Status one year after fertility assessment and counselling in women of reproductive age-a qualitative study.
Over the past 50 years women and men have postponed family formation in high-income societies. Fertility assessment and counselling has been suggested as a method to reduce delayed childbearing and its consequences. This study explored women's perceptions of how attending a fertility assessment intervention influenced their decisions and choices regarding family formation and childbearing. Follow-up data from a longitudinal semi-structured qualitative interview study including 20 women aged 35-40 years seeking individual fertility counselling at the Fertility Assessment and Counselling Clinic at Rigshospitalet, Copenhagen, Denmark. The interviews were conducted one year after their consultation. Data were analysed by qualitative content analysis. The women perceived an increase in their knowledge after they had attended the counselling. The women saw the counselling as a catalyst for change-they changed their behaviour and relationship status. The women stopped thinking about the pros and cons of childbearing and acted instead. The women did not experience any regrets about acting. Some of the women felt that they were still in limbo as they were still in doubt concerning childbearing. The consultation had not given them an answer with a clear deadline in terms of delaying attempts to become pregnant, and this frustrated them. Our study highlights the impact of a fertility assessment and counselling intervention which included a perceived increase in knowledge. The clinic allows for an individualized approach to fertility awareness which is necessary given the unique nature of childbearing decisions.


Hemodynamics During Dialysis and Changes in Cognitive Performance.
Hemodialysis (HD) patients are at increased risk for cognitive impairment. Blood pressure (BP) fluctuations during HD may affect cerebral perfusion and subsequently cognitive function. Examine and provide information on the relationship between intradialytic hemodynamics and cognitive outcomes over a 1-year period. HD patients without diagnosed dementia who were 50 years old or older were given a neurocognitive battery at baseline and at 1-year follow-up. Over the 1-year period, we collected demographic and laboratory data, as well as dialytic BP and ultrafiltration rate (UFR) measurements and tested the association between changes in cognitive test scores and intradialytic hemodynamics, adjusting for demographic and clinical variables. Thirty-nine participants enrolled in the study and 32 remained at 1-year follow-up. The mean (SD) age was 66.8 (10.0) years. Hypertension was present in 100% and diabetes mellitus in 47% of the cohort. The average change in systolic BP from predialysis to postdialysis was -9.9 (16.3) mmHg, and average maximum drop in systolic BP during dialysis was 27.9 (10.2) mmHg. Overall, the cognitive test scores did not have significant changes from baseline to 1 year. In our linear regression analysis there was no association between the BP measures and cognitive changes, although UFR was associated with change in performance on a test of executive functioning. In prevalent HD patients, cognitive function was generally stable over a 1-year period, and there was no association with intradialytic hemodynamic variables.


Antibody response to Pseudomonas aeruginosa exoproducts in cancer patients.
We analyzed sera from 28 patients with various types of malignancies for the occurrence of antibodies against exotoxin A of Pseudomonas aeruginosa and two Pseudomonas proteases. A total of 27 of these individuals were colonized or infected with P. aeruginosa at one time or another during the study, whereas the remaining patient was colonized with four non-P. aeruginosa species of Pseudomonas. Sera were obtained from several of these patients before P. aeruginosa colonization or infection of these individuals was detected, which provided an opportunity to evaluate their responsiveness to pseudomonal exoproducts as they acquired the organism. Exotoxin A was purified from culture supernatant fluids of strain PA-103, and the two proteases were purified from an isolate of strain JR3, a highly proteolytic strain originally recovered from the sputum of a cystic fibrosis patient. Antibodies to the exotoxin A and the two proteases were detected in these sera, and sera which contained relatively high antibody levels to exotoxin A afforded mice complete protection against lethal challenges with this substance. Statistical analyses showed that patients infected with P. aeruginosa had consistently higher antibody levels (P less than 0.005) to the exoproducts than patients who were colonized with this organism. Also, patients colonized with P. aeruginosa possessed significantly higher antibody levels (P less than 0.003) to these three exoproducts than uninfected, hospitalized patients. Parke-Davis type 1 was the strain most commonly isolated from these patients (46%), but colonization or infection due to this organism usually resulted in the production of low levels of antibody to Pseudomonas exoproducts. However, infections with Parke-David type 7 organisms were always associated with intermediate- and high-responder sera to exotoxin A. These results indicated that potentially toxic products were elaborated during the course of cancer-related colonization and infection with P. aeruginosa.


Profiling gene expression to distinguish the likely active diazotrophs from a sea of genetic potential in marine sediments.
Nitrogen (N) cycling microbial communities in marine sediments are extremely diverse, and it is unknown whether this diversity reflects extensive functional redundancy. Sedimentary denitrifiers remove significant amounts of N from the coastal ocean and diazotrophs are typically regarded as inconsequential. Recently, N fixation has been shown to be a potentially important source of N in estuarine and continental shelf sediments. Analysis of expressed genes for nitrite reductase (nirS) and a nitrogenase subunit (nifH) was used to identify the likely active denitrifiers and nitrogen fixers in surface sediments from different seasons in Narragansett Bay (Rhode Island, USA). The overall diversity of diazotrophs expressing nifH decreased along the estuarine gradient from the estuarine head to an offshore continental shelf site. Two groups of sequences related to anaerobic sulphur/iron reducers and sulphate reducers dominated libraries of expressed nifH genes. Quantitative polymerase chain reaction (qPCR) and quantitative reverse transcription polymerase chain reaction (qRT-PCR) data shows the highest abundance of both groups at a mid bay site, and the highest nifH expression at the head of the estuary, regardless of season. Several potential environmental factors, including water temperature, oxygen concentration and metal contamination, may influence the abundance and nifH expression of these two bacterial groups.


A General Approach to Achieving Stability and Safe Behavior in Distributed Robotic Architectures.
This paper proposes a unified energy-based modeling and energy-aware control paradigm for robotic systems. The paradigm is inspired by the layered and distributed control system of organisms, and uses the fundamental notion of energy in a system and the energy exchange between systems during interaction. A universal framework that models actuated and interacting robotic systems is proposed, which is used as the basis for energy-based and energy-limited control. The proposed controllers act on certain energy budgets to accomplish a desired task, and decrease performance if a budget has been depleted. These budgets ensure that a maximum amount of energy can be used, to ensure passivity and stability of the system. Experiments show the validity of the approach.


Microbiota innate stimulation is a prerequisite for T cell spontaneous proliferation and induction of experimental colitis.
Little is known about how the microbiota regulates T cell proliferation and whether spontaneous T cell proliferation is involved in the pathogenesis of inflammatory bowel disease. In this study, we show that stimulation of innate pathways by microbiota-derived ligands and antigen-specific T cell stimulation are both required for intestinal inflammation. Microbiota-derived ligands promoted spontaneous T cell proliferation by activating dendritic cells (DCs) to produce IL-6 via Myd88, as shown by the spontaneous proliferation of T cells adoptively transferred into specific pathogen-free (SPF) RAG-/- mice, but not in germfree RAG-/- mice. Reconstitution of germfree RAG-/- mice with cecal bacterial lysate-pulsed DCs, but not with IL-6-/- or Myd88-/- DCs, restored spontaneous T cell proliferation. CBir1 TCR transgenic (CBir1 Tg) T cells, which are specific for an immunodominant microbiota antigen, induced colitis in SPF RAG-/- mice. Blocking the spontaneous proliferation of CBir1 Tg T cells by co-transferring bulk OT II CD4+ T cells abrogated colitis development. Although transferred OT II T cells underwent spontaneous proliferation in RAG-/- mice, the recipients failed to develop colitis because of the lack of cognate antigen in the intestinal lumen. Collectively, our data demonstrate that induction of colitis requires both spontaneous proliferation of T cells driven by microbiota-derived innate signals and antigen-specific T cell proliferation.


Design, Synthesis, and Structure-Activity Relationship Studies of Novel GPR88 Agonists (4-Substituted-phenyl)acetamides Based on the Reversed Amide Scaffold.
The development of synthetic agonists for the orphan receptor GPR88 has recently attracted significant interest, given the promise of GPR88 as a novel drug target for psychiatric and neurodegenerative disorders. Examination of structure-activity relationships of two known agonist scaffolds 2-PCCA and 2-AMPP, as well as the recently resolved cryo-EM structure of 2-PCCA-bound GPR88, led to the design of a new scaffold based on the "reversed amide" strategy of 2-AMPP. A series of novel (4-substituted-phenyl)acetamides were synthesized and assessed in cAMP accumulation assays as GPR88 agonists, which led to the discovery of several compounds with better or comparable potencies to 2-AMPP. Computational docking studies suggest that these novel GPR88 agonists bind to the same allosteric site of GPR88 that 2-PCCA occupies. Collectively, our findings provide structural insight and SAR requirement at the allosteric site of GPR88 and a new scaffold for further development of GPR88 allosteric agonists.


Expanded eggshell procedure combined with closing-opening technique (a modified vertebral column resection) for the treatment of thoracic and thoracolumbar angular kyphosis.
The purpose of this study was to determine the efficacy of a modified vertebral column resection for the treatment of thoracolumbar angular kyphosis. A total of 13 patients (8 male, 5 female) with thoracolumbar kyphosis (kyphotic angle > 60°) were included in this study (Group A). There were 3 patients with failure of spinal formation (Type 1 deformity), 6 patients with old thoracic or lumbar compression fracture, and 4 patients with old spinal tuberculosis (including 1 case of T3-5 vertebral malunion). The average preoperative kyphotic angle was 67.3° (range 62°-75°). Each patient underwent an expanded eggshell procedure combined with the closing-opening technique for the treatment of thoracolumbar angular kyphosis. Sixteen patients who were previously treated with a closing-opening wedge osteotomy in the same spine classification group (kyphotic angle > 60°) were used as a control group (Group B). In Group A, the average (± SD) operative time was 400 ± 60 minutes, and the average blood loss was 960 ± 120 ml. There were no surgery-related complications observed during or after the operations. The average local kyphotic angle was 20.3° (range 18°-24.5°), and the average correction rate was 68.7%. In Group B, the average operative time was 470 ± 90 minutes, and the average blood loss was 2600 ± 1600 ml (range 1200-8200 ml). There were segmental vessels and spinal canal venous plexus injury in 1 case, spinal cord injury in 1 case, dural tearing in 2 cases, pleural rupture in 2 cases, and hemothorax and pneumothorax in 1 case. Each patient had more than 2 years of follow-up. At the latest follow-up examination, the average regional kyphotic angle was 19.9° ± 9.1° (range 19°-34°), and there was no significant loss of correction (p > 0.05). There was greater blood loss and a higher complication rate in Group B than in Group A (p < 0.05). An expanded eggshell procedure combined with the closing-opening technique for the treatment of thoracolumbar angular kyphosis resulted in significant reduction of the kyphotic angle, few complications, and good follow-up results. However, a larger series of patients and long-term follow-up results is still required to verify the effectiveness and safety of this method.


Communication and teamwork in patient care: how much can we learn from aviation?
To identify evidence on the role of assertiveness and teamwork and the application of aviation industry techniques to improve patient safety for inpatient obstetric care. Studies limited to research with humans in English language retrieved from CINAHL, PubMed, Social Science Abstracts, and Social Sciences Citation Index, and references from reviewed articles. A total of 13 studies were reviewed, including 5 studies of teamwork, communication, and safety attitudes in aviation; 2 studies comparing these factors in aviation and health care; and 6 studies of assertive behavior and decision making by nurses. Studies lacking methodological rigor or focusing on medication errors and deviant behavior were excluded. Pilot attitudes regarding interpersonal interaction on the flight deck predicted effective performance and were amenable to behavior-based training to improve team performance. Nursing knowledge was inconsistently accessed in decision making. Findings regarding nurse assertiveness were mixed. Adaptation of training concepts and safety methods from other fields will have limited impact on perinatal safety without an examination of the contextual experiences of nurses and other health care providers in working to prevent patient harm.


Race, African ancestry, and Helicobacter pylori infection in a low-income United States population.
Gastric cancer incidence in African Americans is twice that of whites, and differing prevalence of Helicobacter pylori strain-specific isolates may help explain the disparity. Serum levels of antibodies to each of 15 H. pylori proteins were assessed using multiplex serology for a sample of 689 African American and white participants from the Southern Community Cohort Study. African and European admixture was estimated using a panel of 276 ancestry genetic markers, with "low," "medium," and "high" categories of African ancestry defined as <85%, 85% to 95%, and ≥95%. The majority (79%) of our study population were sero-positive for H. pylori. African American race was associated with a two- to sixfold increased odds for sero-positivity to eight H. pylori proteins, including the cancer-associated virulence constituents CagA [odds ratio (OR), 6.4; 95% CI, 4.5-9.1], and VacA (OR, 2.3; 95% CI, 1.5-3.5). Compared to whites, African Americans of low, medium, and high African ancestry had 1.6-, 4.1-, and 5.2-fold increased odds of sero-positivity to H. pylori, primarily related to CagA sero-positive strains, for which increasing African ancestry led to 2.5-, 9.6-, and 13.1-fold increased odds. Among African Americans alone, compared to those of low African ancestry, African Americans of medium and high African ancestry had 2.5- and 3.4-fold increased odds of sero-positivity to H. pylori, and 3.5- and 4.9-fold increased odds of CagA sero-positive H. pylori strains. Host genetic variation and/or lifestyle factors associated with African ancestry contribute to the likelihood of infection with H. pylori, particularly its virulent strains, in this low-income U.S. southern population. Our findings that low-income African Americans of high African ancestry have a particularly high prevalence of antibodies against H. pylori provides a framework for further research into better detection and prevention of gastric cancer in this population.


From Secretion in Pichia pastoris to Application in Apple Juice Processing: Exo-Polygalacturonase from Sporothrix schenckii 1099-18.
Polygalacturonases are a group of enzymes under pectinolytic enzymes related to enzymes that hydrolyse pectic substances. Polygalacturonases have been used in various industrial applications such as fruit juice clarification, retting of plant fibers, wastewater treatment drinks fermentation, and oil extraction. The study was evaluated at the heterologous expression, purification, biochemical characterization, computational modeling, and performance in apple juice clarification of a new exo-polygalacturonase from Sporothrix schenckii 1099-18 (SsExo-PG) in Pichia pastoris. Recombinant DNA technology was used in this study. Two different pPIC9K plasmids were constructed with native signal sequence-ssexo-pg and alpha signal sequence-ssexo-pg separately. Protein expression and purification performed after plasmids transformed into the Pichia pastoris. Biochemical and structural analyses were performed by using pure SsExo-PG. The purification of SsExo-PG was achieved using a Ni-NTA chromatography system. The enzyme was found to have a molecular mass of approximately 52 kDa. SsExo-PG presented as stable at a wide range of temperature and pH values, and to be more storage stable than other commercial pectinolytic enzyme mixtures. Structural analysis revealed that the catalytic residues of SsExo- PG are somewhat similar to other Exo-PGs. The KM and kcat values for the degradation of polygalacturonic acid (PGA) by the purified enzyme were found to be 0.5868 μM and 179 s-1, respectively. Cu2+ was found to enhance SsExo-PG activity while Ag2+ and Fe2+ almost completely inhibited enzyme activity. The enzyme reduced turbidity up to 80% thus enhanced the clarification of apple juice. SsExo-PG showed promising performance when compared with other commercial pectinolytic enzyme mixtures. The clarification potential of SsExo-PG was revealed by comparing it with commercial pectinolytic enzymes. The following parameters of the process of apple juice clarification processes showed that SsExo-PG is highly stable and has a novel performance.


Comparison of long-term outcomes in STEMI and NSTE-ACS after coronary stent placement: an analysis in a real world BMS and DES population.
The prognostic difference between STEMI and NSTE-ACS after coronary stent placement remains unclear. We aimed to compare the short- and long-term event rates in patients presenting with ST-segment elevation myocardial infarction (STEMI) and non-ST-segment elevation acute coronary syndrome (NSTE-ACS) after percutaneous coronary intervention (PCI) with either bare-metal stents (BMS) or drug-eluting stents (DES). Between 2000 and 2005 a total of 1749 STEMI and 1921 NSTE-ACS patients received either a BMS or DES in consecutive real world cohorts. Descriptive statistics and multivariate survival analyses were applied to compare the event rates in STEMI and NSTE-ACS during 4 years follow-up. NSTE-ACS patients had significantly higher clinical and angiographic risk profiles at baseline and were treated with less optimal medical therapy during follow-up. At 4 years follow-up, all-cause mortality was significantly higher in STEMI compared to NSTE-ACS after coronary stent placement (17.4% vs. 14.3%; HR 1.60, 95% CI 1.24-2.07). In a landmark analysis no difference was seen in all-cause mortality among STEMI en NSTE-ACS between 1 month and 4 years follow-up (HR 1.10, 95% CI 0.81-1.51). Cardiac death was more prevalent in STEMI patients, while the 4-year cumulative incidences of any myocardial infarction, any coronary revascularization, target lesion revascularization and definite stent thrombosis were similar in both ACS groups. Patients presenting with STEMI have a worse long-term prognosis compared to NSTE-ACS after coronary stent placement, due to higher short-term death rates. However, after the first month STEMI and NSTE-ACS patients have a comparable long-term survival.


Scalpel versus electrosurgery for major abdominal incisions.
Scalpels or electrosurgery can be used to make abdominal incisions. The potential benefits of electrosurgery may include reduced blood loss, dry and rapid separation of tissue, and reduced risk of cutting injury to surgeons. Postsurgery risks possibly associated with electrosurgery may include poor wound healing and complications such as surgical site infection. To assess the effects of electrosurgery compared with scalpel for major abdominal incisions. The first version of this review included studies published up to February 2012. In October 2016, for this first update, we searched the Cochrane Wounds Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL), Ovid MEDLINE (including In-Process & Other Non-Indexed Citations), Ovid Embase, EBSCO CINAHL Plus, and the registry for ongoing trials (www.clinicaltrials.gov). We did not apply date or language restrictions. Studies considered in this analysis were randomised controlled trials (RCTs) that compared electrosurgery to scalpel for creating abdominal incisions during major open abdominal surgery. Incisions could be any orientation (vertical, oblique, or transverse) and surgical setting (elective or emergency). Electrosurgical incisions were made through major layers of the abdominal wall, including subcutaneous tissue and the musculoaponeurosis (a sheet of connective tissue that attaches muscles), regardless of the technique used to incise the skin and peritoneum. Scalpel incisions were made through major layers of abdominal wall including skin, subcutaneous tissue, and musculoaponeurosis, regardless of the technique used to incise the abdominal peritoneum. Primary outcomes analysed were wound infection, time to wound healing, and wound dehiscence. Secondary outcomes were postoperative pain, wound incision time, wound-related blood loss, and adhesion or scar formation. Two review authors independently carried out study selection, data extraction, and risk of bias assessment. When necessary, we contacted trial authors for missing data. We calculated risk ratios (RR) and 95% confidence intervals (CI) for dichotomous data, and mean differences (MD) and 95% CI for continuous data. The updated search found seven additional RCTs making a total of 16 included studies (2769 participants). All studies compared electrosurgery to scalpel and were considered in one comparison. Eleven studies, analysing 2178 participants, reported on wound infection. There was no clear difference in wound infections between electrosurgery and scalpel (7.7% for electrosurgery versus 7.4% for scalpel; RR 1.07, 95% CI 0.74 to 1.54; low-certainty evidence downgraded for risk of bias and serious imprecision). None of the included studies reported time to wound healing.It is uncertain whether electrosurgery decreases wound dehiscence compared to scalpel (2.7% for electrosurgery versus 2.4% for scalpel; RR 1.21, 95% CI 0.58 to 2.50; 1064 participants; 6 studies; very low-certainty evidence downgraded for risk of bias and very serious imprecision).There was no clinically important difference in incision time between electrosurgery and scalpel (MD -45.74 seconds, 95% CI -88.41 to -3.07; 325 participants; 4 studies; moderate-certainty evidence downgraded for serious imprecision). There was no clear difference in incision time per wound area between electrosurgery and scalpel (MD -0.58 seconds/cm2, 95% CI -1.26 to 0.09; 282 participants; 3 studies; low-certainty evidence downgraded for very serious imprecision).There was no clinically important difference in mean blood loss between electrosurgery and scalpel (MD -20.10 mL, 95% CI -28.16 to -12.05; 241 participants; 3 studies; moderate-certainty evidence downgraded for serious imprecision). Two studies reported on mean wound-related blood loss per wound area; however, we were unable to pool the studies due to considerable heterogeneity. It was uncertain whether electrosurgery decreased wound-related blood loss per wound area. We could not reach a conclusion on the effects of the two interventions on pain and appearance of scars for various reasons such as small number of studies, insufficient data, the presence of conflicting data, and different measurement methods. The certainty of evidence was moderate to very low due to risk of bias and imprecise results. Low-certainty evidence shows no clear difference in wound infection between the scalpel and electrosurgery. There is a need for more research to determine the relative effectiveness of scalpel compared with electrosurgery for major abdominal incisions.


Sleep disturbances in obsessive-compulsive disorder: Association with non-response to repetitive transcranial magnetic stimulation (rTMS).
Background Repetitive transcranial magnetic stimulation (rTMS) is a promising augmentation strategy for treatment-refractory OCD. However, a substantial group still fails to respond. Sleep disorders, e.g. circadian rhythm sleep disorders (CRSD), are highly prevalent in OCD and might mediate treatment response. The aims of the current study were to compare sleep disturbances between OCD patients and healthy subjects as well as between rTMS responders and non-responders, and most importantly to determine sleep-related predictors of rTMS non-response. Methods 22 OCD patients received at least 10 sessions rTMS combined with psychotherapy. Sleep disturbances were measured using questionnaires and actigraphy. Sleep in patients was compared to healthy subjects. Treatment response was defined as >35% reduction on YBOCS. Treatment response prediction models were based on measures of CRSD and insomnia. Results Sleep disturbances were more prevalent in OCD patients than healthy subjects. The OCD group consisted of 12 responders and 10 non-responders. The CRSD model could accurately predict non-response with 83% sensitivity and 63% specificity, whereas the insomnia model could not. Conclusions CRSD is more prevalent in OCD patients than healthy subjects, specifically in rTMS non-responders. Therefore, CRSD may serve as a biomarker for different subtypes of OCD corresponding with response to specific treatment approaches.


Essential health services delivery and quality improvement actions under drought and food insecurity emergency in north-east Uganda.
Essential health services can be disrupted due to several naturally occurring public health emergencies such as drought, flood, earthquake and outbreak of infectious diseases. However, little evidence exists on the status of essential health services delivery under the effect of drought and food insecurity. North-east Uganda is severely affected by prolonged drought that significantly affected the livelihood of the residents. Therefore, we aimed to determine the current status of essential health services and quality improvement (QI) actions in health facilities in north-east Uganda. We used a descriptive cross-sectional study design to assess the availability of essential health service and quality improvement activities in drought and food insecurity affected districts of north-east Uganda. We included a total of 150 health facilities from 15 districts with proportionated multistage sampling method. We interviewed health facilities' managers and services focal persons using structured questionnaire and observation checklist. We used a descriptive statistic to analyze the data with SPSS version 22. A few health facilities (8.7%) had mental health specialist. There was also lack of capacity building training on essential health services. Considerable proportion of health facilities had no non-communicable diseases (38.3%), mental health (47.0%), and basic emergency obstetric care (40.3%) services. Stock out of essential medicines were observed in 20% of health facilities. There was lack of supportive supervision, and poor documentation of QI activities. Essential health service and QI were suboptimal in drought and food insecure emergency affected districts. Human resource deployment (especially mental health specialist), provision of capacity building training, improving non-communicable diseases, mental health and basic emergency obstetric care services are required to improve availability of essential health services. Supporting supply chain management to minimize stock out of medicines, and promoting QI activities are also vital to assure quality of health service in drought and food insecurity affected districts in north-Eastern Uganda.


Breeding increases the efficacy of Chondrostereum purpureum in the sprout control of birch.
We tested whether the pairing of selected isolates could be used to increase the efficiency of a decay fungus Chondrostereum purpureum (Pers. Ex Fr.) Pouzar to control hardwood sprouting in Finland. We paired C. purpureum strains efficient in sprout control or highly active in laccase production, and tested the efficacy of their progeny in spout control experiments. This procedure resulted in a strain with an efficacy superior to that of the parental strains. The mortality of birch (Betula pendula Roth. and B. pubescens Ehrh.) 1 cm in stump diameter was 78%, 56% and 9% for the best progeny, the best parental strain and the control, respectively. Mortality was only slightly higher for B. pendula than for B. pubescens but no significant differences were found between the number or maximum height of stump sprouts. Our results showed that cross breeding of this decay fungus is a good alternative in attempts to produce efficient biocontrol agents against hardwood sprouting.


Solution structure of calcium-bound rat S100B(betabeta) as determined by nuclear magnetic resonance spectroscopy,.
The three-dimensional structure of Ca2+-bound rat S100B(betabeta) has been determined using data from a series of two-dimensional (2D), three-dimensional (3D), and four-dimensional (4D) nuclear magnetic resonance (NMR) experiments. Each S100beta subunit (91 residues) contains four helixes (helix 1, E2-R20; helix 2, K29-N38; helix 3, Q50-D61; and helix 4, F70-A83) and one antiparallel beta-sheet (strand 1, K26-K28; and strand 2, E67-D69) which brings the normal and pseudo EF-hands together. As found previously for rat apo-S100B(betabeta) [Drohat, A. C., et al. (1996) Biochemistry 35, 11577-11588], helixes 1, 1', 4, and 4' associate to form an X-type four-helix bundle at the symmetric dimer interface. Additionally, Ca2+ binding does not significantly change the interhelical angle of helixes 1 and 2 in the pseudo EF-hand (apo, Omega1-2 = 132 +/- 4 degrees; and Ca2+-bound, Omega1-2 = 137 +/- 5 degrees). However, the interhelical angle of helixes 3 and 4 in the normal EF-hand (Omega3-4 = 106 +/- 4 degrees) changed significantly upon the addition of Ca2+ (DeltaOmega3-4 = 112 +/- 5 degrees) and is similar to that of the Ca2+-bound EF-hands in calbindin D9K, calmodulin, and troponin (84 degrees </= Omega </= 128 degrees). Further, the four helixes within each S100beta subunit form a splayed-type four-helix bundle (four perpendicular helixes) as observed in Ca2+-bound calbindin D9K. The large Ca2+-dependent conformational change involving helix 3 exposes a cleft, defined by residues in the hinge region, the C-terminal loop, and helix 3, which is absent in the apo structure. This surface on Ca2+-bound S100B(betabeta) is likely important for target protein binding.


Mitochondrial DNA sequences of primates: tempo and mode of evolution.
We cloned and sequenced a segment of mitochondrial DNA from human, chimpanzee, gorilla, orangutan, and gibbon. This segment is 896 bp in length, contains the genes for three transfer RNAs and parts of two proteins, and is homologous in all 5 primates. The 5 sequences differ from one another by base substitutions at 283 positions and by a deletion of one base pair. The sequence differences range from 9 to 19% among species, in agreement with estimates from cleavage map comparisons, thus confirming that the rate of mtDNA evolution in primates is 5 to 10 times higher than in nuclear DNA. The most striking new finding to emerge from these comparisons is that transitions greatly outnumber transversions. Ninety-two percent of the differences among the most closely related species (human, chimpanzee, and gorilla) are transitions. For pairs of species with longer divergence times, the observed percentage of transitions falls until, in the case of comparisons between primates and non-primates, it reaches a value of 45. The time dependence is probably due to obliteration of the record of transitions by multiple substitutions at the same nucleotide site. This finding illustrates the importance of choosing closely related species for analysis of evolutionary process. The remarkable bias toward transitions in mtDNA evolution necessitates the revision of equations that correct for multiple substitutions at the same site. With revised equations, we calculated the incidence of silent and replacement substitutions in the two protein-coding genes. The silent substitution rate is 4 to 6 times higher than the replacement rate, indicating strong functional constraints at replacement sites. Moreover, the silent rate for these two genes is about 10% per million years, a value 10 times higher than the silent rate for the nuclear genes studied so far. In addition, the mean substitution rate in the three mitochondrial tRNA genes is at least 100 times higher than in nuclear tRNA genes. Finally, genealogical analysis of the sequence differences supports the view that the human lineage branched off only slightly before the gorilla and chimpanzee lineages diverged and strengthens the hypothesis that humans are more related to gorillas and chimpanzees than is the orangutan.


Shock waves in stellar atmospheres and breaking waves on an ocean beach.
The phenomenon of ocean waves breaking on a beach is analogous to shock waves in the atmosphere of a pulsating star. In both cases a velocity discontinuity is clearly present. In stars the upper, expanding layer halts and falls back so as to interact with the rising gas at a shock. Similarly, a bore on a beach reaches its maximum extension before sliding back onto the next incoming wave. Analogous quantities such as the surface gravity of the star and the beach gradient in the ocean have similar effects on the flows and the nature of the discontinuity between them. Phenomena that are not analogous include the thermodynamic properties of the two media. Ocean observations may help solve some problems in shock phenomena associated with stellar pulsation.


Cystatin C- and creatinine-based estimated glomerular filtration rate, vascular disease, and mortality in persons with diabetes in the U.S.
OBJECTIVE Serum cystatin C is an alternative to serum creatinine for estimating glomerular filtration rate (GFR), since cystatin C is less influenced by age and muscle mass. Among persons with diabetes, we compared the performance of GFR estimated using cystatin C (eGFRcys) with that using creatinine (eGFRcr) for the identification of reduced kidney function and its association with diabetes complications. RESEARCH DESIGN AND METHODS We analyzed data from adult participants from the 1999-2002 National Health and Nutrition Examination Survey with available cystatin C (N = 4,457). Kidney function was dichotomized as preserved (eGFR ≥60 mL/min/1.73 m(2)) or reduced (eGFR <60 mL/min/1.73 m(2)) using the 2012 Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) cystatin C and the 2009 CKD-EPI creatinine equations. RESULTS Among 778 persons with diabetes, the prevalence of reduced kidney function was 16.5% using eGFRcr and 22.0% using eGFRcys. More persons with diabetes were reclassified from preserved kidney function by eGFRcr to reduced kidney function by eGFRcys than persons without diabetes (odds ratio 3.1 [95% CI 1.9-4.9], P < 0.001). The associations between lower eGFR and higher prevalence of albuminuria, retinopathy, peripheral arterial disease, and coronary artery disease were robust regardless of filtration marker. Similarly, the risk of all-cause mortality increased with lower eGFRcr and eGFRcys. Only lower eGFRcys was significantly associated with cardiovascular mortality. CONCLUSIONS More persons with diabetes had reduced kidney function by eGFRcys than by eGFRcr, and lower eGFRcys was strongly associated with diabetes complications. Whether eGFRcys is superior to eGFRcr in approximating true kidney function in a diabetic population requires additional study.


Altered tethering of the SspB adaptor to the ClpXP protease causes changes in substrate delivery.
SspB is a dimeric adaptor protein that increases the rate at which ssrA-tagged substrates are degraded by tethering them to the ClpXP protease. Each SspB subunit consists of a folded domain that forms the dimer interface and a flexible C-terminal tail. Ternary delivery complexes are stabilized by three sets of tethering interactions. The C-terminal XB peptide of each SspB subunit binds ClpX, the body of SspB binds one part of the ssrA-tag sequence, and ClpX binds another part of the tag. To test the functional importance of these tethering interactions, we engineered monomeric SspB variants and dimeric variants with different length linkers between the SspB body and the XB peptide and employed substrates with degradation tags that bind ClpX weakly and/or contain extensions between the binding sites for SspB and ClpX. We find that monomeric SspB variants can enhance ClpXP degradation of a subset of substrates, that doubling the number of tethering interactions stimulates degradation via changes in Km and Vmax, and that major alterations in the length of the 48-residue SspB linker cause only small changes in the efficiency of substrate delivery. These results indicate that the properties of the degradation tag and the number of SspB.ClpX tethering interactions are the major factors that determine the extent to which the substrate and ClpX are engaged in ternary delivery complexes.


Acanthamoeba keratitis successfully treated medically.
The first medical cure of a corneal infection due to an Acanthamoeba species is reported. The 44-year-old patient developed a suppurative keratitis associated with an epithelial defect, hypopyon, and secondary glaucoma. Acanthamoeba was confirmed as the causative agent four months after presentation when positive cultures were obtained from the cornea and from the conjunctiva. Sensitivity studies of the isolated organism were performed, and the infection was successfully controlled by treatment with a combination of dibromopropamidine and propamidine isethionate ointment and drops and neomycin drops. Keratoplasty was performed 22 months after onset, and no viable acanthamoebae were present in the resected tissue, though possible cyst remnants were identified by immunofluorescent techniques.


Computed tomogram guided fine-needle aspiration cytology of lung mass with histological correlation: A study in Eastern India.
Fine-needle aspiration cytology (FNAC) is an important and useful investigation, and is considered next to imaging in the rapid diagnosis of pulmonary mass lesion for the last few decades. To assess the role of Computed Tomogram (CT) guided FNAC in pulmonary mass lesions; to analyze the results; and to compare with histopathological findings. The clinical, radiological, and cytological data of 130 patients were prospectively studied who underwent CT guided FNAC from October 2009 through September 2011. Thereafter these patients underwent bronchoscopic/trucut biopsy/lobectomy, whatever clinically indicated. Smears and tissue sections were evaluated simultaneously to reach at a definite diagnosis. Out of 130 cases, we found adequate FNAC smear and histopathology reports only in 124 cases. The age range varied from 35 to 73 years with the peak in the fifth to sixth decades. The benign lesions were 10 (8.07%) and malignant lesions were114 (91.93%) shown by cytology. The most common tumor was adenocarcinoma (51.72%) followed by squamous cell carcinoma (22.41%) and small cell carcinoma 6.89%. Diagnostic accuracy of CT guided FNAC was 95%. Post procedural complications such as hemorrhage and chest pain were minimal and were noted only in three cases. CT guided FNAC of pulmonary masses provides simple, easy, and reliable method for reaching rapid tissue diagnosis with minimal complication.


Mutational analysis of the integral membrane methyltransferase isoprenylcysteine carboxyl methyltransferase (ICMT) reveals potential substrate binding sites.
The eukaryotic integral membrane enzyme isoprenylcysteine carboxyl methyltransferase (ICMT) methylates the carboxylate of a lipid-modified cysteine at the C terminus of its protein substrates. This is the final post-translational modification of proteins containing a CAAX motif, including the oncoprotein Ras, and therefore, ICMT may serve as a therapeutic target in cancer development. ICMT has no discernible sequence homology with soluble methyltransferases, and aspects of its catalytic mechanism are unknown. For example, how both the methyl donor S-adenosyl-l-methionine (AdoMet), which is water-soluble, and the methyl acceptor isoprenylcysteine, which is lipophilic, are recognized within the same active site is not clear. To identify regions of ICMT critical for activity, we combined scanning mutagenesis with methyltransferase assays. We mutated nearly half of the residues of the ortholog of human ICMT from Anopheles gambiae and observed reduced or undetectable catalytic activity for 62 of the mutants. The crystal structure of a distantly related prokaryotic methyltransferase (Ma Mtase), which has sequence similarity with ICMT in its AdoMet binding site but methylates different substrates, provides context for the mutational analysis. The data suggest that ICMT and Ma MTase bind AdoMet in a similar manner. With regard to residues potentially involved in isoprenylcysteine binding, we identified numerous amino acids within transmembrane regions of ICMT that dramatically reduced catalytic activity when mutated. Certain substitutions of these caused substrate inhibition by isoprenylcysteine, suggesting that they contribute to the isoprenylcysteine binding site. The data provide evidence that the active site of ICMT spans both cytosolic and membrane-embedded regions of the protein.


Elongation of surface plasmon polariton propagation length without gain.
We have demonstrated that an addition of highly concentrated rhodamine 6G chloride dye to the PMMA film adjacent to a silver film can cause 30% elongation of the propagation length of surface Plasmon polaritons (SPPs). The possibility to elongate the SPP propagation length without optical gain opens a new technological dimension to low-loss nanoplasmonic and metamaterials.


Reducing anchor loss in micromechanical extensional mode resonators.
In this work, we propose a novel method to increase the quality factor of extensional mode micromechanical resonators. The proposed resonator topology is suitable for integration in a silicon-based process to fabricate micromechanical filters and oscillators. It is a half-wavelength-long strip excited longitudinally by electrostatic forces, and it is isolated from the substrate by alternating with bars of a quarter wavelength long. This structure causes a large impedance mismatch between the resonator and the substrate and hence reduces the anchor loss considerably. The performance of the resonator is determined by finite element simulations. We introduce an equivalent electrical circuit to predict the performance of the resonator. The electrical model gives results consistent with the finite element simulations. The proposed resonator is expected to have a very small anchor loss resulting in a very high Q.


The development and historical context of the Datta short laryngoscope handle.
The hormonal, physiologic, and anatomic changes of pregnancy have a number of significant anesthetic implications, including the potential for difficulties and failures in tracheal intubation. The American Society of Anesthesiology closed claims database in the 1970s observed that maternal deaths were involved in 30% of all obstetrics claims, most stemming from difficulty with intubation or ventilation. In the late 1970s, Dr. Sanjay Datta, MBBS, an obstetric anesthesiologist at Brigham and Women's Hospital (Boston, MA), observed a number of differences in the practice of obstetric anesthesia in the United States when compared with his prior experiences in the United Kingdom and Canada. Dr. Datta perceived that parturients within North America had a higher body mass index. In addition, he observed an increased rate of cesarean delivery and general anesthesia use. These differences led him to evaluate ways in which the laryngoscope itself could be altered to improve the ease of intubation of parturients; this led to the development of the short laryngoscope handle. The genesis of the Datta short laryngoscope handle, and the accompanying historical context, will be explored.


A comprehensive preclinical assessment of late-term imaging markers of radiation-induced brain injury.
Cranial radiotherapy (CRT) is an important part of brain tumor treatment, and although highly effective, survivors suffer from long-term cognitive side effects. In this study we aim to establish late-term imaging markers of CRT-induced brain injury and identify functional markers indicative of cognitive performance. Specifically, we aim to identify changes in executive function, brain metabolism, and neuronal organization. Male Sprague Dawley rats were fractionally irradiated at 28 days of age to a total dose of 30 Gy to establish a radiation-induced brain injury model. Animals were trained at 3 months after CRT using the 5-choice serial reaction time task. At 12 months after CRT, animals were evaluated for cognitive and imaging changes, which included positron emission tomography (PET) and magnetic resonance imaging (MRI). Cognitive deficit with signs of neuroinflammation were found at 12 months after CRT in irradiated animals. CRT resulted in significant volumetric changes in 38% of brain regions as well as overall decrease in brain volume and reduced gray matter volume. PET imaging showed higher brain glucose uptake in CRT animals. Using MRI, irradiated brains had an overall decrease in fractional anisotropy, lower global efficiency, increased transitivity, and altered regional connectivity. Cognitive measurements were found to be significantly correlated with six image features that included myelin integrity and local organization of the neural network. These results demonstrate that CRT leads to late-term morphological changes, reorganization of neural connections, and metabolic dysfunction. The correlation between imaging markers and cognitive deficits can be used to assess late-term side effects of brain tumor treatment and evaluate efficacy of new interventions.


Characterization of strong fiber Bragg gratings using an applied thermal chirp and iterative algorithm.
Coupling coefficients of various grating types and strengths are calculated from measurements of the complex reflectivity using an applied thermal chirp and optical frequency domain reflectometry (OFDR). The complex reflectivity is then utilized by a layer peeling algorithm to determine the coupling coefficient of the thermally chirped grating. A guess of the temperature profile enables the coupling coefficient of the unchirped grating to be estimated. An iterative algorithm is then used to converge on the exact coupling coefficient, employing an error minimization method applied to the reflectivity spectra. This technique removes the need for a reference grating while preserving the spatial resolution obtained with the initial OFDR measurement. Successful reconstruction of gratings with integrated |κ|L ~ 9.0 are demonstrated with a spatial resolution of less than 100 μm.


Real-World Impact of Immune Checkpoint Inhibitors in Metastatic Uveal Melanoma.
Uveal melanoma (UM) is the most common intraocular malignancy in adults and shows a high rate of metastatic spread. As randomized clinical trials with immune checkpoint inhibitors (ICI) have not been performed in patients with metastatic UM, we analyzed the real-world outcomes in a nationwide population-based study. Clinical data of patients with UM were extracted from the Danish Metastatic Melanoma database, a nationwide database containing unselected records of patients diagnosed with metastatic melanoma in Denmark. Survival before (pre-ICI, n = 32) and after (post-ICI, n = 94) the approval of first-line treatment with ICI was analyzed. A partial response to first-line treatment was observed in 7% of patients treated with anti-programmed cell death protein (PD)-1 monotherapy and in 21% with combined anti-cytotoxic T lymphocyte antigen (CTLA)-4 plus anti-PD-1 therapy. Median progression-free survival was 2.5 months for patients treated in the pre-ICI era compared to 3.5 months in the post-ICI era (hazard ratio (HR) 0.43; 95% confidence interval (CI) 0.28-0.67; p < 0.001). The estimated one-year overall survival rate increased from 25.0% to 41.9% and the median overall survival improved from 7.8 months to 10.0 months, respectively (HR 0.52; 95% CI 0.34-0.79; p = 0.003). Thus, the introduction of ICI as first-line treatment appears to have significantly improved the real-world survival of patients with metastatic UM, despite relatively low response rates compared to cutaneous melanoma. With the lack of therapies proven effective in randomized trials, these data support the current treatment with ICI in patients with metastatic UM.


Cost-sharing effects on adherence and persistence for second-generation antipsychotics in commercially insured patients.
To assess the relationship between patient cost-sharing (e.g., copayments or coinsurance) and adherence and persistence to second-generation (atypical) antipsychotic (SGA) medications. A retrospective, observational study of adults aged 18-64 years with schizophrenia or bipolar disorder (n = 7,910) who initiated SGA medications with employer-sponsored insurance in the 2003-2006 MarketScan Commercial Claims and Encounters Database. Adherence was defined as percent of days covered in each calendar quarter. Persistence was defined as days from initiation of SGA to the first 90-day gap in medication on-hand. Generalized Estimating Equations were used to determine the effects of cost-sharing on adherence to SGA medications based on patient-quarter data. A Cox proportional hazards model with patient cost-sharing as a time-varying covariate estimated the effects on persistence with SGA medication. Higher cost-sharing was associated with a lower likelihood of adherence. When compared to plans with cost-sharing below $10, adherence rates were approximately 27% lower for patients in plans with SGA cost-sharing of $50 and above and about 10% lower for patients in plans with cost-sharing between $30 and $50. In both cases, the reduction in adherence was significant. Higher cost-sharing was also associated with a shorter time to discontinuation (HR: 1.028; 95% CI [1.006-1.051]). High SGA cost-sharing appears to be a financial barrier to SGA medication compliance, especially when cost-sharing levels exceeded $30. Our findings have implications for health plans, employers, and policymakers who have, or are, contemplating establishing cost-sharing tiers for SCA medications for commercially insured patients with serious mental illnesses.


Different distributions of hepatitis C virus genotypes among HIV-infected patients with acute and chronic hepatitis C according to interleukin-28B genotype.
The C allele of the single nucleotide polymorphism rs12979860, located near the interleukin-28B (IL-28B) gene, has a strong impact on hepatitis C virus (HCV) treatment response, as well as on spontaneous viral clearance. In patients with chronic hepatitis C (CHC), genotype CC carriers harbour HCV genotype 3 more commonly than those with non-CC genotypes. The aim of this study was to compare the HCV genotype distributions, according to IL-28B genotype, in HIV-infected patients with CHC and those with acute hepatitis C (AHC). The rs12979860 genotype was determined by polymerase chain reaction (PCR) in two subpopulations of HIV-infected patients. The first consisted of 80 German patients with AHC. The second consisted of 476 patients with CHC, belonging to one German and two Spanish cohorts. In the AHC group, 31 (81.6%) rs12979860 CC carriers were infected with HCV genotype 1 or 4 vs. 32 (76.2%) among non-CC carriers (P=0.948). In patients with CHC, among those with the CC genotype, 119 (54.6%) were infected with HCV genotype 1 or 4 and 99 (45.4%) with genotype 2 or 3, whereas in the subset with non-CC genotypes, 200 (77.5%) harboured HCV genotype 1 or 4 and 58 (22.5%) genotype 2 or 3 (P<0.001). Among HIV-infected patients with CHC, those bearing the IL-28B genotype CC were more commonly infected with genotype 3 than subjects with non-CC genotypes, whereas in HIV-infected subjects with AHC this finding was not obtained. These results strongly suggest that the protective effect of the CC genotype against evolution to CHC is mainly exerted in patients infected with HCV genotype 1 or 4.


The neurology of aviation, underwater, and space environments.
It can be seen that aspects of different environments can have adverse effects on normal, healthy nervous systems. Aerospace and underwater neurology consultants are often required to evaluate aviators, divers, or astronauts who have neurologic problem to determine whether they should be granted a waiver. In general, these considerations include course of the disorder (static, progressive, or paroxysmal), potential for sudden incapacitation that may compromise safety, predictability of course of the disorder, ability to monitor disease, and potential adverse effects of medications. A background in the toxic effects of an abnormal environment on the nervous system can be used to make better clinical judgements when considering effects of exposing someone with an abnormal nervous system to such environmental stressors.


Hyperactivation of the mammalian degenerin MDEG promotes caspase-8 activation and apoptosis.
Intracellular calcium overload plays a critical role in numerous pathological syndromes such as heart failure, brain ischemia, and stroke. Hyperactivation of the acid-sensing ion channels including degenerin/epithelial amiloride-sensitive sodium (DEG/ENaC) channels has been shown to elevate intracellular calcium and cause subsequent neuronal cell death that is independent of the canonical Egl-1/Ced-9/Ced-4/Ced-3 apoptotic pathway in Caenorhabditis elegans. In mammalian cells, hyperactivation of the DEG/ENaC channels can also lead to cell death, although the underlying mechanism remains largely unknown. Here, we use a tetracycline-inducible system to express the hyperactivation mutant of a mammalian DEG/ENaC channel protein, MDEG G430F, in murine kidney epithelial cells deficient in the key mitochondrial apoptotic proteins Bax and Bak. Remarkably, expression of MDEG G430F induces increased intracellular calcium, reactive oxygen species (ROS) production, and cell death. The MDEG G430F-induced cell death is blocked by the intracellular calcium chelator 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (acetoxymethyl ester), ROS scavengers, and the caspase inhibitor z-VAD-fmk (where z and fmk are benzyloxycarbonyl and fluoromethyl ketone). Mechanistically, the intracellular calcium overload and ROS increase lead to the inhibition of proteasomal and autophagic protein degradation, which promotes the accumulation of protein aggregates containing caspase-8 and subsequent caspase-8 activation. As protein aggregation upon the inhibition of proteasomal and autophagic degradation pathways is mediated by the ubiquitin-binding protein SQSTM1/p62 and the autophagy-related protein LC3, silencing of p62 and LC3 protects cells from MDEG G430F-induced cell death. Our results uncover a new mechanism of caspase-8-mediated apoptosis induced by intracellular calcium overload that is dependent on the autophagy-related proteins LC3 and p62 upon hyperactivation of DEG/ENaC channels.


A multicenter study of women's self-reported reproductive health after spinal cord injury.
Little attention has been given to women's reproductive health issues in the disabled population. This study documents the unique reproductive health conditions, complications, and behaviors in women with spinal cord injury (SCI). A total of 472 women at least 18 years of age who were at least 1 year post-SCI. Their average age at injury was 32 years. An extensive questionnaire regarding gynecologic, sexual, obstetric, and menopausal health issues was developed and piloted. The questionnaire was then administered by a trained woman health care clinician to women who agreed to participate in the study. Private outpatient clinics at 10 regional model SCI systems of care. Women reported similar gynecologic problems in both preinjury and postinjury time periods. Exceptions were urinary tract infections and vaginal yeast infections. Sexually transmitted infections appear to be less common after injury, but the difference was not statistically significant. The number of hysterectomies was similar both before and after injury, but reasons differed greatly. Women with SCI were less likely to have routine mammograms. They reported similar preventive practices such as performing self-breast examinations and obtaining Papanicolaou smears. Almost 14% of women with SCI became pregnant after injury (101 pregnancies). Complications from pregnancy, labor, and delivery were reported to be more frequent in their postinjury than in their preinjury obstetric experiences. They tended to have babies of lower birth weight and with more complications at time of delivery. Of the sample, 87% reported having sexual intercourse before injury, with only 67% having intercourse after injury. Years postinjury and level of injury were predictive of intercourse; extent of injury was not. Experience of orgasms and methods of contraception varied among the two groups. There were significant complaints of dysreflexia and bladder incontinence with sexual function. Menopause after injury was reported by 14.6% of the women. Postinjury menopausal symptoms were of low frequency, but more than those reported by women who had undergone menopause before injury. Only 19% of women who had menopause after SCI were placed on hormone replacement therapy. Almost one third of the women who had menopause after injury reported new bone fractures. This study illustrates the unique reproductive health concerns of women with SCI. Many pregnancy, labor, and delivery experiences in these women are different. Sexual activity and function have several disability-related consequences and the effects of menopause are still unknown, but may be more problematic than for able-bodied women.


Identifying vascular access complications among ESRD patients in Europe. A prospective, multicenter study.
The pilot project of the Research Board of the EDTNA/ERCA was aimed at reviewing the management of vascular access (VA) in European dialysis centres (see August 1999 NN&I). In Part 1 of this study, centre policies related to VA management were investigated. In Part 2, detailed in this article, individual patients were prospectively followed for one year to identify VA-related complications. A cohort of 1,380 adult patients randomly selected from 47 centres in 16 European countries was followed for one year using a computerised data collection system. Data were collected at baseline after 6 and 12 months and each time a VA complication occurred. At the start of the observation period, 77% of patients had a native arteriovenous (AV) fistula, 10% had an AV graft, and 13% had a catheter for access. A total of 489 complications were noted during the one-year period. Most frequently observed were thrombosis, stenosis, infection, bleeding, and flow problems. Hospitalisation (mean duration = 6.2 days) was required in 39% of patients with complications, and 29% of complications resulted in a definitive loss of VA. Complications were more frequently observed in patients with catheters (27%) and AV grafts (37%) compared to those with an AV fistula (15%). The risk of thrombosis was more than four times higher and the risk of bleeding was more than six times higher if an AV graft was used compared to an AV fistula. Catheters presented an eightfold increased risk for patients of developing infections and access flow problems. [figure: see text] This study revealed the high complication rate in VA and strengthened the notion that the AV fistula is the superior access.


Functional Foods: A Promising Strategy for Restoring Gut Microbiota Diversity Impacted by SARS-CoV-2 Variants.
Natural herbs and functional foods contain bioactive molecules capable of augmenting the immune system and mediating anti-viral functions. Functional foods, such as prebiotics, probiotics, and dietary fibers, have been shown to have positive effects on gut microbiota diversity and immune function. The use of functional foods has been linked to enhanced immunity, regeneration, improved cognitive function, maintenance of gut microbiota, and significant improvement in overall health. The gut microbiota plays a critical role in maintaining overall health and immune function, and disruptions to its balance have been linked to various health problems. SARS-CoV-2 infection has been shown to affect gut microbiota diversity, and the emergence of variants poses new challenges to combat the virus. SARS-CoV-2 recognizes and infects human cells through ACE2 receptors prevalent in lung and gut epithelial cells. Humans are prone to SARS-CoV-2 infection because their respiratory and gastrointestinal tracts are rich in microbial diversity and contain high levels of ACE2 and TMPRSS2. This review article explores the potential use of functional foods in mitigating the impact of SARS-CoV-2 variants on gut microbiota diversity, and the potential use of functional foods as a strategy to combat these effects.


Boot camp: a novel intensive approach to voice therapy.
It is well known in the disciplines of neurobiology, exercise physiology, motor learning, and psychotherapy that desirable learning and behavior changes occur primarily from practice that involves high-intensity overload, variability, and specificity of training. We propose a novel treatment approach called intensive short-term voice therapy that uses these practice parameters for recalcitrant dysphonia. Intensive short-term voice therapy involves multiple sessions with a variety of clinicians, incorporating multiple simultaneous therapeutic approaches. The intensive short-term voice therapy approach is characterized by voice therapy for 1-4 successive days each with an average of 5 hours of therapy and five clinicians. This form of intensive voice therapy provides rigorous practice, involving not only overload but also opportunities for specificity and individuality thereby facilitating better transfer of learned skills. This article discusses the conceptual, theoretical, and practical foundations of this novel therapy approach.


Type III CRISPR-Cas: beyond the Cas10 effector complex.
Type III CRISPR-Cas loci encode some of the most abundant, yet complex, immune systems of prokaryotes. They are composed of a Cas10 complex that uses an RNA guide to recognize transcripts from bacteriophage and plasmid invaders. Target recognition triggers three activities within this complex: ssDNA degradation, synthesis of cyclic oligoadenylates (cOA) that act as second messengers to activate CARF-domain effectors, and cleavage of target RNA. This review covers recent research in type III CRISPR-Cas systems that looked beyond the activity of the canonical Cas10 complexes towards: (i) ancillary nucleases and understanding how they provide defense by sensing cOA molecules; (ii) ring nucleases and their role in regulating cOA production; and (iii) CRISPR-associated proteases, including the function of the Craspase complex in a transcriptional response to phage infection.


Alkaline Phosphatase for Estimating the Time since Deposition of Blood Fingerprints by Scanning Electrochemical Microscopy.
Blood is one of the most frequent and valuable traces encountered at crime scenes, where knowing the time since deposition (TSD) of bloodstains tremendously assists forensic experts to screen out crime-related evidence and aids in the reconstruction of the event sequence. Although increasing proof-of-concept methodologies for investigating the TSD of bloodstains have been reported, there is still no accepted strategy in forensic practice as the aging mechanism involves complex components, leading to the inaccuracy of the estimation results. Herein, an endogenous biomarker of alkaline phosphatase (ALP) was chosen to investigate the TSD by scanning electrochemical microscopy (SECM). Results demonstrate that the ALP activity acquired via SECM lateral scan assay exhibited a clear decrease over time, and a similar trend was observed on both poly(vinylidene fluoride) (PVDF) membrane and glass, with the aging kinetics on PVDF membrane being faster than glass. By means of quantitatively calculating the flux of generated p-aminophenol (PAP), we established the aging curve and realized the TSD estimation of blood fingerprints (BFPs) that was unable to be distinguished via optical measurements. Intriguingly, the as-obtained estimation accuracy ranged from 74.6 to 93.7%, proving the possibility of using an ALP biomarker and SECM. More appealingly, the predicted TSDs were capable of accurately differentiating the deposition sequence of overlapping BFPs, which was hardly achieved by optical means. Therefore, this proof-of-concept strategy demonstrates the value of SECM as a forensic tool and opens possibilities for revealing multidimensional information about crime.


In persons with CIDP, auto-HSCT can be conducted fully on an outpatient basis and induces significant clinical responses: A prospective study in a single center.
Chronic inflammatory demyelinating polyneuropathy (CIDP) is one of the world's most common treatable neuropathy which usually responds to immunosuppressive treatment. Autologous hematopoietic stem cell transplantation (aHSCT) is an intense way of inducing immunosuppression. We analyze the evolution of CIDP patients treated with aHSCT in our center. Between 2018 and 2023, persons with CIDP were prospectively autografted employing the "Mexican method" to conduct grafts on an outpatient basis, employing cyclophosphamide 200 mg/Kg and rituximab 1000 mg. The protocol is registered in ClinicalTrials.gov identifier NCT02674217. In our center 21 autologous transplant cases were completed in 2018-2023. Seven patients provided data to assess the efficacy of the procedure. Positive responses (stabilization and/or improvement) were observed in all seven patients: Five reported improvements in the Inflammatory Neuropathy Cause and Treatment (INCAT) score and one reported stabilization. In the Inflammatory Rasch-Built Overall Disability Scale (I-RODS) score. Median INCAT score was 5 (range 1-9), whereas median I-RODS score was 24 (range 11-29). Five patients (71%) reported improvement in the INCAT score, one reported stabilization and one informed worsening; concerning the I-RODS score 5 (71%) informed improvement, whereas two reported stabilization. aHSCT conducted fully in an outpatient basis, employing the conditioning regimen of the "Mexican method" appears to be a feasible therapeutic option for persons with CIDP. Additional studies are needed to confirm these observations.


Factors associated with morbidity and mortality in feedlot calves: the Bruce County beef project, year two.
The results of the second year of the project confirmed most of the major findings from the initial year. Feeding cornsilage, particularly as the major roughage in the first month after arrival was associated with excess mortality. Mixing of cattle from different sources and vaccinating against respiratory disease appeared to be the most important additional factors that increased mortality rates. Delaying vaccination at least two days postarrival may have prevented the negative effects of vaccination but only in calves fed cornsilage. Morbidity rates were highly variable among farms but were positively correlated with mortality rates and treatment costs. The occurrence of infectious thromboembolic meningoencephalitis appeared to share some of the same risk factors as mortality; whereas, urolithiasis did not. Water deprivation may be a risk factor in the occurrence of urolithiasis. Fibrinous pneumonia was again the most frequent cause of death. Relative to year one, infectious thromboembolic meningoencephalitis increased in frequency and only one death was attributed to bovine virus diarrhea.


The sheep growth hormone gene polymorphism and its effects on milk traits.
Growth hormone (GH) is encoded by the GH gene, which may be single copy or duplicate in sheep. The two copies of the sheep GH gene (GH1/GH2-N and GH2-Z) were entirely sequenced in one 106 ewes of Sarda breed, in order to highlight sequence polymorphisms and investigate possible association between genetic variants and milk traits. Milk traits included milk yield, fat, protein, casein and lactose percentage. We evidenced 75 nucleotide changes. Transcription factor binding site prediction revealed two sequences potentially recognised by the pituitary-specific transcription factor POU1FI at the GH1/GH2-N gene, which were lost at the promoter of GH2-Z, which might explain the different tissues of expression of GH1/GH2-N (pituitary) and GH2-Z (placenta). Significant differences in milk traits were observed among genotypes at polymorphic loci only for the GH2-Z gene. Sheep with homozygote genotype ss748770547 CC had higher fat percentage (P < 0.01) than TT. SNP ss748770547 was part of a potential transcription factor binding site for C/EBP alpha (CCAAT/Enhancer Binding Protein), which is involved in the regulation of adipogenesis and adipoblast differentiation. SNP ss748770547, located in the GH2-Z gene 5' flanking region, may be a causal mutation affecting milk fat content. These findings might contribute to the knowledge of the sheep GH locus and might be useful in selection processes in sheep.


Comparative genomic hybridization and mutation analyses of sporadic schwannomas.
Schwannomas of the vestibular nerve are the striking characteristics of neurofibromatosis type 2 (NF2), an autosomal dominant hereditary disease. The NF2 gene on 22q12 has been isolated as the gene responsible for NF2. Previous studies have reported that 60% of sporadic schwannomas showed inactivation of the NF2 gene, but genetic alterations of remaining 40% tumors remain elusive. Moreover, detailed genetic alterations of this tumor remain an open question. In this study, we analyzed genomic alterations in 17 sporadic schwannomas using comparative genomic hybridization (CGH). Loss of chromosome 22q, including the NF2 locus, was the only notable abnormality (5/17, 29%). Further, we performed fluorescence in situ hybridization analysis with a genomic BAC clone harboring the NF2 gene and found that the 5 tumors with loss detected by CGH as well as three cases without such a detectable loss by CGH, or a total, 8/17 (47%), showed loss of the NF2 locus. Mutation search by PCR-SSCP followed by direct sequencing revealed that 71% (12/17) of the tumors had one or two mutations in the NF2 gene. Our analyses disclosed that 14 (82%) of 17 tumors had structural alteration of NF2; among these 14 cases, 9 (64%) had two inactivating mutations in the NF2 gene, either a somatic mutation in one allele coupled with loss of the other allele or two independent somatic mutations. Our present results suggested that (i) most of the sporadic schwannomas have two-hit mutations in the NF2 gene, and (ii) NF2 is the only major causative gene in the genesis of schwannomas that is activated or inactivated by copy number alterations.


Anatomy and Histochemistry of the Vegetative System of Brachystele guayanensis (Lindl.) Schltr. (Orchidaceae), a Potential Medicinal Species.
The orchid genus Brachystele Schltr. (Orchidoideae, Cranichideae, Spiranthinae) comprises 20 species distributed from Mexico to Argentina, with 10 species found in Brazil. Anatomical studies of Orchidoideae Lindl. have been scarce, and the anatomy and histochemistry of Brachystele are still largely unknown. In this study, we conducted a characterization of the vegetative organs of B. guayanensis (Lindl.) Schltr. using standard anatomical and histochemical microtechniques. In this study, we provide the first information about the anatomy and histochemistry of Brachystele. The studied species was observed to display anatomical characters commonly found in the vegetative organs of representatives of the Cranichideae tribe (e.g., uniseriate epidermis; homogeneous mesophyll with 6-11 layers; rhizomes with rings of fibers; vascular bundles in the form of "^" or "v"; fleshy roots with uniseriate velamen, simple trichomes, and spiranthosomes). Others can be interpreted as adaptive strategies conditioned by the environment and their terrestrial life form (e.g., cuticle thickness; amphistomatic leaves; roots with reduced velamen compared to the cortex (18-20 layers); and raphides). In this study, cataphylls, and the presence of spiranthosomes in leaves, including stomatal guard cells, as well as alkaloids in these structures, are anatomically described for the first time in Orchidaceae. The presence of hyphae and pelotons in the stem of B. guayanensis is described for the first time in Cranichideae. Histochemical tests confirmed the presence of lignin, proteins, and alkaloids, the lipidic nature of the cuticle, starch grains stored in spiranthosomes, and the composition of the raphides. Alkaloids were observed in abundance, particularly in the roots, suggesting a potential role in defense against pathogens and herbivores, as well as potential medicinal activities, as seen in phylogenetically related groups to Brachystele.


A review of neighborhood effects and early child development: How, where, and for whom, do neighborhoods matter?
This paper describes a scoping review of 42 studies of neighborhood effects on developmental health for children ages 0-6, published between 2009 and 2014. It focuses on three themes: (1) theoretical mechanisms that drive early childhood development, i.e. how neighborhoods matter for early childhood development; (2) dependence of such mechanisms on place-based characteristics i.e. where neighborhood effects occur; (3) dependence of such mechanisms on child characteristics, i.e. for whom is development most affected. Given that ecological systems theories postulate diverse mechanisms via which neighborhood characteristics affect early child development, we specifically examine evidence on mediation and/or moderation effects. We conclude by discussing future challenges, and proposing recommendations for analyses that utilize ecological longitudinal population-based databases.


Effects of venovenous extracorporeal membrane oxygenation on cerebral oxygenation in hypercapnic ARDS.
Extracorporeal membrane oxygenation (ECMO) is increasingly used in ARDS patients with hypoxemia and/or severe hypercapnia refractory to conventional treatment strategies. However, it is associated with severe intracranial complications, e.g. ischemic or hemorrhagic stroke. The arterial carbon dioxide partial pressure (PaCO2) is one of the main determinants influencing cerebral blood flow and oxygenation. Since CO2 removal is highly effective during ECMO, reduction of CO2 may lead to alterations in cerebral perfusion. We report on the variations of cerebral oxygenation during the initiation period of ECMO treatment in a patient with hypercapnic ARDS, which may partly explain the findings of ischemic and/or hemorrhagic complications in conjunction with ECMO.


Novel protein-DNA interactions associated with increased immunoglobulin transcription in response to antigen plus interleukin-5.
Although much has been learned about basal levels of immunoglobulin (Ig) transcription, the regulatory effects of cytokines and antigen (Ag) upon Ig expression in lymphocytes have not been fully characterized. We previously reported that Ag plus interleukin-5 (IL-5) caused increased steady-state Ig mRNA levels in Ag-specific cell lines. In this study, we have identified a region between -250 and -125 bp 5' of the Ig transcription start site that is necessary for the induction of increased mu mRNA levels by Ag plus IL-5. Mobility shift and UV cross-linking studies indicated that IL-5 plus Ag induced increased protein binding to this region. Furthermore, this sequence was found to be closely related to another A + T-rich sequence at -525 bp 5' of the transcription start site. Both sequences exhibited similar B-cell-specific and inducible protein binding. Our data suggest that treatment with IL-5 plus Ag induces several DNA-binding proteins, some of which may participate in increasing Ig transcription above basal levels by binding to sequences 5' of the octamer motif.


Quantitative methods for evaluating the efficacy of thalamic deep brain stimulation in patients with essential tremor.
Deep brain stimulation (DBS) of the thalamus is a safe and efficient method for treatment of disabling tremor in patient with essential tremor (ET). However, successful tremor suppression after surgery requires careful selection of stimulus parameters. Our aim was to examine the possible use of certain quantitative methods for evaluating the efficacy of thalamic DBS in ET patients in clinical practice, and to compare these methods with traditional clinical tests. We examined 22 patients using the Essential Tremor Rating Scale (ETRS) and quantitative assessment of tremor with the stimulator both activated and deactivated. We used an accelerometer (CATSYS tremor Pen) for quantitative measurement of postural tremor, and a eurythmokinesimeter (EKM) to evaluate kinetic tremor in a rapid pointing task. The efficacy of DBS on tremor suppression was prominent irrespective of the method used. The agreement between clinical rating of postural tremor and tremor intensity as measured by the CATSYS tremor pen was relatively high (rs = 0.74). The agreement between kinetic tremor as assessed by the ETRS and the main outcome variable from the EKM test was low (rs = 0.34). The lack of agreement indicates that the EKM test is not comparable with the clinical test. Quantitative methods, such as the CATSYS tremor pen, could be a useful complement to clinical tremor assessment in evaluating the efficacy of DBS in clinical practice. Future studies should evaluate the precision of these methods and long-term impact on tremor suppression, activities of daily living (ADL) function and quality of life.


Electroporation of Chlorella vulgaris to enhance biomethane production.
This research investigated the feasibility of using electroporation (EP) as a pretreatment method for algal biomass used as feedstock for anaerobic digestion. The results showed that pretreating algal biomass with EP significantly improved the soluble COD (SCOD), increasing it to more than 830% at 28 kWh/m(3) treatment intensity (TI). Besides TI, culture conditions also affected the performance of the EP process. On the basis of SCOD, a sample pH of 7.0 and cell concentration of 13.2g/L were found to be optimal for the EP process. Despite a direct relationship between TI and ionic strength (IS), SCOD decreased with increasing IS. At 35 kWh/m(3) TI, bio-CH4 production increased by as high as 110%. It was also observed that lower TI levels resulted in high rates of gain per energy input compared to higher degrees of treatment.


Agent-based simulations of China inbound tourism network.
Based on the results of a large-scale survey, we construct an agent-based network model for the China independent inbound tourism system and, by the approach of numerical simulation, investigate the responses of the tourist behaviors to perturbations in different scenarios, including the closure of a tourism city, the opening of a new port city in western China, and the increase of the tourism attractiveness of a specific city. Numerical results show that: (1) the closure of a single city in general will affect the tourist visitations of many other cities and, comparing to the non-port cities, the overall visitation volume of the system is more influenced by closing a port city; (2) the opening of a new port city in western China will attract more tourists to the western cities, but has a negligible impact on either the overall visitation volume or the imbalanced tourist distribution; and (3) the increase of the tourism attractiveness of a non-port (port) city normally increases (decreases) the overall visitation volume, yet there are exceptions due to the spillover effect. Furthermore, by increasing the tourism attractiveness of a few cities simultaneously, we investigate also the strategy of multiple-city-upgrade in tourism development. We find that the overall tourist volume is better improved by upgrading important non-port cities that are geographically distant from each other. The study reveals the rich dynamic inherent in complex tourism network, and the findings could be helpful to the development and management of China inbound tourism.


Neurogenic bladder in spinal cord injury patients.
Neurogenic bladder dysfunction due to spinal cord injury poses a significant threat to the well-being of patients. Incontinence, renal impairment, urinary tract infection, stones, and poor quality of life are some complications of this condition. The majority of patients will require management to ensure low pressure reservoir function of the bladder, complete emptying, and dryness. Management typically begins with anticholinergic medications and clean intermittent catheterization. Patients who fail this treatment because of inefficacy or intolerability are candidates for a spectrum of more invasive procedures. Endoscopic managements to relieve the bladder outlet resistance include sphincterotomy, botulinum toxin injection, and stent insertion. In contrast, patients with incompetent sphincters are candidates for transobturator tape insertion, sling surgery, or artificial sphincter implantation. Coordinated bladder emptying is possible with neuromodulation in selected patients. Bladder augmentation, usually with an intestinal segment, and urinary diversion are the last resort. Tissue engineering is promising in experimental settings; however, its role in clinical bladder management is still evolving. In this review, we summarize the current literature pertaining to the pathology and management of neurogenic bladder dysfunction in patients with spinal cord injury.


Molecular Basis of Gut Microbiome-Associated Colorectal Cancer: A Synthetic Perspective.
A significant challenge toward studies of the human microbiota involves establishing causal links between bacterial metabolites and human health and disease states. Certain strains of commensal Escherichia coli harbor the 54-kb clb gene cluster which codes for small molecules named precolibactins and colibactins. Several studies suggest colibactins are genotoxins and support a role for clb metabolites in colorectal cancer formation. Significant advances toward elucidating the structures and biosynthesis of the precolibactins and colibactins have been made using genetic approaches, but their full structures remain unknown. In this Perspective we describe recent synthetic efforts that have leveraged biosynthetic advances and shed light on the mechanism of action of clb metabolites. These studies indicate that deletion of the colibactin peptidase ClbP, a modification introduced to promote accumulation of precolibactins, leads to the production of non-genotoxic pyridone-based isolates derived from the diversion of linear biosynthetic intermediates toward alternative cyclization pathways. Furthermore, these studies suggest the active genotoxins (colibactins) are unsaturated imines that are potent DNA damaging agents, thereby confirming an earlier mechanism of action hypothesis. Although these imines have very recently been detected in bacterial extracts, they have to date confounded isolation. As the power of "meta-omics" approaches to natural products discovery further advance, we anticipate that chemical synthetic and biosynthetic studies will become increasingly interdependent.


Monocyte-derived dendritic cells from Crohn patients show differential NOD2/CARD15-dependent immune responses to bacteria.
Three common mutations in the NOD2/CARD15 gene are strongly associated with Crohn's disease (CD). NOD2 is an intracellular receptor of muramyl dipeptide (MDP), a component of peptidoglycan present in the cell wall of gram-positive (G+) and gram-negative (G-) bacteria. We generated monocyte-derived dendritic cells (MoDCs) from CD patients mutated or not for CARD15 (n = 53) or from healthy donors (n = 12) and analyzed their activation in response to live Salmonella typhimurium as a model of pathogenic G- bacteria. MoDCs carrying the L1007fs mutation, although phenotypically activated by bacteria, produced a significantly reduced amount of tested cytokines. MoDCs carrying R702W or compound G908R/R702W NOD2 mutations displayed an increased basal level of IL-8 release. After a bacterial encounter, these cells were phenotypically activated and produced levels of cytokines similar to healthy controls. Interestingly, although L1007fs/WT mutations conferred reduced production of cytokines, including IL-12, these cells were perfectly capable of inducing T-cell polarization toward the Th1 phenotype. NOD2 mutations affect the basal characteristics of MoDCs and their response to G- bacteria differently. MoDCs could be involved in CD onset because they have defects in releasing inflammatory cytokines and in polarizing T-cell responses.


Epigenetic regulation of multiple tumor-related genes leads to suppression of breast tumorigenesis by dietary genistein.
Breast cancer is one of the most lethal diseases in women; however, the precise etiological factors are still not clear. Genistein (GE), a natural isoflavone found in soybean products, is believed to be a potent chemopreventive agent for breast cancer. One of the most important mechanisms for GE inhibition of breast cancer may involve its potential in impacting epigenetic processes allowing reversal of aberrant epigenetic events during breast tumorigenesis. To investigate epigenetic regulation for GE impedance of breast tumorigenesis, we monitored epigenetic alterations of several key tumor-related genes in an established breast cancer transformation system. Our results show that GE significantly inhibited cell growth in a dose-dependent manner in precancerous breast cells and breast cancer cells, whereas it exhibited little effect on normal human mammary epithelial cells. Furthermore, GE treatment increased expression of two crucial tumor suppressor genes, p21(WAF1) (p21) and p16(INK4a) (p16), although it decreased expression of two tumor promoting genes, BMI1 and c-MYC. GE treatment led to alterations of histone modifications in the promoters of p21 and p16 as well as the binding ability of the c-MYC-BMI1 complex to the p16 promoter contributing to GE-induced epigenetic activation of these tumor suppressor genes. In addition, an orally-fed GE diet prevented breast tumorigenesis and inhibited breast cancer development in breast cancer mice xenografts. Our results suggest that genistein may repress early breast tumorigenesis by epigenetic regulation of p21 and p16 by impacting histone modifications as well as the BMI1-c-MYC complex recruitment to the regulatory region in the promoters of these genes. These studies will facilitate more effective use of soybean product in breast cancer prevention and also help elucidate the mechanisms during the process of early breast tumorigenesis.


Mini Review - Phenolics for skin photo-aging.
Photo-aging is one of the foremost problems caused by generation of reactive oxygen species when skin is exposed on UV irradiation. In view of that, generation of reactive oxygen species intermingle with proteins, DNA, saccharides and fatty acids triggering oxidative mutilation and effects are in the appearance of distressed cell metabolism, morphological and ultra-structural changes, mistreat on the routes and revisions in the demarcation, propagation and skin apoptosis living cells which leads to photo-aging. Plant phenolics are universally found in both edible and inedible plants and have extended substantial interest as photo-protective for human skin due to their antioxidant activities. The objective of this review is to highlight the use of plant phenolics for their antioxidant activities against photo-aging.


Locus coeruleus neurons and sympathetic nerves: activation by cutaneous sensory afferents.
The effects of mechanical and thermal cutaneous sensory stimulation on the activity of central norepinephrine (NE) neurons in the locus coeruleus (LC) and on peripheral sympathetic nerve activity (NE-SNA) in a renal branch of the splanchnic nerve were studied, using electrophysiological techniques in the anesthetized rat. Noxious, mechanical sensory stimulation caused a parallel and virtually identical change in central NE-LC activity and peripheral NE-SNA and both neuronal systems showed a biphasic excitation-inhibition response. Both non-noxious and noxious thermal sensory stimulation above 36 degrees C caused a parallel increase in NE-LC firing rate and NE-SNA, changes that were accompanied by increases in arterial blood pressure and heart rate. The increases in central and peripheral NE neuronal activity were linearly correlated. However, the two neuronal systems differed in their ability to adapt during prolonged noxious thermal stimulation. NE-LC neurons adapted completely and returned to baseline firing rate within 5 min during ongoing noxious thermal stimulation. In contrast, an increase in NE-SNA remained throughout the stimulation period. In recent studies in this laboratory, blood volume depletion was found associated with a marked NE-LC activation. In contrast to the present results during prolonged noxious stimulation, there was no attenuation of the NE-LC activation during prolonged hemorrhage. Taken in conjunction, these data indicate a discriminatory capacity of the NE-LC system in monitoring sensory or autonomic stimuli. Thus, only those stimuli of imperative importance for the animal were found to be associated with continuing robust NE-LC discharge.


HSPC111 governs breast cancer growth by regulating ribosomal biogenesis.
Activation of c-Myc plays a decisive role in the development of many human cancers. As a transcription factor, c-Myc facilitates cell growth and proliferation by directly transcribing a multitude of targets, including rRNAs and ribosome proteins. However, how to elucidate the deregulation of rRNAs and ribosome proteins driven by c-Myc in cancer remains a significant challenge and thus warrants close investigation. In this report, a crucial role for the HSPC111 (NOP16) multiprotein complex in governing ribosomal biogenesis and tumor growth was determined. It was discovered that enhanced HSPC111 expression paralleled the upregulation of c-Myc and was directly regulated by c-Myc in breast cancer cells. Knockdown of HSPC111 dramatically reduced the occurrence of tumorigenesis in vivo, and largely restrained tumor cell growth in vitro and in vivo. In stark contrast, HSPC111 overexpression significantly promoted tumor cell growth. Biochemically, it was demonstrated that RNA 3'-phosphate cyclase (RTCD1/RTCA) interacted with HSPC111, and RTCD1 was involved in the HSPC111 multiprotein complex in regulating rRNA production and ribosomal biogenesis. Moreover, HSPC111 and RTCD1 synergistically modulated cell growth and cellular size through commanding rRNA synthesis and ribosome assembly coupled to protein production. Finally, overall survival analysis revealed that concomitant upregulation of HSPC111 and RTCD1 correlated with the worst prognosis in a breast cancer cohort. Inhibition of HSPC111-dependent ribosomal biosynthesis and protein synthesis is a promising therapeutic strategy to diminish breast cancer tumor progression.


Genome-wide and paternal diversity reveal a recent origin of human populations in North Africa.
The geostrategic location of North Africa as a crossroad between three continents and as a stepping-stone outside Africa has evoked anthropological and genetic interest in this region. Numerous studies have described the genetic landscape of the human population in North Africa employing paternal, maternal, and biparental molecular markers. However, information from these markers which have different inheritance patterns has been mostly assessed independently, resulting in an incomplete description of the region. In this study, we analyze uniparental and genome-wide markers examining similarities or contrasts in the results and consequently provide a comprehensive description of the evolutionary history of North Africa populations. Our results show that both males and females in North Africa underwent a similar admixture history with slight differences in the proportions of admixture components. Consequently, genome-wide diversity show similar patterns with admixture tests suggesting North Africans are a mixture of ancestral populations related to current Africans and Eurasians with more affinity towards the out-of-Africa populations than to sub-Saharan Africans. We estimate from the paternal lineages that most North Africans emerged ∼15,000 years ago during the last glacial warming and that population splits started after the desiccation of the Sahara. Although most North Africans share a common admixture history, the Tunisian Berbers show long periods of genetic isolation and appear to have diverged from surrounding populations without subsequent mixture. On the other hand, continuous gene flow from the Middle East made Egyptians genetically closer to Eurasians than to other North Africans. We show that genetic diversity of today's North Africans mostly captures patterns from migrations post Last Glacial Maximum and therefore may be insufficient to inform on the initial population of the region during the Middle Paleolithic period.


The cytolytic T lymphocyte response to the murine cytomegalovirus. I. Distinct maturation stages of cytolytic T lymphocytes constitute the cellular immune response during acute infection of mice with the murine cytomegalovirus.
Limiting dilution (LD) analysis with two modifications, the expansion and the restimulation LD assay, led to the detection and quantification of two distinct in vivo maturation stages within the lineage of virus-specific self-restricted CTL after infection of mice with the murine cytomegalovirus (MCMV). A low frequency set, representing an average of 15% of the specifically activated CTL-P in a draining lymph node, generated virus-specific lytic activity in the absence of antigen, solely under expansion conditions provided by growth and differentiation interleukins. These cells were considered to be active and were denoted antigen-independent or interleukin-receptive CTL-P (IL-CTL-P). A high frequency set required additional antigen in vitro to generate functionally active clones, and therefore the cells were termed antigen-dependent. Both sets are present in vivo simultaneously at the peak of the acute immune response and represent antigen-activated cells because their existence strictly depends on a preceding priming event. IL-CTL-P disappear quickly after acute infection and are absent during the memory state. It is proposed that the isolation of IL-CTL-P could serve to detect viral antigen expression during persistent and/or recurrent herpes virus infections.


Should radiotherapy after primary systemic therapy be administered with the same recommendations made for operable breast cancer patients who receive surgery as first treatment? A critical review.
Primary systemic therapy is not only used in patients with locally advanced inoperable non-metastatic breast cancer but also for operable stage II and III cancer aimed at breast conservation. The indications for local-regional radiotherapy for patients who receive primary systemic therapy are still evolving. The purpose of this article is to provide a comprehensive discussion of how primary systemic therapy in operable breast cancer patients could affect the indications of radiotherapy to optimize local-regional treatment. An overview of available literature data regarding neoadjuvant treatment and radiotherapy is analyzed and discussed. Considering the variability of data on this issue, an appropriate approach could still be to tailor treatment decision to the individual clinical case.


Parkinson's Disease Society Brain Bank, London: overview and research.
The UK Parkinson's Disease Society Brain Bank receives tissue from patients with Parkinson's disease and a variety of different movement disorders. Half of the brain is used for full neuropathological examination prior to allocation for specific research projects. Clinical misdiagnosis occurs in a significant proportion of cases and clinico-pathological correlation provides valuable information for disease recognition. With the expanding number of other specialist brain banks there is a need for agreement on diagnostic criteria. Furthermore, awareness of different methods of tissue handling is essential.


The ABCD matrix in graded index tapers used for beam expansion and compression.
A new form is proposed for the ABCD matrix in a graded index taper with a large variation in cross section such as might be used for single-mode beam expansion. Expressions are given for the loss of power from the fundamental mode and the coupling efficiency between fibers when two tapers are used in an expanded beam connector. Exact solutions are found for linear tapers and for a class of tapers with zero slope ends. The distinction between adiabatic and nonadiabatic tapers is made clear from the functional form of the matrix in the linear case. Comparisons are made with previously published results and the effect of taper shape on the coupling efficiency is discussed.