and corrosion products based on aluminium oxide with both tetrahedrally (840 cm-1) and octahedrally (600 cm-1) coordinated Al3+, and with some hydroxyl group content (3075, 1420, 1164 cm-1). Remarkable is the prominent presence of structural motifs as in γ-AlH3 (1045, 1495 cm-1). The tail contains predominantly aluminium oxide with octahedrally coordinated Al3+ and in addition carbonate (1100 cm-1) and aluminium chloride (347 cm-1). Video recordings of the active filigree show hydrogen evolution inside the active head and a very fast precipitation of corrosion products. Re-dissolution, transport and re-formation of the corrosion products is also observed, accompanying start-stop-cycles of the propagation of FFC; this mechanism leads to wavy surface morphologies by lifting of certain coating areas after the passage of the corrosion front as evidenced by 3D optical profilometer analysis. When exposed to the acidic head conditions for a sufficient time, the initiation of other forms of localised corrosion, such as pitting, is possible, which in turn facilitates further propagation of the filament. The in situ detection of hydride which transforms into the typical aluminium corrosion products in due course points to a prominent role of hydride as intermediate in the aqueous corrosion of aluminium.


Building a shared vision for an online cancer survivorship community.
In order to achieve comprehensive, closed-loop care for cancer survivors, new strategies are needed to bring together patients, providers, and support services in local communities. To address this challenge, an online community for cancer survivorship was envisioned and designed collaboratively by cancer survivors, family members, community professionals, and informatics researchers in middle Tennessee. The vision developed by the community members serves as a foundation for medical informatics systems to build capacity in local communities to improve cancer care and social support. Using ecological systems theory and social capital as theoretical frameworks, key themes are identified for the future of communication and collaboration in cancer survivorship.


TRICARE; certain survivors of deceased active duty members; and adoption intermediaries. Final rule.
This rule confirms as final a January 2007 interim final rule which implements two provisions of the National Defense Authorization Act for Fiscal Year 2006 (NDAA FY06). First, Section 715 of the NDAA FY06 extends the time frame certain dependents of active duty service members (ADSM) who die while on active duty for more than 30 days shall receive TRICARE medical benefits at active duty dependent payment rates. Second, Section 592 of the NDAA FY06 modifies the requirement for those intermediaries who provide adoption placements. Additionally, this final rule makes an administrative clarification to the following two eligibility provisions: those placed in the legal custody of a member or former member; and those placed in the home of a member or former member in anticipation of adoption. This clarification makes a distinction between the two groups and specifies that for placement into legal custody by court order, the court order must be for a period of 12 consecutive months.


Inhibition of compensatory lung growth in endothelial nitric oxide synthase-deficient mice.
Pneumonectomy results in rapid compensatory growth of the remaining lung and also leads to increased flow and shear stress, which are known to stimulate endothelial nitric oxide synthase (eNOS). Nitric oxide is an essential mediator of vascular endothelial growth factor-induced angiogenesis, which should necessarily occur during compensatory lung growth. Thus our hypothesis is that eNOS is critical for compensatory lung growth. To test this, left pneumonectomy was performed in eNOS-deficient mice (eNOS-/-), and compensatory growth of the right lung was characterized throughout 14 days postpneumonectomy and compared with wild-type pneumonectomy and sham controls. Compensatory lung growth was severely impaired in eNOS-/- mice, as demonstrated by significant reductions in lung weight index, lung volume index, and volume of respiratory region. Also, pneumonectomy-induced increases in alveolar surface density and cell proliferation were prevented in eNOS-/- mice, indicating that eNOS plays a role in alveolar hyperplasia. Compensatory lung growth was also impaired in wild-type mice treated with the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester. Together, these results indicate that eNOS is critical for compensatory lung growth.


Biweekly paclitaxel (Taxol) and cisplatin in breast and ovarian cancer.
Two trials are being conducted to evaluate paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) combined with cisplatin in previously treated and untreated breast and ovarian cancer. Preliminary results are presented. The objectives of these nonrandomized trials are (1) to determine the toxicity of paclitaxel/cisplatin in a biweekly schedule, (2) to establish the maximum tolerated dose of paclitaxel in combination with a fixed dose of cisplatin (60 mg/m2), (3) to determine the feasibility of repeated biweekly administrations, and (4) to evaluate the efficacy of the combination in these diseases. In the breast cancer study, 22 patients have been enrolled to date and eight patients have completed treatment. Dose-limiting neutropenia, which occurred with the starting dose of paclitaxel (90 mg/m2) followed by 60 mg/m2 cisplatin, has precluded any attempts to escalate the paclitaxel dose. Overall, the regimen has been well tolerated at the doses just described. There has been little grade III and no grade IV nonhematologic toxicity. Among 16 patients currently evaluable for response, four had a complete response and II had a partial response, for an overall response rate of 94%. In the ovarian cancer study, 14 patients have been enrolled thus far. Paclitaxel/cisplatin appears to be well tolerated, although it is still too early to assess response rates or to define the MTD. Patients continue to be accrued in both trials.


Establishment and maintenance of a persistent infection of L132 cells by human coronavirus strain 229E.
A persistent infection by human coronavirus 229E (HCV/229E) was established in a human continuous cell line (L132). Following the initial infection with stock HCV/229E, several cultures were established of which two (HV1 and HV4) have been maintained by continuous passage for two years. These cultures have shed high titres of infectious virus continuously into the supernatant fluid since their initiation. The persistently infected cells were resistant to homologous super-infection but supported polio virus replication to normal titres. Preliminary tests indicated that 50-100 percent of the cells contain virus. Neither interferon nor reverse transcriptase could be detected in these cultures and the presence of defective interfering particles could not be demonstrated. VH1 and VH4 coronaviruses, isolated from these persistently infected cultures (HV) and identified by 229E antiserum neutralization, were more cytocidal than the parent virus as judged by plaque characteristics and CPE, however they were indistinguishable on the basis of density, EM morphology, and genome size. Present evidence indicated that temperature plays an important but as yet undetermined role in the establishment and maintenance of stable 229E persistently infected cell cultures.


Single nucleotidic polymorphism 844 A->G of FCAR is not associated with IgA nephropathy in Caucasians.
IgA nephropathy is characterized by a high heterogeneity of clinical expression with 10-30% of patients progressing to end-stage renal failure. The gene of the FcαRI or CD89 presents a single-nucleotide polymorphism responsible for a proinflammatory phenotype of neutrophils in vitro and ex vivo. The aim of our study was to assess whether this CD89 polymorphism 844 A->G is (i) a marker of disease susceptibility and/or (ii) associated with a more severe prognosis. All patients diagnosed with IgA nephropathy and for whom DNA frozen sample was available were included in this European monocentric retrospective analysis and compared to a cohort of healthy volunteers. Allelic discrimination was performed by real-time quantitative polymerase chain reaction (Applied Biosystems™). We first compared the distribution of A and G alleles between patients and volunteers and then studied the relationships between alleles and renal survival, histological score, proteinuria and renal function at diagnosis. Seven hundred and twenty-six patients were analyzed for the study of susceptibility and 425 in the association study. The presence of the G allele was not associated with the occurrence of IgA nephropathy (χ(2) test 0.57, ns). Likewise, renal survival and the criteria for disease activity at time of diagnosis were not affected by the presence of the G allele. No significant association between 844 A->G CD89 polymorphism and the expression of the IgA nephropathy in Caucasians exists. This result does, however, not preclude the implication of other CD89 polymorphisms neither the possibility for a role of CD89 in the pathogenesis of IgA nephropathy.


Antigen-specific TGF-β-induced regulatory T cells secrete chemokines, regulate T cell trafficking, and suppress ongoing autoimmunity.
The ability to regulate ongoing inflammation using regulatory T cells (Tregs) is under intense investigation. Strategies to induce and expand Ag-specific Tregs are being developed, and whether various types of Tregs are suppressive in the inflammatory conditions associated with ongoing disease needs to be determined. In this study, we report that TGF-β-induced Tregs (iTregs) and expanded Tregs specific for a major self-Ag in autoimmune gastritis suppress inflammation and associated pathology when administered late in the process of ongoing disease. Transferred iTregs localized to the stomach, maintained Foxp3 and suppressor functions, and engaged several distinct mechanisms to alleviate disease progression. In addition to suppressing the production of inflammatory cytokines in the stomach and preventing the destruction of parietal cells, we show that iTregs secrete numerous chemokines and regulate both iTreg and effector T cell trafficking into the stomach. These data support efforts to use iTregs in therapies to treat autoimmunity and inflammatory diseases and provide novel insight into the biological mechanisms of iTreg-mediated immune suppression.


Internally quenched peptides for the study of lysostaphin: An antimicrobial protease that kills Staphylococcus aureus.
Lysostaphin (EC. 220.127.116.11) is a protein secreted by Staphylococcus simulans biovar staphylolyticus and has been shown to be active against methicillin resistant S. aureus (MRSA). The design and synthesis of three internally quenched substrates for lysostaphin based on the peptidoglycan crossbridges of S. aureus, and their use in fluorescence resonance energy transfer (FRET) assays is reported. These substrates enabled the gathering of information about the endopeptidase activity of lysostaphin and the effect that mutations have on its enzymatic ability. Significant problems with the inner filter effect and substrate aggregation were encountered; their minimisation and the subsequent estimation of the kinetic parameters for the interaction of lysostaphin with the substrates is described, as well as a comparison of substrates incorporating two FRET pairs: Abz-EDDnp and DABCYL-EDANS. In addition to this, the points of cleavage caused by lysostaphin in Abz-pentaglycine-EDDnp have been determined by HPLC and mass spectrometry analysis to be between glycines 2 and 3(approximately 60%) and glycines 3 and 4 (approximately 40%).


Ionic current driven by a viscosity gradient.
Gradients of voltage, pressure, temperature, and salinity can transport objects in micro- and nanofluidic systems by well-known mechanisms. This paper explores the dynamics of particles in a viscosity gradient with numerical simulations. The different stochastic rules used to integrate the random motion of Brownian particles affect the steady-state distribution of particles in a diffusivity gradient. Importantly, the simulations illuminate the important role that the boundary conditions play, disallowing a steady-state flux when the boundary conditions mimic those of a closed container, but allowing flux when they mimic electrodes. These results provide an interpretation for measurements of a steady ionic current flowing between electrodes separated by a nanofluidic channel with a liquid viscosity gradient.


Childhood overweight and obesity and timing of puberty in boys and girls: cohort and sibling-matched analyses.
Early puberty is a risk indicator for adult diseases. Identification of modifiable causes of earlier puberty is, therefore, warranted. We estimate the association between childhood body mass index (BMI) and pubertal timing in a cohort study and in a sibling-matched study to adjust for unobserved time-stable confounders shared within families. For the cohort study, 11 046 of 22 439 (49%) invited children, born 2000-203, from the Danish National Birth Cohort (DNBC) had information on childhood BMI at 7 years and self-reported, half-yearly puberty information from 11 years on Tanner stages, menarche, voice break, first ejaculation, acne, and axillary hair. For the sibling-matched study, 1700 brothers and sisters were included among 86 820 live-born singletons from the DNBC. Childhood overweight (85th ≤ BMI < 95th percentile) and obesity (BMI ≥ 95th percentile) were associated with earlier age attaining the pubertal milestones in a dose-dependent manner in boys and girls. When modelling all pubertal milestones simultaneously, the pubertal milestones were attained earlier in: overweight boys: -3.1 [95% confidence interval (CI): -4.5, -1.7] months, overweight girls: -5.5 (95% CI: -7.1, -3.9) months, obese boys: -3.5 (95% CI: -5.1, -2.0) months, obese girls: -5.2 (95% CI: -7.1, -3.4) months compared with normal weight (BMI < 85th percentile) children. In the sibling-matched study, higher BMI was associated with earlier age at attaining most pubertal milestones in girls, but only a tendency toward earlier pubertal timing was observed in boys. Childhood overweight and obesity were associated with earlier pubertal timing even after adjustment for unobserved time-stable confounders shared within families.


Organization and operation of a human breast milk bank.
Breast milk has replaced proprietary formulas as an important feeding for the premature and sick neonate. Over 30 milk banks have been established in the United States to provide breast milk to high-risk neonates. Planning a milk bank poses many problems as no uniform standards for quality control exist. A format for developing a breast milk bank and the protocol for donor screening, collection techniques, transport and storage of milk are presented. Rationale supporting each section of the protocol is also included.


Molecular Modeling in Drug Design.
This Special Issue contains thirteen articles that provide a vivid snapshot of the state-of-the-art of molecular modeling in drug design, illustrating recent advances and critically discussing important challenges [...].


Three cases of primary sclerosing cholangitis associated with ulcerative colitis; diagnostic usefulness of magnetic resonance cholangiopancreatography.
Primary sclerosing cholangitis is often accompanied by inflammatory bowel disease in western countries. However, the incidence of primary sclerosing cholangitis in patients with ulcerative colitis appears to be much lower in Japan. Between 1980 and 1998, a total of 402 patients with ulcerative colitis were seen in our department. The patients were evaluated by abdominal ultrasonography, endoscopic retrograde cholangiopancreatography, and/or magnetic resonance cholangiopancreatography when persisting abnormalities of biochemical findings suggested the presence of hepatobiliary diseases. Of the 402 patients with ulcerative colitis, 3 patients with primary sclerosing cholangitis were found. There were 2 men and 1 woman. One patient had left-sided colitis while 2 had total colitis. Magnetic resonance cholangiopancreatography was done in 2 of these 3 patients and demonstrated diagnostic features of primary sclerosing cholangitis. All 3 patients had intra- and extrahepatic involvement by primary sclerosing cholangitis. One male patient died due to progressive hepatic failure. The other male patient was treated with ursodeoxycholic acid, but serum alkaline phosphatase level remained above the normal range. The female patient maintained normal serum alkaline phosphatase levels without specific medication. Magnetic resonance cholangiopancreatography is the most safe and convincing tool for the diagnosis of coexistent primary sclerosing cholangitis in the patients with ulcerative colitis.


Immunogenetics of cytokine genes in parthenium dermatitis: a review.
Parthenium dermatitis is a chronic immuno-inflammatory, distressing skin disease and is mediated by activated T-lymphocyte which is primarily manifested on the exposed sites of the face, neck, hand and flexures. Parthenium hysterophorus is ubiquitous, hence it is diffi-cult to avoid the aero-allergenic antigen parthenin, responsible for the contact dermatitis. The pathogenesis of parthenium dermatitis is characterized by infiltration of T-lymphocytes into challenged skin sites and the development of a cutaneous inflammation due to altered regulatory network of pro and anti-inflammatory cytokines. Regulation of inflammatory events perpetuated by cytokines continues to complicate efforts to analyze both the function of individual cytokine and the influence of candidate gene polymorphism on expression and disease severity. The genetic polymorphisms in these cytokines are significantly affecting immunological parameters and, subsequently, modulation and polarization of immune responses. This review has focused mainly on understanding of the mechanisms of genetic susceptibility of cytokine genes in this disease and, further, this process is likely to achieve significant advances in the diagnosis and management of parthenium dermatitis.


Digestion of starch and fibre carbohydrates in peas by adult cockerels.
The digestion of starch and fibre by adult cockerels fed peas which were whole, ground, autoclaved, heated, dehulled, cooked or supplemented with a cellulose degrading enzyme was studied. The starch in ground peas that had been autoclaved, heated or dehulled was slightly, though not significantly, better digested than the starch in peas that had been ground only. Feeding peas whole greatly reduced starch digestibility. Cooking failed to improve starch digestibility as retrograded starch produced in the cooking process was not digested. The more finely ground the foodstuff, the better the in vitro enzymatic hydrolysis of starch. Starch digestibility correlated well (r2 = 0.80) with the true metabolisable energy values obtained for the different pea treatments. In experiment 1 cockerels digested on average 0.22 of the pea fibre from the different forms of peas. A significant decrease in xylose digestion was observed when birds were fed dehulled peas, birds excreting more xylose than they ingested. In experiment 2, cockerels digested on average 0.38 of the pea fibre from different forms of peas. A reduction in fibre digestion was observed when birds were fed cooked peas as a consequence of a decreased digestibility of all monosaccharide residues. A slight increase in fibre digestion was observed when peas were augmented with 'cellulase', entirely because of an increase in xylose digestion.


Predictors of need for noninvasive ventilation during respiratory tract infections in medically stable, non-ventilated subjects with amyotrophic lateral sclerosis.
Acute lower respiratory infections can impair muscle strength in patients with amyotrophic lateral sclerosis (ALS). When associated with an increase in load on the respiratory system, this situation may precipitate hypercapnic respiratory failure in non-ventilated patients with ALS. The aim of this study was to determine whether a clinical or functional parameter can predict the need for noninvasive ventilation (NIV) during an acute respiratory infection for medically stable, non-ventilated patients with ALS. This was a prospective study involving all non-ventilated subjects with ALS admitted due to an acute respiratory infection to a respiratory care unit from a tertiary hospital. Thirty-two non-ventilated subjects with ALS were admitted to our respiratory care unit due to an acute respiratory infection: 60.72 ± 10.54 y, 13 males, 23 with spinal onset, FVC of 1.58 ± 0.83 L, FVC of 56.21 ± 23.15% of predicted, peak cough flow of 3.41 ± 1.77 L/s, maximum insufflation capacity of 1.87 ± 0.94 L, revised Amyotrophic Lateral Sclerosis Functional Rating Scale score of 22.80 ± 8.83, and Norris bulbar score of 23.48 ± 12.14. Fifteen subjects required NIV during the episode. Logistic regression analysis showed that the only predictors of need for NIV were percent-of-predicted FVC (odds ratio of 1.06, 95% CI 1.01-1.11, P = .02) and peak cough flow (odds ratio of 2.57, 95% CI 1.18-5.59, P = .02). In medically stable, non-ventilated patients with ALS, measurement of percent-of-predicted FVC and peak cough flow can predict the need for NIV during an acute lower respiratory tract infection.


Medium perfusion enables engineering of compact and contractile cardiac tissue.
We hypothesized that functional constructs with physiological cell densities can be engineered in vitro by mimicking convective-diffusive oxygen transport normally present in vivo. To test this hypothesis, we designed an in vitro culture system that maintains efficient oxygen supply to the cells at all times during cell seeding and construct cultivation and characterized in detail construct metabolism, structure, and function. Neonatal rat cardiomyocytes suspended in Matrigel were cultured on collagen sponges at a high initial density (1.35 x 10(8) cells/cm(3)) for 7 days with interstitial flow of medium; constructs cultured in orbitally mixed dishes, neonatal rat ventricles, and freshly isolated cardiomyocytes served as controls. Constructs were assessed at timed intervals with respect to cell number, distribution, viability, metabolic activity, cell cycle, presence of contractile proteins (sarcomeric alpha-actin, troponin I, and tropomyosin), and contractile function in response to electrical stimulation [excitation threshold (ET), maximum capture rate (MCR), response to a gap junctional blocker]. Interstitial flow of culture medium through the central 5-mm-diameter x 1.5-mm-thick region resulted in a physiological density of viable and differentiated, aerobically metabolizing cells, whereas dish culture resulted in constructs with only a 100- to 200-microm-thick surface layer containing viable and differentiated but anaerobically metabolizing cells around an acellular interior. Perfusion resulted in significantly higher numbers of live cells, higher cell viability, and significantly more cells in the S phase compared with dish-grown constructs. In response to electrical stimulation, perfused constructs contracted synchronously, had lower ETs, and recovered their baseline function levels of ET and MCR after treatment with a gap junctional blocker; dish-grown constructs exhibited arrhythmic contractile patterns and failed to recover their baseline MCR levels.


Antigenic studies on coronavirus. I. Identification of the structural antigens of human coronavirus, strain 229E.
Antigenic analysis of human coronavirus, strain 229E (HCV/229E), using a microimmunodiffusion technique, has resulted in the detection of six virion antigens. Comparison of the effect of several different virus-disrupting agents has shown that sodium deoxycholate or Triton X-100 were the best for HCV/229E disruption. Of the six coronavirion antigens, three were identified as virus specific and the remainder as host antigens, which were present as either integrated or nonspecifically adsorbed host components. One of the virus-specific antigens was identified as the internal nucleoprotein (229E/RNP). On the basis of immunodiffusion reactions with isolated 229E/RNP it was concluded that human convalescent sera reacted specifically with the 229E/RNP antigen.


The R-I-M-E method for evaluation of medical students on an obstetrics and gynecology clerkship.
The purpose of this study was to implement and assess the R-I-M-E (Reporter, Interpreter, Manager, Educator) system as a method for evaluation for medical students during the obstetrics and gynecology core clerkship. With the R-I-M-E method (which is an objective framework for evaluation), mid clerkship review sessions are held during which residents verbally assess each student on competencies that include problem identification and reasoning skills, communication skills, physical examination skills, written documentation, and professional attributes. The clerkship director provides timely feedback to the students. Residents and students were surveyed to assess their attitudes about the R-I-M-E method. Most of the attitudes of residents and students were positive about the R-I-M-E evaluation system. The directors feel that R-I-M-E method allows more meaningful, complete, and timely feedback to students, especially in areas that are traditionally difficult to assess, such as professionalism. The R-I-M-E method is an effective evaluation tool for medical students during an obstetrics and gynecology clerkship and is well received by teachers and learners.


Analysis on Fungal Diversity in Rhizosphere Soil of Continuous Cropping Potato Subjected to Different Furrow-Ridge Mulching Managements.
Knowledge about fungi diversity following different planting patterns could improve our understanding of soil processes and thus help us to develop sustainable management strategies. The objective of this study was to determine the impact of different furrow-ridge mulching techniques on fungal diversity in rhizosphere soil under continuous cropping system. The investigated treatments were: flat plot without mulch (CK); flat plot with mulch (T1); on-ridge planting with full mulch (T2); on-furrow planting with full mulch (T3); on-ridge planting with half mulch (T4); and on-furrow planting with half mulch (T5). NGS (Illumina) methods and ITS1 sequences were used in monitoring fungi diversity of the potato rhizosphere soil. The fungi diversity in the rhizosphere soil was ranked in the order T5 > T2 > T4 > T1 > CK at the early growth stage and T2 > T3 > T1 > T4 > CK at the late growth stage of potato. The fungal communities found in the rhizosphere soil were Ascomycota, Zygomycota, Basidiomycota, Chytridiomycota, and other unidentified fungal communities. Among the fungal community in the rhizosphere soil, Ascomycota was found to be dominant fungi population, with the highest percentage (89%) in the T5 soil whereas the T2 soils had the lowest percentage (67%). The Fusarium abundance in fully-mulched treated soils was higher than in half-mulched treated soil. The dominant genus in the T4 soil was Mortierella, whereas lower populations (1-2%) of Scutellinia, Cryphonectria, Acremonium, and Alternaria were found in that treatment. Among the eumycetes, the dominant fungal class in all treated soils was the Sordariomycetes, which ranged from 57 to 85% in T2 and T5 soils, respectively. The Fusarium percentages in half-mulched treated soils (T4 and T5) were 55 and 28% lower than that of complete mulched treated soils (T2 and T3), respectively. The cluster analysis results showed that, CK, T4, and T5 treated soils and T1, T2, and T3 treated soils had similarities in microbial compositions, respectively. Potato tuber yield was greater under the on-ridge planting with full mulch (T2) treated soil, followed by on-ridge planting with half-mulch (T4) treated soil. The rhizosphere soil under the on-ridge planting with full-mulch (T2) soil had the highest fungal diversity, suggesting that this management was the best environment for fungi, whereas the on-ridge planting with half-mulch (T4) soil had the minimum abundance of Fusarium.


Disposition of 14C-alpha-cyclodextrin in germ-free and conventional rats.
The absorption, disposition, metabolism, and excretion of uniformly (14)C-labeled alpha-cyclodextrin ((14)C-alpha-CD) was examined in four separate experiments with Wistar rats. In Experiment 1, (14)C-alpha-CD (25 microCi, 50 mg/kg bw) was administered intravenously to four male and four female conventional rats. In Experiment 2, (14)C-alpha-CD (25 microCi, 200 mg/kg bw) was given by gavage to four male and four female germ-free rats. In Experiments 3 and 4, (14)C-alpha-CD was given to groups of four male and four female conventional rats by gavage at different dose levels (100 microCi, 200 mg/kg bw; 25 microCi, 200 and 100 mg/kg bw). In all experiments, (14)C was measured in respiratory CO(2), urine, and feces over periods of 24-48 h, and in the contents of the gastrointestinal tract, blood, main organs, and residual carcass at termination of the experiments. The chemical identity of the (14)C-labeled compounds was examined by HPLC in blood (Experiment 1), urine (Experiments 1-4), feces (Experiments 2-4), and samples of intestinal contents (Experiments 2 and 4). Recovered (14)C was expressed as percentage of the administered dose. Experiment 1 showed that intravenously administered alpha-CD is excreted rapidly with urine. During the first 2h after dosing, plasma (14)C levels decreased rapidly (t(1/2), 26 and 21 min in male and female rats, respectively). About 13% of the administered (14)C dose (range 4.6-30.6) was detected in the feces, respiratory CO(2), organs, and carcass at the end of the experiment, i.e., 24 h after dosing. The presence of about 1.9% in the intestinal contents and feces suggests that a certain fraction of systemic alpha-CD is eliminated with the bile or saliva. Conclusive evidence, either positive or negative, for a hydrolysis and further metabolism of a small fraction of the administered alpha-CD by the enzymes of the mammalian body could not be gained from this experiment. Upon oral administration of (14)C-alpha-CD to germ-free rats (Experiment 2), about 1.3% of the label expired as CO(2) within 24 h. In the urine collected from 0 to 8 h after dosing, (14)C-alpha-CD was the only radiolabeled compound detected. The amounts of alpha-CD detected in the urine suggest that on average about 1% of an oral dose is absorbed in rats during small-intestinal passage. In conventional rats (Experiments 3 and 4), a delayed appearance of respiratory (14)CO(2) was observed which is attributed to the non-digestibility of alpha-CD and its subsequent microbial fermentation in the cecum and colon. In the urine collected at 4 h after dosing, a small amount of unchanged (14)C-alpha-CD was detected which confirms that about 1% of the ingested alpha-CD is absorbed intact and is excreted via the kidneys. No (14)C-alpha-CD was found in the feces. It is concluded from the data that ingested (14)C-alpha-CD is not digested in the small intestine of rats but is fermented completely by the intestinal microbiota to absorbable short-chain fatty acids. Therefore, the metabolism of alpha-CD resembles closely that of resistant starch or other fermentable dietary fibers.


The influence of a digital clinical reasoning test on medical student learning behavior during clinical clerkships.
Recently, a new digital clinical reasoning test (DCRT) was developed to evaluate students' clinical-reasoning skills. Although an assessment tool may be soundly constructed, it may still prove inadequate in practice by failing to function as intended. Therefore, more insight is needed into the effects of the DCRT in practice. Individual semi-structured interviews and template analysis were used to collect and process qualitative data. The template, based on the interview guide, contained six themes: (1) DCRT itself, (2) test debriefing, (3) reflection, (4) practice/workplace, (5) DCRT versus practice and (6) 'other'. Thirteen students were interviewed. The DCRT encourages students to engage more in formal education, self-study and workplace learning during their clerkships, particularly for those who received insufficient results. Although the faculty emphasizes the different purposes of the DCRT (assessment of/as/for learning), most students perceive the DCRT as an assessment of learning. This affects their motivation and the role they assign to it in their learning process. Although students appreciate the debriefing and reflection report for improvement, they struggle to fill the identified knowledge gaps due to the timing of receiving their results. Some students are supported by the DCRT in exhibiting lifelong learning behavior. This study has identified several ways in which the DCRT influences students' learning practices in a way that can benefit their clinical-reasoning skills. Additionally, it stresses the importance of ensuring the alignment of theoretical principles with real-world practice, both in the development and utilization of assessment tools and their content. Further research is needed to investigate the long-term impact of the DCRT on young physicians' working practice.


Slice it hot: acute adult brain slicing in physiological temperature.
Here we present a protocol for preparation of acute brain slices. This procedure is a critical element for electrophysiological patch-clamp experiments that largely determines the quality of results. It has been shown that omitting the cooling step during cutting procedure is beneficial in obtaining healthy slices and cells, especially when dealing with highly myelinated brain structures from mature animals. Even though the precise mechanism whereby elevated temperature supports neural health can only be speculated upon, it stands to reason that, whenever possible, the temperature in which the slicing is performed should be close to physiological conditions to prevent temperature related artifacts. Another important advantage of this method is the simplicity of the procedure and therefore the short preparation time. In the demonstrated method adult mice are used but the same procedure can be applied with younger mice as well as rats. Also, the following patch clamp experiment is performed on horizontal cerebellar slices, but the same procedure can also be used in other planes as well as other posterior areas of the brain.


Cytochrome P450 reaction-phenotyping: an industrial perspective.
It is now widely accepted that the fraction of the dose metabolized by a given drug-metabolizing enzyme is one of the major factors governing the magnitude of a drug interaction and the impact of a polymorphism on (total) drug clearance. Therefore, most pharmaceutical companies determine the enzymes involved in the metabolism of a new chemical entity (NCE) in vitro, in conjunction with human data on absorption, distribution, metabolism and excretion. This so called reaction-phenotyping, or isozyme-mapping, usually involves the use of multiple reagents (e.g., recombinant proteins, liver subcellular fractions, enzyme-selective chemical inhibitors and antibodies). For the human CYPs, reagents are readily available and in vitro reaction-phenotyping data are now routinely included in most regulatory documents. Ideally, the various metabolites have been definitively identified, incubation conditions have afforded robust kinetic analyses, and well characterized (high quality) reagents and human tissues have been employed. It is also important that the various in vitro data are consistent (e.g., scaled turnover with recombinant CYP proteins, CYP inhibition and correlation data with human liver microsomes) and enable an integrated in vitro CYP reaction-phenotype. Results of the in vitro CYP reaction-phenotyping are integrated with clinical data (e.g., human radiolabel and drug interaction studies) and a complete package is then submitted for regulatory review. If the NCE receives market approval, information on key routes of clearance and their associated potential for drug-drug interactions are included in the product label. The present review focuses on in vitro CYP reaction-phenotyping and the integration of data. Relatively simple strategies enabling the design and prioritization of follow up clinical studies are also discussed.


A possible defective estimation of antineutrophil cytoplasmic antibodies in systemic lupus erythematosus due to the coexistence of periodontitis: preliminary observations.
The objectives of this study is to determine if periodontitis-related ANCA hinder the accurate estimation of this kind of autoantibodies in systemic lupus erythematosus (SLE), due to the frequent coexistence of SLE and periodontitis, and the high incidence of antineutrophil cytoplasmic antibodies (ANCA) in this periodontal condition. Thirty SLE, thirty periodontitis lacking systemic involvement patients, and twenty healthy controls were utilized in this study. The periodontal condition and the presence of ANCA in sera of all individuals was carefully evaluated. For ANCA determination an EIA assay was utilized, directed to a neutrophil granular extract and six neutrophil granule proteins. Sixty percent of SLE patients had periodontitis, and sixty-five percent were ANCA positive. Eighty three percent of all ANCA cases were coexisting with periodontitis. A significant association (p > 0.005) between periodontitis and ANCA was found (Chi Square Test). Fifty percent of the patients with periodontitis lacking systemic involvement were ANCA positive. The results obtained in this study suggest that the figures of ANCA previously reported for SLE, might be overestimated due to the inadvertent presence of periodontitis.


Advances toward tissue engineering for the treatment of stress urinary incontinence.
Suburethral pubovaginal sling placement is a common surgical procedure for the treatment of stress urinary incontinence. A wide variety of graft materials is available, each associated with inherent desirable and undesirable characteristics and complications. In this article, we discuss the rationale for and application of small intestinal submucosa (SIS) in lower urinary tract tissue engineering, with emphasis on the use of SIS as a suitable and biologically compatible sling material. In addition, we discuss exciting research regarding the engineering of true functional sphincter reconstruction using this biologic scaffold and pre-seeded muscle cells.


Carbon-centered radical addition to O=C of amides or esters as a route to C-O bond formations.
Among various types of radical reactions, the addition of carbon radicals to unsaturated bonds is a powerful tool for constructing new chemical bonds, in which the typical applied unsaturated substrates include alkenes, alkynes and imines. Carbonyl is perhaps the most common unsaturated group in nature. This work demonstrates a novel C-O bond formation through carbon-centered radical addition to the carbonyl oxygen of amide or ester, in which amide and ester groups are easily activated through the radical process. EPR spectroscopy and radical clock experiments support the radical process for this transformation, and density functional theory (DFT) calculations support the possibility of carbon-centered radical addition to the carbonyl oxygen of amides or esters.


Characterization of lymphocyte responsiveness in early experimental syphilis. II. Nature of cellular infiltration and Treponema pallidum distribution in testicular lesions.
The nature of the cellular infiltration and the distribution of Treponema pallidum during rabbit testicular infection were examined by immunofluorescence and light microscopy. Low numbers of treponemes are demonstrable in the perivascular regions on day 3 post-infection. On days 6, 10, and 13, large numbers of organisms are found in the interstitial spaces. The treponemes do not appear to invade the walls or lumina of the seminiferous tubules, although tubular atrophy is obvious. On days 17, 24 and 31, treponemes are no longer identifiable by immunofluorescence in infected testicles. The cellular infiltration, which is apparent on day 6, reaches its peak on day 13, corresponding to the amount of swelling observed grossly. The infiltrate is primarily lymphocytic, but macrophages are also observed during the peak cellular response. These cells are located, as are the treponemes, in the interstitial spaces. The lymphocytes are demonstrated by specific immunofluorescence to be predominantly T cells. The peak T cell infiltration in the infected testicle is followed rapidly by the disappearance of the organisms from that organ. Thus, it is postulated that infiltration by specifically sensitized T cells results in the clearance of large numbers of T. pallidum from infected tissues.


Potential disruption of seed dispersal in the absence of a native Kauai thrush.
Hawaii has experienced a catastrophic decline in frugivorous native birds coupled with the introduction of non-native species. Puaiohi (Myadestes palmeri), a critically endangered thrush, is the sole extant native songbird capable of dispersing fleshy fruited plants in the rainforest of Kauai island, Hawaii. As this species has declined to occupy a small proportion of its original range, a suite of largely omnivorous non-native birds have been introduced to this region, including the common and widespread Japanese White-eye (Zosterops japonicus). This reshuffling of the bird community could have long-term implications for plant community composition if introduced birds incompletely replace the ecological role of native species. The objective of this study was to evaluate the potential consequences of the local extirpation of Puaiohi for seed dispersal. Specifically, we compared the diet of Puaiohi and Japanese White-eye, vegetation characteristics, and seed rain at sites with and without Puaiohi in the Na Pali-Kona Forest Reserve on the island of Kauai. We found high overlap in the composition of seeds consumed by the two bird species, but differences in the characteristics of seeds consumed; Japanese White-eye appeared more likely to consume smaller seeded species compared with Puaiohi. Sites with Puaiohi received substantially higher seed rain during the study period, despite no significant differences in overall fruit abundance. Our results suggest that non-native birds are unlikely to completely replace the seed dispersal services provided by Puaiohi. If Puaohi continue to be rare and range restricted, we predict a shift in plant community composition through an increase in non-native and small-seeded plants, and possible dispersal failure of other native species. Our findings lend further support to efforts to conserve Puaiohi across its current and former range, and to consider introductions to other suitable areas to ensure the persistence not only of the species and but also its functional role in Hawaii's montane ecosystems.


Laser removal of pigmented and vascular lesions.
Twenty years of laser dermatology has resulted in current technology which allows variable spot sizes, different wavelengths, and a variety of effective cooling devices. These developments have made the treatment of cutaneous pigmented and vascular lesions safe and efficacious by targeting selected chromophores while minimizing damage to the surrounding tissue. Vascular lesions are targeted by a variety of wavelength lasers including the KTP (532 nm), pulsed dye (585-595 nm), and the Nd:YAG (1064 nm) laser systems. Pigmented lesions may be treated with a wide variety of lasers due to the broad absorption spectrum of melanin. Intense pulsed light (IPL), with its widely adjustable parameters, has established itself as a useful adjunctive for the treatment of a variety of pigmented and vascular lesions. The purpose of this review article is to present the current treatment options for the common aesthetic complaints of pigmented and vascular lesions.


Cetirizine vs placebo in chronic idiopathic urticaria--a double blind randomised cross-over study.
The efficacy of cetirizine dihydrochloride, a new H1-antagonist claimed to have minimal sedative property was evaluated in 28 patients with chronic idiopathic urticaria. Cetirizine 10 mg and placebo were compared in a double-blind cross-over trial. Response was measured by patients with daily charts on itch and wheals. Response was also assessed by patients and investigators while on cetirizine and placebo on an analogue scale. The mean analogue score by the investigator while on cetirizine (64.6 mm) was better than while on placebo (47.0 mm) (p = 0.008). Reduction of itch with cetirizine was significantly greater than with placebo. The mean itch scores for placebo and cetirizine were 21.2 and 14.5 respectively (p = 0.01) showing a significant improvement of itch by cetirizine. The mean score for wheal response for placebo and cetirizine were 21.0 and 16.16 respectively (p = 0.07 NS) showing a reduction of wheal from cetirizine was greater than placebo but the difference was not statistically significant. The patients' mean visual analogue score for placebo was 38.4 and for cetirizine was 57.5 (p = 0.006). Cetirizine was significantly better in controlling symptoms of chronic urticaria than placebo. Five of the 28 (17.9%) patients reported mild drowsiness while taking cetirizine. Two patients reported drowsiness while on placebo. Tolerance was reported as good in 85%, moderate in 10% and bad in 5% according to patients' assessment, while tolerance was excellent and good in 80%, moderate in 15% of patients according to investigator's assessment.


Fear conditioning and extinction: influence of sex and menstrual cycle in healthy humans.
Although sex differences have been demonstrated in behavioral paradigms of fear conditioning, the findings have been inconsistent, and fear extinction has been little studied. The present study investigated the influence of sex and menstrual cycle phase on the recall of fear extinction. Three groups of healthy adult participants were studied: women at 2 different phases of the menstrual cycle (early follicular [early cycle] and late follicular [midcycle]) and men. Participants underwent a 2-day fear conditioning and extinction protocol. The paradigm entailed habituation, fear conditioning, and extinction learning on Day 1 and extinction recall and fear renewal on Day 2. Skin conductance served as the dependent variable. During fear acquisition on Day 1, men showed significantly larger conditioned responses relative to women; early cycle and midcycle women did not differ. No significant group differences were found during extinction learning. On Day 2, men and early cycle women expressed greater extinction memory than midcycle women. These data confirm sex differences in conditioned fear acquisition and suggest that midcycle hormones attenuate extinction recall.


Sclerosing encapsulating peritonitis: complication of laparoscopic cholecystectomy.
Sclerosing encapsulating peritonitis (SEP) is a rare cause of intestinal obstruction. It is difficult to make a definite preoperative diagnosis, and most cases are diagnosed at the time of laparotomy. It is usually of unknown origin, although, at times, it may be seen secondary to a variety of conditions. Spillage of bile and gallstones at laparoscopic cholecystectomy is an unusual cause of SEP and has not been reported in literature, to date. Contrast-enhanced computed tomography of the abdomen revealed small-bowel loops congregated to the center of the abdomen encased by a soft-tissue density mantle with loculated fluid in the interloop bowel location. Excision of the sac and adhesiolysis was done in our patient for recurrent episodes of intestinal obstruction, who recovered well in the postoperative period.


Distribution, determinants, and prognostic value of gamma-glutamyltransferase for all-cause mortality in a cohort of construction workers from southern Germany.
Serum gamma-glutamyltransferase (GGT) is commonly measured as a marker of hepatobiliary disorders in clinical practice, but little is known about its distribution and prognostic value for all-cause mortality. Distribution and determinants of serum GGT levels were assessed among 8,043 construction workers ages 25-64 who underwent occupational health examinations in six centers in Southern Germany from 1986 to 1988. Study participants were followed for all-cause mortality until 1994. Serum GGT levels were considerably higher in this cohort than among male employees examined in a national survey conducted during the same period. The factors most strongly related to serum GGT were self-reported alcohol consumption, body mass index, diabetes, and hypertension, but relations of GGT levels were also found with nationality, occupation, and smoking. There was a strong dose-response relation between serum GGT levels and all-cause mortality (P value for trend < 0.001). Compared with men with GGT levels below 15 U/liter (measured at 25 degrees C), relative risks (95% CI) were 1.46 (0.86-2.49), 1.78 (1.08-2.94), 2.09 (1.26-3.45), and 3.44 (2.20-5.38) for men with GGT levels of 15-19, 20-29, 30-49, and > or = 50 U/liter, respectively. This relation was reduced but not eliminated by control for body mass index, diabetes, hypertension, alcohol consumption, and other covariates in multivariable analysis. Serum GGT is a strong risk indicator of all-cause mortality.


Synthesis and monoamine transporter affinity of new 2beta-carbomethoxy-3beta-[aryl or heteroaryl]phenyltropanes.
A series of 16 new 2beta-carbomethoxy-3beta-[aryl or heteroaryl]phenyltropane derivatives was synthesized and evaluated for binding to monoamine transporters. Most of the compounds exhibited nanomolar affinity for the serotonin transporter (SERT). Four compounds presented a particularly attractive pharmacological profile, with very high SERT affinity (K(i) 0.15-0.5 nM) and selectivity versus the dopamine transporter of 25- to 77-fold.


Gadofosveset-enhanced magnetic resonance imaging of human carotid atherosclerotic plaques: a proof-of-concept study.
To investigate the potential of gadofosveset-enhanced MR imaging for the characterization of human carotid atherosclerotic plaques. Sixteen (9 symptomatic, 7 asymptomatic) patients with 70% to 99% carotid stenosis (according to NASCET criteria) were included (13 men, 3 women, mean age 67.6 years). All patients underwent baseline precontrast MR imaging of the carotid plaque. Immediately after completion of the baseline examination, 0.03 mmol/kg gadofosveset was administered. At 24 hours postinjection, the acquisition was repeated. Twelve patients were scheduled for carotid endarterectomy. Carotid endarterectomy specimens were HE-, CD31-, CD68-, and albumin-stained to correlate signal enhancement with plaque composition, intraplaque microvessel density, and macrophage and albumin content. A random intercept model was used to compare signal enhancement between symptomatic and asymptomatic patients, adjusting for size of various plaque components. This study was approved by the institutional medical ethics committee. All participants gave written informed consent. Signal enhancement (SE) of the plaque was significantly higher in symptomatic patients compared with asymptomatic patients (median log SE 0.182 vs. -0.109, respectively, P < 0.001). A positive association (as expressed by a regression coefficient beta = 0.0035) was found between signal enhancement on the log scale and intraplaque albumin content (P = 0.038). There was no association between signal enhancement and various other plaque components. In this study, the potential of gadofosveset-enhanced human carotid plaque MR imaging for identification of high-risk plaques was demonstrated. Signal enhancement of the plaque after administration of gadofosveset was associated with differences in intraplaque albumin content. Although promising, we emphasize that these results are based on a small patient population. Larger prospective studies are warranted.


Outcome of Buried Versus Exposed Kirchner Wires in Terms of Infection in Fractures of Phalanges and Metacarpal Bones of Hand.
Introduction and objective The fracture of hand bones is very common among manual hand workers and a fractured hand imparts a great effect on a person's productivity both socioeconomically and from a body image point of view. The most common method of hand fractures fixation is with the help of Kirschner wires. Kirchner wires can be inserted in exposed or in buried manner. There are a few studies that provide a comparative analysis of rate of infection between these two techniques. This study aimed to assess the rate of infection in buried versus exposed Kirschner (K)-wires for hand fractures. Material and method The study was designed as a randomized controlled trial with consecutive non-random sampling. It was conducted in the Department of Plastic Surgery, Holy Family Hospital, Rawalpindi, Pakistan, and lasted from June to December 2019. Blinding was not possible as both the operating surgeon and patient were aware of the procedure being done; however, the assessor was blinded and was not aware which group got which treatment. Total 122 patients with fractures of metacarpals and phalanges of hand were included in the study and were divided into two groups with 61 patients in each. Group A was treated with buried K-wires and group B with exposed K-wires. The patients were followed for one month for the outcomes in terms of infection in the patients. Results Group A had 24 females (39.3%) and 37 males (60.7%). Group B had 16 females (26.2%) and 45 males (73.8%). In group A, nine (14.8%) patients had ages between 10 and 20 years, 18 (29.5%) patients between 21 and 30 years, 14 (23.0%) patients between 31 and 40 years, 11 (18.0%) patients between 41 and 50 years, and nine (14.8%) were between 51 and 60 years. The mean duration of surgery was 35.16 minutes for group A and 27.30 minutes for group B. Based on modified Oppenheim scoring system for pin site infection, out of 61 patients, seven (11.5%) with buried K-wires while 14 (23%) with exposed K-wires developed pin site infection. Conclusion Rate of infection is low in buried K-wires as compared to exposed K-wires though not statistically significant (p>0.05) for the fractures of metacarpals and phalangeal fractures of hand.


Comparison of Flexible Ureterorenoscope Quality of Vision: An In Vitro Study.
Flexible ureterorenoscopy (fURS) is one of the best solutions for treatment of renal calculi <2 cm and for upper urinary tract urothelial carcinoma conservative treatment. An adequate quality of vision is mandatory to help surgeon get better outcomes. No studies have been done, to our knowledge, about what fURS in the market has the best quality of vision. Seven different fURS were used to compare the image quality (Lithovue, Olympus V, Olympus V2, Storz Flex XC-in White Light and in Clara+Chroma mode-Wolf Cobra Vision, Olympus P6, and Storx Flex X2). Two standardized grids to evaluate contrast and image definition and three stones of different composition were filmed in four standardized different scenarios. These videos were shown to 103 subjects (51 urologists and 52 nonurologists) who had to evaluate them with a rating scale from 1 (very bad) to 5 (very good). No difference in terms of scores was observed for sex of the participants. Digital (D) ureterorenoscopes were rated better than fiber optics (FOs) ureterorenoscopes. Overall, Flex XC White Light and XC Clara+Chroma image quality resulted steadily better than other fURS (p < 0.0001). Olympus V generally provided a vision better than Lithovue. Cobra Vision and Olympus V2 had superimposable values that were significantly lower than Lithovue's ones. Olympus P6 and Storz X2 offered a low quality of vision compared to the others. In the medium simulating bleeding, Olympus V and V2 significantly improved their scores of 12% and 8.1%, contrary to rest of the ureterorenoscopes. D ureterorenoscopes have a better image quality than FO ones. The only disposable ureterorenoscope tested was comparable to the majority of other D ureterorenoscopes. The best image quality was provided by Storz D ureterorenoscopes, being Clara Chroma the favorite Spies Mode, according to literature.


Indirubin Treatment of Lipopolysaccharide-Induced Mastitis in a Mouse Model and Activity in Mouse Mammary Epithelial Cells.
Indirubin is a Chinese medicine extracted from indigo and known to be effective for treating chronic myelogenous leukemia, neoplasia, and inflammatory disease. This study evaluated the in vivo anti-inflammatory activity of indirubin in a lipopolysaccharide- (LPS-) induced mouse mastitis model. The indirubin mechanism and targets were evaluated in vitro in mouse mammary epithelial cells. In the mouse model, indirubin significantly attenuated the severity of inflammatory lesions, edema, inflammatory hyperemia, milk stasis and local tissue necrosis, and neutrophil infiltration. Indirubin significantly decreased myeloperoxidase activity and downregulated the production of tumor necrosis factor-α, interleukin-1β (IL-1β), and IL-6 caused by LPS. In vitro, indirubin inhibited LPS-stimulated expression of proinflammatory cytokines in a dose-dependent manner. It also downregulated LPS-induced toll-like receptor 4 (TLR4) expression and inhibited phosphorylation of LPS-induced nuclear transcription factor-kappa B (NF-κB) P65 protein and inhibitor of kappa B. In addition to its effect on the NF-κB signaling pathway, indirubin suppressed the mitogen-activated protein kinase (MAPK) signaling by inhibiting phosphorylation of extracellular signal-regulated kinase (ERK), P38, and c-jun NH2-terminal kinase (JNK). Indirubin improved LPS-induced mouse mastitis by suppressing TLR4 and downstream NF-κB and MAPK pathway inflammatory signals and might be a potential treatment of mastitis and other inflammatory diseases.


Computerised decision to reduce inappropriate medication in the elderly: a systematic review with meta-analysis protocol.
Life expectancy continues to increase in developed countries. Elderly people are more likely to consume more medications and become vulnerable to age-related changes in drugs' pharmacokinetics and pharmacodynamics. Recent studies have identified opportunities and barriers for deprescribing potentially inappropriate medications. It has already been demonstrated that computerised decision support systems can reduce physician orders for unnecessary tests. We will systematically review the available literature to understand if computerised decision support is effective in reducing the use of potentially inappropriate medications, thus having an impact on health outcomes. A systematic review will be conducted using MEDLINE, CENTRAL, EMBASE and Web of Science databases, as well as the grey literature assessing the effectiveness of computer decision support interventions in deprescribing inappropriate medication, with an impact on health outcomes in the elderly. The search will be performed during January and February 2018. Two reviewers will conduct articles' screening, selection and data extraction, independently and blind to each other. Eligible sources will be selected after discussing non-conformities. All extracted data from the included articles will be assessed based on studies' participants, design and setting, methodological quality, bias and any other potential sources of heterogeneity. This review will be conducted and reported in adherence to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement of quality for reporting systematic reviews and meta-analyses. As a systematic review, this research is exempt from ethical approval. We intend to publish the full article in a related peer-reviewed journal and present it at international conferences. CRD42017067021.


Development of a versatile enzyme immunoassay for non-invasive assessment of glucocorticoid metabolites in a diversity of taxonomic species.
Endocrinology is a useful tool for conservation biologists and animal managers, and measuring glucocorticoids can help understand biological mechanisms associated with species decline and animal welfare. The current study describes the development and optimization of a glucocorticoid enzyme immunoassay (EIA) to non-invasively assess adrenal activity in a variety of taxa. The antiserum (CJM006) was raised in rabbits to a corticosterone-3-CMO-BSA immunogen and used in a standard competitive EIA system. However, the EIA initially produced results with unacceptably high inter-assay variation, attributed to consistent patterns observed within the optical density of developing plates. To determine the cause of this variability, a number of factors were examined using synthetic corticosterone standard and endogenous faecal extract, including: plate type (Nunc MaxiSorp® II versus Immulon IB plates); the use of non-specific secondary antibody; type (artificial versus natural) and presence (light versus dark) of light during incubation; plate loading temperature (4°C versus room temperature); and substrate reagent temperature (4°C versus room temperature). Results indicated that variability was associated with plate location effects, which were not initially detected because control samples were always run in the same positions across plates. Light and temperature were the two major factors that affected EIA reliability. For this assay, the standard protocol required slight modification, with the optimal protocol using Nunc MaxiSorp® plates, room temperature substrate reagents and dark incubation conditions. Following optimization, this EIA was then validated biochemically for 38 species, through parallel displacement curves and interference assessment tests of faecal and urine samples. Additionally, biological validation was performed opportunistically in a subset of species, with use of this EIA demonstrating significant elevations in faecal glucocorticoid metabolites following potentially challenging events. In summary, this glucocorticoid EIA cross-reacts with excreted glucocorticoid metabolites across a wide range of taxa, including ungulates, primates, felids, birds, rodents and amphibians. We conclude that when used with optimal reagent and incubation conditions, this EIA will be useful for non-invasive monitoring of adrenal activity in a wide range of wildlife species.


Metallothionein from Wild Populations of the African Catfish Clarias gariepinus: From Sequence, Protein Expression and Metal Binding Properties to Transcriptional Biomarker of Metal Pollution.
Anthropogenic pollution with heavy metals is an on-going concern throughout the world, and methods to monitor release and impact of heavy metals are of high importance. With a view to probe its suitability as molecular biomarker of metal pollution, this study has determined a coding sequence for metallothionein of the African sharptooth catfish Clarias gariepinus. The gene product was recombinantly expressed in Escherichia coli in presence of Zn(II), Cd(II), or Cu, and characterised by Electrospray Ionisation Mass Spectrometry and elemental analysis. C. gariepinus MT displays typical features of fish MTs, including 20 conserved cysteines, and seven bound divalent cations (Zn(II) or Cd(II)) when saturated. Livers from wild C. gariepinus fish collected in all three seasons from four different sites on the Kafue River of Zambia were analysed for their metal contents and for MT expression levels by quantitative PCR. Significant correlations were found between Zn and Cu levels and MT expression in livers, with MT expression clearly highest at the most polluted site, Chililabombwe, which is situated in the Copperbelt region. Based on our findings, hepatic expression of MT from C. gariepinus may be further developed as a major molecular biomarker of heavy metal pollution resulting from mining activities in this region.


Characterization of DC-STAMP+ Cells in Human Bone Marrow.
Osteoclasts (OC), specialized cells derived from monocytes, maintain skeletal homeostasis under normal conditions but degrade bone in patients with rheumatoid (RA) and psoriatic arthritis (PsA). Monocytes initially develop in the bone marrow (BM), circulate in peripheral blood, and differentiate into distinct cell types with diverse functions. Imaging studies in (RA) patients and murine arthritis models demonstrate that bone marrow edema detected on MRI is the result of enhanced myelopoiesis which precedes the development of bone erosions detected on plain radiographs several years later. A major knowledge gap, however, is whether OC develop in the BM and circulate to the joint and if the differentiation to OC takes place in the joint space in response to differentiation signals such as RANKL and TNF. We have previously demonstrated that osteoclast precursors (OCP) are increased in the circulaton of patients with RA and PsA. We showed that DC-STAMP (Dendritic Cell-Specific Transmembrane protein), a 7-pass transmembrane protein expressed on the surface of monocytes, is essential for cell-to-cell fusion during OC differentiation and is a valid biomarker of OCP. Herein, we examined OCP in human bone marrow and identified one novel subset of DC-STAMP+CD45intermediate monocytes which was absent in the blood. We also found that OCPs reside in human BM with a higher frequency than in the peripheral blood. These findings support the notion that the BM is a major reservoir of circulating OCPs. In addition, we demonstrated that a higher frequency of DC-STAMP+ cells in the BM have detectable intracellular IFN-γ, IL-4 and IL-17A than DC-STAMP+ cells circulating in the peripheral blood. Finally, the frequency of DC-STAMP+ monocytes and T cells is signficantly higher in PsA BM compared to healthy controls, suggesting an enhanced myelopoiesis is a central event in inflammatory arthritis.


Myotonic dystrophy type I in childhood Long-term evolution in patients surviving the neonatal period.
In a retrospective study, 32 patients with myotonic dystrophy, including congenital (n=17) and infantile/juvenile forms (n=15) were studied during a long follow-up lasting 7-28 years (median: 17 years). The clinical presentation was extremely variable; however, a continuum did exist between severe and less severe congenital forms, and later-onset forms, without genotype-phenotype correlation. We observed some unusual presentations, such as 3 cases of isolated club-feet during the neonatal period, and 7 patients (23%) with a completely isolated mental deficiency, language delay and school failure, who only completed the clinical picture several years later. Wechsler scale testing was performed in all cases, and repeated with 8 patients. It demonstrated a decrease in intellectual abilities in 5 patients, suggesting the possibility of a degenerative cerebral process occurring in these children. This decrease has also been reported in some adult cases. This study illustrates the extremely heterogeneous clinical presentation of myotonic dystrophy in childhood.


Pituitary-specific transcription factor (Pit-1) binding site in the human renin gene 5'-flanking DNA stimulates promoter activity in placental cell primary cultures and pituitary lactosomatotropic cell lines.
Renin gene expression is limited to a number of specific tissues, including the kidney, adrenal glands, reproductive organs (of particular relevance to this study, the placenta), and the pituitary gland. In the present study, we investigated the human renin (hRen) 5'-flanking DNA sequences required to drive the expression of a luciferase reporter gene in placental and pituitary cells and in two cell lines, 293 and JEG-3, which have been proposed as model systems with which to study transcriptional regulation of renin genes. The activities of specific sequences in the hRen 5'-flanking DNA sequences in human placental cell primary cultures were very similar to those that we previously reported in pituitary cells, suggesting the involvement of common promoter elements and related transcription factors. Accordingly, the binding site for the pituitary-specific transcription factor (Pit-1) was the major determinant of renin promoter activity in both pituitary and placental cells. Gel mobility shift analysis showed a placental nuclear factor with a gel mobility different from that of Pit-1. However, Northern blot analysis failed to demonstrate abundant Pit-1-related mRNAs in renin-expressing cultures of chorionic and decidual cells, suggesting that the placental factor is not closely related to Pit-1. Although a factor from 293 cells also bound to the Pit-1 site, it had gel mobility shift characteristics different from Pit-1 and the placental factor. Moreover, the low promoter activity in 293 cells was independent of this site or, indeed, of sequences upstream from the TATA box. In JEG-3 cells, renin 5'-flanking DNA sequences showed virtually no transcriptional activity.(ABSTRACT TRUNCATED AT 250 WORDS)


Investigating the Asymmetrical Roles of Syllabic and Phonemic Awareness in Akshara Processing.
In this study, we examine the relative contributions of syllabic awareness, phonemic awareness, and oral vocabulary knowledge in early akshara reading ability from Grades 1 through 5. The performance of 488 students in two states of South India, Karnataka (Kannada language) and Andhra Pradesh (Telugu language), was measured. Results from a commonality analysis indicate that there was an increasing independent contribution of syllabic awareness to Kannada and Telugu decoding through the five grades, but the unique contribution of phonemic awareness steadily declined through the five grades, as it became subsumed within syllabic awareness. The contribution of oral vocabulary knowledge did not present a clear pattern across the five grades. This study builds on a growing body of literature on the akshara orthographies to shed light on the precise nature of the developmental asymmetry in the dual syllabic and phonemic representation in akshara reading.


Septins are involved at the early stages of macroautophagy in S. cerevisiae.
Autophagy is a conserved cellular degradation pathway wherein double-membrane vesicles called autophagosomes capture long-lived proteins, and damaged or superfluous organelles, and deliver them to the lysosome for degradation. Septins are conserved GTP-binding proteins involved in many cellular processes, including phagocytosis and the autophagy of intracellular bacteria, but no role in general autophagy was known. In budding yeast, septins polymerize into ring-shaped arrays of filaments required for cytokinesis. In an unbiased genetic screen and in subsequent targeted analysis, we found autophagy defects in septin mutants. Upon autophagy induction, pre-assembled septin complexes relocalized to the pre-autophagosomal structure (PAS) where they formed non-canonical septin rings at PAS. Septins also colocalized with autophagosomes, where they physically interacted with the autophagy proteins Atg8 and Atg9. When autophagosome degradation was blocked in septin-mutant cells, fewer autophagic structures accumulated, and an autophagy mutant defective in early stages of autophagosome biogenesis (atg1Δ), displayed decreased septin localization to the PAS. Our findings support a role for septins in the early stages of budding yeast autophagy, during autophagosome formation.This article has an associated First Person interview with the first author of the paper.


[Progress and prospect of synthetic biodegradable polymers for bone repair and reconstruction].
To review the latest researches of synthetic biodegradable polymers for bone repair and reconstruction, to predict the progress of bone substitute materials and bone tissue engineering scaffolds in future. The literature concerning synthetic biodegradable polymers as bone substitute materials or bone tissue engineering scaffolds was collected and discussed. Aliphatic polyester, polyanhydride, polyurethane and poly (amino acids) were the most extensively studied synthetic biodegradable polymers as bone substitutes and the scaffolds. Each polymer was of good biological safety and biocompatibility, and the degradation products were nontoxic to human body. The mechanical properties and degradation rate of the polymers could be adjusted by the type or number of the monomers and different synthetic methods. Therefore, the polymers with suitable mechanical strength and degradation rate could be produced according to the different requirements for bone grafting. Preliminary studies in vivo showed their favorable capacity for bone repair. The synthetic biodegradable polymers, especially the copolymers, composite materials and those carrying bone growth factors are expected to be the most promising and ideal biomaterials for bone repair and reconstruction.


CRISPR-Cas9 Targeted Mutagenesis Leads to Simultaneous Modification of Different Homoeologous Gene Copies in Polyploid Oilseed Rape (Brassica napus).
In polyploid species, altering a trait by random mutagenesis is highly inefficient due to gene redundancy. We have stably transformed tetraploid oilseed rape (Brassica napus) with a CRISPR-Cas9 construct targeting two ALCATRAZ (ALC) homoeologs. ALC is involved in valve margin development and, thus, contributes to seed shattering from mature fruits. Knocking out ALC would increase shatter resistance to avoid seed loss during mechanical harvest. We obtained a transgenic T1 plant with four alc mutant alleles by the use of a single target sequence. All mutations were stably inherited to the T2 progeny. The T2 generation was devoid of any wild-type alleles, proving that the underlying T1 was a nonchimeric double heterozygote. T-DNA and ALC loci were not linked, as indicated by random segregation in the T2 generation. Hence, we could select double mutants lacking the T-DNA already in the first offspring generation. However, whole-genome sequencing data revealed at least five independent insertions of vector backbone sequences. We did not detect any off-target effects in two genome regions homologous to the target sequence. The simultaneous alteration of multiple homoeologs by CRISPR-Cas9 mutagenesis without any background mutations will offer new opportunities for using mutant genotypes in rapeseed breeding.


Image analysis of nuclear/cytoplasmic ratio in cervical smears to discriminate three grades of cervical intraepithelial neoplasia.
To suggest which system of cytologic classification (Bethesda System or World Health Organization [WHO]) of cervical lesions is more adequate for the Public Health Service of São Paulo State, Brazil, based on morphometric study with the Jandel Videoanalysis (JAVA) System. The study comprised groups of typical smears: cervical intraepithelial neoplasia (CIN 1) separated into two subgroups: cytopathic effects of human papillomavirus associated or not with dyskaryosis, CIN 2 and CIN 3. The JAVA system of image analysis was used to determine the nuclear/cytoplasmic (N/C) ratio in abnormal cells from each group. Significant differences were detected between the three grades of CIN and between the two subgroups of CIN 1. Although image analysis is not applicable to large-scale population screening of cervical smears, its use in the present study favored the three-grade cytologic classification (WHO). In addition, the three-grade classification offers the clinician more options for treatment. Considering the clinical-laboratory characteristics of our public health service, the three-grade classification is more adequate.


Secondary use of hospital information system data for safe bedside radiography in terms of patient factors.
The purpose of this study was to use patient data gathered by a hospital information system (HIS) to improve the safe performance of bedside radiography. Hierarchical cluster analysis was used to investigate the factors of hospitalised patients who had undergone radiography in the X-ray room or at the bedside. Logistic regression analysis was then performed to quantify patient factors and calculate the probability of undergoing general radiography or bedside radiography. Patients were grouped into six clusters by hierarchical cluster analysis on the basis of their factors. We found a remarkable difference between clusters for the ratio of bedside radiography. Results indicated that "types of transportation" and "level of mobility" related to the ratio of bedside radiography. Logistic regression analysis of the associations between the probability of undergoing bedside radiography and patient factors indicated that type of transportation and level of mobility were highly correlated with bedside radiography or general radiography. Our results suggested that the secondary use of HIS data for the quantitative evaluation of patient factors and implementation of those quantitative values in medical records may be useful for the safe performance of bedside radiography as well as providing a method of decision support for doctors to order bedside radiography.


Transplantation of kidneys from living related donors. Comparison with cadaveric kidney transplantation.
In a series of 271 transplantations of renal allografts, performed over 10 years, the rates of graft survival, patient survival, and morbidity in the recipients of allografts from living related donors (47 allografts) have been compared with those in the recipients of cadaveric allografts (224 allografts). The one-year graft survival rates were 88% for allografts from living related donors (100%, if these were HLA-identical) and 55% for cadaveric allografts, while the patient survival rates were 97% and 87%, respectively, in the same period. Morbidity rates (expressed as the number of days spent in hospital) for recipients of allografts from living related donors were approximately 50% of those for recipients of cadaveric grafts. Complications in the living related donors were minimal, and acceptable. It is concluded that transplantation of allografts from living related donors has many advantages over transplantation of cadaveric kidneys, and is a valuable adjunct to a cadaveric renal transplantation programme. Greater use of living related kidney donors should be encouraged in Australia.