Intravenous formulation of Panax notoginseng root extract: human pharmacokinetics of ginsenosides and potential for perpetrating drug interactions.
XueShuanTong, a lyophilized extract of Panax notoginseng roots (Sanqi) for intravenous administration, is extensively used as add-on therapy in the treatment of ischemic heart and cerebrovascular diseases and comprises therapeutically active ginsenosides. Potential for XueShuanTong-drug interactions was determined; the investigation focused on cytochrome P450 (CYP)3A induction and organic anion-transporting polypeptide (OATP)1B inhibition. Ginsenosides considerably bioavailable for drug interactions were identified by dosing XueShuanTong in human subjects and their interaction-related pharmacokinetics were determined. The CYP3A induction potential was determined by repeatedly dosing XueShuanTong for 15 days in human subjects and by treating cryopreserved human hepatocytes with circulating ginsenosides; midazolam served as a probe substrate. Joint inhibition of OATP1B by XueShuanTong ginsenosides was assessed in vitro, and the data were processed using the Chou-Talalay method. Samples were analyzed by liquid chromatography/mass spectrometry. Ginsenosides Rb1, Rd, and Rg1 and notoginsenoside R1 were the major circulating XueShuanTong compounds; their interaction-related pharmacokinetics comprised compound dose-dependent levels of systemic exposure and, for ginsenosides Rb1 and Rd, long terminal half-lives (32‒57 and 58‒307 h, respectively) and low unbound fractions in plasma (0.8%‒2.9% and 0.4%‒3.0%, respectively). Dosing XueShuanTong did not induce CYP3A. Based on the pharmacokinetics and inhibitory potency of the ginsenosides, XueShuanTong was predicted to have high potential for OATP1B3-mediated drug interactions (attributed chiefly to ginsenoside Rb1) suggesting the need for further model-based determination of the interaction potential for XueShuanTong and, if necessary, a clinical drug interaction study. Increased awareness of ginsenosides' pharmacokinetics and XueShuanTong-drug interaction potential will help ensure the safe use of XueShuanTong and coadministered synthetic drugs.


Surgical treatment of distal ulnar artery aneurysm.
In the past 7 years, we have encountered six patients with finger ischemia as a result of digital artery occlusion associated with seven distal ulnar artery aneurysms. Our experience with the management of these patients forms the basis of this report. All patients were men, with a mean age of 29 years, and all experienced repetitive trauma to the involved upper extremity. Each patient presented with the acute onset of cool and painful digits, with no previous history of cold sensitivity or Raynaud's syndrome. None of the patients had any serologic or clinical evidence of autoimmune disease. Angiography revealed occlusion of the ulnar artery on the affected side in two patients and patent ulnar artery aneurysms in the remaining five patients. There was occlusion of multiple common and proper digital arteries in all patients. One patient with bilateral ulnar artery aneurysms underwent operative repair consisting of aneurysm excision and replacement with autogenous vein grafts from the lower extremity. All patients have improved symptoms, and the grafts remained patent over a mean follow-up of 24 months (range: 13 to 57 months). Based on these results, we recommend that excision and grafting be considered for patients with symptomatic patent ulnar artery aneurysms. Selected patients with thrombosed aneurysms with ongoing digital ischemia may also benefit from surgical intervention.


Adult HIV care resources, management practices and patient characteristics in the Phase 1 IeDEA Central Africa cohort.
Despite recent advances in the management of HIV infection and increased access to treatment, prevention, care and support, the HIV/AIDS epidemic continues to be a major global health problem, with sub-Saharan Africa suffering by far the greatest humanitarian, demographic and socio-economic burden of the epidemic. Information on HIV/AIDS clinical care and established cohorts' characteristics in the Central Africa region are sparse. A survey of clinical care resources, management practices and patient characteristics was undertaken among 12 adult HIV care sites in four countries of the International Epidemiologic Databases to Evaluate AIDS Central Africa (IeDEA-CA) Phase 1 regional network in October 2009. These facilities served predominantly urban populations and offered primary care in the Democratic Republic of Congo (DRC; six sites), secondary care in Rwanda (two sites) and tertiary care in Cameroon (three sites) and Burundi (one site). Despite some variation in facility characteristics, sites reported high levels of monitoring resources, including electronic databases, as well as linkages to prevention of mother-to-child HIV transmission programs. At the time of the survey, there were 21,599 HIV-positive adults (median age=37 years) enrolled in the clinical cohort. Though two-thirds were women, few adults (6.5%) entered HIV care through prevention of mother-to-child transmission services, whereas 55% of the cohort entered care through voluntary counselling and testing. Two-thirds of patients at sites in Cameroon and DRC were in WHO Stage III and IV at baseline, whereas nearly all patients in the Rwanda facilities with clinical stage information available were in Stage I and II. WHO criteria were used for antiretroviral therapy initiation. The most common treatment regimen was stavudine/lamivudine/nevirapine (64%), followed by zidovudine/lamivudine/nevirapine (19%). Our findings demonstrate the feasibility of establishing large clinical cohorts of HIV-positive individuals in a relatively short amount of time in spite of challenges experienced by clinics in resource-limited settings such as those in this region. Country differences in the cohort's site and patient characteristics were noted. This information sets the stage for the development of research initiatives and additional programs to enhance adult HIV care and treatment in Central Africa.


Type 1 fimbriae, a colonization factor of uropathogenic Escherichia coli, are controlled by the metabolic sensor CRP-cAMP.
Type 1 fimbriae are a crucial factor for the virulence of uropathogenic Escherichia coli during the first steps of infection by mediating adhesion to epithelial cells. They are also required for the consequent colonization of the tissues and for invasion of the uroepithelium. Here, we studied the role of the specialized signal transduction system CRP-cAMP in the regulation of type 1 fimbriation. Although initially discovered by regulating carbohydrate metabolism, the CRP-cAMP complex controls a major regulatory network in Gram-negative bacteria, including a broad subset of genes spread into different functional categories of the cell. Our results indicate that CRP-cAMP plays a dual role in type 1 fimbriation, affecting both the phase variation process and fimA promoter activity, with an overall repressive outcome on fimbriation. The dissection of the regulatory pathway let us conclude that CRP-cAMP negatively affects FimB-mediated recombination by an indirect mechanism that requires DNA gyrase activity. Moreover, the underlying studies revealed that CRP-cAMP controls the expression of another global regulator in Gram-negative bacteria, the leucine-responsive protein Lrp. CRP-cAMP-mediated repression is limiting the switch from the non-fimbriated to the fimbriated state. Consistently, a drop in the intracellular concentration of cAMP due to altered physiological conditions (e.g. growth in presence of glucose) increases the percentage of fimbriated cells in the bacterial population. We also provide evidence that the repression of type 1 fimbriae by CRP-cAMP occurs during fast growth conditions (logarithmic phase) and is alleviated during slow growth (stationary phase), which is consistent with an involvement of type 1 fimbriae in the adaptation to stress conditions by promoting biofilm growth or entry into host cells. Our work suggests that the metabolic sensor CRP-cAMP plays a role in coupling the expression of type 1 fimbriae to environmental conditions, thereby also affecting subsequent attachment and colonization of host tissues.


Foxa3 induces goblet cell metaplasia and inhibits innate antiviral immunity.
Goblet cell metaplasia accompanies common pulmonary disorders that are prone to recurrent viral infections. Mechanisms regulating both goblet cell metaplasia and susceptibility to viral infection associated with chronic lung diseases are incompletely understood. We sought to identify the role of the transcription factor FOXA3 in regulation of goblet cell metaplasia and pulmonary innate immunity. FOXA3 was identified in airways from patients with asthma and chronic obstructive pulmonary disease. We produced transgenic mice conditionally expressing Foxa3 in airway epithelial cells and developed human bronchial epithelial cells expressing Foxa3. Foxa3-regulated genes were identified by immunostaining, Western blotting, and RNA analysis. Direct binding of FOXA3 to target genes was identified by chromatin immunoprecipitation sequencing correlated with RNA sequencing. FOXA3 was highly expressed in airway goblet cells from patients with asthma and chronic obstructive pulmonary disease. FOXA3 was induced by either IL-13 or rhinovirus. Foxa3 induced goblet cell metaplasia and enhanced expression of a network of genes mediating mucus production. Paradoxically, FOXA3 inhibited rhinovirus-induced IFN production, IRF-3 phosphorylation, and IKKε expression and inhibited viral clearance and expression of genes required for antiviral defenses, including MDA5, RIG-I, TLR3, IRF7/9, and nuclear factor-κB. FOXA3 induces goblet cell metaplasia in response to infection or Th2 stimulation. Suppression of IFN signaling by FOXA3 provides a plausible mechanism that may serve to limit ongoing Th1 inflammation during the resolution of acute viral infection; however, inhibition of innate immunity by FOXA3 may contribute to susceptibility to viral infections associated with chronic lung disorders accompanied by chronic goblet cell metaplasia.


Codon usage regulates human KRAS expression at both transcriptional and translational levels.
KRAS and HRAS are highly homologous oncogenic Ras GTPase family members that are mutated in a wide spectrum of human cancers. Despite having high amino acid identity, KRAS and HRAS have very different codon usage biases: the HRAS gene contains many common codons, and KRAS is enriched for rare codons. Rare codons in KRAS suppress its protein expression, which has been shown to affect both normal and cancer biology in mammals. Here, using HRAS or KRAS expression in different human cell lines and in vitro transcription and translation assays, we show that KRAS rare codons inhibit both translation efficiency and transcription and that the contribution of these two processes varies among different cell lines. We observed that codon usage regulates mRNA translation efficiency such that WT KRAS mRNA is poorly translated. On the other hand, common codons increased transcriptional rates by promoting activating histone modifications and recruitment of transcriptional coactivators. Finally, we found that codon usage also influences KRAS protein conformation, likely because of its effect on co-translational protein folding. Together, our results reveal that codon usage has multidimensional effects on protein expression, ranging from effects on transcription to protein folding in human cells.


Feasibility of Aerobic Exercise and Tai-Chi Interventions in Advanced Lung Cancer Patients: A Randomized Controlled Trial.
A majority of lung cancer patients are diagnosed at advanced stages. Although there is considerable evidence of the benefits of aerobic exercise and tai-chi for lung cancer patients, little is known about the comparative effectiveness of the 2 exercise modes in advanced lung cancer patients. To explore the feasibility and preliminary effects of aerobic exercise and tai-chi interventions on survival and well-being among advanced lung cancer patients. In an assessor-blinded, exploratory randomized controlled trial, 30 advanced lung cancer patients were randomized to an aerobic exercise group, a tai-chi group (both attending 12-week, twice-weekly supervised sessions), or a self-management control group (receiving written exercise guidelines). The primary outcomes focused on feasibility including intervention completion, exercise adherence, and adverse events, while the secondary outcomes addressed preliminary effects and included 1-year survival, cancer symptoms (Pittsburgh Sleep Quality Index, Hospital Anxiety and Depression Score, Brief Fatigue Inventory), quality of life (EORTC QLQ-C30, QLQ-LC13), physical performance (6-minute walk test, up-and-go, sit-to-stand, 1-leg standing), activity levels (actigraph), and circadian rhythms (salivary cortisol). Intervention feasibility was established with a satisfactory completion rate at post-intervention for the aerobic exercise group (80%) and the tai-chi group (78%). The tai-chi group attained higher adherence than the exercise group in terms of attendance in supervised sessions (89% vs 75% of scheduled classes) and self-practice (225% vs 87% of the prescribed amount). Higher adherence to self-practice in the tai-chi group remained at the 6-month follow-up (81% vs 38% of the prescribed amount). No adverse event as a result of the intervention was reported. Effect-related outcomes did not show statistically significant changes in any group, except an improvement post-intervention in the up-and-go (-2.26, 95% CI: -4.04, -0.48) and sit-to-stand tests (4.52, 95% CI: 2.19, 6.85) in the aerobic exercise group. The findings support the feasibility of aerobic exercise and tai-chi interventions in advanced lung cancer patients. A future study with a larger sample from multiple sites is recommended to confirm the comparative effects of the 2 exercise interventions relative to the self-management group and to enhance the generalizability of the findings.


Defective equipment and tractor-trailer crash involvement.
The role of defective equipment in large truck crashes on interstate highways in Washington State was investigated using a case-control study design. For each large truck involved in a crash, three trucks were randomly selected from the traffic stream at the same time and place as the crash, but one week later both crash and comparison trucks were inspected by Commercial Vehicle Enforcement officers of the Washington State Patrol. The effects of truck equipment condition, truck operating characteristics (carrier type, carrier operation, and truck load), and driver characteristics (driver age, hours of driving) on crash involvement were analyzed by comparing their relative frequency among crash-involved and comparison sample tractor-trailers. A logistic regression model was used to estimate the adjusted odds ratio for each factor. Overall, 77% of tractor-trailers in crashes and 66% of those not involved in crashes had defective equipment warranting a citation. Forty-one percent in crashes had defective equipment warranting taking the truck out of service, and 31% not in crashes had these defects. Brake defects were the most common type and were found in 56% of tractor-trailers in crashes; steering equipment defects were found in 21%. The relative risk of crash involvement for trucks with brake defects was about one and one-half times that for trucks without brake defects. For trucks with steering defects, the relative risk of crash involvement was at least twice that for trucks with steering defects, the relative risk of crash involvement was at least twice that for trucks without defects, and the risk increased substantially for trucks with out-of-service steering defects.


Micronutrient Deficiency Independently Predicts Adverse Health Outcomes in Patients With Heart Failure.
Despite growing evidence on the important role of micronutrients in prognosis of heart failure (HF), there has been limited research that micronutrient deficiency predicts health outcomes in patients with HF. The aim of this study was to determine whether micronutrient deficiency independently predicts adverse health outcomes. A total of 113 consecutive outpatients with HF completed a 3-day food diary to measure intake of 15 micronutrients. The Computer Aided Nutrition Analysis Program for Professionals was used to analyze the food diaries and determine dietary micronutrient deficiencies. Patients completed the Minnesota Living With HF Questionnaire to assess health-related quality of life (HRQoL) and were followed up for 1 year to determine cardiac-related hospitalization or cardiac death. Hierarchical multiple linear regressions and Cox proportional hazard regressions were used to determine whether micronutrient deficiencies predicted health outcomes. Fifty-eight patients (51%) had at least 3 micronutrient deficiencies (range, 0-14). Calcium, magnesium, and vitamin D were the most common micronutrient deficiencies. Micronutrient deficiency was independently associated with worse HRQoL (β = .187, P = .025) in hierarchical multiple linear regression. Thirty-nine patients were hospitalized or died during 1-year follow-up because of cardiac problems. The number of micronutrient deficiencies independently predicted cardiac event-free survival (hazard ratio, 1.14; 95% confidence interval, 1.02-1.28). These findings show that micronutrient deficiency independently predicted poor HRQoL and earlier cardiac event-free survival in patients with HF. Further research is needed to provide for specific dietary guidelines for better health outcomes in HF patients.


Demonstration of glycated insulin in human diabetic plasma and decreased biological activity assessed by euglycemic-hyperinsulinemic clamp technique in humans.
The presence and biological significance of circulating glycated insulin has been evaluated by high-pressure liquid chromatography (HPLC), electrospray ionization mass spectrometry (ESI-MS), radioimmunoassay (RIA), receptor binding, and hyperinsulinemic-euglycemic clamp techniques. ESI-MS analysis of an HPLC-purified plasma pool from four male type 2 diabetic subjects (HbA(1c) 8.1 +/- 0.2%, plasma glucose 8.7 +/- 1.3 mmol/l [means +/- SE]) revealed two major insulin-like peaks with retention times of 14-16 min. After spectral averaging, the peak with retention time of 14.32 min exhibited a prominent triply charged (M+3H)(3+) species at 1,991.1 m/z, representing monoglycated insulin with an intact M(r) of 5,970.3 Da. The second peak (retention time 15.70 min) corresponded to native insulin (M(r) 5,807.6 Da), with the difference between the two peptides (162.7 Da) representing a single glucitol adduct (theoretical 164 Da). Measurement of glycated insulin in plasma of type 2 diabetic subjects by specific RIA gave circulating levels of 10.1 +/- 2.3 pmol/l, corresponding to approximately 9% total insulin. Biological activity of pure synthetic monoglycated insulin (insulin B-chain Phe(1)-glucitol adduct) was evaluated in seven overnight-fasted healthy nonobese male volunteers using two-step euglycemic-hyperinsulinemic clamps (2 h at 16.6 micro g x kg(-1) x min(-1), followed by 2 h at 83.0 micro g x kg(-1) x min(-1); corresponding to 0.4 and 2.0 mU x kg(-1) x min(-1)). At the lower dose, the exogenous glucose infusion rates required to maintain euglycemia during steady state were significantly lower with glycated insulin (P < 0.01) and approximately 70% more glycated insulin was required to induce a similar rate of insulin-mediated glucose uptake. Maximal responses at the higher rates of infusion were similar for glycated and control insulin. Inhibitory effects on endogenous glucose production, insulin secretion, and lipolysis, as indicated by measurements of C-peptide, nonesterified free fatty acids, and glycerol, were also similar. Receptor binding to CHO-T cells transfected with human insulin receptor and in vivo metabolic clearance revealed no differences between glycated and native insulin, suggesting that impaired biological activity is due to a postreceptor effect. The present demonstration of glycated insulin in human plasma and related impairment of physiological insulin-mediated glucose uptake suggests a role for glycated insulin in glucose toxicity and impaired insulin action in type 2 diabetes.


Use of vaginal electrical resistance to diagnose oestrus, dioestrus and early pregnancy in synchronized tropically adapted beef heifers.
The primary objective of this study was to determine whether a single measurement of intravaginal electrical resistance (VER), using the commercially available Ovatec probe, can discriminate between dioestrus and oestrus in Bos indicus females, which had been treated to synchronize oestrus. Santa Gertrudis heifers (n = 226) received one of three oestrous synchronization treatments: double PGF(2alpha) 10 days apart, 8-day controlled internal drug release (CIDR) treatment or CIDR pre-synchronization + PGF(2alpha) 10 days after CIDR removal. The heifers were inseminated within 12 h following observed oestrus, or, if not observed, at a fixed time approximately 80 h, following the last synchronization treatment. They were palpated per rectum for signs of pregnancy 9 weeks after artificial insemination (AI). Vaginal electrical resistance measurements were taken at the completion of synchronization treatments (presumed dioestrus), immediately prior to AI (oestrus), and then at 3 and 9 weeks post-AI. Mean VER differed between presumed dioestrus and oestrus (113.7 vs 87.4, p < 0.001). The area under the receiver operating characteristics (ROC) curve was 0.925, indicating that VER was highly discriminatory between dioestrus and oestrus. Vaginal electrical resistance at time of AI was negatively associated with odds of conception when all inseminations were included in the analyses [odds ratio (OR) = 0.97; 95% CI 0.95-1.00; p = 0.018], but not when fixed time AIs were excluded (OR = 1.00; 95% CI 0.97-1.03; p = 0.982). Mean VER readings differed between pregnant and non-pregnant animals at both 3 weeks (120.5 vs 96.7, p < 0.001) and 9 weeks (124.0 vs 100.3, p < 0.001) post-AI. However, 3- and 9-week VER measurements were not highly discriminatory between pregnancy and non-pregnancy (area under ROC curve = 0.791 and 0.736, respectively). Mean VER at time of AI for animals diagnosed in oestrus differed between each of the oestrous synchronization treatments (84.7, 73.6 and 78.9, groups 1-3 respectively, p < 0.001). These findings suggest that measurement of VER may improve accuracy of oestrus diagnoses when selecting cattle for AI following oestrous synchronization programmes involving tropically adapted cattle.


Regulation of cyclooxygenase 2 mRNA stability by the mitogen-activated protein kinase p38 signaling cascade.
A tetracycline-regulated reporter system was used to investigate the regulation of cyclooxygenase 2 (Cox-2) mRNA stability by the mitogen-activated protein kinase (MAPK) p38 signaling cascade. The stable beta-globin mRNA was rendered unstable by insertion of the 2, 500-nucleotide Cox-2 3' untranslated region (3' UTR). The chimeric transcript was stabilized by a constitutively active form of MAPK kinase 6, an activator of p38. This stabilization was blocked by SB203580, an inhibitor of p38, and by two different dominant negative forms of MAPK-activated protein kinase 2 (MAPKAPK-2), a kinase lying downstream of p38. Constitutively active MAPKAPK-2 was also able to stabilize chimeric beta-globin-Cox-2 transcripts. The MAPKAPK-2 substrate hsp27 may be involved in stabilization, as beta-globin-Cox-2 transcripts were partially stabilized by phosphomimetic mutant forms of hsp27. A short (123-nucleotide) fragment of the Cox-2 3' UTR was necessary and sufficient for the regulation of mRNA stability by the p38 cascade and interacted with a HeLa protein immunologically related to AU-rich element/poly(U) binding factor 1.


Treating allergic rhinitis in pregnancy. Safety considerations.
Allergic rhinitis affects approximately one-third of women of childbearing age. As a result, symptoms ranging from sneezing and itching to severe nasal obstruction may require pharmacotherapy. However, product labels state that medications for allergic rhinitis should be avoided during pregnancy due to lack of fetal safety data, even though the majority of the agents have human data which refute these notions. We present a systematic and critical review of the medical literature on the use of pharmacotherapy for the management of allergic rhinitis during pregnancy. Electronic databases and other literature sources were searched to identify observational controlled studies focusing on the rate of fetal malformations in pregnant women exposed to agents used to treat allergic rhinitis and related diseases compared with controls. Immunotherapy and intranasal sodium cromoglycate (cromolyn) and beclo-methasone would be considered as first-line therapy, both because of their lack of association with congenital abnormalities and their superior efficacy to other agents. First-generation (e.g. chlorpheniramine) and second-generation (e.g. cetirizine) antihistamines have not been incriminated as human teratogens. However, first-generation antihistamines are favoured over their second generation counterparts based on their longevity, leading to more conclusive evidence of safety. There are no controlled trials with loratadine and fexofenadine in human pregnancy. Oral, intranasal and ophthalmic decongestants (e.g. pseudoephedrine, phenylephrine and oxymetazoline, respectively) should be considered as second-line therapy, although further studies are needed to clarify their fetal safety. No human reproductive studies have been reported with the ophthalmic antihistamines ketorolac and levocabastine, although preliminary data reported suggest no association between pheniramine and congenital malformations. There are no documented epidemiological studies with intranasal corticosteroids (e.g. budesonide, fluticasone propionate, mometasone) during pregnancy; however, inhaled corticosteroids (e.g. beclomethasone) have not been incriminated as teratogens and are commonly used by pregnant women who have asthma. In summary, women with allergic rhinitis during pregnancy can be treated with a number of pharmacological agents without concern of untoward effects on their unborn child. Although the choice of agents in part should be based on evidence of fetal safety, issue of efficacy needs to be addressed in order to optimally manage this condition.


Clinical findings, treatment, and outcome in 11 dairy heifers with breakdown injury due to interosseous medius muscle rupture.
To describe the diagnosis, treatment, and prognosis of fetlock breakdown due to interosseus medius muscle rupture in cattle. Retrospective clinical study. Dairy heifers with unilateral or bilateral interosseus medius muscle rupture (n = 11). Breakdown injury due to rupture of the interosseus medius muscle was documented clinically, radiographically, and ultrasonographically. Breakdown was bilateral in 4 heifers (3 in forelimbs, 1 in hind limbs) and unilateral in 7 (all hind limbs). One heifer with severe bilateral hind limb breakdown was slaughtered and the remaining 10 were treated by transfixation pin cast (1 heifer), box rest (n = 2), and/or a cast and splint (7). Hyperextension of the fetlock and hyperflexion of the proximal interphalangeal joints during weight bearing were characteristic for interosseus muscle breakdown. Ultrasonographically, the origin and body of the interosseus muscle and the branches to the sesamoid bones were primarily affected by the rupture. Conservative treatment was successful (used for their intended purpose) in 8 of 9 heifers with a median lifespan of 32 months after discharge from the clinic (range 6-83). Rupture of the interosseus medius muscle in young cattle may be more common than previously suggested in the literature. Imaging with ultrasound allowed more detailed localization of lesions of the musculo-tendinous structure. Interosseus medius muscle rupture had a favorable prognosis when treated conservatively in these heifers.


Linkage of Whole Genome Sequencing, Epidemiological, and Clinical Data to Understand the Genetic Diversity and Clinical Outcomes of Shigella flexneri among Men Who Have Sex with Men in England.
The public health value of whole genome sequencing (WGS) for Shigella spp. in England has been limited by a lack of information on sexual identity and behavior. We combined WGS data with other data sources to better understand Shigella flexneri transmission in men who have sex with men (MSM). WGS data for all S. flexneri isolates referred to the national reference laboratory were linked to i) clinical and behavioral data collected in seven of 21 health regions in England using a standardized exposure questionnaire and, ii) national HIV surveillance data. We included 926 S. flexneri isolates, of which 43.0% (n = 398) fell phylogenetically within two domestically circulating clades associated with genotypic markers of azithromycin resistance. Approximately one third of isolates in these clades were from people living with HIV, primarily acquired through sex between men. 182 (19.7%) isolates had linked questionnaire data; 88% (84/95) of MSM isolates fell phylogenetically within the domestically circulating clades, while 92% (72/78) of isolates from other cases fell within lineages linked with travel to high-risk regions. There was no evidence of sustained transmission between networks of MSM and the wider community. MSM were more likely to be admitted to hospital and receive antimicrobials. Our study emphasizes the importance of sex between men as a major route of transmission for S. flexneri. Combined WGS, epidemiological and clinical data provide unique insights that can inform contact tracing, clinical management and the delivery of targeted prevention activities. Future studies should investigate why MSM experience more severe clinical outcomes. IMPORTANCE Within the last 2 decades there have been an increasing number of Shigella spp. outbreaks among men who have sex with men (MSM) worldwide. In 2015, Public Health England (PHE) introduced routine whole genome sequencing (WGS) for the national surveillance of Shigella spp. However, the lack of information on sexual identity and behavior has hindered interpretation. Our study illustrates the power of linking WGS data with epidemiological, behavioral, and clinical data. We provide unique population-level insights into different transmission networks that can inform the delivery of appropriate public health interventions and patient management. Furthermore, we describe and compare clinical characteristics and outcomes of S. flexneri infection in MSM and other exposure groups. We found that MSM were more likely to be admitted to hospital and receive antimicrobials, indicating that their infections were potentially more severe. The exact reasons for this are unclear and require further exploration.


Characteristics of HIV-infected childbearing women in Barbados.
To describe the demographic profile, social and family characteristics, and lifestyle traits of HIV-infected childbearing women in the Caribbean nation of Barbados in comparison to a control group of HIV-negative women. Data for this report were drawn from the Pediatrics HIV Surveillance Program of the Queen Elizabeth Hospital in Barbados. The data covered all HIV-infected women in the country who delivered between 1986-2000, with similar data coming from a control group of HIV-negative childbearing women. Routine information recorded during antenatal care was obtained from the women's case records. Additional data were collected from interviews with the women. There were 182 HIV-infected women who delivered during the study period, and a group of 202 childbearing women served as controls. In comparison to the control group, the HIV-infected women were younger, more often multiparous, and more likely to have been unemployed at the time of their pregnancy. The HIV-infected women also had had an earlier onset of sexual activity, had had more sexual partners during their lifetime, and were more likely to be involved with an older sexual partner. At the time of giving birth most of the HIV-infected women were asymptomatic for AIDS and were living with either their parents (mother or father or both) or the baby's father. In addition, at the time of their six-weeks-postnatal visit, the large majority of the HIV-infected women were involved in caring for their children. The proportion of HIV-infected women who were diagnosed prior to childbirth increased significantly over the study period, rising from 25% during 1986-1990 to 82% during 1996-2000. Slightly over one-fifth of the HIV-infected women had had one or more subsequent pregnancies after they had learned that they were infected. The early age of sexual activity as well as repeated pregnancies, especially from different and older partners, may have contributed significantly to both vertical and horizontal HIV transmission in Barbados. Future studies of HIV incidence and its trend among childbearing women could be important for monitoring the HIV epidemic in this country. Many of the HIV-infected childbearing women in our study were unemployed, sick, and had multiple children. Therefore, to help them to plan for and cope with the disease and also the care of their children beyond the perinatal period, there is a need to provide the women with repeated counseling with continued follow-up and, where necessary, additional economic, social, and medical support.


Child-caregiver interaction in two remote Indigenous Australian communities.
This paper reports on a study in two remote multilingual Indigenous Australian communities: Yakanarra in the Kimberley region of Western Australia and Tennant Creek in the Barkly region of the Northern Territory. In both communities, processes of language shift are underway from a traditional language (Walmajarri and Warumungu, respectively) to a local creole variety (Fitzroy Valley Kriol and Wumpurrarni English, respectively). The study focuses on language input from primary caregivers to a group of preschool children, and on the children's productive language. The study further highlights child-caregiver interactions as a site of importance in understanding the broader processes of language shift. We use longitudinal data from two time-points, approximately 2 years apart, to explore changes in adult input over time and developmental patterns in the children's speech. At both time points, the local creole varieties are the preferred codes of communication for the dyads in this study, although there is some use of the traditional language in both communities. Results show that for measures of turn length (MLT), there are notable differences between the two communities for both the focus children and their caregivers. In Tennant Creek, children and caregivers use longer turns at Time 2, while in Yakanarra the picture is more variable. The two communities also show differing trends in terms of conversational load (MLT ratio). For measures of morphosyntactic complexity (MLU), children and caregivers in Tennant Creek use more complex utterances at Time 2, while caregivers in Yakanarra show less complexity in their language at that time point. The study's findings contribute to providing a more detailed picture of the multilingual practices at Yakanarra and Tennant Creek, with implications for understanding broader processes of language shift. They also elucidate how children's language and linguistic input varies diachronically across time. As such, we contribute to understandings of normative language development for non-Western, non middle-class children in multilingual contexts.


Deriving spatially explicit water uses from land use change modelling results in four river basins across Europe.
The objective of this study is to provide spatially distributed water use maps at a high spatial and thematic resolution as detailed input data for further modelling purposes. The maps were derived on the basis of official water statistics and land use maps to represent the current conditions in four European river basins affected by water scarcity (Adige, Ebro, Evrotas and Sava) and allow setting up reference scenarios. The present land use distribution was modelled based on CORINE data with the land use change model iCLUE. Modelling the land use allows to create dynamic land and water use maps adapted to the needs of eventual scenario analyses compared to using only observed land use maps. The availability of several CORINE datasets allowed calibrating and validating the results of the iCLUE model carrying out a three map comparison. Sectoral water uses were attributed to different land use classes and by this means located in space. Both the location and the magnitude of urban and agricultural water uses can be derived from the final maps. The created maps together with the corresponding land use data provide a coherent set of information crucial to most environmental modelling activities and often missing at this spatial and thematic resolution. This work also aims at visualizing and validating the water use statistics provided by official institutions such as the River Basin Management Plans. The results show that in some cases they are not consistent and underline the importance of harmonised data collection regarding water statistics, as otherwise comparisons within one study area and with others are hampered. This study is embedded in the EU-FP7 GLOBAQUA project which analyses the effects of stressors, such as changes of land and water use, on aquatic ecosystems in areas suffering water scarcity.


Tunable filter-based multispectral imaging for detection of blood stains on construction material substrates part 2: realization of rapid blood stain detection.
Based on the blood stain detection method and criteria established in part 1 of this article, we combine and organize all necessary tasks to realize the multispectral imaging-based rapid blood stain detection system. To rapidly detect blood stains on the test surface, the developed system automatically captures the spectral images, extracts their spectral data, determines the positions of blood stains, and accurately highlights the positions of blood stains on the display. To achieve such a system, several tasks are newly introduced, including adjustment of camera exposure times to prevent image saturation or excessive darkness, the search for the sampled clean positions of the substrate to determine the substrate reflectance spectrum, and suitable detection procedures and proper arrangement of criteria to eliminate unnecessary calculations. Parallel processes between image capturing and blood stain identification help shorten the time for blood stain identifications despite a large amount of spectral data to be processed. The developed system can identify blood against several other reddish brown stains on several substrates. The measured average identification times on different test surfaces range from only 23.3 to 28.7 s, including the image capturing process.


Prospects of RNAi and microRNA-based therapies for hepatitis C.
RNA interference (RNAi) represents a promising new approach to combat viral infections, and recent developments in the field of gene therapy have increased the feasibility of clinical applications. to explore the utility of RNAi for the treatment of the ultimately life-threatening liver disease caused by hepatitis C virus (HCV), which affects approximately 170 million people worldwide. A review of current developments in liver-directed gene delivery and the potential application of RNAi for the treatment of HCV. In addition, the involvement of microRNAs (miRNA) in HCV infection and the potential therapeutic implications are emphasized. RNAi technologies have fuelled rapid progress in the basic understanding of HCV biology and revealed numerous new viral and host-cell factors as potential targets for therapy. Together with the improvement of gene delivery technology and the discovery of the critical role of miRNA in HCV infection, RNAi and miRNA-based antiviral strategies hold great promise for the future.


Innovation Procurement in Health Systems: Exploring Practice and Lessons Learned.
As rising healthcare costs continue to challenge the sustainability of global health systems, there has been a strategic shift toward a focus on value, which considers the outcomes and value of healthcare delivery relative to the costs of care delivery. A unique feature of this focus on value has influenced a shift in procurement whereby health organizations are advancing the procurement of innovative solutions to achieve defined outcomes that overcome challenges such as the quality, safety and cost of care delivery. In this paper, we report on the implementation of three innovation procurement models in four Ontario healthcare organizations. These case studies provide evidence of the value and impact of innovation procurement approaches emerging from the four healthcare organizations. Three models of innovation procurement are described in the four cases, along with qualitative analysis of experiences and outcomes for both the organizations and the participating vendors. Evidence of the value and impact of procuring innovative solutions to address health organization challenges offers insights and new approaches to leveraging public procurement methodologies to achieve value and impact for health systems.


Genomics and the evolution of aminoacyl-tRNA synthesis.
Translation is the process by which ribosomes direct protein synthesis using the genetic information contained in messenger RNA (mRNA). Transfer RNAs (tRNAs) are charged with an amino acid and brought to the ribosome, where they are paired with the corresponding trinucleotide codon in mRNA. The amino acid is attached to the nascent polypeptide and the ribosome moves on to the next codon. Thus, the sequential pairing of codons in mRNA with tRNA anticodons determines the order of amino acids in a protein. It is therefore imperative for accurate translation that tRNAs are only coupled to amino acids corresponding to the RNA anticodon. This is mostly, but not exclusively, achieved by the direct attachment of the appropriate amino acid to the 3'-end of the corresponding tRNA by the aminoacyl-tRNA synthetases. To ensure the accurate translation of genetic information, the aminoacyl-tRNA synthetases must display an extremely high level of substrate specificity. Despite this highly conserved function, recent studies arising from the analysis of whole genomes have shown a significant degree of evolutionary diversity in aminoacyl-tRNA synthesis. For example, non-canonical routes have been identified for the synthesis of Asn-tRNA, Cys-tRNA, Gln-tRNA and Lys-tRNA. Characterization of non-canonical aminoacyl-tRNA synthesis has revealed an unexpected level of evolutionary divergence and has also provided new insights into the possible precursors of contemporary aminoacyl-tRNA synthetases.


Primary functional and aesthetic restoration of the fingernail in distal fingertip amputations with the eponychial flap.
Fingertip injuries often result in fingernail defects. Reconstruction of this structure is important for adequate functional and aesthetic results. This study evaluates the eponychial flap reconstruction technique in 45 fingertip amputations with loss of more than half the fingernail. In 33 cases the procedure was performed in combination with a palmar island flap for pulp reconstruction. Average follow up was 5 months. All eponychial flaps healed uneventfully. In 44 cases, the pulp volume was restored without nail growth disturbance. Five complications (9%) were observed (pain, soft nail, and nail deformity). Only one hook nail deformity required reoperation. All patients were satisfied with the aesthetic and functional outcome. We found eponychial flap fingernail reconstruction effective even for injuries proximal to the lunula and have extended the indication for this technique to very proximal fingernail defects. Eponychial flap reconstruction is a simple, safe, and time-effective technique without donor site morbidity. Simultaneous reconstruction of dorsal and palmar injuries should both be performed primarily resulting in the restoration of a satisfying fingertip.


Effects of ionic strength and surface charge on protein adsorption at PEGylated surfaces.
PEGylated Nb2O5 surfaces were obtained by the adsorption of poly(L-lysine)-g-poly(ethylene glycol) (PLL-g-PEG) copolymers, allowing control of the PEG surface density, as well as the surface charge. PEG (MW 2 kDa) surface densities between 0 and 0.5 nm(-2) were obtained by changing the PEG to lysine-mer ratio in the PLL-g-PEG polymer, resulting in net positive, negative and neutral surfaces. Colloid probe atomic force microscopy (AFM) was used to characterize the interfacial forces associated with the different surfaces. The AFM force analysis revealed interplay between electrical double layer and steric interactions, thus providing information on the surface charge and on the PEG layer thickness as a function of copolymer architecture. Adsorption of the model proteins lysozyme, alpha-lactalbumin, and myoglobin onto the various PEGylated surfaces was performed to investigate the effect of protein charge. In addition, adsorption experiments were performed over a range of ionic strengths, to study the role of electrostatic forces between surface charges and proteins acting through the PEG layer. The adsorbed mass of protein, measured by optical waveguide lightmode spectroscopy (OWLS), was shown to depend on a combination of surface charge, protein charge, PEG thickness, and grafting density. At high grafting density and high ionic strength, the steric barrier properties of PEG determine the net interfacial force. At low ionic strength, however, the electrical double layer thickness exceeds the thickness of the PEG layer, and surface charges "shining through" the PEG layer contribute to protein interactions with PLL-g-PEG coated surfaces. The combination of AFM surface force measurements and protein adsorption experiments provides insights into the interfacial forces associated with various PEGylated surfaces and the mechanisms of protein resistance.


Engineering of protein secretion in yeast: strategies and impact on protein production.
Yeasts combine the ease of genetic manipulation and fermentation of a microorganism with the capability to secrete and modify foreign proteins according to a general eukaryotic scheme. Their rapid growth, microbiological safety, and high-density fermentation in simplified medium have a high impact particularly in the large-scale industrial production of foreign proteins, where secretory expression is important for simplifying the downstream protein purification process. However, secretory expression of heterologous proteins in yeast is often subject to several bottlenecks that limit yield. Thus, many studies on yeast secretion systems have focused on the engineering of the fermentation process, vector systems, and host strains. Recently, strain engineering by genetic modification has been the most useful and effective method for overcoming the drawbacks in yeast secretion pathways. Such an approach is now being promoted strongly by current post-genomic technology and system biology tools. However, engineering of the yeast secretion system is complicated by the involvement of many cross-reacting factors. Tight interdependence of each of these factors makes genetic modification difficult. This indicates the necessity of developing a novel systematic modification strategy for genetic engineering of the yeast secretion system. This mini-review focuses on recent strategies and their advantages for systematic engineering of yeast strains for effective protein secretion.


Development and validation of a MEDLINE search filter/hedge for degenerative cervical myelopathy.
Degenerative cervical myelopathy (DCM) is a common condition with many unmet clinical needs. Pooled analysis of studies is an important tool for advancing medical understanding. This process starts with a systematic search of the literature. Identification of studies in DCM is challenged by a number of factors, including non-specific terminology and index terms. Search filters or HEDGEs, are search strings developed and validated to optimise medical literature searches. We aimed to develop a search filter for DCM for the MEDLINE database. The diagnostic test assessment framework of a "development dataset" and seperate "validation dataset" was used. The development dataset was formed by hand searching four leading spinal journals (Spine, Journal of Neurosurgery Spine, Spinal Cord and Journal of Spinal Disorders and Techniques) in 2005 and 2010. The search filter was initially developed focusing on sensitivity and subsequently refined using NOT functions to improve specificity. One validation dataset was formed from DCM narrative and systematic review articles and the second, articles published in April of 1989, 1993, 1997, 2001, 2005, 2009, 2013 and 2017 retrieved via the search MeSH term 'Spine'. Metrics of sensitivity, specificity, precision and accuracy were used to test performance. Hand searching identified 77/1094 relevant articles for 2005 and 55/1199 for 2010. We developed a search hedge with 100% sensitivity and a precision of 30 and 29% for the 2005 and 2010 development datasets respectively. For the selected time periods, EXP Spine returned 2113 publications and 30 were considered relevant. The search filter identified all 30 relevant articles, with a specificity of 94% and precision of 20%. Of the 255 references listed in the narrative index reviews, 225 were indexed in MEDLINE and 165 (73%) were relevant articles. All relevant articles were identified and accuracy ranged from 67 to 97% over the three reviews. Of the 42 articles returned from 3 recent systematic reviews, all were identified by the filter. We have developed a highly sensitive hedge for the research of DCM. Whilst precision is similarly low as other hedges, this search filter can be used as an adjunct for DCM search strategies.


Toxoplasma gondii impairs memory in infected seniors.
Almost 30% of humans present a Toxoplasma gondii positive antibody status and its prevalence increases with age. The central nervous system is the main target. However, little is known about the influence of asymptomatic i.e. latent Toxoplasmosis on cognitive functions in humans. To investigate neurocognitive dysfunctions in asymptomatic older adults with T. gondii positive antibody status a double-blinded neuropsychological study was conducted. The participants were classified from a population-based sample (N=131) of healthy participants with an age of 65 years and older into two groups with 42 individuals each: Toxoplasmosis positive (T-pos; IgG>50 IU/ml) and Toxoplasmosis negative (T-neg; IgG=0 IU/ml). The outcome measures were a computer-based working-memory test (2-back) and several standardized psychometric tests of memory and executive cognitive functions. T-pos seniors showed an impairment of different aspects of memory. The rate of correctly detected target symbols in a 2-back task was decreased by nearly 9% (P=0.020), corresponding to a performance reduction of about 35% in working memory relative to the T-neg group. Moreover, T-pos seniors had a lower performance in a verbal memory test, both regarding immediate recall (10% reduction; P=0.022), delayed recognition (6%; P=0.037) and recall from long-term memory assessed by the word fluency tests (12%; P=0.029). In contrast, executive functions were not affected. The effects remained mostly unchanged after controlling for medication. The impairment of memory functions in T-pos seniors was accompanied by a decreased self-reported quality of life. Because of the high prevalence of asymptomatic Toxoplasmosis and an increasing population of older adults this finding is of high relevance for public health.


Efficacy and safety of LCI699 for hypertension: a meta-analysis of randomized controlled trials and systematic review.
This study reviews the available data from randomized controlled trials on efficacy and safety of LCI699, a novel inhibitor of aldosterone synthase, as treatment of hypertension. We performed a meta-analysis of phase II randomized, controlled trials comparing the efficacy/safety of LCI699 with placebo in hypertension patients. For this purpose, PubMed, Embase, Cochrane Library database, ISI-Science Citation Index, and the Chinese Biomedicine Literature Database were searched until August 2013. The available data on mean sitting systolic blood pressure (MSSBP), mean sitting diastolic blood pressure (MSDBP), adverse effects, renin-angiotensin-aldosterone system biomarkers (RAASB) and adrenocorticotropic hormone-stimulated cortisol concentration (AHSC) were collected. All data were analyzed using Review Manager, version 5.2. The present study finally included three randomized controlled trials, comprising of 623 patients in total. The daily use of ≥ 1 mg LCI699 was associated with a significant reduction of MSSBP (Weighted mean difference/WMD = -8.80, 95% CI: -11.31 to -5.68, p < 0.00001, I2 = 0%) and MSDBP (WMD = -4.94, 95% CI: -7.49 to -2.40, p = 0.00001, I2 = 9%). Adverse reactions occurred in 73 of the 139 patients (52.51%) treated with LCI699 and in 34 of the 63 patients (53.96%) treated with placebo. Pooled meta-analysis showed that the use of LCI699 was associated with no increased risk of side effects compared with placebo (RR = 0.90; 95% CI: 0.68 to 1.18, p = 0.43, I2 = 0%). Suppression of plasma aldosterone was measured at all doses of LCI699 treatment groups. LCI699 suppressed the ACTH-stimulated cortisol response in a dose- and time-dependent manner. Current evidence indicates that the novel aldosterone inhibitor LCI699 is an effective and well-tolerated antihypertensive agent that lowers plasma aldosterone concentration and produces a mild ACTH-stimulated cortisol response suppressive effect.


Validation study of a new transit time ultrasonic flow probe for continuous great vessel measurements.
Continuous measurement of cardiac output is important during experimental and clinical cardiac surgery as an indicator of ventricular function. Previous flow probes underestimated flow secondary to position and flow (S-series probes; Transonic Systems, Inc., Ithaca, NY), required frequent calibrations (electromagnetic), and were cumbersome to use. The new A-series probe (ASP) by Transonic Systems, Inc., uses a new X method of ultrasonic illumination insensitive to perturbations in flow. The ASPs were found to be accurate during in vitro studies, but have not been validated in vivo. Six anesthetized pigs were instrumented for right atrium to left atrium bypass, and ASPs were placed on the ascending aorta and pulmonary artery. Baseline measurements included aortic (Ao) and pulmonic flow (P), and thermodilution (Td) cardiac output. Animals then were placed on right heart bypass, and flow was randomly varied from 1 to 6 L/min, and Ao flow was recorded. In addition, ASPs were rotated and their direction reversed. After data collection, the occlusive roller pump (RP) was calibrated using a timed collection method. Calibrated RP flows were plotted versus ASP flows, and regression was applied. There was no difference between mean Ao, P, and Td cardiac outputs at baseline. In addition, changes in position and direction of the probe did not affect measurement of flow. The ASPs showed a highly linear correlation with RP ([r = 0.98, p < 0.01] ASP[L/min] = 0.98 RP-0.032). During laminar flow states, ASPs are accurate and insensitive to position on the great vessels.


Complications of Third Molar Extraction: A retrospective study from a tertiary healthcare centre in Oman.
This retrospective study aimed to investigate complications associated with the extraction of third molars at a tertiary healthcare centre in Oman. All consecutive patients who underwent extraction of one or more impacted third molars under general anaesthesia at Sultan Qaboos University Hospital, Muscat, Oman, between January 2007 and December 2017 were included. Age, gender, indication for extraction, teeth removed, procedure and complications were recorded. A total of 1,116 third molars (56% mandibular and 44% maxillary) were extracted and the majority (67.7%) were from female patients. The mean age at extraction was 24 ± 5 years and most patients (77.7%) were 20-29 years old. The intraoperative and postoperative complication rates were 3.7% and 8.3%, respectively. The intraoperative complications included tuberosity fracture (1.2%), root fracture (1.1%), bleeding (0.7%), soft tissue injury (0.5%) and adjacent tooth damage (0.2%). Postoperative complications were sensory nerve injuries (7.2%), swelling/pain/trismus (0.6%) and dry socket (0.5%). Nerve injury was temporary in 41 patients and permanent in four cases. A statistically significant relationship was observed between those aged 30-39 years and dry socket (P = 0.010) as well as bone removal and all postoperative complications (P = 0.001). Most complications resulting from third molar extractions were minor and within the reported ranges in the scientific literature. However, increased age and bone removal were associated with a higher risk of complications. These findings may help to guide treatment planning, informed consent and patient education.


Geometrically pinned magnetic domain wall for multi-bit per cell storage memory.
Spintronic devices currently rely on magnetic switching or controlled motion of domain walls (DWs) by an external magnetic field or a spin-polarized current. Controlling the position of DW is essential for defining the state/information in a magnetic memory. During the process of nanowire fabrication, creating an off-set of two parts of the device could help to pin DW at a precise position. Micromagnetic simulation conducted on in-plane magnetic anisotropy materials shows the effectiveness of the proposed design for pinning DW at the nanoconstriction region. The critical current for moving DW from one state to the other is strongly dependent on nanoconstricted region (width and length) and the magnetic properties of the material. The DW speed which is essential for fast writing of the data could reach values in the range of hundreds m/s. Furthermore, evidence of multi-bit per cell memory is demonstrated via a magnetic nanowire with more than one constriction.


Corrected mean-field models for spatially dependent advection-diffusion-reaction phenomena.
In the exclusion-process literature, mean-field models are often derived by assuming that the occupancy status of lattice sites is independent. Although this assumption is questionable, it is the foundation of many mean-field models. In this work we develop methods to relax the independence assumption for a range of discrete exclusion-process-based mechanisms motivated by applications from cell biology. Previous investigations that focused on relaxing the independence assumption have been limited to studying initially uniform populations and ignored any spatial variations. By ignoring spatial variations these previous studies were greatly simplified due to translational invariance of the lattice. These previous corrected mean-field models could not be applied to many important problems in cell biology such as invasion waves of cells that are characterized by moving fronts. Here we propose generalized methods that relax the independence assumption for spatially inhomogeneous problems, leading to corrected mean-field descriptions of a range of exclusion-process-based models that incorporate (i) unbiased motility, (ii) biased motility, and (iii) unbiased motility with agent birth and death processes. The corrected mean-field models derived here are applicable to spatially variable processes including invasion wave-type problems. We show that there can be large deviations between simulation data and traditional mean-field models based on invoking the independence assumption. Furthermore, we show that the corrected mean-field models give an improved match to the simulation data in all cases considered.


Li(0.8)Mg(2.1)B2O5F: the first borate fluoride with magnesium-oxygen-fluorine octahedral chains.
The first alkali metal and magnesium mixed borate fluoride, Li(0.8)Mg(2.1)B2O5F, was prepared by the high-temperature solution method. It crystallizes in the monoclinic space group P2(1)/n (no. 14) with lattice constants a = 3.0881(4) Å, b = 13.6020(2) Å, c = 9.6563(1) Å, β = 91.744(9)°, Z = 4. In the structure, a special magnesium-oxygen-fluorine octahedral chain is found in borates for the first time. The octahedral chains are linked into a three-dimensional framework containing tunnels with (1)(∞)[LiO4](7-) chains and isolated B2O5 groups filling in. The differences of octahedral chains in Li(0.8)Mg(2.1)B2O5F and magnesium inosilicate minerals are discussed. The TG-DSC curves, IR spectrum and UV-Vis-NIR diffuse reflectance spectrum were measured.


Technique for multiplying the repetition rates of periodic trains of pulses by means of a temporal self-imaging effect in chirped fiber gratings.
We show that a temporal effect that is equivalent to the spatial self-imaging (Talbot) effect applies to the reflection of periodic signals from linearly chirped fiber gratings. The effect can be used for multiplying the repetition frequency of a given periodic pulse train without distorting the individual pulse characteristics. The practical limit on the frequency-multiplication factor depends only on the temporal width of the individual pulse. Thus we demonstrate that a suitable combination of well-known techniques for short-pulse generation, such as pulse mode locking, and the technique proposed here allows us to obtain short-pulse trains with ultrahigh repetition rates (in the terahertz regime). Results from simulations show good agreement with those predicted by theory.


A Consensus on the Diagnosis and Treatment of Acromegaly Comorbidities: An Update.
The aim of the Acromegaly Consensus Group was to revise and update the consensus on diagnosis and treatment of acromegaly comorbidities last published in 2013. The Consensus Group, convened by 11 Steering Committee members, consisted of 45 experts in the medical and surgical management of acromegaly. The authors received no corporate funding or remuneration. This evidence-based consensus was developed using the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) system to describe both the strength of recommendations and the quality of evidence following critical discussion of the current literature on the diagnosis and treatment of acromegaly comorbidities. Acromegaly Consensus Group participants conducted comprehensive literature searches for English-language papers on selected topics, reviewed brief presentations on each topic, and discussed current practice and recommendations in breakout groups. Consensus recommendations were developed based on all presentations and discussions. Members of the Scientific Committee graded the quality of the supporting evidence and the consensus recommendations using the GRADE system. Evidence-based approach consensus recommendations address important clinical issues regarding multidisciplinary management of acromegaly-related cardiovascular, endocrine, metabolic, and oncologic comorbidities, sleep apnea, and bone and joint disorders and their sequelae, as well as their effects on quality of life and mortality.


Cost implications of implementation of pathogen-inactivated platelets.
Pathogen inactivation (PI) is a new approach to blood safety that may introduce additional costs. This study identifies costs that could be eliminated, thereby mitigating the financial impact. Cost information was obtained from five institutions on tests and procedures (e.g., irradiation) currently performed, that could be eliminated. The impact of increased platelet (PLT) availability due to fewer testing losses, earlier entry into inventory, and fewer outdates with a 7-day shelf life were also estimated. Additional estimates include costs associated with managing (1) special requests and (2) test results, (3) quality control and proficiency testing, (4) equipment acquisition and maintenance, (5) replacement of units lost to positive tests, (6) seasonal or geographic testing, and (7) health department interactions. All costs are mean values per apheresis PLT unit in USD ($/unit). The estimated test costs that could be eliminated are $71.76/unit and a decrease in transfusion reactions corresponds to $2.70/unit. Avoiding new tests (e.g., Babesia and dengue) amounts to $41.80/unit. Elimination of irradiation saves $8.50/unit, while decreased outdating with 7-day storage can be amortized to $16.89/unit. Total potential costs saved with PI is $141.65/unit. Costs are influenced by a variety of factors specific to institutions such as testing practices and the location in which such costs are incurred and careful analysis should be performed. Additional benefits, not quantified, include retention of some currently deferred donors and scheduling flexibility due to 7-day storage. While PI implementation will result in additional costs, there are also potential offsetting cost reductions, especially after 7-day storage licensing.


Skin thymic stromal lymphopoietin initiates Th2 responses through an orchestrated immune cascade.
Thymic stromal lymphopoietin (TSLP) has emerged as a key initiator in Th2 immune responses, but the TSLP-driven immune cascade leading to Th2 initiation remains to be delineated. Here, by dissecting the cellular network triggered by mouse skin TSLP in vivo, we uncover that TSLP-promoted IL-4 induction in CD4(+) T cells in skin-draining lymph nodes is driven by an orchestrated 'DC-T-Baso-T' cascade, which represents a sequential cooperation of dendritic cells (DCs), CD4(+) T cells and basophils. Moreover, we reveal that TSLP-activated DCs prime naive CD4(+) T cells to produce IL-3 via OX40L signalling and demonstrate that the OX40L-IL-3 axis has a critical role in mediating basophil recruitment, CD4(+) T-cell expansion and Th2 priming. These findings thus add novel insights into the cellular network and signal axis underlying the initiation of Th2 immune responses.


A tool to facilitate clinical biomarker studies--a tissue dictionary based on the Human Protein Atlas.
The complexity of tissue and the alterations that distinguish normal from cancer remain a challenge for translating results from tumor biological studies into clinical medicine. This has generated an unmet need to exploit the findings from studies based on cell lines and model organisms to develop, validate and clinically apply novel diagnostic, prognostic and treatment predictive markers. As one step to meet this challenge, the Human Protein Atlas project has been set up to produce antibodies towards human protein targets corresponding to all human protein coding genes and to map protein expression in normal human tissues, cancer and cells. Here, we present a dictionary based on microscopy images created as an amendment to the Human Protein Atlas. The aim of the dictionary is to facilitate the interpretation and use of the image-based data available in the Human Protein Atlas, but also to serve as a tool for training and understanding tissue histology, pathology and cell biology. The dictionary contains three main parts, normal tissues, cancer tissues and cells, and is based on high-resolution images at different magnifications of full tissue sections stained with H & E. The cell atlas is centered on immunofluorescence and confocal microscopy images, using different color channels to highlight the organelle structure of a cell. Here, we explain how this dictionary can be used as a tool to aid clinicians and scientists in understanding the use of tissue histology and cancer pathology in diagnostics and biomarker studies.


Compressibility of ferrofluids: Towards a better understanding of structural properties.
This paper addresses a computational method aimed at obtaining the isothermal compressibility of ferrofluids by means of molecular dynamics (MD) simulations. We model ferrofluids as a system of dipolar soft spheres and carry out MD simulations in the NPT ensemble. The obtained isothermal compressibility computed via volume fluctuations provides us with a strong evidence that dipolar interactions lead to a higher compressibility of dipolar soft sphere systems: the stronger the dipolar interactions, the bigger is the deviation of the compressibility from the one of a system with no dipoles. Furthermore, we use the isothermal compressibility to calculate the structure factor of ferrofluids at low values of wave vectors, i.e. in the range where it is difficult to predict its behaviour because of a problem with accounting for long-range particle correlations that give the main contribution to the structure factor in this range. Our approach based on the interpolation of the structure factor and the computed isothermal compressibility allows us to obtain the smooth structure factor in the range of low wave vectors and the reliable fractal dimension of the clusters formed in the system.


Improved thermostability and PCR efficiency of Thermococcus celericrescens DNA polymerase via site-directed mutagenesis.
The Thermococcus celericrescens (Tcel) DNA polymerase gene, which contains a 2328-bp open reading frame that encodes 775 amino acid residues, was expressed in the Escherichia coli strain Rosetta(DE3)pLysS. The expressed enzyme was purified through heat treatment, HisTrap™ HP column chromatography and then HiTrap™ SP HP column chromatography. Tcel DNA polymerase has poor thermostability and PCR efficiency compared to those of other family B DNA polymerases. To improve thermostability and PCR efficiency, mutant Tcel DNA polymerases were created via site-directed mutagenesis. Specifically, we targeted the A752 residue for enhanced thermostability and the N213 residue for improved PCR efficiency. The mutant Tcel DNA polymerases all showed enhanced PCR efficiency and thermostability compared to those of the wild-type Tcel DNA polymerase. Specifically, the double mutant TcelA752K/N213D DNA polymerase had an approximately three-fold increase in thermostability over that of the wild-type enzyme and amplified a long 10-kb PCR product in an extension time of 2min. However, there was a small change in the 3'→5' exonuclease activity compared with that of the wild-type Tcel DNA polymerase, even though the mutation is in the ExoII motif. The double mutant TcelA752K/N213D DNA polymerase had a 2.6-fold lower error rate compared to that of Taq DNA polymerase. It seems that the double mutant TcelA752K/N213D DNA polymerase can be used in LA (long and accurate) PCR.


Scaling the size of perimetric stimuli reduces variability and returns constant thresholds across the visual field.
The conventional stimulus for standard automated perimetry is fixed in size, giving elevated contrast thresholds and reduced test reliability in the periphery. Here, we test the hypothesis that appropriate scaling of the size of perimetric stimuli will return fixed thresholds and reduced variability across the visual field. We derived frequency-of-seeing (FOS) curves in five healthy subjects at central (3 degrees) and peripheral (27 degrees) locations with a method of constant stimuli (MOCS) using a desktop LCD display. FOS curves for a Goldmann III (GIII) stimulus were compared with those for size scaled spots. To consider clinical translation, we tested a further five healthy subjects (22-24 years) with the Melbourne Rapid Fields (MRF) tablet perimeter at several locations spanning 1 degree to 25 degrees from fixation, deriving FOS curves (MOCS) and also conducting repeated adaptive clinical thresholding to assess intra- and interobserver variability. We found that GIII contrast thresholds were significantly elevated in the periphery compared with the parafovea, with concomitant reduction of FOS slope. Using appropriately size scaled spots, threshold and slope differences between these locations were significantly reduced. FOS data collected with the tablet perimeter confirmed that size scaling confers broad equivalence of the shape of the FOS curve across the visual field. Repeated adaptive thresholding with size scaled stimuli gave relatively constant intra-observer variability across the visual field, which compares favorably with published normative data obtained with the GIII stimulus. The reduced variability will improve signal-to-noise ratio for correct classification of normal visual field test results, whereas the lower contrast thresholds yield greater dynamic range, which should improve the ability to reliably monitor moderate defects.


Profiling of atherosclerotic lesions by gene and tissue microarrays reveals PCSK6 as a novel protease in unstable carotid atherosclerosis.
Carotid plaque instability is a major cause of ischemic stroke, but detailed knowledge about underlying molecular pathways is still lacking. Here, we evaluated large-scale transcriptomic and protein expression profiling in a biobank of carotid endarterectomies followed by characterization of identified candidates, as a platform for discovery of novel proteins differentially regulated in unstable carotid lesions. Genes highly upregulated in symptomatic versus asymptomatic plaques were selected from Affymetrix microarray analyses (n=127 plaques), and tissue microarrays constructed from 34 lesions were assayed for 21 corresponding proteins by immunohistochemistry. Quantification of stainings demonstrated differential expression of CD36, CD137, and DOCK7 (P<0.05) in unstable versus stable lesions and the most significant upregulation of a proprotein convertase, PCSK6 (P<0.0001). Increased expression of PCSK6 in symptomatic lesions was verified by quantitative real-time polymerase chain reaction (n=233), and the protein was localized to smooth muscle α-actin positive cells and extracellular matrix of the fibrous cap by immunohistochemistry. PCSK6 expression positively correlated to genes associated with inflammation, matrix degradation, and mitogens in microarrays. Stimulation of human carotid smooth muscle cells in vitro with cytokines caused rapid induction of PCSK6 mRNA. Using a combination of transcriptomic and tissue microarray profiling, we demonstrate a novel approach to identify proteins differentially expressed in unstable carotid atherosclerosis. The proprotein convertase PCSK6 was detected at increased levels in the fibrous cap of symptomatic carotid plaques, possibly associated with key processes in plaque rupture such as inflammation and extracellular matrix remodeling. Further studies are needed to clarify the role of PCSK6 in atherosclerosis.


Enhanced optical guiding of colloidal particles using a supercontinuum light source.
We demonstrate enhanced optical guiding distances for microscopic particles using a supercontinuum light beam. The enhanced spectral bandwidth of the source leads to an elongated focal region. As a result we obtain a significant radial gradient force and axial radiation pressure force over a longer distance when compared to a monochromatic Gaussian beam. The guiding distances of up to 3mm that are observed for micron-sized particles with the supercontinuum beam are approximately twice those observed using continuous wave and femtosecond laser sources when considering beams of equivalent diameter. This guiding scheme is expected to be applicable to colloidal particles, biological cells and cold atom ensembles.


Upregulation of DNA methyltransferase-mediated gene silencing, anchorage-independent growth, and migration of colon cancer cells by interleukin-6.
Inflammatory bowel disease is characterized by chronic inflammation which predisposes to colorectal cancer. The mechanisms by which inflammation promotes tumorigenesis are not fully known. We aimed to investigate the links between colonic inflammation and tumorigenesis via epigenetic gene silencing. Colon cancer specimens were assessed for the expression of DNA methyltransferase-1 (DNMT-1) using immunohistochemistry. Colorectal carcinoma cell lines were assessed for DNMT1 expression, methylcytosine content, promoter methylation, gene expression, and tumorigenesis in response to interleukin (IL)-6. DNMT1 was expressed at higher levels in both the peritumoral stroma and tumor in inflammatory bowel disease-associated cancers compared with sporadic colon cancers. IL-6 treatment of colon cancer cells resulted in an increase in DNMT1 expression, independent of de novo gene expression. IL-6 increased the methylation of promoter regions of genes associated with tumor suppression, adhesion, and apoptosis resistance. Expression of a subset of these genes was downregulated by IL-6, an effect that was prevented by preincubation with 5-azadeoxycytidine, a DNMT1 inhibitor. Anchorage-independent growth and migration of colon cancer cells was also increased by IL-6 in a 5-azadeoxycytidine-sensitive manner. Our results indicate that DNMT-mediated gene silencing may play a role in inflammation-associated colon tumorigenesis.


Gene-Xpert: Diagnosis of Pulmonary Tuberculosis in a Sputum Smear Negative Patient.
This case report has tried to highlight the ease and benefit of Gene-Xpert testing in difficult to diagnose patient with sputum smear negative pulmonary tuberculosis. Early treatment of tuberculosis is usually delayed by lack of rapid and accurate diagnostic modalities, especially in resource-limited settings like ours. Gene-Xpert is a rapid test based on real time PCR assay and molecular technology for the detection of Mycobacterium tuberculosis. It is highly sensitive tool and enables simultaneous detection of rifampicin resistance within short period of time i,e. <2hrs. It has distinct advantage of providing same-day diagnosis which could potentially limit loss to follow up during diagnostic evaluation of smear negative tuberculosis patients. Keywords: Gene-Xpert; pulmonary tuberculosis; sputum microscopy.


Development of a flow cytometry live cell assay for the screening of inhibitors of hepatitis C virus (HCV) replication.
In this study, we established a flow cytometry live cell-based assay that permits the screening of hepatitis C virus (HCV) inhibitors. Specifically, we created a stable cell line, which harbors a subgenomic replicon encoding an NS5A-YFP fusion protein. This system allows direct measurement of YFP fluorescence in live hepatoma cells in which the HCV replicon replicates. We demonstrated that this stable fluorescent system permits the rapid and sensitive quantification of HCV replication inhibition by direct-acting antiviral agents (DAA) including protease and NS5A inhibitors and host-targeting antiviral agents (HTA) including cyclophilin inhibitors. This flow cytometry-based live cell assay is well suited for multiple applications such as the evaluation of HCV replication as well as antiviral drug screening.


Examination of Treatment Satisfaction Instruments in Psoriasis: 2017 Results from the Psoriasis Working Group of the International Dermatology Outcome Measures (IDEOM).
In dermatology clinical trials, assessment of patients' treatment satisfaction is crucial but often lacking. To address this need, IDEOM's Psoriasis Working Group seeks to evaluate, develop, and validate treatment satisfaction instruments for the psoriasis population. The Psoriasis Working Group aimed to determine (1) factors affecting psoriasis patients' satisfaction with their therapies, (2) adequacy of two commonly used generic treatment satisfaction instruments in reflecting the psoriasis patients' perspective, and (3) whether a need exists to develop a new treatment satisfaction instrument. Patient perspectives on satisfaction with treatment efficacy, safety, convenience, and overall satisfaction were elicited.Stakeholders were presented with information regarding the feasibility and content validity of two generic treatment instruments, the Treatment Satisfaction Questionnaire for Medication (TSQM) and the Treatment Satisfaction with Medicines Questionnaire (SATMED-Q). We conducted a nominal group discussion and survey to determine whether stakeholders considered these instruments feasible and adequate to address treatment satisfaction for psoriasis therapies. Forty-five stakeholders participated in the nominal group discussion and survey. 53% of participants voted that the TSQM and SATMED-Q are not adequate and that we should create a new dermatology-specific treatment satisfaction instrument. Patients and other stakeholders also provided feedback on aspects of treatment satisfaction important to them. These include speed of onset and durability of therapeutic effect of a medication, permanence of side effects, and convenience of administering the medication. Stakeholders, including patients and providers, determined that generic treatment satisfaction questionnaires are not adequate to evaluate treatment satisfaction in psoriasis patients.


The Application of Contrast Media for In Vivo Feature Enhancement in X-Ray Computed Tomography of Soil-Grown Plant Roots.
The use of in vivo X-ray microcomputed tomography (μCT) to study plant root systems has become routine, but is often hampered by poor contrast between roots, soil, soil water, and soil organic matter. In clinical radiology, imaging of poorly contrasting regions is frequently aided by the use of radio-opaque contrast media. In this study, we present evidence for the utility of iodinated contrast media (ICM) in the study of plant root systems using μCT. Different dilutions of an ionic and nonionic ICM (Gastrografin 370 and Niopam 300) were perfused into the aerial vasculature of juvenile pea plants via a leaf flap (Pisum sativum). The root systems were imaged via μCT, and a variety of image-processing approaches used to quantify and compare the magnitude of the contrast enhancement between different regions. Though the treatment did not appear to significantly aid extraction of full root system architectures from the surrounding soil, it did allow the xylem and phloem units of seminal roots and the vascular morphology within rhizobial nodules to be clearly visualized. The nonionic, low-osmolality contrast agent Niopam appeared to be well tolerated by the plant, whereas Gastrografin showed evidence of toxicity. In summary, the use of iodine-based contrast media allows usually poorly contrasting root structures to be visualized nondestructively using X-ray μCT. In particular, the vascular structures of roots and rhizobial nodules can be clearly visualized in situ.


The 'other' coral symbiont: Ostreobium diversity and distribution.
Ostreobium is an endolithic algal genus thought to be an early-diverging lineage of the Bryopsidales (Ulvophyceae, Chlorophyta). Ostreobium can live in low-light conditions on calcium carbonate substrata in tropical conditions. It is best known as a symbiont of corals, where it lives deep within the animal skeleton and exchanges nitrogen and carbon, as well as providing nutrients and photoassimilates. In contrast to the relatively well-studied role of the photosynthetic zooxanthellae symbionts in coral (Symbiodinium), Ostreobium phylogeny, diversity and distribution are all poorly understood. Here, we describe the phylogenetic position and diversity of Ostreobium based on plastid 16S ribosomal DNA (rDNA), 18S rDNA and rbcL genes from a nuclear genome survey and complete plastid genome, and determined its environmental diversity and distribution by screening the publicly available environmental data for those genes. The results shed light on the phylogeny and the ecology of the 'other' coral symbiont.


Carcinoma of the gallbladder in ulcerative colitis and primary sclerosing cholangitis. Report of two cases.
The authors present the cases of two patients with carcinoma of the gallbladder complicating chronic ulcerative colitis. Both patients had concomitant primary sclerosing cholangitis. Twelve such cases of gallbladder carcinoma have been reported in the literature. The presence of primary sclerosing cholangitis in patients with ulcerative colitis is associated with malignancy of the extrahepatic biliary tree. It is suggested that, if colectomy is necessary, the liver should be biopsied and a cholecystectomy performed if the gallbladder is deemed abnormal.


Coherent THz Emission Enhanced by Coherent Synchrotron Radiation Wakefield.
We demonstrate that emission of coherent transition radiation by a ∼1 GeV energy-electron beam passing through an Al foil is enhanced in intensity and extended in frequency spectral range, by the energy correlation established along the beam by coherent synchrotron radiation wakefield, in the presence of a proper electron optics in the beam delivery system. Analytical and numerical models, based on experimental electron beam parameters collected at the FERMI free electron laser (FEL), predict transition radiation with two intensity peaks at ∼0.3 THz and ∼1.5 THz, and extending up to 8.5 THz with intensity above 20 dB w.r.t. the main peak. Up to 80-µJ pulse energy integrated over the full bandwidth is expected at the source, and in agreement with experimental pulse energy measurements. By virtue of its implementation in an FEL beam dump line, this work promises dissemination of user-oriented multi-THz beamlines parasitic and self-synchronized to EUV and x-ray FELs.


Impact of Covid-19 on farming systems in Europe through the lens of resilience thinking.
Resilience is the ability to deal with shocks and stresses, including the unknown and previously unimaginable, such as the Covid-19 crisis. This paper assesses (i) how different farming systems were exposed to the crisis, (ii) which resilience capacities were revealed and (iii) how resilience was enabled or constrained by the farming systems' social and institutional environment. The 11 farming systems included have been analysed since 2017. This allows a comparison of pre-Covid-19 findings and the Covid-19 crisis. Pre-Covid findings are from the SURE-Farm systematic sustainability and resilience assessment. For Covid-19 a special data collection was carried out during the early stage of lockdowns. Our case studies found limited impact of Covid-19 on the production and delivery of food and other agricultural products. This was due to either little exposure or the agile activation of robustness capacities of the farming systems in combination with an enabling institutional environment. Revealed capacities were mainly based on already existing connectedness among farmers and more broadly in value chains. Across cases, the experience of the crisis triggered reflexivity about the operation of the farming systems. Recurring topics were the need for shorter chains, more fairness towards farmers, and less dependence on migrant workers. However, actors in the farming systems and the enabling environment generally focused on the immediate issues and gave little real consideration to long-term implications and challenges. Hence, adaptive or transformative capacities were much less on display than coping capacities. The comparison with pre-Covid findings mostly showed similarities. If challenges, such as shortage of labour, already loomed before, they persisted during the crisis. Furthermore, the eminent role of resilience attributes was confirmed. In cases with high connectedness and diversity we found that these system characteristics contributed significantly to dealing with the crisis. Also the focus on coping capacities was already visible before the crisis. We are not sure yet whether the focus on short-term robustness just reflects the higher visibility and urgency of shocks compared to slow processes that undermine or threaten important system functions, or whether they betray an imbalance in resilience capacities at the expense of adaptability and transformability. Our analysis indicates that if transformations are required, e.g. to respond to concerns about transnational value chains and future pandemics from zoonosis, the transformative capacity of many farming systems needs to be actively enhanced through an enabling environment.


Laminin-111 peptide C16 regulates invadopodia activity of malignant cells through β1 integrin, Src and ERK 1/2.
Laminin peptides influence tumor behavior. In this study, we addressed whether laminin peptide C16 (KAFDITYVRLKF, γ1 chain) would increase invadopodia activity of cells from squamous cell carcinoma (CAL27) and fibrosarcoma (HT1080). We found that C16 stimulates invadopodia activity over time in both cell lines. Rhodamine-conjugated C16 decorates the edge of cells, suggesting a possible binding to membrane receptors. Flow cytometry showed that C16 increases activated β1 integrin, and β1 integrin miRNA-mediated depletion diminishes C16-induced invadopodia activity in both cell lines. C16 stimulates Src and ERK 1/2 phosphorylation, and ERK 1/2 inhibition decreases peptide-induced invadopodia activity. C16 also increases cortactin phosphorylation in both cells lines. Based on our findings, we propose that C16 regulates invadopodia activity over time of squamous carcinoma and fibrosarcoma cells, probably through β1 integrin, Src and ERK 1/2 signaling pathways.


Measurement of LRRK2 and Ser910/935 phosphorylated LRRK2 in peripheral blood mononuclear cells from idiopathic Parkinson's disease patients.
A significant number of autosomal dominantly inherited Parkinson's disease (PD) cases are due to mutations in the leucine-rich repeat kinase 2 (LRRK2) gene. In cells, these pathogenic mutations have a number of differing effects on LRRK2 enzymatic activity and protein stability. In particular, five of the six described pathogenic LRRK2 mutations ablate the constitutive phosphorylation of LRRK2 on Ser910 and Ser935, two residues required for binding of LRRK2 to 14-3-3 proteins. This suggests a potential pathogenic role for these residues. However, LRRK2 kinase inhibitors, which have shown early promise as neuroprotective agents, also ablate the phosphorylation of Ser910 and Ser935. Additionally, LRRK2 is phosphorylated on Ser910 and Ser935 following activation of the inflammatory toll-like receptor pathway and inflammatory cytokines are often increased in PD patients. Whether LRRK2 protein or phosphorylation is altered in idiopathic PD is unknown. We therefore measured LRRK2 protein and its phosporylation in peripheral blood mononuclear cells (PBMCs) from 33 idiopathic Parkinson's disease patients and 27 age-matched controls. We found no significant difference in total LRRK2 protein levels in PBMCs from PD patients compared to controls. Furthermore, total LRRK2 protein expression was not effected by age, disease duration, disease severity or levodopa medication. The amount of phosphorylation on LRRK2 at both Ser910 and Ser935 correlated highly with total LRRK2 levels and was also unchanged in PD patients. Therefore, changes in LRRK2 Ser910/Ser935 phosphorylation in PBMCs are unlikely to contribute to idiopathic Parkinson's disease or be of utility as a disease biomarker. However, the invariance of Ser910 and Ser935 phosphorylation in PD PBMC's suggests that these residues could be used as pharmacodynamic biomarkers for the effectiveness of LRRK2 kinase inhibitors in patients.


Biallelic TBCD Mutations Cause Early-Onset Neurodegenerative Encephalopathy.
We describe four families with affected siblings showing unique clinical features: early-onset (before 1 year of age) progressive diffuse brain atrophy with regression, postnatal microcephaly, postnatal growth retardation, muscle weakness/atrophy, and respiratory failure. By whole-exome sequencing, we identified biallelic TBCD mutations in eight affected individuals from the four families. TBCD encodes TBCD (tubulin folding co-factor D), which is one of five tubulin-specific chaperones playing a pivotal role in microtubule assembly in all cells. A total of seven mutations were found: five missense mutations, one nonsense, and one splice site mutation resulting in a frameshift. In vitro cell experiments revealed the impaired binding between most mutant TBCD proteins and ARL2, TBCE, and β-tubulin. The in vivo experiments using olfactory projection neurons in Drosophila melanogaster indicated that the TBCD mutations caused loss of function. The wide range of clinical severity seen in this neurodegenerative encephalopathy may result from the residual function of mutant TBCD proteins. Furthermore, the autopsied brain from one deceased individual showed characteristic neurodegenerative findings: cactus and somatic sprout formations in the residual Purkinje cells in the cerebellum, which are also seen in some diseases associated with mitochondrial impairment. Defects of microtubule formation caused by TBCD mutations may underlie the pathomechanism of this neurodegenerative encephalopathy.


MRI-guided stereotactic laser corpus callosotomy for epilepsy: distinct methods and outcomes.
Several small series have described stereotactic MRI-guided laser interstitial thermal therapy for partial callosotomy of astatic and generalized tonic-clonic (GTC) seizures, especially in association with Lennox-Gastaut syndrome. Larger case series and comparison of distinct stereotactic methods for stereotactic laser corpus callosotomy (SLCC), however, are currently lacking. The objective of this study was to report seizure outcomes in a series of adult patients with epilepsy following anterior, posterior, and complete SLCC procedures and to compare the results achieved with a frameless stereotactic surgical robot versus direct MRI guidance frames. The authors retrospectively reviewed sequential adult epilepsy surgery patients who underwent SLCC procedures at a single institution. They describe workflows, stereotactic errors, percentage disconnection, hospitalization durations, adverse events, and seizure outcomes after performing anterior, posterior, and complete SLCC procedures using a frameless stereotactic surgical robot versus direct MRI guidance platforms. Thirteen patients underwent 15 SLCC procedures. The median age at surgery was 29 years (range 20-49 years), the median duration of epilepsy was 21 years (range 9-48 years), and median postablation follow-up was 20 months (range 4-44 months). Ten patients underwent anterior SLCC with a median 73% (range 33%-80%) midsagittal length of callosum acutely ablated. Following anterior SLCC, 6 of 10 patients achieved meaningful (> 50%) reduction of target seizures. Four patients underwent posterior (completion) SLCC following prior anterior callosotomy, and 1 patient underwent complete SLCC as a single procedure; 3 of these 5 patients experienced meaningful reduction of target seizures. Overall, 8 of 10 patients in whom astatic seizures were targeted and treated by anterior and/or posterior SLCC experienced meaningful improvement. SLCC procedures with direct MRI guidance (n = 7) versus a frameless surgical robot (n = 8) yielded median radial accuracies of 1.1 mm (range 0.2-2.0 mm) versus 2.4 mm (range 0.6-6.1 mm; p = 0.0011). The most serious adverse event was a clinically significant intraparenchymal hemorrhage in a patient who underwent the robotic technique. This is the largest reported series of SLCC for epilepsy to date. SLCC provides seizure outcomes comparable to open surgery outcomes reported in the literature. Direct MRI guidance is more accurate, which has the potential to reduce the risks of SLCC. Methodological advancements and larger studies are needed.


NEDD4-2 associates with gamma(c) and regulates its degradation rate.
Interleukin-2 (IL-2) is a cytokine that regulates proliferation, differentiation and survival of various lymphoid cell subsets. Its actions are mediated through its binding to the IL-2 receptor which is composed of three subunits (IL-2Ralpha, IL-2Rbeta and gamma(c)). Only beta and gamma(c) have been shown to transduce intra cellular signals. The gamma(c) chain is shared by the interleukin-2, 4, 7, 9, 15 and 21 receptors, and is essential for lymphocyte functions. The regulation of gamma(c) expression level is therefore critical for the ability of cells to respond to these cytokines. In the present work, we show that the IL-2R constitutively associates with the ubiquitin ligase NEDD4-2, and to a lesser extent NEDD4-1. We identified the specific binding site on gamma(c). And we show that the loss of NEDD4 association on gamma(c) is accompanied by a dramatic increase of the half-life of the receptor subunit.


A unique set of centrosome proteins requires pericentrin for spindle-pole localization and spindle orientation.
Majewski osteodysplastic primordial dwarfism type II (MOPDII) is caused by mutations in the centrosome gene pericentrin (PCNT) that lead to severe pre- and postnatal growth retardation. As in MOPDII patients, disruption of pericentrin (Pcnt) in mice caused a number of abnormalities including microcephaly, aberrant hemodynamics analyzed by in utero echocardiography, and cardiovascular anomalies; the latter being associated with mortality, as in the human condition. To identify the mechanisms underlying these defects, we tested for changes in cell and molecular function. All Pcnt(-/-) mouse tissues and cells examined showed spindle misorientation. This mouse phenotype was associated with misdirected ventricular septal growth in the heart, decreased proliferative symmetric divisions in brain neural progenitors, and increased misoriented divisions in fibroblasts; the same phenotype was seen in fibroblasts from three MOPDII individuals. Misoriented spindles were associated with disrupted astral microtubules and near complete loss of a unique set of centrosome proteins from spindle poles (ninein, Cep215, centriolin). All these proteins appear to be crucial for microtubule anchoring and all interacted with Pcnt, suggesting that Pcnt serves as a molecular scaffold for this functionally linked set of spindle pole proteins. Importantly, Pcnt disruption had no detectable effect on localization of proteins involved in the cortical polarity pathway (NuMA, p150(glued), aPKC). Not only do these data reveal a spindle-pole-localized complex for spindle orientation, but they identify key spindle symmetry proteins involved in the pathogenesis of MOPDII.


Congenital contractural arachnodactyly and femoral fracture in a newborn infant: a causal relationship or a coincidence?
Congenital contractural arachnodactyly (CCA, Beals syndrome) is an autosomal-dominant connective tissue disorder characterized by multiple flexion contractures, arachnodactyly, severe kyphoscoliosis, abnormal pinnae, and muscular hypoplasia. Although it is a connective tissue disorder affecting bone structure and formation, coexistence of bony fractures in CCA patients have not been reported before. In this article we report a newborn infant diagnosed with CCA who developed a femoral fracture possibly due to abnormal bone structure and birth injury in spite of cesarean delivery.