Low secondary transmission of 2009 pandemic influenza A (H1N1) in households following an outbreak at a summer camp: relationship to timing of exposure.
Following an outbreak of 2009 pandemic influenza A (H1N1) at a residential summer camp for boys aged 10-16 years, we assessed secondary household transmission of the novel virus after their return home. Of 212 study participants who attended camp, 49 had confirmed or probable influenza for a primary attack rate of 23%. Of 87 exposed household contacts who did not attend camp, only three instances of probable transmission were observed, for a household secondary attack rate of 3·5%. All secondary cases occurred in households where the ill camp attendee returned home 1 day after onset of illness, with an attack rate of 14·3% in household contacts in this category. Returning home after peak infectivity to others and advanced warning prior to reintegration of sick individuals into the household probably contributed to the overall low secondary attack rate observed.


An epigenetic marker panel for detection of lung cancer using cell-free serum DNA.
We investigated the feasibility of detecting aberrant DNA methylation of some novel and known genes in the serum of lung cancer patients. To determine the analytic sensitivity, we examined the tumor and the matched serum DNA for aberrant methylation of 15 gene promoters from 10 patients with primary lung tumors by using quantitative methylation-specific PCR. We then tested this 15-gene set to identify the more useful DNA methylation changes in the serum of a limited number of lung cancer patients and controls. In an independent set, we tested the six most promising genes (APC, CDH1, MGMT, DCC, RASSF1A, and AIM1) for further elucidation of the diagnostic application of this panel of markers. Promoter hypermethylation of at least one of the genes studied was detected in all 10 lung primary tumors. In majority of cases, aberrant methylation in serum DNA was accompanied by methylation in the matched tumor samples. In the independent set, using a single gene that had 100% specificity (DCC), 35.5% (95% CI: 25-47) of the 76 lung cancer patients were correctly identified. For patients without methylated DCC, addition of a logistic regression score that was based on the five remaining genes improved sensitivity from 35.5% to 75% (95% CI: 64-84) but decreased the specificity from 100% to 73% (95% CI: 54-88). This approach needs to be evaluated in a larger test set to determine the role of this gene set in early detection and surveillance of lung cancer.


Sensitive and selective quantification of free and total malondialdehyde in plasma using UHPLC-HRMS.
Quantification of malondialdehyde (MDA) as a marker of lipid peroxidation is relevant for many research fields. We describe a new sensitive and selective method to measure free and total plasmatic MDA using derivatization with 2,4-dinitrophenylhydrazine (DNPH) and ultra-HPLC-high-resolution MS. Free and total MDA were extracted from minute sample amounts (10 μl) using acidic precipitation and alkaline hydrolysis followed by acidic precipitation, respectively. Derivatization was completed within 10 min at room temperature, and the excess DNPH discarded by liquid-liquid extraction. Quantification was achieved by internal standardization using dideuterated MDA as internal standard. The method's lowest limit of quantification was 100 nM and linearity spanned greater than three orders of magnitude. Intra- and inter-day precisions for total MDA were 2.9% and 3.0%, respectively, and those for free MDA were 12.8% and 24.9%, respectively. Accuracy was 101% and 107% at low and high concentrations, respectively. In human plasma, free MDA levels were 120 nM (SD 36.26) and total MDA levels were 6.7 μM (SD 0.46). In addition, we show the applicability of this method to measure MDA plasma levels from a variety of animal species, making it invaluable to scientists in various fields.


The aqueous extract of a popular herbal nutrient supplement, Angelica sinensis, protects mice against lethal endotoxemia and sepsis.
Despite recent advances in antibiotic therapy and intensive care, sepsis remains a widespread problem in critically ill patients. The high mortality from sepsis is in part mediated by bacterial endotoxin, which stimulates macrophages/monocytes to sequentially release early (e.g., tumor necrosis factor, interleukin-1, and interferon-gamma) and late [e.g., high mobility group box 1 protein (HMGB1)] proinflammatory cytokines. Our discovery of HMGB1 as a late mediator of lethal systemic inflammation has initiated a new field of investigation for the development of experimental therapeutics. A popular Chinese herb, Angelica sinensis (also known as Dang Gui or Dong Quai) has been used traditionally for treating women with gynecological disorders (such as dysmenorrheal and hot flashes). Here we examined the effect of Angelica sinensis extract on endotoxin-induced HMGB1 release in vitro, and explored its therapeutic potential in animal models of lethal endotoxemia and sepsis [induced by cecal ligation and puncture (CLP)] in vivo. We demonstrated that a low-molecular-weight (<10 kDa) fraction of A. sinensis extract significantly attenuated endotoxin-induced HMGB1 release in part through interfering with its cytoplasmic translocation in macrophage cultures. Prophylactic administration of an aqueous extract of A. sinensis significantly attenuated systemic HMGB1 accumulation in vivo, and conferred a dose-dependent protection against lethal endotoxemia. Furthermore, delayed administration of A. sinensis extract beginning 24 h after CLP attenuated systemic HMGB1 accumulation, and significantly rescued mice from lethal sepsis. Taken together, these data suggest that A. sinensis contains water-soluble components that exert protective effects against lethal endotoxemia and experimental sepsis in part by attenuating systemic accumulation of a late proinflammatory cytokine, HMGB1.


Association of CYP2A6 gene deletion with cigarette smoking status in Japanese adults.
Genetic variation of CYP2A6 is shown to alter nicotine metabolism. This study was developed to investigate the genetic influence of the whole deletion-allele of CYP2A6 on active and passive smoking behavior. Two hundred and forty Japanese adults, who visited Aichi Cancer Center as outpatients, were genotyped for the wild-type (CYP2A6*1A, CYP2A6*1B) and the whole deletion-type (CYP2A6*4C) polymorphism of CYP2A6. Information about active and passive smoking status was obtained by a self-administered questionnaire. Genetic influence of CYP2A6 polymorphism on smoking behavior was evaluated using the Mantel extension test. The frequency of the deletion allele was 18%. All 8 subjects carrying two deletion alleles had no smoking habit, and the homozygous deletion genotype showed a tendency to correlate with active smoking status after adjustment for sex and age (p = 0.054). However, the proportion of never smokers among heterozygous subjects was almost the same as among subjects carrying no deletion allele (54% and 58%, respectively). Furthermore, CYP2A6 genotypes were correlated neither with the number of cigarettes smoked per day nor with the age at starting smoking (p = 0.364 and 0.880, respectively). Among never smokers, CYP2A6 genotypes were not correlated with exposure to passive smoking at home or in the workplace (p = 0.623 and 0.484, respectively). Despite the possible protection against active smoking behavior in subjects homozygous for the deletion allele, the CYP2A6 polymorphism has only a limited impact on public health because no protective effect was found in heterozygous subjects.


Comparative Effectiveness of Usual Care With or Without Chiropractic Care in Patients with Recurrent Musculoskeletal Back and Neck Pain.
Chiropractic care is a popular alternative for back and neck pain, with efficacy comparable to usual care in randomized trials. However, the effectiveness of chiropractic care as delivered through conventional care settings remains largely unexplored. To evaluate the comparative effectiveness of usual care with or without chiropractic care for patients with chronic recurrent musculoskeletal back and neck pain. Prospective cohort study using propensity score-matched controls. Using retrospective electronic health record data, we developed a propensity score model predicting likelihood of chiropractic referral. Eligible patients with back or neck pain were then contacted upon referral for chiropractic care and enrolled in a prospective study. For each referred patient, two propensity score-matched non-referred patients were contacted and enrolled. We followed the participants prospectively for 6 months. Main outcomes included pain severity, interference, and symptom bothersomeness. Secondary outcomes included expenditures for pain-related health care. Both groups' (N = 70 referred, 139 non-referred) pain scores improved significantly over the first 3 months, with less change between months 3 and 6. No significant between-group difference was observed. (severity - 0.10 (95% CI - 0.30, 0.10), interference - 0.07 (- 0.31, 0.16), bothersomeness - 0.1 (- 0.39, 0.19)). After controlling for variances in baseline costs, total costs during the 6-month post-enrollment follow-up were significantly higher on average in the non-referred versus referred group ($1996 [SD = 3874] vs $1086 [SD = 1212], p = .034). Adjusting for differences in age, gender, and Charlson comorbidity index attenuated this finding, which was no longer statistically significant (p = .072). We found no statistically significant difference between the two groups in either patient-reported or economic outcomes. As clinical outcomes were similar, and the provision of chiropractic care did not increase costs, making chiropractic services available provided an additional viable option for patients who prefer this type of care, at no additional expense.


A pilot study of a randomized controlled trial of yoga as an intervention for PTSD symptoms in women.
Posttraumatic stress disorder (PTSD) is a debilitating condition that affects approximately 10% of women in the United States. Although effective psychotherapeutic treatments for PTSD exist, clients with PTSD report additional benefits of complementary and alternative approaches such as yoga. In particular, yoga may downregulate the stress response and positively impact PTSD and comorbid depression and anxiety symptoms. We conducted a pilot study of a randomized controlled trial comparing a 12-session Kripalu-based yoga intervention with an assessment control group. Participants included 38 women with current full or subthreshold PTSD symptoms. During the intervention, yoga participants showed decreases in reexperiencing and hyperarousal symptoms. The assessment control group, however, showed decreases in reexperiencing and anxiety symptoms as well, which may be a result of the positive effect of self-monitoring on PTSD and associated symptoms. Between-groups effect sizes were small to moderate (0.08-0.31). Although more research is needed, yoga may be an effective adjunctive treatment for PTSD. Participants responded positively to the intervention, suggesting that it was tolerable for this sample. Findings underscore the need for future research investigating mechanisms by which yoga may impact mental health symptoms, gender comparisons, and the long-term effects of yoga practice.


Perspectives on the Social, Physical, and Emotional Impact of Living With Perthes' Disease in Children and Their Family: A Mixed Methods Study.
Aim. To determine the social, physical, and emotional impact of living with Perthes' disease on affected children and their family (caregivers). Patients and Methods. Through a mixed methods approach, we interviewed 18 parents and explored the perspectives of 12 children affected by Perthes' disease (mean = 7.1 years, SD = ±4.1 years) using a survey tool. Thematic analysis of parents' interviews provided an insight into disease-specific factors influencing patients and family's daily life activities. Using the childhood survey tool, good and bad day scores were analyzed using MANOVA (multivariate analysis of variance). Results. Thematic analysis of the parent interviews (main themes n = 4) identified a marked effect of the disease on many facets of the child's life, particularly pain and the impact on sleep, play, and school attendance. In addition, the interviews identified a negative effect on the family life of the parents and siblings. Children indicated that activities of daily living were affected even during "good days" (P < .05), but pain was the key limiting factor. Conclusion. Perthes' disease negatively affects the social, physical, and emotional well-being of children and their family. These findings provide outcome domains that are important to measure in day-to-day care and add in-depth insight into the challenges caused by this disease for health care professionals involved in clinical management.


Patterns of recurrence after laparoscopic versus open abdominal radical hysterectomy in patients with cervical cancer: a propensity-matched analysis.
Recent evidence has suggested that laparoscopic radical hysterectomy is associated with an increased risk of recurrence in comparison with open abdominal radical hysterectomy. The aim of our study was to identify patterns of recurrence after laparoscopic and open abdominal radical hysterectomy for cervical cancer. This a retrospective multi-institutional study evaluating patients with recurrent cervical cancer after laparoscopic and open abdominal surgery performed between January 1990 and December 2018. Inclusion criteria were: age ≥18 years old, radical hysterectomy (type B or type C), no recurrent disease, and clinical follow-up >30 days. The primary endpoint was to evaluate patterns of first recurrence following laparoscopic and open abdominal radical hysterectomy. The secondary endpoint was to estimate the effect of the primary surgical approach (laparoscopy and open surgery) in post-recurrence survival outcomes (event-free survival and overall survival). In order to reduce possible confounding factors, we applied a propensity-matching algorithm. Survival outcomes were estimated using the Kaplan-Meier model. A total of 1058 patients were included in the analysis (823 underwent open abdominal radical hysterectomy and 235 patients underwent laparoscopic radical hysterectomy). The study included 117 (14.2%) and 35 (14.9%) patients who developed recurrent cervical cancer after open or laparoscopic surgery, respectively. Applying a propensity matched comparison (1:2), we reduced the population to 105 patients (35 vs 70 patients with recurrence after laparoscopic and open radical hysterectomy). Median follow-up time was 39.1 (range 4-221) months and 32.3 (range 4-124) months for patients undergoing open and laparoscopic surgery, respectively. Patients undergoing laparoscopic radical hysterectomy had shorter progression-free survival than patients undergoing open abdominal surgery (HR 1.98, 95% CI 1.32 to 2.97; p=0.005). Patients undergoing laparoscopic radical hysterectomy were more likely to develop intrapelvic recurrences (74% vs 34%; p<0.001) and peritoneal carcinomatosis (17% vs 1%; p=0.005) than patients undergoing open surgery. Patients undergoing laparoscopic radical hysterectomy are at higher risk of developing intrapelvic recurrences and peritoneal carcinomatosis. Further evidence is needed in order to corroborate our findings.


RADVAN: a randomised phase 2 trial of WBRT plus vandetanib for melanoma brain metastases - results and lessons learnt.
Brain metastases occur in up to 75% of patients with advanced melanoma. Most are treated with whole-brain radiotherapy (WBRT), with limited effectiveness. Vandetanib, an inhibitor of vascular endothelial growth factor receptor, epidermal growth factor receptor and rearranged during transfection tyrosine kinases, is a potent radiosensitiser in xenograft models. We compared WBRT with WBRT plus vandetanib in the treatment of patients with melanoma brain metastases. In this double-blind, multi-centre, phase 2 trial patients with melanoma brain metastases were randomised to receive WBRT (30 Gy in 10 fractions) plus 3 weeks of concurrent vandetanib 100 mg once daily or placebo. The primary endpoint was progression-free survival in brain (PFS brain). The main study was preceded by a safety run-in phase to confirm tolerability of the combination. A post-hoc analysis and literature review considered barriers to recruiting patients with melanoma brain metastases to clinical trials. Twenty-four patients were recruited, six to the safety phase and 18 to the randomised phase. The study closed early due to poor recruitment. Median PFS brain was 3.3 months (90% confidence interval (CI): 1.6-5.6) in the vandetanib group and 2.5 months (90% CI: 0.2-4.8) in the placebo group (P=0.34). Median overall survival (OS) was 4.6 months (90% CI: 1.6-6.3) and 2.5 months (90% CI: 0.2-7.2), respectively (P=0.54). The most frequent adverse events were fatigue, alopecia, confusion and nausea. The most common barrier to study recruitment was availability of alternative treatments. The combination of WBRT plus vandetanib was well tolerated. Compared with WBRT alone, there was no significant improvement in PFS brain or OS, although we are unable to provide a definitive result due to poor accrual. A review of barriers to trial accrual identified several factors that affect study recruitment in this difficult disease area.


EPR and ENDOR analysis of Fe3+ impurity centers in fluoroelpasolite lattices.
Fe(3+) ions in hexagonal and cubic fluoroelpasolite crystals (A(1)(2)B(I)M(III)F(6)) have been investigated in a combined Electron Paramagnetic Resonance (EPR) and Electron Nuclear Double Resonance (ENDOR) study. A detailed analysis of the ENDOR spectra for the nearest (19)F and (23)Na shells in X (9.5 GHz) and Q band (34 GHz) allowed the complex EPR spectra to be disentangled and to determine the spin Hamiltonian parameters for the various S = 5/2 Fe(3+) centres. W-band (95 GHz) EPR measurements as a function of temperature were performed to provide unambiguous evidence about the absolute signs of the Zero Field Splitting (ZFS) and SuperHyperFine (SHF) parameters for Fe(3+) in Cs(2)NaAlF(6) as already determined from the ENDOR work. It could be concluded that all principal (19)F hyperfine values were positive, in agreement with earlier assignments in the literature for related systems. A comparative analysis of the (19)F SHF data for Fe(3+) at a perfectly octahedral site in the cubic crystal, and at two slightly trigonally distorted environments in the hexagonal crystals, indicates that the metal-to-ligand distance changes upon doping. The obtained set of parameters concerning one defect in various analogous environments can furthermore be used to test different methods of theoretical calculations for ZFS and SHF values.


Regulation of osteoclastogenesis by Simon extracts composed of caffeic acid and related compounds: successful suppression of bone destruction accompanied with adjuvant-induced arthritis in rats.
Simon extracts are vitamin K(1)-rich food materials extracted from the leaves of the Simon sweet potato. Although vitamin K is known to stimulate bone formation, we postulated that Simon extracts also contain unknown biological compounds having the ability to regulate bone resorption. Here we prepared the vitamin K-free fraction from the Simon extracts and investigated the ability of this fraction on the differentiation of osteoclasts. A remarkable inhibitory effect of osteoclastogenesis was observed when osteoclast precursors were treated with this fraction in rat bone marrow culture systems as well as in a pure differentiation system using murine osteoclast precursor cell line. The vitamin K-free Simon extracts markedly suppressed severe bone destruction mediated by abundant osteoclasts associated with adjuvant-induced arthritis in rats. High performance liquid chromatography (HPLC) analysis revealed that the vitamin K-free Simon extracts contained three types of low molecular weight inhibitors for osteoclastogenesis; caffeic acid, chlorogenic acids and isochlorogenic acids. Among these substances, caffeic acid showed the most powerful inhibitory effects on osteoclastogenesis. Caffeic acid significantly suppressed expression of NFATc1, a key transcription factor for the induction of osteoclastogenesis. Our current study enlightened a high utility of the Simon extracts and their chemical components as effective regulators for bone resorption accompanied with inflammation and metabolic bone diseases.


Colorectal-cancer incidence and mortality with screening flexible sigmoidoscopy.
The benefits of endoscopic testing for colorectal-cancer screening are uncertain. We evaluated the effect of screening with flexible sigmoidoscopy on colorectal-cancer incidence and mortality. From 1993 through 2001, we randomly assigned 154,900 men and women 55 to 74 years of age either to screening with flexible sigmoidoscopy, with a repeat screening at 3 or 5 years, or to usual care. Cases of colorectal cancer and deaths from the disease were ascertained. Of the 77,445 participants randomly assigned to screening (intervention group), 83.5% underwent baseline flexible sigmoidoscopy and 54.0% were screened at 3 or 5 years. The incidence of colorectal cancer after a median follow-up of 11.9 years was 11.9 cases per 10,000 person-years in the intervention group (1012 cases), as compared with 15.2 cases per 10,000 person-years in the usual-care group (1287 cases), which represents a 21% reduction (relative risk, 0.79; 95% confidence interval [CI], 0.72 to 0.85; P<0.001). Significant reductions were observed in the incidence of both distal colorectal cancer (479 cases in the intervention group vs. 669 cases in the usual-care group; relative risk, 0.71; 95% CI, 0.64 to 0.80; P<0.001) and proximal colorectal cancer (512 cases vs. 595 cases; relative risk, 0.86; 95% CI, 0.76 to 0.97; P=0.01). There were 2.9 deaths from colorectal cancer per 10,000 person-years in the intervention group (252 deaths), as compared with 3.9 per 10,000 person-years in the usual-care group (341 deaths), which represents a 26% reduction (relative risk, 0.74; 95% CI, 0.63 to 0.87; P<0.001). Mortality from distal colorectal cancer was reduced by 50% (87 deaths in the intervention group vs. 175 in the usual-care group; relative risk, 0.50; 95% CI, 0.38 to 0.64; P<0.001); mortality from proximal colorectal cancer was unaffected (143 and 147 deaths, respectively; relative risk, 0.97; 95% CI, 0.77 to 1.22; P=0.81). Screening with flexible sigmoidoscopy was associated with a significant decrease in colorectal-cancer incidence (in both the distal and proximal colon) and mortality (distal colon only). (Funded by the National Cancer Institute; PLCO ClinicalTrials.gov number, NCT00002540.).


Matrix Gla protein deficiency causes arteriovenous malformations in mice.
Arteriovenous malformations (AVMs) in organs, such as the lungs, intestine, and brain, are characteristic of hereditary hemorrhagic telangiectasia (HHT), a disease caused by mutations in activin-like kinase receptor 1 (ALK1), which is an essential receptor in angiogenesis, or endoglin. Matrix Gla protein (MGP) is an antagonist of BMPs that is highly expressed in lungs and kidneys and is regulated by ALK1. The objective of this study was to determine the role of MGP in the vasculature of the lungs and kidneys. We found that Mgp gene deletion in mice caused striking AVMs in lungs and kidneys, where overall small organ size contrasted with greatly increased vascularization. Mechanistically, MGP deficiency increased BMP activity in lungs. In cultured lung epithelial cells, BMP-4 induced VEGF expression through induction of ALK1, ALK2, and ALK5. The VEGF secretion induced by BMP-4 in Mgp-/- epithelial cells stimulated proliferation of ECs. However, BMP-4 inhibited proliferation of lung epithelial cells, consistent with the increase in pulmonary vasculature at the expense of lung tissue in the Mgp-null mice. Similarly, BMP signaling and VEGF expression were increased in Mgp-/- mouse kidneys. We therefore conclude that Mgp gene deletion is what we believe to be a previously unidentified cause of AVMs. Because lack of MGP also causes arterial calcification, our findings demonstrate that the same gene defect has drastically different effects on distinct vascular beds.


Morning-evening variation in human brain metabolism and memory circuits.
It has been posited that a critical function of sleep is synaptic renormalization following a net increase in synaptic strength during wake. We hypothesized that wake would alter the resting-state functional organization of the brain and increase its metabolic cost. To test these hypotheses, two experiments were performed. In one, we obtained morning and evening resting-state functional MRI scans to assess changes in functional brain organization. In the second experiment, we obtained quantitative positron emission tomography measures of glucose and oxygen consumption to assess the cost of wake. We found selective changes in brain organization. Most prominently, bilateral medial temporal regions were locally connected in the morning but in the evening exhibited strong correlations with frontal and parietal brain regions involved in memory retrieval. We speculate that these changes may reflect aspects of memory consolidation recurring on a daily basis. Surprisingly, these changes in brain organization occurred without increases in brain metabolism.


Intensive observation of toxic side effects after several-year of cyclosporin treatment in kidney transplant patient.
In this work we are going to show results of intensive observation of adverse reactions of cyclosporine therapy during 18 months. The research was applied on 30 patients with kidney transplant. The medium time of kidney transplant survival was 9,7+/-2,3 years, with time span of 6 to 15 years. All the patients were subjects to several years' cyclosporine treatment, which was applied on a daily basis with a dosage of 2 to 5 mg/kg of body weight. The concentration of cyclosporine in blood was measured once a month. The concentration of cyclosporine in blood in 19 patients was in referent values of 122,50 nag/ml up to 280,50 nag/ml of blood. In 4 of the patients the concentration was heightened up to 370 to 538 nag/ml (X=766,37 nag/ml), and in 7 patients cyclosporine was below normal dosage down to 30,78 to 96,30 nag/ml in blood (x=77,12 nag/ml). We noticed these toxic side effects: increased values of systolic and diastolic arterial blood pressure in 5 patients, neurotoxic tremor effects in 4 patients, hyperplasia gingival and hirsute in 1 patient each.


Hepatic abscess linked to oral actinomycetes: a case report.
Organ abscesses are rare, life-threatening complications that can be caused by bacteremia from oral infections. Metastatic infection is a well-established concept. Dental and periapical infections can cause infections in distant organs and tissues. The frequency of these systemic infections and systemic diseases is open to debate, as some patients are more susceptible to infections than others. This article presents the case report of a 52-year-old woman who was hospitalized with a hepatic abscess after a routine periodontal maintenance procedure. The patient had poor oral health, involving several nonrestorable teeth, multiple failed endodontic treatments, and asymptomatic chronic periapical pathologies. Her dental history included previous diagnoses of moderate generalized chronic periodontitis and advanced localized periodontitis. It was possible that bacteremia developed during her most recent dental treatment, leading to the hepatic abscess. Systemic antibiotic therapy, drainage of the hepatic abscess, and oral rehabilitation resulted in complete recovery.


Novel genital alphapapillomaviruses in baboons (Papio hamadryas anubis) with cervical dysplasia.
Genital Alphapapillomavirus (αPV) infections are one of the most common sexually transmitted human infections worldwide. Women infected with the highly oncogenic genital human papillomavirus (HPV) types 16 and 18 are at high risk for development of cervical cancer. Related oncogenic αPVs exist in rhesus and cynomolgus macaques. Here the authors identified 3 novel genital αPV types (PhPV1, PhPV2, PhPV3) by PCR in cervical samples from 6 of 15 (40%) wild-caught female Kenyan olive baboons (Papio hamadryas anubis). Eleven baboons had koilocytes in the cervix and vagina. Three baboons had dysplastic proliferative changes consistent with cervical squamous intraepithelial neoplasia (CIN). In 2 baboons with PCR-confirmed PhPV1, 1 had moderate (CIN2, n = 1) and 1 had low-grade (CIN1, n = 1) dysplasia. In 2 baboons with PCR-confirmed PhPV2, 1 had low-grade (CIN1, n = 1) dysplasia and the other had only koilocytes. Two baboons with PCR-confirmed PhPV3 had koilocytes only. PhPV1 and PhPV2 were closely related to oncogenic macaque and human αPVs. These findings suggest that αPV-infected baboons may be useful animal models for the pathogenesis, treatment, and prophylaxis of genital αPV neoplasia. Additionally, this discovery suggests that genital αPVs with oncogenic potential may infect a wider spectrum of non-human primate species than previously thought.


Stimulation of erythropoiesis by thalidomide in multiple myeloma patients: its influence on FasL, TRAIL and their receptors on erythroblasts.
The expression of proteins of the tumor necrosis factor (TNF) family on erythroblasts was measured during thalidomide treatment in 29 patients with multiple myeloma (MM). A clinical response was observed in 17 patients (58.6%) and haemoglobin concentration increased in 22 patients (75.9%). The expression of FasL, Fas, TNF-related apoptosis-inducing ligand (TRAIL) and TRAIL-R1 on erythroblasts decreased significantly during thalidomide treatment. Additional in vitro studies revealed that the apoptosis of erythroblasts and the expression of FasL, TRAIL, TRAIL-R1 and TRAIL-R2 was lower in cultures with thalidomide than in control cultures. Altogether our results suggest that thalidomide may stimulate erythropoiesis in MM patients by decreasing the expression of TNF-like ligands/receptors on erythroblasts.


Efficacy of resveratrol to supplement oral nifedipine treatment in pregnancy-induced preeclampsia.
Preeclampsia (PE) is a complication affecting pregnant women worldwide, which usually manifests as severe maternal hypertension. Resveratrol (RESV), a naturally existing polyphenol, is known to exhibit beneficial effects in cardiovascular disease including hypertension. We evaluated the outcome of treatment combining oral nifedipine (NIFE) and RESV against PE. Using a randomized group assignment, 400 PE patients were enrolled and received oral treatments of either NIFE + RESV or NIFE + placebo. Primary endpoints were defined as time to control blood pressure and time before a new hypertensive crisis. Secondary endpoints were defined as the number of doses needed to control blood pressure, maternal and neonatal adverse effects. Compared with the NIFE + placebo group, the time needed to control blood pressure was significantly reduced in NIFE + RESV group, while time before a new hypertensive crisis was greatly delayed in NIFE + RESV group. The number of treatment doses needed to control blood pressure was also categorically lower in NIFE + RESV group. No differences in maternal or neonatal adverse effects were observed between the two treatment groups. Our data support the potential of RESV as a safe and effective adjuvant of oral NIFE to attenuate hypertensive symptoms among PE patients.


DNA vaccine encoding peptide P10 against experimental paracoccidioidomycosis induces long-term protection in presence of regulatory T cells.
Paracoccidioidomycosis is a granulomatous systemic mycosis endemic in Brazil and other Latin America countries. A DNA vaccine encoding the immunoprotective peptide 10 (P10) significantly reduced the fungal burden in mice when given prior to or after intratracheal challenge with Paracoccidioides brasiliensis. Presently, the generation/expansion of CD4+ CD44hi memory T cells as well as Foxp3+ Treg cells in mice immunized with the DNA vaccine (pcDNA3-P10) before and after infection with P. brasiliensis was investigated. Memory CD4+ CD44hi T cells simultaneously with Foxp3+ Treg cells increased in the spleens and lungs of pcDNA3-P10 immunized mice on day 0, 30, 60 and 120 postinfection. Histopathology of the lung tissue showed minimal inflammation in immunized mice compared with the unimmunized group, suggesting a role for regulatory T cells in controlling the immunopathology. The DNA vaccine shows that the repeated immunization generates memory cells and regulatory T cells that replace the initially protective pro-inflammatory T cells conferring a long term protection while preserving the integrity of the infected tissue.


Regional differences in the abiotic environment contribute to genomic divergence within a wild tomato species.
The wild currant tomato Solanum pimpinellifolium inhabits a wide range of abiotic habitats across its native range of Ecuador and Peru. Although it has served as a key genetic resource for the improvement of domestic cultivars, little is known about the genetic basis of traits underlying local adaptation in this species, nor what abiotic variables are most important for driving differentiation. Here we use redundancy analysis (RDA) and other multivariate statistical methods (structural equation modelling [SEM] and generalized dissimilarity modelling [GDM]) to quantify the relationship of genomic variation (6,830 single nucleotide polymorphisms [SNPs]) with climate and geography, among 140 wild accessions. RDA, SEM and GDM each identified environment as explaining more genomic variation than geography, suggesting that local adaptation to heterogeneous abiotic habitats may be an important source of genetic diversity in this species. Environmental factors describing temporal variation in precipitation and evaporative demand explained the most SNP variation among accessions, indicating that these forces may represent key selective agents. Lastly, by studying how SNP-environment associations vary throughout the genome (44,064 SNPs), we mapped the location and investigated the functions of loci putatively contributing to climatic adaptations. Together, our findings indicate an important role for selection imposed by the abiotic environment in driving genomic differentiation between populations.


Adenylation and S-methylation of cysteine by the bifunctional enzyme TioN in thiocoraline biosynthesis.
The antitumor agent thiocoraline is a nonribosomally biosynthesized bisintercalator natural product, which contains in its peptidic backbone two S-methylated l-cysteine residues. S-Methylation occurs very rarely in nature, and is observed extremely rarely in nonribosomal peptide scaffolds. We have proposed that during thiocoraline biosynthesis, TioN, a stand-alone adenylation domain interrupted by the S-adenosyl-l-methionine binding region of a methyltransferase enzyme, is capable of performing two functions: the adenylation and S-methylation of l-cysteine. Herein, by preparation of knockouts of TioN and its MbtH-like protein partner TioT, we confirmed their role in thiocoraline biosynthesis. We also co-expressed recombinant TioN and TioT and biochemically investigated three potential pathways involving activation, methylation, and loading of l-cysteine onto the TioN partner thiolation domain, TioS(T4). The valuable insights gained into the pathway(s) followed for the production of S-Me-l-Cys-S-TioS(T4) will serve as a guide for the development of novel engineered interrupted adenylation enzymes for combinatorial biosynthesis.


Characterization of mitral valve anterior leaflet perfusion patterns.
Although previous histologic studies have demonstrated the presence of blood vessels in the anterior mitral leaflet (AML) and second-order chordae (SC), little is known of the pattern of leaflet perfusion. Hence, the pattern and source of AML perfusion was investigated in an ovine model. Fluorescein angiograms were obtained in 17 ovine hearts immediately after heparinization and cardioplegic arrest, using non-selective left coronary artery (LCA) and selective left anterior descending (LAD), proximal, mid- and distal left circumflex (LCx) perfusion. Serial photographs using a flash/filter system to optimize fluorescence were obtained through a left atriotomy. The proximal half of the AML was seen to be richly vascularized. A loop of vessels was consistently observed in the mitral annulus and AML; these vessels ran along the annulus, extended to the sites of SC insertion, and created anastomoses between these insertions. The SC contributed to the AML perfusion and the anastomotic loop. Selective perfusion of the LAD or proximal LCx artery (ligated before the first obtuse marginal artery) did not perfuse the AML (n = 6). Perfusion of the mid- and distal LCx (n = 7) consistently supplied the AML via SC insertion sites and annular branches. The ovine AML is perfused by vessels that run through the SC and annulus simultaneously, and then create a communicating arcade in the leaflet. These vessels originate from the mid- and distal portions of the LCx. A loss of perfusion as a result of microvascular disease could have adverse implications. Derangements in the extensive vascular component of the mitral valve could be an important contributing factor to valve disease.


Human protein reference database and human proteinpedia as discovery resources for molecular biotechnology.
In the recent years, research in molecular biotechnology has transformed from being small scale studies targeted at a single or a small set of molecule(s) into a combination of high throughput discovery platforms and extensive validations. Such a discovery platform provided an unbiased approach which resulted in the identification of several novel genetic and protein biomarkers. High throughput nature of these investigations coupled with higher sensitivity and specificity of Next Generation technologies provided qualitatively and quantitatively richer biological data. These developments have also revolutionized biological research and speed of data generation. However, it is becoming difficult for individual investigators to directly benefit from this data because they are not easily accessible. Data resources became necessary to assimilate, store and disseminate information that could allow future discoveries. We have developed two resources--Human Protein Reference Database (HPRD) and Human Proteinpedia, which integrate knowledge relevant to human proteins. A number of protein features including protein-protein interactions, post-translational modifications, subcellular localization, and tissue expression, which have been studied using different strategies were incorporated in these databases. Human Proteinpedia also provides a portal for community participation to annotate and share proteomic data and uses HPRD as the scaffold for data processing. Proteomic investigators can even share unpublished data in Human Proteinpedia, which provides a meaningful platform for data sharing. As proteomic information reflects a direct view of cellular systems, proteomics is expected to complement other areas of biology such as genomics, transcriptomics, molecular biology, cloning, and classical genetics in understanding the relationships among multiple facets of biological systems.


Conditions triggering local incident reviews in UK hospital maternity units: A national survey.
In countries, such as the UK, where maternal deaths are rare, reviews of other severe complications of pregnancy and the puerperium can provide an additional perspective to help learn lessons to improve future care. The objective of this survey was to identify the types of incidents which triggered local reviews in the UK, in order to inform national safety reporting guidance. A national descriptive survey. UK. Consultant-led maternity units. Seventy-one per cent of maternity units provided an incident review trigger list. The conditions included were classified by two assessors. Incidents that were listed by at least 5% of maternity units were reported and compared with incidents recommended for review by the Royal College of Obstetricians and Gynaecologists (RCOG). The conditions covered were highly variable, although those recommended by the RCOG were most highly represented. The most commonly listed conditions that had not been recommended for review by the RCOG included inadequate staffing levels (70%), cardiac arrest (69%) and maternal sepsis (64%). Substantial variation exists in the types of incident listed for review by maternity units in the UK. Importantly, some units are not reviewing cases of severe infective complications even though this is a current major concern. Future guidance concerning local serious incident review processes should include how the list of conditions triggering a review should be managed in the light of changing clinical and safety priorities.


Perception and production of object-related grasping in 6-month-olds.
In this study, 6-month-olds' perception of an object-related human grasping action was compared with their level of grasping performance using a within-participants design. In the action perception task, infants were presented with the video of an actor's grasping movement toward an occluded target object. Subsequently, an expected and an unexpected final state of this grasping movement were presented simultaneously, and infants' looking times were measured. In the action production task, infants were presented with three graspable objects. Infants' grasping behavior was coded to be either palmar or thumb-opposite grasping. Results indicate that infants who were already able to perform a thumb-opposite grasp differentiated between the two final states in the action perception task by looking longer toward the unexpected final state. In contrast, infants who showed only palmar grasps looked equally long toward both final states. This finding supports the assumption that action perception and action control are already closely related in infants as young as 6 months.


Sulfonylurea receptor 1 mutations that cause opposite insulin secretion defects with chemical chaperone exposure.
The beta-cell ATP-sensitive potassium (K(ATP)) channel composed of sulfonylurea receptor SUR1 and potassium channel Kir6.2 serves a key role in insulin secretion regulation by linking glucose metabolism to cell excitability. Mutations in SUR1 or Kir6.2 that decrease channel function are typically associated with congenital hyperinsulinism, whereas those that increase channel function are associated with neonatal diabetes. Here we report that two hyperinsulinism-associated SUR1 missense mutations, R74W and E128K, surprisingly reduce channel inhibition by intracellular ATP, a gating defect expected to yield the opposite disease phenotype neonatal diabetes. Under normal conditions, both mutant channels showed poor surface expression due to retention in the endoplasmic reticulum, accounting for the loss of channel function phenotype in the congenital hyperinsulinism patients. This trafficking defect, however, could be corrected by treating cells with the oral hypoglycemic drugs sulfonylureas, which we have shown previously to act as small molecule chemical chaperones for K(ATP) channels. The R74W and E128K mutants thus rescued to the cell surface paradoxically exhibited ATP sensitivity 6- and 12-fold lower than wild-type channels, respectively. Further analyses revealed a nucleotide-independent decrease in mutant channel intrinsic open probability, suggesting the mutations may reduce ATP sensitivity by causing functional uncoupling between SUR1 and Kir6.2. In insulin-secreting cells, rescue of both mutant channels to the cell surface led to hyperpolarized membrane potentials and reduced insulin secretion upon glucose stimulation. Our results show that sulfonylureas, as chemical chaperones, can dictate manifestation of the two opposite insulin secretion defects by altering the expression levels of the disease mutants.


Prognostic factors for finger interphalangeal joint osteoarthritis: a systematic review.
Radiographic hand OA affects one in five adults. Symptomatic hand OA can result in functional impairment, pain and reduced quality of life. A prevalent form of hand OA is IP joint OA, however prognostic factors for IP joint OA remain poorly understood. This systematic review aimed to identify prognostic factors for IP joint OA, and to summarize the diagnostic criteria for IP joint OA in prognostic studies. EMBASE, MEDLINE, Scopus and The Cochrane Library were searched from inception until 19 February 2020 (PROSPERO CRD42019116782). Eligible studies described diagnostic criteria defining IP joint OA, and assessed potential prognostic factors for IP joint OA. Risk of bias was assessed using a modified Quality in Prognosis Studies (QUIPS) tool and a best evidence synthesis was used. Eighteen studies were included (risk of bias: eight high, three moderate, seven low). All defined OA radiographically, and three studies incorporated clinical symptoms into their definition of OA. Forty-nine potential prognostic factors were assessed. Eight were prognostic: older age in women, female gender (both moderate evidence); family history of Heberden's nodes, Kashin-Beck disease, older age in men, dental occupation in men, finger fracture, parity (all limited evidence). Higher BMI in women (limited evidence) was prognostic for symptomatic radiographic OA. No prognostic factors for symptomatic OA were identified. IP joint OA is most commonly defined radiographically, yet criteria were heterogeneous. Eight prognostic factors for radiographic IP joint OA and one for symptomatic radiographic IP joint OA were identified, all with limited or moderate evidence. Further studies on causality and on prognostic pathways are needed.


Structural Changes Correlated with Magnetic Spin State Isomorphism in the S2 State of the Mn4CaO5 Cluster in the Oxygen-Evolving Complex of Photosystem II.
The Mn4CaO5 cluster in Photosystem II catalyzes the four-electron redox reaction of water oxidation in natural photosynthesis. This catalytic reaction cycles through four intermediate states (Si, i = 0 to 4), involving changes in the redox state of the four Mn atoms in the cluster. Recent studies suggest the presence and importance of isomorphous structures within the same redox/intermediate S-state. It is highly likely that geometric and electronic structural flexibility play a role in the catalytic mechanism. Among the catalytic intermediates that have been identified experimentally thus far, there is clear evidence of such isomorphism in the S2 state, with a high-spin (5/2) (HS) and a low spin (1/2) (LS) form, identified and characterized by their distinct electron paramagnetic resonance (EPR spectroscopy) signals. We studied these two S2 isomers with Mn extended X-ray absorption fine structure (EXAFS) and absorption and emission spectroscopy (XANES/XES) to characterize the structural and electronic structural properties. The geometric and electronic structure of the HS and LS S2 states are different as determined using Mn EXAFS and XANES/XES, respectively. The Mn K-edge XANES and XES for the HS form are different from the LS and indicate a slightly lower positive charge on the Mn atoms compared to the LS form. Based on the EXAFS results which are clearly different, we propose possible structural differences between the two spin states. Such structural and magnetic redox-isomers if present at room temperature, will likely play a role in the mechanism for water-exchange/oxidation in photosynthesis.


Recent changes to the governance and accountability of the regulated health professions in Ontario.
Recent legislative changes to the governance and accountability of the health professions in Ontario have seen greater government involvement in the self-regulated health professions. The mandate of all self-regulatory bodies for the health professions in Ontario is protection of the public. Self-regulating professions monitor and enforce standards for entry to practice and continuing competence. Nurse leaders in education and clinical practice must be aware of these changes to the regulatory structure for health professions in Ontario. This paper describes the concept of self-regulation and its legislative history within the health professions. Nurse leaders have an important role in ensuring that team members, patients and the public are aware of the meaning of and rationale for self-regulation and the primacy of protecting the public interest.


Bim-mediated deletion of antigen-specific CD8 T cells in patients unable to control HBV infection.
HBV-specific CD8(+) T cells are critical for a successful immune response to HBV infection. They are markedly diminished in number in patients who fail to control the virus, but the mechanisms resulting in their depletion remain ill defined. Here, we dissected the defective HBV-specific CD8(+) T cell response associated with chronic HBV infection by gene expression profiling. We found that HBV-specific CD8(+) T cells from patients with different clinical outcomes could be distinguished by their patterns of gene expression. Microarray analysis revealed that overlapping clusters of functionally related apoptotic genes were upregulated in HBV-specific CD8(+) T cells from patients with chronic compared with resolved infection. Further analysis confirmed that levels of the proapoptotic protein Bcl2-interacting mediator (Bim) were upregulated in HBV-specific CD8(+) T cells from patients with chronic HBV infection. Blocking Bim-mediated apoptosis enhanced recovery of HBV-specific CD8(+) T cells both in culture and directly ex vivo. Consistent with evidence that Bim mediates apoptosis of CD8(+) T cells expressing low levels of CD127 (IL-7R), the few surviving HBV-specific CD8(+) T cells were CD127(hi )and had elevated levels of the antiapoptotic protein Mcl1, suggesting they were amenable to IL-7-mediated rescue from apoptosis. We therefore postulate that Bim-mediated attrition of HBV-specific CD8(+) T cells contributes to the inability of these cell populations to persist and control viral replication.


Bioresorbable vascular scaffolds in coronary chronic total occlusions: clinical, vasomotor and optical coherence tomography findings at three-year follow-up (ABSORB-CTO study).
Percutaneous treatment of coronary chronic total occlusion (CTO) is associated with important arterial remodelling after successful recanalisation. Bioresorbable vascular scaffold (BVS) implantation in CTO has been shown to be feasible and can be of interest when the scaffold achieves complete bioresorption. We sought to evaluate the three-year results in terms of clinical events, serial imaging outcomes and vasomotion response in CTO lesions successfully treated with BVS. A total of 33 patients (35 CTO lesions) successfully treated with BVS were included in the ABSORB-CTO study. Clinical outcomes, quantitative coronary angiography (QCA) and optical coherence tomography (OCT) assessment were analysed at one- and three-year follow-up. Vasomotion examination was also performed at three years. At three years, cumulative target vessel reocclusion was observed in four lesions (11.4%). By QCA, the in-scaffold segment presented a lumen loss of 0.23±0.46 mm compared with the baseline procedure (p=0.001) and a lumen gain of 0.05±0.29 mm between one and three years (p=0.220). By OCT, the mean neointimal area stenosis progressed from 11.45% at 12 months to 17.10% at 36 months (p<0.001) and mean scaffold area increased continuously at 12 (+12%; p<0.001) and at 36 months (+14.85%; p=0.001). Late acquired incomplete scaffold apposition (LAISA) observed at 12 months in three patients was completely undetectable at three years. Most cases responded to endothelium-dependent vasomotor stimuli (69%). Vasoconstriction to acetylcholine was the predominant response (45%). Successful recanalisation of coronary CTO with BVS implantation is associated with favourable clinical and imaging outcomes. Despite vessel motility restoration, successfully treated CTOs remain with signs of endothelial dysfunction.


Pathophysiology of duchenne muscular dystrophy: current hypotheses.
Duchenne muscular dystrophy is a devastating inherited neuromuscular disorder that affects one in 3300 live male births. Although the responsible gene and its product, dystrophin, have been characterized for more than 15 years, and a mouse model (mdx) has been developed, comprehensive understanding of the mechanism leading from the absence of dystrophin to the muscular degeneration is still debated. First, dystrophin is considered a key structural element in the muscle fiber, and the primary function of the dystrophin-associated protein complex is to stabilize plasma membrane, although a role of signaling is still possible. Mechanically induced damage through eccentric contractions puts a high stress on fragile membranes and provokes micro-lesions that could eventually lead to loss of calcium homeostasis, and cell death. Altered regeneration, inflammation, impaired vascular adaptation, and fibrosis are probably downstream events that take part in the muscular dystrophy and that probably vary a lot along species (i.e., mdx mice), probands within families, stressing the importance of epigenic factors. Because no etiologic therapy is available for Duchenne muscular dystrophy, a better understanding of the primary and downstream mechanisms could prove useful for producing new adjuvant treatments. All pathophysiologic mechanisms are reviewed together with perspectives on management.


Evaluation of core and surface body temperatures, prevalence, onset, duration and severity of hot flashes in men after bilateral orchidectomy for prostate cancer.
To assess the prevalence, onset, duration and severity of hot flashes in men after bilateral orchidectomy (BO) for prostate cancer, to evaluate body temperature changes during hot flashes and to determine whether an elevated temperature within a few days after BO can be caused by deprivation of androgen. Patients (n = 101) were questioned about the characteristics of their hot flashes after BO for prostate cancer. A subgroup of these men (n = 17) were instructed to record their oral and forehead temperatures during and at fixed intervals between hot flashes daily for 4 weeks. The mean age was 71.6 years, mean follow-up after BO was 33.2 months. Hot flashes were reported by 87 men (86%) with previous spontaneous remission in 9 (10%). The median time between BO and the onset of hot flashes was 21 days (range 1-730), median number of hot flashes 3 per day (range 1-20), and median duration was 120 seconds (range 5 to 1800). There was no significant difference between median oral (36.4(o) C) and forehead (36.0(o) C) temperature in the normal state, but during hot flashes the median forehead temperature (37.0(o) C) was higher than the oral temperature (36.5(o) C) (p = 0.0004). Both median oral and forehead temperatures were higher during hot flashes (36.5(o) C and 37.0(o) C) than in the normal state (36.4(o) C and 36.0(o) C, respectively) (p < 0.0001). During hot flashes, the oral temperature was 38(o) C to 40(o) C in only 3.2% of 593 readings in 17 patients. The median oral and forehead temperatures are higher during hot flashes than in normal periods. Oral temperature elevation > 38(o) C within days after a BO is unlikely to be the result of androgen deprivation alone.


Comparing molecular methods for early detection and serotyping of enteroviruses in throat swabs of pediatric patients.
Enteroviruses include over 100 serotypes and usually cause self-limited infections with non-specific symptoms in children, with the exceptions of polioviruses and enterovirus 71 which frequently cause neurologic complications. Therefore, early detection and serotyping of enteroviruses are critical in clinical management and disease surveillance. Traditional methods for detection and serotyping of enteroviruses are virus isolation and immunofluorescence assay, which are time-consuming. In this study, we compare virus isolation and two molecular tests for detection and serotyping of enteroviruses in clinical samples. One hundred and ten throat swabs were collected from pediatric outpatients with enterovirus-like illnesses (hand-foot-mouth disease, herpangina, and non-specific febrile illness). Virus isolation was conducted using multiple cell lines and isolated viruses were serotyped using immunofluorescent assay. In the molecular tests, a semi-nested RT-PCR and a novel CODEHOP platform were used to detect the 5'UTR and VP1 genes of enteroviruses, respectively. Amplified nucleotides were sequenced and genotyped. Among the 110 cases, 39(35%), 52(47%), and 46(42%) were tested positive with these three tests, respectively. Using the consensus results of these three tests as the gold standard, agreement of the VP1 CODEHOP test was 96%, which is higher than those of the virus isolation (89%) and the 5'-UTR test (88%). The VP1 CODEHOP test also has the best performance on serotyping confirmed with serum neutralization tests. The VP1 CODEHOP test performed well for detection and serotyping of enteroviruses in clinical specimens and could reduce unnecessary hospitalization cares during enterovirus seasons.


Methodological Factors in Determining Risk of Dementia After Transient Ischemic Attack and Stroke: (II) Effect of Attrition on Follow-Up.
Cognitive outcomes in cohorts and trials are often based only on face-to-face clinic assessment. However, cognitive impairment is strongly associated with increased morbidity and mortality, leading to substantial loss to clinic follow-up. In the absence of previous population-based data, we determined the effect of such attrition on measured risk of dementia after transient ischemic attack and stroke. Patients with transient ischemic attack or stroke prospectively recruited (2002-2007) into the Oxford Vascular (OXVASC) study had baseline clinical/cognitive assessment and follow-up to 2014. Dementia was diagnosed through face-to-face clinic interview, supplemented by home visits and telephone assessment in patients unable to attend clinic and by hand-searching of primary care records in uncontactable patients. Of 1236 patients (mean age/SD, 75.2/12.1 years; 582 men), 527 (43%) died by 5-year follow-up. Follow-up assessment rates (study clinic, home visit, or telephone) of survivors were 947 in 1026 (92%), 857 in 958 (89%), 792 in 915 (87%), and 567 in 673 (84%) at 1, 6, 12 months and 5 years. Dementia developed in 260 patients, of whom 110 (42%; n=50 primary care records, n=49 home visit, and n=11 telephone follow-up) had not been available for face-to-face clinic follow-up at the time of diagnosis. The 5-year cumulative incidence of postevent dementia was 29% (26%-32%) overall but was only 17% (14% to 19%) in clinic assessed versus 45% (39%-51%) in nonclinic-assessed patients (P difference<0.001). Exclusion of patients unavailable for clinic follow-up reduces the measured risk of postevent dementia. Use of multiple follow-up methods, including home visits, telephone assessments, and consent, to access primary care records substantially increases ascertainment of longer-term dementia outcomes.


SpoIIE mutants of Bacillus subtilis comprise two distinct phenotypic classes consistent with a dual functional role for the SpoIIE protein.
Mutations in the spoIIE locus of Bacillus subtilis block sporulation at the stage of asymmetric septation and prevent compartment-specific activation of the transcription factor delta F. Recent ultrastructural studies of spoIIE mutants led to the conclusion that cells blocked at the stage of asymmetric septation form primarily thick septal structures similar to those formed at the mid-cell site during vegetative growth, although in an earlier study Piggot (J. Bacteriol. 114:1241-1253, 1973) clearly detected a more complex range of phenotypes. We have examined the phenotypes of six spoIIE mutants, including one example of the previously studied null type, spoIIE21. We confirmed that the spoIIE21 mutant and two other null mutants exhibit the classic thick-septum phenotype. However, two of the missense mutants, the spoIIE64 and spoIIE71 mutants, were found to display a strikingly different phenotype characterized by the presence of only thin asymmetric septa, frequently at both polar positions, as noted by Piggot. This phenotype is essentially identical to those of spoIIA (delta F) and spoIIG (delta E) null mutants, which also form sporulation septa that appear structurally normal at the level of electron microscopy. Despite the formation of apparently normal asymmetric septa, spoIIE64 and spoIIE71 mutants are fully defective in activation of delta F-dependent gene expression. These results indicate that the functional roles performed by SpoIIE in septum assembly and sigma factor regulation are distinct and separable.


Factors influencing parental decision about genetics evaluation for their deaf or hard-of-hearing child.
To identify factors that are associated with why parents of deaf children who have had GJB2/GJB6 testing as part of a genetics research study do or do not take their children for genetics evaluation. Self-administered questionnaire was completed by parents of a deaf child participating in a GJB2/GJB6 testing study. A total of 30 parents (representing 24 children) completed the questionnaire; 11 of 24 children (46%) underwent a genetics evaluation. Compared with parents who did not take their child for a genetics evaluation, those who did were more likely to (1) have supportive pediatricians, (2) feel it was important or would be helpful to their child, (3) recall the recommendation for evaluation, (4) have family members who wanted the child to have an evaluation, and (5) be Hispanic or Asian. Genetic test results, knowledge of genetics evaluation, psychosocial factors, language concerns, or structural factors were not substantively associated with attending a genetics evaluation. Parental perceptions, family environment, and pediatricians play a role in decisions regarding genetics evaluation. Because genetic testing for deafness likely will occur outside of traditional genetics clinics and without comprehensive genetics evaluation, efforts to increase pediatricians' awareness of the usefulness of genetics evaluation may be essential to ensure appropriate care for deaf and hard-of-hearing children as recommended by the American College of Medical Genetics.


Isolation of airborne oxacillin-resistant Staphylococcus aureus from culturable air samples of urban residences.
Culturable single-stage impactor samples were collected onto nutrient agar in kitchen and bedroom areas of eight urban and four suburban residences in Philadelphia, Pennsylvania. Staphylococcus aureus colonies were identified by replica plating of the original impactor samples onto Chapman Stone medium followed by isolation of up to eight colonies for coagulase testing. Kirby-Bauer disk diffusion method was utilized to evaluate S. aureus resistance to both oxacillin and cefaclor. The median concentrations of total culturable bacteria observed in bedrooms and trash areas were 300 CFU/m(3) and 253 CFU/m(3), respectively. Median culturable Staphylococcus spp. concentrations in bedrooms and trash areas were 142 CFU/m(3) and 204 CFU/m(3), respectively. A total of 148 individual S. aureus colonies were isolated and tested for antibiotic resistance. Cefaclor resistance was encountered among only 6 of the 148 (4%) colonies. Nearly one-quarter of all S. aureus isolates tested displayed resistance (n = 30) or intermediate resistance (n = 5) to oxacillin. Twenty-six percent (n = 20) of trash area isolates and 21% (n = 15) of bedroom isolates displayed resistance or intermediate resistance to oxacillin. The median difference in percent resistance between trash and bedroom areas was 10% (p = 0.1). Results suggest that there may be a systematic difference in bacterial populations between downtown and suburban residences. Storage of household waste and handling of food may contribute to presence of the organism in the air of residences.


Molecular analysis of the putative inactivation particle in the inactivation gate of brain type IIA Na+ channels.
Fast Na+ channel inactivation is thought to involve binding of phenylalanine 1489 in the hydrophobic cluster IFM in L(III-IV) of the rat brain type IIA Na+ channel. We have analyzed macroscopic and single channel currents from Na+ channels with mutations within and adjacent to hydrophobic clusters in L(III-IV). Substitution of F1489 by a series of amino acids disrupted inactivation to different extents. The degree of disruption was closely correlated with the hydrophilicity of the amino acid at position 1489. These mutations dramatically destabilized the inactivated state and also significantly slowed the entry into the inactivated state, consistent with the idea that F1489 forms a hydrophobic interaction with a putative receptor during the fast inactivation process. Substitution of a phe residue at position 1488 or 1490 in mutants lacking F1489 did not restore normal inactivation, indicating that precise location of F1489 is critical for its function. Mutations of T1491 disrupted inactivation substantially, with large effects on the stability of the inactivated state and smaller effects on the rate of entry into the inactivated state. Mutations of several other hydrophobic residues did not destabilize the inactivated state at depolarized potentials, indicating that the effects of mutations at F1489 and T1491 are specific. The double mutant YY1497/8QQ slowed macroscopic inactivation at all potentials and accelerated recovery from inactivation at negative membrane potentials. Some of these mutations in L(III-IV) also affected the latency to first opening, indicating coupling between L(III-IV) and channel activation. Our results show that the amino acid residues of the IFM hydrophobic cluster and the adjacent T1491 are unique in contributing to the stability of the inactivated state, consistent with the designation of these residues as components of the inactivation particle responsible for fast inactivation of Na+ channels.


Fluid Optimisation in Emergency Laparotomy (FLO-ELA) Trial: study protocol for a multi-centre randomised trial of cardiac output-guided fluid therapy compared to usual care in patients undergoing major emergency gastrointestinal surgery.
Postoperative morbidity and mortality in patients undergoing major emergency gastrointestinal surgery are a major burden on healthcare systems. Optimal management of perioperative intravenous fluids may reduce mortality rates and improve outcomes from surgery. Previous small trials of cardiac-output guided haemodynamic therapy algorithms in patients undergoing gastrointestinal surgery have suggested this intervention results in reduced complications and a modest reduction in mortality. However, this existing evidence is based mainly on elective (planned) surgery, with little evaluation in the emergency setting. There are fundamental clinical and pathophysiological differences between the planned and emergency surgical setting which may influence the effects of this intervention. A large definitive trial in emergency surgery is needed to confirm or refute the potential benefits observed in elective surgery and to inform widespread clinical practice. The FLO-ELA trial is a multi-centre, parallel-group, open, randomised controlled trial. 3138 patients aged 50 and over undergoing major emergency gastrointestinal surgery will be randomly allocated in a 1:1 ratio using minimisation to minimally invasive cardiac output monitoring to guide protocolised administration of intra-venous fluid, or usual care without cardiac output monitoring. The trial intervention will be carried out during surgery and for up to 6 h postoperatively. The trial is funded through an efficient design call by the National Institute for Health and Care Research Health Technology Assessment (NIHR HTA) programme and uses existing routinely collected datasets for the majority of data collection. The primary outcome is the number of days alive and out of hospital within 90 days of randomisation. Participants and those delivering the intervention will not be blinded to treatment allocation. Participant recruitment started in September 2017 with a 1-year internal pilot phase and is ongoing at the time of publication. This will be the largest contemporary randomised trial examining the effectiveness of perioperative cardiac output-guided haemodynamic therapy in patients undergoing major emergency gastrointestinal surgery. The multi-centre design and broad inclusion criteria support the external validity of the trial. Although the clinical teams delivering the trial interventions will not be blinded, significant trial outcome measures are objective and not subject to detection bias. ISRCTN 14729158. Registered on 02 May 2017.


Environmental epigenetics and allergic diseases: recent advances.
Significant strides in the understanding of the role of epigenetic regulation in asthma and allergy using both epidemiological approaches as well as experimental ones have been made. This review focuses on new research within the last 2 years. These include advances in determining how environmental agents implicated in airway disease can induce epigenetic changes, how epigenetic regulation can influence T helper cell differentiation and T regulatory cell production, and new discoveries of epigenetic regulation associated with clinical outcomes.


Plastic compression of a collagen gel forms a much improved scaffold for ocular surface tissue engineering over conventional collagen gels.
We compare the use of plastically compressed collagen gels to conventional collagen gels as scaffolds onto which corneal limbal epithelial cells (LECs) are seeded to construct an artificial corneal epithelium. LECs were isolated from bovine corneas (limbus) and seeded onto either conventional uncompressed or novel compressed collagen gels and grown in culture. Scanning electron microscopy (SEM) results showed that fibers within the uncompressed gel were loose and irregularly ordered, whereas the fibers within the compressed gel were densely packed and more evenly arranged. Quantitative analysis of LECs expansion across the surface of the two gels showed similar growth rates (p > 0.05). Under SEM, the LECs, expanded on uncompressed gels, showed a rough and heterogeneous morphology, whereas on the compressed gel, the cells displayed a smooth and homogeneous morphology. Transmission electron microscopy (TEM) results showed the compressed scaffold to contain collagen fibers of regular diameter and similar orientation resembling collagen fibers within the normal cornea. TEM and light microscopy also showed that cell-cell and cell-matrix attachment, stratification, and cell density were superior in LECs expanded upon compressed collagen gels. This study demonstrated that the compressed collagen gel was an excellent biomaterial scaffold highly suited to the construction of an artificial corneal epithelium and a significant improvement upon conventional collagen gels.


A comparative study of barrier membranes as graft protectors in the treatment of localized bone defects. An experimental study in a canine model.
Guided bone regeneration is a predictable and well-documented surgical approach for the treatment of deficient alveolar ridges prior to endosseous implant placement. The purpose of this study was to compare a new resorbable membrane (GORE RESOLUT ADAPT Regenerative Membrane, i.e. 67% glycolide (PGA) : 33% trimethyline carbonate (TMC)) with Bio-Gide, a resorbable collagen membrane. Five canines were used in the study. Three saddle-type osseous defects were created bilaterally in edentulous areas of the mandible. The defects were filled with assayed, canine demineralized freeze-dried bone (DFDB) in a thermoplastic gelatin matrix. Using a randomized block design, four sites were covered with PGA : TMC membranes of four different porosities, one site was covered with a collagen membrane and one site consisted of DFDB alone (control). At 3 months, the animals were euthanized and the mandibles were removed en bloc for laboratory processing. A total of 30 sites were reviewed microradiographically and underwent histomorphometric analysis for bone regeneration, soft tissue presence and remaining graft material. All sites exhibited uneventful healing. A significantly higher percentage of bone regeneration was seen in the sites protected by the PGA : TMC membrane. A higher component of soft tissue was visible beneath the collagen membrane as compared with the PGA : TMC membrane. The control sites exhibited noticeable deformation of the regenerated bone secondary to collapse of the overlying periosteum. The authors conclude that the PGA : TMC membrane protected the DFDB-filled defect and allowed a greater amount of bone regeneration than the defect protected by the collagen membrane or the control.


Infant feeding experiences among teen mothers in North Carolina: Findings from a mixed-methods study.
Adolescent mothers in the U.S. are much less likely to initiate breastfeeding than older mothers, and teens who do initiate breastfeeding tend to breastfeed for shorter durations. The purpose of this mixed-methods study is to investigate breastfeeding practices, barriers and facilitators among adolescent mothers ages 17 and younger. Quantitative descriptive analyses are conducted using data from the North Carolina Pregnancy Risk Assessment Monitoring System (PRAMS). The population-based sample comprises 389 teens ages 13-17 giving birth to a live born infant in North Carolina in 2000 - 2005 and in 2007. Qualitative analyses are based on in-depth interviews with 22 Black, White and Hispanic teen mothers residing in rural and urban areas of North Carolina conducted between November 2007 and February 2009. In quantitative analyses, 52% (196 of 389) of North Carolina teen mothers initiated breastfeeding, but half of those who initiated breastfeeding (92/196) stopped within the first month postpartum. Hispanic teens (44/52 or 89%) were much more likely than Black (61/159 or 41%) or White teens (87/164 or 52%) to initiate breastfeeding and to continue for a longer duration. Nearly sixty two percent (29/52) of Hispanic respondents breastfed for greater than four weeks as compared to 16% (29/159) of Black respondents and 26% (39/164) of White respondents. Common barriers to breastfeeding initiation and continuation included not liking breastfeeding, returning to school, nipple pain, and insufficient milk. Qualitative data provided context for the quantitative findings, elucidating the barriers and facilitators to breastfeeding from the teens' perspective and insight into the ways in which breastfeeding support to teens could be enhanced. The large number of adolescents ceasing breastfeeding within the first month points to the need for more individualized follow-up after hospital discharge in the first few days postpartum, to address common technical challenges and to provide assistance managing the transition back to school. Provision of an extra home visit or outpatient visit for teens within the first few days following hospital discharge, and advocacy to make schools more compatible with breastfeeding, could potentially help teens who desire to breastfeed to successfully continue. These interventions warrant further research to test their effectiveness among adolescents.


An organelle-specific protein landscape identifies novel diseases and molecular mechanisms.
Cellular organelles provide opportunities to relate biological mechanisms to disease. Here we use affinity proteomics, genetics and cell biology to interrogate cilia: poorly understood organelles, where defects cause genetic diseases. Two hundred and seventeen tagged human ciliary proteins create a final landscape of 1,319 proteins, 4,905 interactions and 52 complexes. Reverse tagging, repetition of purifications and statistical analyses, produce a high-resolution network that reveals organelle-specific interactions and complexes not apparent in larger studies, and links vesicle transport, the cytoskeleton, signalling and ubiquitination to ciliary signalling and proteostasis. We observe sub-complexes in exocyst and intraflagellar transport complexes, which we validate biochemically, and by probing structurally predicted, disruptive, genetic variants from ciliary disease patients. The landscape suggests other genetic diseases could be ciliary including 3M syndrome. We show that 3M genes are involved in ciliogenesis, and that patient fibroblasts lack cilia. Overall, this organelle-specific targeting strategy shows considerable promise for Systems Medicine.


Controllable curvature from planar polymer sheets in response to light.
The ability to change shape and control curvature in 3D structures starting from planar sheets can aid in assembly and add functionality to an object. Herein, we convert planar sheets of shape memory polymers (SMPs) into 3D objects with controllable curvature by dictating where the sheets shrink. Ink patterned on the surface of the sheet absorbs infrared (IR) light, resulting in localized heating, and the material shrinks locally wherever the temperature exceeds the activation temperature, Ta. We introduce two different mechanisms for controlling curvature within SMP sheets. The 'direct' mechanism uses localized shrinkage to induce curvature only in regions patterned with ink. The 'indirect' mechanism uses localized shrinkage in regions patterned with ink to induce curvature in neighboring regions without ink through a balance of internal stresses. Finite element analysis predicts the final shape of the polymer sheets with excellent qualitative agreement with experimental studies. Results from this study show that curvature can be controlled by the distribution and darkness of the ink pattern on the polymer sheet. Additionally, we utilize the direct and indirect curvature mechanisms to demonstrate the formation and actuation of gripper devices, which represent the potential utility of this approach.


Stimulated X-ray Raman Imaging of Conical Intersections.
The conical intersection dynamics of thiophenol is studied theoretically using the stimulated X-ray Raman imaging (SXRI) technique. SXRI employs a hard X-ray narrowband/broadband hybrid probe field and provides a real-time and real-space image of the passage through conical intersections. The signal, calculated using the minimal-coupling radiation/matter Hamiltonian, carries the phase information, and the real-space image of the transition charge density can be reconstructed by its Fourier transform. The two conical intersections (S2/S1 (11ππ*/1πσ*) and S1/S0 (1πσ*/S0)) can be distinguished and identified by the diffraction patterns in the level crossing regimes.


A common polymorphism in the brain-derived neurotrophic factor gene (BDNF) modulates human cortical plasticity and the response to rTMS.
The brain-derived neurotrophic factor gene (BDNF) is one of many genes thought to influence synaptic plasticity in the adult brain and shows a common single nucleotide polymorphism (BDNF Val66Met) in the normal population that is associated with differences in hippocampal volume and episodic memory. It is also thought to influence possible synaptic changes in motor cortex following a simple motor learning task. Here we extend these studies by using new non-invasive transcranial magnetic stimulation (TMS) and transcranial direct current stimulation (TDCS) techniques that directly test the excitability and plasticity of neuronal circuits in human motor cortex in subjects at rest. We investigated whether the susceptibility to TMS probes of plasticity is significantly influenced by the BDNF polymorphism. Val66Met carriers were matched with Val66Val individuals and tested on the following protocols: continuous and intermittent theta burst TMS; median nerve paired associative stimulation; and homeostatic plasticity in the TDCS/1 Hz rTMS model. The response of Met allele carriers differed significantly in all protocols compared with the response of Val66Val individuals. We suggest that this is due to the effect of BNDF on the susceptibility of synapses to undergo LTP/LTD. The circuits tested here are implicated in the pathophysiology of movement disorders such as dystonia and are being assessed as potential new targets in the treatment of stroke. Thus the polymorphism may be one factor that influences the natural response of the brain to injury and disease.


Nicotine-induced skeletal muscle vasodilation is mediated by release of epinephrine from nerve terminals.
To determine the role of sympathetic innervation on nicotine-induced alterations in peripheral (hindlimb) blood flow in the pentobarbital-anesthetized dog, one hindlimb was acutely denervated and remained attached to the body by only the femoral artery and vein, whereas the contralateral limb remained innervated and intact. Measurements were made of aortic pressure, femoral artery and venous pressures, femoral artery flow, and plasma catecholamine levels during intravenous systemic infusion of nicotine. The response to nicotine (9-36 micrograms.kg-1.min-1) on the denervated side was a transient increase followed by a persistent decrease in flow and increase in vascular resistance. The response in the innervated limb was a large increase in blood flow and decrease in vascular resistance. The vasoconstrictor and vasodilator responses could be abolished by pretreatment with both propranolol and phentolamine. The vasodilator response could not be abolished by cholinergic or histaminergic receptor antagonism. Hexamethonium abolished all systemic and peripheral responses to nicotine. Desipramine selectively abolished the vasodilator response in the innervated hindlimb. The vasodilator and vasoconstrictor responses could be mimicked with systemic or local administration of epinephrine. We conclude that, in the hindlimbs of dogs, nicotine stimulates the release of epinephrine from nerve terminals and/or tissue stores to activate beta 2-adrenoceptors and promote vasodilation in skeletal muscle of innervated preparations.


Circulating Tumor Cells (CTC) Are Associated with Defects in Adaptive Immunity in Patients with Inflammatory Breast Cancer.
Circulating tumor cells (CTCs) play a crucial role in tumor dissemination and are prognostic in primary and metastatic breast cancer. Peripheral blood (PB) immune cells contribute to an unfavorable microenvironment for CTC survival. This study aimed to correlate CTCs with the PB T-cell immunophenotypes and functions of patients with inflammatory breast cancer (IBC). This study included 65 IBC patients treated at the MD Anderson Cancer Center. PB was obtained from patients prior to starting a new line of chemotherapy for CTCs enumeration by CellSearch(®), and T cell phenotype and function by flow cytometry; the results were correlated with CTCs and clinical outcome. At least 1 CTC (≥1) or ≥5 CTCs was detected in 61.5% or 32.3% of patients, respectively. CTC count did not correlate with total lymphocytes; however, patients with ≥1 CTC or ≥5 CTCs had lower percentages (%) of CD3+ and CD4+ T cells compared with patients with no CTCs or <5 CTCs, respectively. Patients with ≥1 CTC had a lower percentage of T-cell receptor (TCR)-activated CD8+ T cells synthesizing TNF-α and IFN-γ and a higher percentage of T-regulatory lymphocytes compared to patients without CTCs. In multivariate analysis, tumor grade and % CD3+ T-cells were associated with ≥1 CTC, whereas ≥5 CTC was associated with tumor grade, stage, % CD3+ and % CD4+ T cells, and % TCR-activated CD8 T-cells synthesizing IL-17. IBC patients with CTCs in PB had abnormalities in adaptive immunity that could potentially impact tumor cell dissemination and initiation of the metastatic cascade.


The serotonin theory of depression: a systematic umbrella review of the evidence.
The serotonin hypothesis of depression is still influential. We aimed to synthesise and evaluate evidence on whether depression is associated with lowered serotonin concentration or activity in a systematic umbrella review of the principal relevant areas of research. PubMed, EMBASE and PsycINFO were searched using terms appropriate to each area of research, from their inception until December 2020. Systematic reviews, meta-analyses and large data-set analyses in the following areas were identified: serotonin and serotonin metabolite, 5-HIAA, concentrations in body fluids; serotonin 5-HT1A receptor binding; serotonin transporter (SERT) levels measured by imaging or at post-mortem; tryptophan depletion studies; SERT gene associations and SERT gene-environment interactions. Studies of depression associated with physical conditions and specific subtypes of depression (e.g. bipolar depression) were excluded. Two independent reviewers extracted the data and assessed the quality of included studies using the AMSTAR-2, an adapted AMSTAR-2, or the STREGA for a large genetic study. The certainty of study results was assessed using a modified version of the GRADE. We did not synthesise results of individual meta-analyses because they included overlapping studies. The review was registered with PROSPERO (CRD42020207203). 17 studies were included: 12 systematic reviews and meta-analyses, 1 collaborative meta-analysis, 1 meta-analysis of large cohort studies, 1 systematic review and narrative synthesis, 1 genetic association study and 1 umbrella review. Quality of reviews was variable with some genetic studies of high quality. Two meta-analyses of overlapping studies examining the serotonin metabolite, 5-HIAA, showed no association with depression (largest n = 1002). One meta-analysis of cohort studies of plasma serotonin showed no relationship with depression, and evidence that lowered serotonin concentration was associated with antidepressant use (n = 1869). Two meta-analyses of overlapping studies examining the 5-HT1A receptor (largest n = 561), and three meta-analyses of overlapping studies examining SERT binding (largest n = 1845) showed weak and inconsistent evidence of reduced binding in some areas, which would be consistent with increased synaptic availability of serotonin in people with depression, if this was the original, causal abnormaly. However, effects of prior antidepressant use were not reliably excluded. One meta-analysis of tryptophan depletion studies found no effect in most healthy volunteers (n = 566), but weak evidence of an effect in those with a family history of depression (n = 75). Another systematic review (n = 342) and a sample of ten subsequent studies (n = 407) found no effect in volunteers. No systematic review of tryptophan depletion studies has been performed since 2007. The two largest and highest quality studies of the SERT gene, one genetic association study (n = 115,257) and one collaborative meta-analysis (n = 43,165), revealed no evidence of an association with depression, or of an interaction between genotype, stress and depression. The main areas of serotonin research provide no consistent evidence of there being an association between serotonin and depression, and no support for the hypothesis that depression is caused by lowered serotonin activity or concentrations. Some evidence was consistent with the possibility that long-term antidepressant use reduces serotonin concentration.


Insights into Sex Chromosome Evolution and Aging from the Genome of a Short-Lived Fish.
The killifish Nothobranchius furzeri is the shortest-lived vertebrate that can be bred in the laboratory. Its rapid growth, early sexual maturation, fast aging, and arrested embryonic development (diapause) make it an attractive model organism in biomedical research. Here, we report a draft sequence of its genome that allowed us to uncover an intra-species Y chromosome polymorphism representing-in real time-different stages of sex chromosome formation that display features of early mammalian XY evolution "in action." Our data suggest that gdf6Y, encoding a TGF-β family growth factor, is the master sex-determining gene in N. furzeri. Moreover, we observed genomic clustering of aging-related genes, identified genes under positive selection, and revealed significant similarities of gene expression profiles between diapause and aging, particularly for genes controlling cell cycle and translation. The annotated genome sequence is provided as an online resource (http://www.nothobranchius.info/NFINgb).


Implementing a voluntary, nonregulatory approach to nitrogen management in Tampa Bay, FL: a public/private partnership.
Participants in the Tampa Bay Estuary Program have agreed to adopt nitrogen-loading targets for Tampa Bay based on the water-quality and related light requirements of underwater seagrasses. Based on modeling results, it appears that light levels can be maintained at necessary levels by "holding the line" at existing nitrogen loadings; however, this goal may be difficult to achieve given the 20% increase in the watershed"s human population and associated 7% increase in nitrogen loading that are projected to occur over the next 20 years. To address the long-term management of nitrogen sources, a nitrogen management consortium of local electric utilities, industries, and agricultural interests, as well as local governments and regulatory agency representatives, has developed a consortium action plan to address the target load reduction needed to "hold the line" at 1992 to 1994 levels. To date, implemented and planned projects collated in the Consortium Action Plan meet and exceed the agreed-upon nitrogen-loading reduction goal. An example of the success of the private partnership aspect of this program can be seen in three phosphate fertilizer mining and manufacturing companies with facilities located on Tampa Bay. These companies are participants in the Estuary Program and the Nitrogen Management Consortium to provide support and input for a program that advocates voluntary, nonregulatory cooperation to reach environmental goals.


Methamphetamine acts on subpopulations of neurons regulating sexual behavior in male rats.
Methamphetamine (Meth) is a highly addictive stimulant. Meth abuse is commonly associated with the practice of sexual risk behavior and increased prevalence of Human Immunodeficiency Virus and Meth users report heightened sexual desire, arousal, and sexual pleasure. The biological basis for this drug-sex nexus is unknown. The current study demonstrates that Meth administration in male rats activates neurons in brain regions of the mesolimbic system that are involved in the regulation of sexual behavior. Specifically, Meth and mating co-activate cells in the nucleus accumbens core and shell, basolateral amygdala, and anterior cingulate cortex. These findings illustrate that in contrast to current belief drugs of abuse can activate the same cells as a natural reinforcer, that is sexual behavior, and in turn may influence compulsive seeking of this natural reward.


Latent Order in High-Angle Grain Boundary of GaN.
We report the existence of latent order during core relaxation in the high-angle grain boundaries (GBs) of GaN films using atomic-resolution scanning transmission electron microscopy and ab initio density functional theory calculations. Core structures in the high-angle GBs are characterized by two pairs of Ga-N bonds located next to each other. The core type correlates strongly with the bond angle differences. We identify an order of core relaxation hidden in the high-angle GBs by further classifying the 5/7 atom cores into a stable 5/7 core (5/7(S)) and a metastable 5/7 core (5/7(M)). This core-type classification indicates that metastable cores can exist at real high-angle GBs under certain circumstances. Interestingly, 5/7(M) exhibits distinct defect states compared to 5/7(S), despite their similar atomic configurations. We investigate the reconstruction of defect states observed in 5/7(M) by analyzing the real-space wave functions. An inversion occurred between two localized states during the transition from 5/7(S) to 5/7(M). We suggest an inversion mechanism to explain the formation of new defect states in 5/7(M).


Climate extremes, land-climate feedbacks and land-use forcing at 1.5°C.
This article investigates projected changes in temperature and water cycle extremes at 1.5°C of global warming, and highlights the role of land processes and land-use changes (LUCs) for these projections. We provide new comparisons of changes in climate at 1.5°C versus 2°C based on empirical sampling analyses of transient simulations versus simulations from the 'Half a degree Additional warming, Prognosis and Projected Impacts' (HAPPI) multi-model experiment. The two approaches yield similar overall results regarding changes in climate extremes on land, and reveal a substantial difference in the occurrence of regional extremes at 1.5°C versus 2°C. Land processes mediated through soil moisture feedbacks and land-use forcing play a major role for projected changes in extremes at 1.5°C in most mid-latitude regions, including densely populated areas in North America, Europe and Asia. This has important implications for low-emissions scenarios derived from integrated assessment models (IAMs), which include major LUCs in ambitious mitigation pathways (e.g. associated with increased bioenergy use), but are also shown to differ in the simulated LUC patterns. Biogeophysical effects from LUCs are not considered in the development of IAM scenarios, but play an important role for projected regional changes in climate extremes, and are thus of high relevance for sustainable development pathways.This article is part of the theme issue 'The Paris Agreement: understanding the physical and social challenges for a warming world of 1.5°C above pre-industrial levels'.


Perceived need for substance use treatment among young women from disadvantaged communities in Cape Town, South Africa.
Initiation of treatment for substance use disorders is low among young women from disadvantaged communities in Cape Town, South Africa. Yet little is known about the factors that influence perceived need for treatment (a determinant of treatment entry) within this population. Baseline data on 720 young, drug-using women, collected as part of a randomized field experiment were analyzed to identify predisposing, enabling and health need factors associated with perceived need for treatment. Overall, 46.0% of our sample perceived a need for treatment. Of these participants, 92.4% wanted treatment for their substance use problems but only 50.1% knew where to access services. In multivariable logistic regression analyses, we found significant main effects for ethnicity (AOR = 1.54, 95% CI = 1.05-1.65), income (AOR = 0.96, 95% CI = 0.93-0.99), anxiety (AOR = 1.22, 95% CI = 1.05-1.45), and not having family members with drug problems (AOR = 1.45, 95% CI = 1.05-2.04) on perceived need for treatment. When the sample was stratified by methamphetamine use, income (AOR = 0.87, 95% CI = 0.79-0.96), awareness of treatment services (AOR =1.84, 95% CI = 1.03-3.27), anxiety (AOR =1.41, 95% CI = 1.06-1.87) and physical health status (AOR = 6.29, 95% CI = 1.56-25.64) were significantly associated with perceived need for treatment among those who were methamphetamine-negative. No variables were significantly associated with perceived need for treatment among participants who were methamphetamine-positive. A sizeable proportion of young women who could benefit from substance use treatment do not believe they need treatment, highlighting the need for interventions that enhance perceived need for treatment in this population. Findings also show that interventions that link women who perceive a need for treatment to service providers are needed. Such interventions should address barriers that limit young women's use of services for substance use disorders.


Molecular mechanism of actin-dependent retrograde flow in lamellipodia of motile cells.
In motile, eukaryotic cells, a variety of cell-associated material (collectively termed here as 'particles') continuously flows, relative to the substratum, from the front to the back of the extreme margin of the cell (termed the 'lamellipodium'). This retrograde particle flow, occurs both over the surface of, and inside the lamellipodium. Force to drive retrograde particle flow in lamellipodia is dependent on actin filaments, but the precise mechanism of force generation, and function of the flow is generally not well understood. Actin filaments themselves, in lamellipodia of most motile cell types studied also flow retrograde relative to the substratum. This actin flow, in Aplysia bag cell neuronal growth cones, is known to be driven by activity of a myosin. In these growth cones, retrograde flow of cell surface-attached particles is coupled to retrograde actin flow. In Aplysia, force from retrograde actin flow may limit certain types of growth cone motility. In other motile cell types, such as keratocytes and fibroblasts, the mechanism of retrograde particle flow and function of retrograde actin flow in lamellipodia is poorly understood. For these cell types, recent data provide a basis for proposing alternative actin-based mechanisms to drive retrograde particle flow in lamellipodia. One mechanism is based on activity of a putative pointed end- directed actin motor, and the other on tension-driven surface lipid flow. Here I will review recent advances that have been made in determining the molecular mechanism of force generation to drive retrograde particle flow relative to the substratum in lamellipodia of motile cells. I will address the function of retrograde actin flow in lamellipodia, and apparent differences between Aplysia and other motile cell types.