[Cerebral hemodynamics in the context of differential diagnosis and management of headache in children].
Using transcranial Doppler ultrasonography, cerebral hemodynamics has been studied in 140 children, aged 6-16 years, with different types of headache in the interictal period. According to a type of headache the patients have been divided into groups as follows: migraine, headache of tension and unspecified headache with arterial hypotension, arterial hypertension and normal blood pressure. The predominance of different changes of cerebral hemodynamics was found in patients compared to the control group. These changes were heterogeneous within the type of headache. All types of headache, with exception of headache with arterial hypertension, were characterized by low vascular tone of the cerebral arteries and increased bloodstream velocity. A high percent of cerebral venous defects was observed in all the groups. These disturbances should be taken into account in prescription of vasoactive drugs to children with headache.


Evaluation of Clinical Parameters in Implant Maintenance Phase for Prevention of Peri-implantitis.
Peri-implant mucositis and peri-implantitis are one of the common biological complications affecting implant success. The present study aimed to evaluate various clinical parameters during implant maintenance phase. The study included patients undergoing implant maintenance phase for 1-year follow-up. The study consists of a total of forty individuals with age ranging from 35 to 65 years. They were further categorized into two subgroups on the basis of their history, i.e., Group 1: patients with no history of periodontitis before implant placement and Group 2: patients with a history of periodontitis before implant placement. Among the selected patients, a total of 98 implants were studied. All were individually evaluated for clinical parameters such as gingival index, pocket probing depth (PPD), and bleeding on probing (BOP). All the data obtained were tabulated and analyzed using statistical software SPSS version 18.0 for Windows (SPSS Inc., Chicago, USA). Quantitative analysis was done using t-test and Mann-Whitney U-test. The mean age of the patients in Group 1 and Group 2 was 58.6 and 62.8 years, respectively, with not much gender difference. The mean plaque index for Group 1 was 0.17 ± 0.20, while for Group 2, it was 0.24 ± 0.14. The mean PPD and mean BOP for Group 1 came to be 2.60 ± 0.42 and 0.42 ± 0.15, respectively, whereas for Group 2, it was 4.08 ± 0.30 and 0.39 ± 0.48, respectively. Only PPD was found to be statistically significantly different between both the groups. Group 1 showed 2.0% peri-implantitis, whereas Group 2 showed 28% peri-implantitis. Due to increased prevalence of peri-implantitis cases with the increase in usage of implants, it becomes imperative to look up to the etiological factors and contributing factors so that the incidence of these can be minimized.


An animal model to evaluate the function and regulation of the adaptively evolving stress protein SEP53 in oesophageal bile damage responses.
Squamous epithelium in mammals has evolved an atypical stress response involving down-regulation of the classic HSP70 protein and induction of sets of proteins including one named SEP53. This atypical stress response might be due to the unusual environmental pressures placed on squamous tissue. In fact, SEP53 plays a role as an anti-apoptotic factor in response to DNA damage induced by deoxycholic acid stresses implicated in oesophageal reflux disease. SEP53 also has a genetic signature characteristic of an adaptively and rapidly evolving gene, and this observation has been used to imply a role for SEP53 in immunity. Physiological models of squamous tissue are required to further define the regulation and function of SEP53. We examined whether porcine squamous epithelium would be a good model to study SEP53, since this animal suffers from a bile-reflux disease in squamous oesophageal tissue. We have (1) cloned and sequenced the porcine SEP53 locus from porcine bacterial artificial chromosome genomic DNA, (2) confirmed the strikingly divergent nature of the C-terminal portion of the SEP53 gene amongst mammals, (3) discovered that a function of the conserved N-terminal domain of the gene is to maintain cytoplasmic localisation, and (4) examined SEP53 expression in normal and diseased porcine pars oesophagea. SEP53 expression in porcine tissue was relatively confined to gastric squamous epithelium, consistent with its expression in normal human squamous epithelium. Immunohistochemical staining for SEP53 protein in normal and damaged pars oesophagea demonstrated significant stabilisation of SEP53 protein in the injured tissue. These results suggest that porcine squamous epithelium would be a robust physiological model to examine the evolution and function of the SEP53 stress pathway in modulating stress-induced responses in squamous tissue.


Associations of preoperative serum high-density lipoprotein cholesterol and low-density lipoprotein cholesterol levels with the prognosis of ovarian cancer.
The effect of serum lipids on ovarian cancer is controversial. We conducted this study to evaluate the prognostic value of preoperative plasma lipid profile in patients with ovarian cancer. The medical records of 156 epithelial ovarian cancer patients who underwent surgical resection in our department were retrospectively reviewed and analyzed. Serum lipids profiles, including total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglyceride (TG), apolipoprotein A-I (apoA-I), apolipoprotein B (apoB) and clinicopathologic data, were analyzed. Cox proportional hazards regression analyses and Kaplan-Meier method were performed to evaluate the overall survival (OS) and progression-free survival (PFS). Multivariable Cox regression analysis found that preoperative higher LDL-C level was significantly associated with worse OS (HR 2.088, 95% CI 1.052-4.147, p = 0.035), whereas higher HDL-C level showed significant association with better PFS (HR 0.491, 95% CI 0.247-0.975, p = 0.042). Further Kaplan-Meier survival analysis demonstrated that OS was longer for patients with low levels of LDL-C (< 2.76 mmol/L) compared to those with high levels of LDL-C (≥ 2.76 mmol/L) (P = 0.028), and PFS was better for patients with high levels of HDL-C (≥ 1.19 mmol/L) compared to those with low levels of HDL-C (< 1.19 mmol/L) (P = 0.001). Preoperative HDL-C and LDL-C levels are significant predictors of clinical outcome in patients with epithelial ovarian cancer.


Overview of anal fistula and systematic review of ligation of the intersphincteric fistula tract (LIFT).
Anal fistula management has long been a challenge for surgeons. Presently, no technique exists that is ideal for treating all types of anal fistula, whether simple or complex. A higher incidence of poor sphincter function and recurrence after surgery has encouraged the development of a new sphincter-sparing procedure, ligation of the intersphincteric fistula tract (LIFT), first described by Van der Hagen et al. in 2006. We assessed the safety, feasibility, success rate, and continence of LIFT as a sphincter-saving procedure. A literature search of articles in electronic databases published from January 2006 to August 2012 was performed. Analysis followed Preferred Reporting Items for Systematic Reviews recommendations. All LIFT-related articles published in the English language were included. We excluded case reports, abstracts, letters, non-English language articles, and comments. The procedure was described in detail as reported by Rojanasakul. Thirteen original studies, including 435 patients, were reviewed. The most common fistula procedure type was transsphincteric (92.64 %). The overall median operative time was 39 (±20.16) min. Eight authors performed LIFT as a same-day surgery, whereas the others admitted patients to the hospital, with an overall median stay of 1.25 days (range 1-5 days). Postoperative complications occurred in 1.88 % of patients. All patients remained continent postoperatively. The overall mean length of follow-up was 33.92 (±17.0) weeks. The overall mean healing rate was 81.37 (±16.35) % with an overall mean healing period of 8.15 (±5.96) weeks. Fistula recurrence occurred in 7.58 % of patients. LIFT represents a new, easy-to-learn, and inexpensive sphincter-sparing procedure that provides reasonable results. LIFT is safe and feasible, with favorable short- and long-term outcomes. However, additional prospective randomized studies are required to confirm these findings.


Swan-Ganz catheter use in trauma patients can be reduced without negatively affecting outcomes.
The use of pulmonary artery catheter (PAC) is controversial. The purpose of this study was to document the changing pattern of PAC use and to determine its effect on outcome. The use of PAC was analyzed in patients ≥16 years old admitted to the surgical intensive care unit (SICU) over a 9-year period starting in 2000. Patients with SICU length of stay exceeding 30 days were excluded. For the evaluation of PAC's effect on outcome, PAC and no-PAC patients were matched utilizing propensity scores. During the 9-year study period, a total of 5,192 trauma patients were admitted to the SICU. Of these, 426 (8.2%) were <16 years old, and 174 (3.4%) had a SICU length of stay that exceeded 30 days. For the remaining 4,592 patients, the mean ± SD age was 39.5 ± 18.8 years, and the mean ISS was 19.9 ± 12.4. PAC was utilized in 19.5% (n = 896) of all the patients admitted to the SICU. The trend for PAC use decreased significantly over the years (P value for trend <0.001), from 38.6% in the year 2000 to 4.2% in the year 2008. This decrease was noted in all age and injury severity groups of patients. The overall mortality, however, remained at the same levels (P value for trend = 0.111). Patients managed with a PAC were significantly older, more severely injured, more frequently injured by a blunt mechanism, and were admitted more often in a hypotensive or comatose status. In the early part of the study, the PAC was utilized on the first day and for 4.1 days on average. In the later part of the study, however, the PAC was used on the second day and for a shorter period of time (3 days on average, P < 0.001). In the matched study population, patients in the PAC group had almost twofold higher odds for death, when compared to the no-PAC group [34.2% vs. 22.5%, Odds Ratio (95% CI): 1.78 (1.42, 2.26), P < 0.001]. Patients younger than 50 years of age who had an ISS ≥16 had worse outcome when managed with a PAC, whereas patients aged 30-69 years with an ISS <16 had a higher survival. The overall complication rate was fivefold higher in patients receiving a PAC [46.3% vs. 14.2%, Odds Ratio (95% CI): 5.22 (4.04, 6.74), P < 0.001]. The use of PAC has decreased almost 10-fold over the last decade at our institution. The PAC is being used later during the ICU course and for a shorter period of time. In a matched population, the use of PAC is associated with a significantly higher mortality and complication rate, but the reason for this association remains uncertain. The use of PAC is invasive and is associated with known complications and financial costs. While the use of PAC maybe useful in a select population, routine and widespread use of the PAC should be avoided.


Environmental metrics for community health improvement.
Environmental factors greatly affect human health. Accordingly, environmental metrics are a key part of the community health information base. We review environmental metrics relevant to community health, including measurements of contaminants in environmental media, such as air, water, and food; measurements of contaminants in people (biomonitoring); measurements of features of the built environment that affect health; and measurements of "upstream" environmental conditions relevant to health. We offer a set of metrics (including unhealthy exposures, such as pollutants, and health-promoting assets, such as parks and green space) selected on the basis of relevance to health outcomes, magnitude of associated health outcomes, corroboration in the peer-reviewed literature, and data availability, especially at the community level, and we recommend ways to use these metrics most effectively.


Oculomotor nerve palsy from posterior communicating artery aneurysm.
The mortality rate for posterior communicating artery (PCA) aneurysm with oculomotor nerve palsy is 33 per 100. The incidence of oculomotor palsy with PCA aneurysm varies from 34% to 56%. The classic presentation of a PCA aneurysm involving the oculomotor nerve is sudden onset of severe unilateral frontal headache, ptosis, a dilated and fixed pupil, and limitations of adduction, depression, and elevation of the eye. Carotid arteriography is diagnostic. This case is presented because of the high probability of being seen first by a primary care physician. Delay in clinical recognition and treatment can have dire consequences for the patient.


Reversible inhibition of protein synthesis in lung by halothane.
Alterations in the synthesis and degradation of proteins were investigated in intact lungs exposed to the volatile anaesthetic halothane. In rat lungs perfused in situ with Krebs-Henseleit bicarbonate buffer containing 4.5% (w/v) bovine serum albumin, 5.6 mM-glucose, plasma concentrations of 19 amino acids and 690 microM-[U-14C]-phenylalanine and equilibrated with O2/N2/CO2 (4:15:1), protein synthesis, calculated based on the specific radioactivity of aminoacyl-tRNA, was inhibited by halothane. The anaesthetic did not affect degradation of lung proteins. The inhibition of protein synthesis was rapid in onset, dose-dependent, and quickly reversible. It did not appear to be associated with overall energy depletion, with non-specific changes in cellular permeability, or with decreased availability of amino acids as substrates for protein synthesis.


Self-reported quality of life in pharmacoresistant temporal lobe epilepsy: correlation with clinical variables and memory evaluation.
This work explores the effects of clinical variables on self-reported quality of life (QoL) in pharmacoresistant temporal lobe epilepsy (TLE), correlating this information with results from the Quality of Life in Epilepsy questionnaire (QOLIE-31) and selective memory tests of the Barcelona Battery and the Rey-Osterrieth figure. We retrospectively analysed the records of 60 TLE patients and correlated patient variables (e.g. gender, aetiology; mesial TLE with hippocampal sclerosis [HS] versus lesional TLE, side of ictal onset, age, age at onset, duration of epilepsy, seizure frequency, and use of AEDs) with selective memory test scores and self-reported QoL. Right ictal onset was associated with lower emotional well-being scores. MTLE-HS patients had lower QOLIE-31 scores for seizure worry, social function, overall QoL, energy/fatigue, cognitive function, and obtained a lower overall score, compared to those with lesional TLE. Older age at epilepsy onset was associated with worse emotional well-being, energy/fatigue, medication effects, and seizure worry outcomes. Higher seizure frequency and older age at time of evaluation were associated with lower cognitive function scores. Generalised seizures were associated with lower scores based on the variables: seizure worry, overall quality of life, emotional well-being, and cognitive function. Regarding memory tests, only visuospatial memory correlated positively with cognitive function score. Patients with MTLE-HS underwent evaluation for pharmacoresistant epilepsy, on average, 10 years later than those with lesional TLE. MTLE-HS, right-sided epileptogenic zone, late onset, and higher seizure frequency were associated with worse QoL. Objective testing revealed specific memory deficits that were not reflected in self-reported QoL scores.


Evidence-based guidelines for the prevention of ventilator-associated pneumonia: results of a knowledge test among European intensive care nurses.
As part of a needs analysis preceding the development of an e-learning platform on infection prevention, European intensive care unit (ICU) nurses were subjected to a knowledge test on evidence-based guidelines for preventing ventilator-associated pneumonia (VAP). A validated multiple-choice questionnaire was distributed to 22 European countries between October 2006 and March 2007. Demographics included nationality, gender, ICU experience, number of ICU beds and acquisition of a specialised degree in intensive care. We collected 3329 questionnaires (response rate 69.1%). The average score was 45.1%. Fifty-five percent of respondents knew that the oral route is recommended for intubation; 35% knew that ventilator circuits should be changed for each new patient; 38% knew that heat and moisture exchangers were the recommended humidifier type, but only 21% knew that these should be changed once weekly; closed suctioning systems were recommended by 46%, and 18% knew that these must be changed for each new patient only; 51% and 57%, respectively, recognised that subglottic drainage and kinetic beds reduce VAP incidence. Most (85%) knew that semi-recumbent positioning prevents VAP. Professional seniority and number of ICU beds were shown to be independently associated with better test scores. Further research may determine whether low scores are related to a lack of knowledge, deficiencies in training, differences in what is regarded as good practice, and/or a lack of consistent policy.


Follow-up of the re-evaluation of indigo carmine (E 132) as a food additive.
Indigo carmine (E 312) was re-evaluated in 2014 by the EFSA Panel on Food Additives and Nutrient sources added to Food (ANS). The ANS Panel confirmed the acceptable daily intake (ADI) of 5 mg/kg body weight (bw) per day for indigo carmine allocated by JECFA (1975). The ANS Panel indicated that the ADI was applicable to a material with a purity of 93% pure colouring and manufactured using processes resulting in comparable residuals as material used in the Borzelleca et al. studies (1985, 1986) and Borzelleca and Hogan (1985) which were the basis for deriving the ADI. The ANS Panel considered that any extension of the ADI to indigo carmine of lower purity and/or manufactured using a different process would require new data to address the adverse effects on the testes observed in the Dixit and Goyal (2013) study. Following a European Commission call for data to submit data to fill the data gaps, an IBO submitted technical and toxicological data. Considering the technical data, the EFSA Panel on Food Additives and Flavourings (FAF Panel) recommended some modifications of the existing EU specifications for E 132, mainly to lower the limits for toxic elements. Considering the toxicological data, an IBO has submitted a 56-day dietary study to address the adverse effects on testes using a material with 88% purity. The results of this study submitted did not confirm the severe adverse effects observed in the Dixit and Goyal study. Considering all the available information, the Panel confirmed the ADI of 5 mg/kg bw per day for indigo carmine (E 132) disodium salts, meeting the proposed revisions of the specifications (85% minimum for the colouring matter). The Panel concluded that there is no safety concern for the use of indigo carmine (E 132) disodium salts at the reported use levels and submitted analytical data.


Tooth eruption in a patient with craniometaphyseal dysplasia: case report.
Craniometaphyseal dysplasia (CMD) is a very rare genetic disorder of bone remodeling caused by osteoclast dysfunction. The clinical and radiographical features of oral findings are presented in a sporadic case of CMD in a child (age 10 years, 7 months). An intraoral examination showed severe malocclusions, including anterior crossbite and deep bite. Furthermore, a radiographic examination showed increased radiopacity of the maxilla and mandibular bones due to hyperostosis and sclerosis of the jaw. There was no root resorption of the canines or molars in the primary dentition, although root formation of the permanent teeth was proceeding. Dental age was calculated to be approximately 1 year, 4 months younger than his chronological age. The eruption speed of the permanent lateral incisors after the gingival emergence was shown to be within normal values, and we discuss whether the canines and premolars in the permanent dentition could erupt or not.


Dietary strategies for improving folate status in institutionalized elderly persons.
The aim of this work was to compare the efficacy of two strategies designed to improve folate status: increasing the intake of vegetables, and the consumption of a folic acid-fortified food. Residents (126) from three old people's homes in the Madrid region (Spain) were studied. To each centre a dietary intervention was assigned to be followed for 6 months: (1) the consumption of margarine fortified with 200 microg folic acid/10 g portion (centre M), (2) increasing the consumption of vegetables to three servings per day (centre V), (3) control (centre C). At the beginning and end of the intervention period the subjects' intakes, serum and erythrocyte concentrations of folate were measured. The use of fortified margarine (centre M) led to a significant increase in folate intake (260.9 microg/d), serum concentration (10.3 (sd 8.3) nmol/l) and erythrocyte concentration (638.4 nmol/l). At centre V the increase in total vegetable intake achieved was very poor; these foods met with very poor acceptance, although the intake of certain vegetables particularly rich in folate improved. Therefore, the intake of this vitamin increased a little (26.7 (sd 33.0) microg/d); erythrocyte folate concentration also increased somewhat (460.5 nmol/l), althought less than centre M. The daily consumption of margarine fortified with folic acid was the more effective strategy for improving the folate status of the study subjects.


MicroRNA: a connecting road between apoptosis and cholesterol metabolism.
Resistance to apoptosis leads to tumorigenesis and failure of anti-cancer therapy. Recent studies also highlight abrogated lipid/cholesterol metabolism as one of the root causes of cancer that can lead to metastatic transformations. Cancer cells are dependent on tremendous supply of cellular cholesterol for the formation of new membranes and continuation of cell signaling. Cholesterol homeostasis network tightly regulates this metabolic need of cancer cells on cholesterol and other lipids. Genetic landscape is also shared between apoptosis and cholesterol metabolism. MicroRNAs (miRNAs) are the new fine tuners of signaling pathways and cellular processes and are known for their ability to post-transcriptionally repress gene expression in a targeted manner. This review summarizes the current knowledge about the cross talk between apoptosis and cholesterol metabolism via miRNAs. In addition, we also emphasize herein recent therapeutic modulations of specific miRNAs and their promising potential for the treatment of deadly diseases including cancer and cholesterol related pathologies. Understanding of the impact of miRNA-based regulation of apoptosis and metabolic processes is still at its dawn and needs further research for the development of future miRNA-based therapies. As both these physiological processes affect cellular homeostasis, we believe that this comprehensive summary of miRNAs modulating both apoptosis and cholesterol metabolism will open uncharted territory for scientific exploration and will provide the foundation for discovering novel drug targets for cancer and metabolic diseases.


Emissions of aldehydes and ketones from a two-stroke engine using ethanol and ethanol-blended gasoline as fuel.
Besides aliphatic gasoline, ethanol-blended gasoline intended for use in small utility engines was recently introduced on the Swedish market. For small utility engines, little data is available showing the effects of these fuels on exhaust emissions, especially concerning aldehydes and ketones (carbonyls). The objective of the present investigation was to study carbonyl emissions and regulated emissions from a two-stroke chain saw engine using ethanol, gasoline, and ethanol-blended gasoline as fuel (0%, 15%, 50%, 85%, and 100% ethanol). The effects of the ethanol-blending level and mechanical changes of the relative air/fuel ratio, lambda, on exhaust emissions was investigated, both for aliphatic and regular gasoline. Formaldehyde, acetaldehyde, and aromatic aldehydes were the most abundant carbonyls in the exhaust. Acetaldehyde dominated for all ethanol-blended fuels (1.2-12 g/kWh, depending on the fuel and lambda), and formaldehyde dominated for gasoline (0.74-2.3 g/kWh, depending on the type of gasoline and lambda). The main effects of ethanol blending were increased acetaldehyde emissions (30-44 times for pure ethanol), reduced emissions of all other carbonyls exceptformaldehyde and acrolein (which showed a more complex relation to the ethanol content), reduced carbon monoxide (CO) and ntirogen oxide (NO) emissions, and increased hydrocarbon (HC) and nitrogen dixodie (NO2) emissions. The main effects of increasing lambda were increased emissions of carbonyls and nitrogen oxides (NOx) and reduced CO and HC emissions. When the two types of gasoline are considered, benzaldehyde and tolualdehyde could be directly related to the gasoline content of aromatics or olefins, but also acrolein, propanal, crotonaldehyde, and methyl ethyl ketone mainly originated from aromatics or olefins, while the main source for formaldehyde, acetaldehyde, acetone, methacrolein, and butanal was saturated aliphatic hydrocarbons.


Minimally invasive lateral approaches for the treatment of spinal tumors: single-position surgery without the "flip".
Although primary tumors of the spine and neural elements are rare, metastatic disease to the spine is quite common. Traditionally, surgical treatment for spinal tumor patients involves open decompression with or without stabilization. The single-position minimally invasive (MIS) lateral approach, which has been recently described over the recent decade, allows simultaneous access to the anterior and posterior columns with the patient positioned in the lateral decubitus position. Herein, we review the application of single-position MIS lateral surgery for the treatment of spinal neoplasm. The aim was to review the evolution, operative technique, outcomes, and complications associated with MIS lateral approaches for spinal tumors. The history of spinal tumor diagnosis and management are reviewed and discussed as well as the author's experience and literature regarding spinal tumor treatment outcome and surgical complications, with particular attention to single-position, MIS lateral approaches. In addition, the author's surgical technique is outlined in detail for thoracic, thoracolumbar and lumbar tumors. Furthermore, there are specific indications and complications associated with the surgical treatment of spinal tumors, and the MIS, single-position lateral approach, when applied appropriately, allows for concurrent access to the anterior and posterior column while mitigating the complications associated with traditional, open posterior-based approaches. In the treatment of spinal neoplasms, the goals of surgery are dictated by a number of tumor-specific and patient-specific factors. Therefore, operative treatment of tumors in the future may be a consolidation of historical surgical techniques and MIS, single-position lateral approaches. Regardless, multidisciplinary management is imperative for the individualized treatment of the patient and optimization of outcome.


The correlation between cognitive impairment and ambulatory blood pressure in patients with cerebral small vessel disease.
The present study was aimed to analyze the correlation between cognitive impairment and ambulatory blood pressure in patients with cerebral small vessel disease (CSVD). 108 patients with CSVD received in our hospital were selected. Assessment of cognitive impairment was by the Montreal Cognitive Assessment (MoCA). 39 cases were established as the impairment group and 69 cases were established as the normal group. 24 h ambulatory blood pressure was monitored, and changes in ambulatory blood pressure parameters between the two groups were compared. Also, the correlation between blood pressure parameters and MoCA score were analyzed. Comparisons of ambulatory systolic blood pressure, ambulatory pulse pressure and the ratios of night blood pressure reduction of patients in both groups showed statistical differences (p < 0.05), while the changes in diastolic blood pressure showed no statistical differences (p > 0.05). The comparison of the blood pressure curves in both groups showed statistical differences (p < 0.05). The ambulatory systolic blood pressure, ambulatory pulse pressure and the ratio of night blood pressure reduction of patients with CSVD showed prominently negative correlations with MoCA score (p < 0.05). Cognitive impairment and the ambulatory blood pressure of patients with CSVD are intimately correlated. The rise of ambulatory systolic blood pressure, pulse pressure, and the decline of blood pressure may represent risk factors for cognitive impairment in patients with CSVD. Improving blood pressure management will reduce the incidence of cognitive impairment caused by CSVD.


Feasibility of EUS-guided hepaticogastrostomy for inoperable malignant hilar biliary strictures.
EUS-guided biliary drainage (EUS-BD) has emerged as a complementary technique for primary drainage or as a rescue technique after failed endoscopic retrograde cholangiography. The objective of this study was to demonstrate the feasibility of EUS-BD for malignant hilar stenosis (MHS), both as an initial and rescue procedure. This study was a retrospective work based on a prospective registry of patients with malignant drainage stenosis of the hilum. For this analysis, only patients who underwent EUS-BD drainage were included. The drainage procedure could be performed by EUS-BD alone or in combination with another technique, for initial drainage or reintervention. Between January 2015 and September 2018, 20 patients were included. The mean patient age was 68 years. Seven patients had primary liver tumors and 13 had obstructions caused by metastasis. Four patients had Type II stenosis, 7 had Type IIIA, 2 had Type IIIb, and 7 had Type IV stenosis. Sixteen patients underwent EUS-guided hepaticogastrostomy (EUS-HGS) for initial drainage and four as reintervention. For initial drainage, 2 patients underwent EUS-HGS alone and 14 underwent EUS-HGS in combination with another technique: 11 combined with endoscopic retrograde cholangiopancreatography (ERCP), 2 with percutaneous transhepatic drainage, and 1 with ERCP and percutaneous transhepatic drainage. The technical success rate for EUS-HGS in the drainage of MHS was 100%, and the clinical success rate was 95%. The mean percentage of liver drained was 84%, with an average 1.7 endoscopic sessions and an average 2.7 protheses. The early complication rate was 35% and the mortality rate was 5%. Five EUS-HGS/ERCP combination drainage procedures were performed in one session and six were performed in two sessions with similar complication rates and percentages of liver segments drained. EUS-BD is a feasible and safe technique for initial drainage and for reintervention procedures. The EUS-HGS/ERCP combination seemed to be useful in cases of complex stenosis and could be performed during the same session or in two sessions.


Does arousal frequency predict daytime function?
Patients with the sleep apnoea/hypopnoea syndrome (SAHS) have impaired daytime function with demonstrable sleepiness and impaired cognition. The hypothesis that brief arousals from sleep cause these daytime impairments was tested. One hundred and fifty patients with sleep disordered breathing were studied prospectively, comparing overnight polysomnography with daytime measures of objective sleepiness, psychological well-being and cognitive performance. Significant, but weak (r2<0.1), relationships were seen between several nocturnal measures (apnoea/hypopnoea index, arousals and desaturation variables) and daytime measures of quality of life, well-being, subjective sleepiness, symptoms and cognitive performance. The only significant relationship between nocturnal variables and objective sleepiness was a very weak correlation (r2<0.05) between the lowest oxygen saturation and mean maintenance of wakefulness test (MWT) result. The MWT was better correlated with daytime function than the multiple sleep latency test. This study shows a lack of strong relationships between conventional nocturnal measures and daytime function in patients with sleep disordered breathing.


The association of a priori and a posterior dietary patterns with the risk of incident stroke in Chinese older people in Hong Kong.
Dietary patterns can be identified by a priori and a posterior approaches. Limited data have related dietary patterns with stroke risk in Chinese population. This study examined stroke risk associated with patterns identified by both approaches. Data on 1,338 Chinese men and 1,397 Chinese women aged >= 65 years participating in a cohort study examining the risk factors for osteoporosis in Hong Kong were analyzed. Baseline dietary data were collected between 2001 and 2003 using a validated food frequency questionnaire. Adherence to a priori dietary patterns including the Mediterranean Diet Score (MDS) and the Dietary Approaches to Stop Hypertension (DASH) was assessed. Factor analysis (FA) identified three a posterior dietary patterns: "vegetables-fruits", "snacks-drinks-milk products", and "meat-fish". Data on incidence of stroke were retrieved from an official database in 2008. Cox regression was used to estimate hazard ratios (HRs) for stroke risk adjusted for potential confounders. There were 156 incident stroke events during a median follow up of 5.7 years. A posterior dietary patterns derived by FA were not associated with risk of incident stroke in either men or women. MDS was inversely associated with risk of incident stroke [Adjusted HR=0.55 (95% CI: 0.31-0.99) of the highest level of MDS compared with the lowest level, ptrend=0.008] in men but not in women. Similar non-significant inverse association was observed between DASH accordance and stroke risk in men [Multivariate HR (95% CI) of the DASH score of >=4.5 = 0.62 (0.38-1.04) compared with the DASH score of <=4, ptrend = 0.068] but not in women. Our findings suggest that higher MDS and possibly higher DASH scores were associated with lower stroke risk in Chinese older people in Hong Kong.


Direct measurement of current-phase relations in superconductor/topological insulator/superconductor junctions.
Proximity to a superconductor is predicted to induce exotic quantum phases in topological insulators. Here, scanning superconducting quantum interference device (SQUID) microscopy reveals that aluminum superconducting rings with topologically insulating Bi2Se3 junctions exhibit a conventional, nearly sinusoidal 2π-periodic current-phase relations. Pearl vortices occur in longer junctions, indicating suppressed superconductivity in aluminum, probably due to a proximity effect. Our observations establish scanning SQUID as a general tool for characterizing proximity effects and for measuring current-phase relations in new materials systems.


Early T Cell Infiltration Correlates with Anti-CTLA4 Treatment Response in Murine Cancer Models.
Immune checkpoint inhibitor (ICI) Abs are a revolutionary class of cancer treatment, but only ∼30% of patients receive a lasting benefit from therapy. Preclinical studies using animals from the same genetic backgrounds, challenged with the same cancer models, also show nonuniform responses. Most mouse studies that have evaluated tumor-infiltrating leukocytes after ICI therapy cannot directly correlate their findings with treatment outcomes, because terminal methods were used to acquire immune infiltrate data. In the present study, we used fine-needle aspiration (a nonterminal sampling method) to collect multiple aspirates over several days from s.c. implanted P815, CT26, and 4T1 mouse cancer models treated with ICI Abs. These aspirates were then analyzed with flow cytometry to directly correlate tumor-infiltrating leukocyte populations with treatment success. We found that the P815 and CT26 models respond well to anti-CTLA4 therapies. Among P815-challenged animals, mice that regressed following anti-CTLA4 treatment showed significant increases in CD8+ T cells on days 3, 5, and 7 and in CD4+ T cells on days 5 and 7 and a decrease in macrophages and monocytes on days 3, 5, and 7 after treatment. Similar results were obtained in the CT26 model on day 11 posttreatment. Our study is the first, to our knowledge, to directly correlate early tumor infiltration of T cells with anti-CTLA4 treatment success, thus providing a mechanistic clue toward understanding why alloidentical mice challenged with identical tumors do not respond uniformly to ICI therapies.


Clinical Use of FSH in Male Infertility.
The established clinical indication for FSH use in male infertility is the treatment of patients with hypogonadotropic hypogonadism for stimulation of spermatogenesis that allows the induction of a clinical pregnancy in the female partner and finally the birth of a healthy child. Several clinical studies with urinary, purified, and recombinant FSH preparations in combination with hCG have demonstrated the high treatment efficacy regarding these clinical endpoints. Shortcomings of this hormone therapy are the long duration of treatment, sometimes longer than 2 years, and the inconvenience of injections every second or third day. However, improvements of therapy might be expected with new hormonal treatment options already available for infertility treatment in the female. FSH use for treatment of patients with normogonadotropic idiopathic infertility and oligozoospermia is still considered experimental in most countries. Recent meta-analyses have shown that FSH can significantly increase pregnancy rates in the female partners of these patients, but the effect-size is relatively low. Therefore, predictive factors for treatment success have to be identified, including FSH pharmacogenetics, to select the right normogonadotropic patients with idiopathic infertility for FSH therapy.


Hetero-Multivalency of Pseudomonas aeruginosa Lectin LecA Binding to Model Membranes.
A single glycan-lectin interaction is often weak and semi-specific. Multiple binding domains in a single lectin can bind with multiple glycan molecules simultaneously, making it difficult for the classic "lock-and-key" model to explain these interactions. We demonstrated that hetero-multivalency, a homo-oligomeric protein simultaneously binding to at least two types of ligands, influences LecA (a Pseudomonas aeruginosa adhesin)-glycolipid recognition. We also observed enhanced binding between P. aeruginosa and mixed glycolipid liposomes. Interestingly, strong ligands could activate weaker binding ligands leading to higher LecA binding capacity. This hetero-multivalency is probably mediated via a simple mechanism, Reduction of Dimensionality (RD). To understand the influence of RD, we also modeled LecA's two-step binding process with membranes using a kinetic Monte Carlo simulation. The simulation identified the frequency of low-affinity ligand encounters with bound LecA and the bound LecA's retention of the low-affinity ligand as essential parameters for triggering hetero-multivalent binding, agreeing with experimental observations. The hetero-multivalency can alter lectin binding properties, including avidities, capacities, and kinetics, and therefore, it likely occurs in various multivalent binding systems. Using hetero-multivalency concept, we also offered a new strategy to design high-affinity drug carriers for targeted drug delivery.


Trajectories of cardiorespiratory fitness in patients with juvenile dermatomyositis.
Previous research demonstrated decreased cardiorespiratory fitness (CRF) in patients with JDM during active disease and remission. However, longitudinal data regarding trajectories of CRF are currently lacking. The objective of this study was to determine trajectories of CRF in patients with both monocyclic and chronic JDM, and to identify potential predictors of these trajectories. Thirty-six patients with JDM [median age (interquartile range) at diagnosis: 8.3 (6.3-15.4) years] treated in our paediatric rheumatology outpatient clinic were included. All patients performed multiple cardiopulmonary exercise tests between 2003 and 2016. Relevant CRF parameters were analysed, including peak oxygen uptake, maximal workload, mechanical efficacy and oxygen uptake at ventilatory anaerobic threshold. We analysed trajectories up to 10 years after diagnosis and determined predictors of CRF outcome parameters by multilevel analyses. Trajectories demonstrated significant declines in CRF during the active phase of the disease with subsequent improvement in CRF during the initial years after diagnosis. However, hereafter no further improvements, and even a decrease, in CRF were observed over time in both monocyclic and chronic subtypes of JDM. We found that a longer disease duration, younger age of onset and higher prednisone dose negatively influence CRF. Patients with both monocyclic and chronic JDM show decreases in long-term CRF trajectories. Longer disease duration, younger age of onset and higher prednisone dose negatively influence CRF. This study stresses the need for regular evaluation of CRF and implementation of (exercise) interventions to improve CRF in patients with JDM, even in monocyclic patients.


Protective effects of memantine against ischemia-reperfusion injury in spontaneously hypertensive rats.
Memantine, an uncompetitive NMDA open-channel blocker, has been shown to be effective in preventing neuronal damage after permanent focal cerebral ischemia. Reperfusion after a long period of ischemia may aggravate the progression of neuronal damage. Those drugs that show protective effects after permanent cerebral ischemia, therefore, might fail to do so against ischemia-reperfusion injury. In this study we evaluated the effects of memantine on brain edema formation and ischemic injury volume after transient cerebral ischemia. Male Spontaneously Hypertensive Rats (SHR) weighing 250-300 g were anesthetized with halothane and subjected to 1 hour of temporary middle cerebral artery occlusion by an intraluminal suture. 20 mg/kg of memantine or saline were injected intraperitoneally 5 min. after the induction of ischemia. Physiological parameters and regional cerebral blood flow were monitored during the surgical procedure. Brain water content and ischemic injury volume were measured with the wet dry method and 2,3,5-triphenyl tetrazolium chloride monohydrate (TTC) staining, respectively, at 24 hours after occlusion. There were no statistically significant differences between the groups regarding physiological parameters during the procedure. Memantine treatment (n=9) reduced the brain water content significantly in the cortex compared to saline treatment (n=8; 83. 1+/-0.7% vs. 84.5+/-1.5%, respectively, p<0.05). The total volume of ischemic brain injury was 300+/-49 mm(3) in the animals treated with saline (n=13). Treatment with 20 mg/kg memantine (n=14) reduced the ischemic injury volume to 233+/-61 mm(3) (P<0.01). These results demonstrate that the harmful effects of recirculation after a period of ischemia can be attenuated by the treatment of memantine, perhaps by its action at the NMDA receptors.


End-stage kidney disease due to Alport syndrome: outcomes in 296 consecutive Australia and New Zealand Dialysis and Transplant Registry cases.
Alport syndrome is a rare inheritable renal disease. Clinical outcomes for patients progressing to end-stage kidney disease (ESKD) are not well described. This study aimed to investigate the characteristics and clinical outcomes of patients from Australia and New Zealand commencing renal replacement therapy (RRT) for ESKD due to Alport syndrome between 1965 and 1995 (early cohort) and between 1996 and 2010 (contemporary cohort) compared with propensity score-matched, RRT-treated, non-Alport ESKD controls. A total of 58 422 patients started RRT during this period of which 296 (0.5%) patients had Alport ESKD. In the early cohort, Alport ESKD was associated with superior dialysis patient survival [adjusted hazard ratio (HR): 0.41, 95% confidence interval (CI): 0.20-0.83, P = 0.01], renal allograft survival (HR: 0.74, 95% CI: 0.54-1.01, P = 0.05) and renal transplant patient survival (HR: 0.43, 95% CI: 0.28-0.66, P < 0.001) compared with controls. In the contemporary cohort, no differences were observed between the two groups for dialysis patient survival (HR: 1.42, 95% CI: 0.65-3.11, P = 0.38), renal allograft survival (HR: 1.01, 95% CI: 0.57-1.79, P = 0.98) or renal transplant patient survival (HR: 0.67, 95% CI: 0.26-1.73, P = 0.41). One Alport patient (0.4%) had post-transplant anti-glomerular basement membrane (anti-GBM) disease. Four female and 41 male Alport patients became parents on RRT with generally good neonatal outcomes. Alport syndrome patients experienced comparable dialysis and renal transplant outcomes to matched non-Alport ESKD controls in the contemporary cohort due to relatively greater improvements in outcomes for non-Alport ESKD patients over time. Post-transplant anti-GBM disease was rare.


Group Sequential Survival Trial Design and Monitoring Using the Log-Rank Test.
For randomized group sequential survival trial designs with unbalanced treatment allocation, the widely used Schoenfeld formula is inaccurate, and the commonly used information time as the ratio of number of events at interim look to the number of events at the end of trial can be biased. In this paper, a sample size formula for the two-sample log-rank test under the proportional hazards model is proposed that provides more accurate sample size calculation for unbalanced survival trial designs. Furthermore, a new information time is introduced for the sequential survival trials such that the new information time is more accurate than the traditional information time when the allocation of enrollments is unbalanced in groups. Finally, we demonstrate the monitoring process using the sequential conditional probability ratio test and compare it with two other well known group sequential procedures. An example is given to illustrate unbalanced survival trial design using available software.


Factors for occurrence of extended-spectrum β-lactamase-producing Escherichia coli in broilers.
To clarify the factors for occurrence of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli in broilers, two flocks (1 day of age) fed a diet with or without antibiotics were kept in a broiler house sanitized with disinfectants. ESBL-producing E. coli, however, was detected at a concentration of over 10(6) CFU/g of feces at 9 days of age to 49 days of age in both broiler flocks. Therefore, this indicated that the antibiotics other than cephalosporins used in this study had no effect due to co-selection on the numbers of ESBL-producing E. coli in broiler feces during this period. When a flock was kept with diet containing antibiotics for 49 days in a laboratory animal room, no ESBL-producing E. coli was detected in the flock. These results suggest that the occurrence of ESBL-producing E. coli may not be related to feeding with antibiotics and that the contamination of broiler houses with ESBL-producing E. coli might be an important factor.


The link between childhood trauma and depression: insights from HPA axis studies in humans.
Childhood trauma is a potent risk factor for developing depression in adulthood, particularly in response to additional stress. We here summarize results from a series of clinical studies suggesting that childhood trauma in humans is associated with sensitization of the neuroendocrine stress response, glucocorticoid resistance, increased central corticotropin-releasing factor (CRF) activity, immune activation, and reduced hippocampal volume, closely paralleling several of the neuroendocrine features of depression. Neuroendocrine changes secondary to early-life stress likely reflect risk to develop depression in response to stress, potentially due to failure of a connected neural circuitry implicated in emotional, neuroendocrine and autonomic control to compensate in response to challenge. However, not all of depression is related to childhood trauma and our results suggest the existence of biologically distinguishable subtypes of depression as a function of childhood trauma that are also responsive to differential treatment. Other risk factors, such as female gender and genetic dispositions, interfere with components of the stress response and further increase vulnerability for depression. Similar associations apply to a spectrum of other psychiatric and medical disorders that frequently coincide with depression and are aggravated by stress. Taken together, this line of evidence demonstrates that psychoneuroendocrine research may ultimately promote optimized clinical care and help prevent the adverse outcomes of childhood trauma.


Reliability of the American Community Survey Estimates of Risk-Adjusted Readmission Rankings for Hospitals Before and After Peer Group Stratification.
Since the transition to the American Community Survey, data uncertainty has complicated its use for policy making and research, despite the ongoing need to identify disparities in health care outcomes. The US Centers for Medicare & Medicaid Services' new, stratified payment adjustment method for its Hospital Readmissions Reduction Program may be able to reduce the reliance on data linkages to socioeconomic survey estimates. To determine whether there are differences in the reliability of socioeconomically risk-adjusted hospital readmission rates among hospitals that serve a disproportionate share of low-income populations after stratifying hospitals into peer group-based classification groups. This cross-sectional study uses data from the 2014 New York State Health Cost and Utilization Project State Inpatient Database for 96 278 hospital admissions for acute myocardial infarction, pneumonia, and congestive heart failure. The analysis included patients aged 18 years and older who were not transferred to another hospital, who were discharged alive, who did not leave the hospital against medical advice, and who were discharged before December 2014. The main outcomes were 30-day hospital readmissions after acute myocardial infarction, pneumonia, and congestive heart failure assessed using hierarchical logistic regression. The mean (SD) age of the patients was 69.6 (16.0) years for the safety-net hospitals and 74.9 (14.7) years for the non-safety-net hospitals; 9382 (48.8%) and 7003 (48.5%) patients, respectively, were female. For safety net designations, 20% (3 of 15) of all evaluations concealed and distorted differences in risk, with factors such as poverty failing to identify similar risk of acute myocardial infarction readmission until unreliable estimates were excluded from the analysis (OR, 1.23 [95% CI, 1.00-1.52], P = .02; vs OR, 1.17 [95% CI, 0.94-1.46], P = .15). By comparison, 2 of the 60 models (3%) for the peer group-based classification altered the association between socioeconomic status and readmission risk, concealing similarities in congestive heart failure readmission when adjusted using high school completion rates (OR, 1.27 [95% CI 1.02-1.58], P = .04; vs OR, 1.23 [95% CI, 0.98-1.53], P = .06) and distorting similarities in pneumonia readmissions when accounting for the proportion of lone-parent families (OR, 1.27 [95% CI, 0.98-1.66], P = .07; vs OR, 1.35 [95% CI, 1.02-1.80], P = .04) between the lowest and highest socioeconomic status hospitals in quartile 1. There was greater precision in socioeconomic adjusted readmission estimates when hospitals were stratified into the new payment adjustment criteria compared with safety net designations. A contributing factor for improved reliability of American Community Survey estimates under the new payment criteria was the merging of patients from low-income neighborhoods with greater homogeneity in survey estimates into groupings similar to those for higher-income patients, whose neighborhoods often exhibit greater estimate variability. Additional efforts are needed to explore the effect of measurement error on American Community Survey-adjusted readmissions using the new peer group-based classification methods.


The PUF family of RNA-binding proteins: does evolutionarily conserved structure equal conserved function?
Drosophila Pumilio (Pum) protein is a founder member of a novel family of RNA-binding proteins, known as the PUF family. The PUF proteins constitute an evolutionarily highly conserved family of proteins present from yeast to humans and plants, and are characterized by a highly conserved C-terminal RNA-binding domain, composed of eight tandem repeats. The conserved biochemical features and genetic function of PUF family members have emerged from studies of model organisms. PUF proteins bind to related sequence motifs in the 3' untranslated region (3'UTR) of specific target mRNAs and repress their translation. Frequently, PUF proteins function asymmetrically to create protein gradients, thus causing asymmetric cell division and regulating cell fate specification. Thus, it was recently proposed that the primordial role of PUF proteins is to sustain mitotic proliferation of stem cells. Here we review the evolution, conserved genetic and biochemical properties of PUF family of proteins, and discuss protein interactions, upstream regulators and downstream targets of PUF proteins. We also suggest that a conserved mechanism of PUF function extends to the newly described mammalian members of the PUF family (human PUM1 and PUM2, and mouse Pum1 and Pum2), that show extensive homology to Drosophila Pum, and could have an important role in cell development, fate specification and differentiation.


Anesthesia modality does not affect clinical outcomes of intra-arterial vasodilator treatment in patients with symptomatic cerebral vasospasms.
Background: Delayed cerebral ischemia and cerebral vasospasm remain the leading causes of poor outcome in survivors of aneurysmal subarachnoid hemorrhage. Refractory cerebral vasospasms can be treated with endovascular vasodilator therapy, which can either be performed in conscious sedation or general anesthesia. The aim of this study is to compare the effect of the anesthesia modality on long-term clinical outcomes in patients undergoing endovascular vasodilator therapy due to cerebral vasospasm and hypoperfusion. Methods: Modified Rankin Scale (mRS) scores were retrospectively analyzed at time of discharge from the hospital and six months after aneurysmal subarachnoid hemorrhage. Additionally, National Institutes of Health Stroke Scale (NIHSS) was assessed 24 hours before, immediately before, immediately after, and 24 hours after endovascular vasodilator therapy, and at discharge and six months. Interventional parameters such as duration of intervention, choice and dosage of vasodilator and number of arteries treated were also recorded. Results: A total of 98 patients were included in this analysis and separated into patients who had interventions in conscious sedation, general anesthesia and a mix of both. Neither mRS at discharge nor at six months showed a significant difference for functionally independent outcomes (mRS 0-2) between groups. NIHSS before endovascular vasodilator therapy was significantly higher in patients receiving interventions in general anesthesia but did not differ anymore between groups six months after the initial bleed. Conclusion: This study did not observe a difference in outcome whether patients underwent endovascular vasodilator therapy in general anesthesia or conscious sedation for refractory cerebral vasospasms. Hence, the choice should be made for each patient individually.


Differential effects of aging and insulin-like growth factor-1 on synapses in CA1 of rat hippocampus.
Aging-related impairments of learning and memory can be ameliorated by 28 days of intracerebroventricular (icv) infusion of insulin-like growth factor-1 (IGF-1) in old rats. The present study investigated whether there is an aging-related synaptic decline in the stratum radiatum of hippocampal CA1 and whether IGF-1 can ameliorate that decline. Five young (4 months), five middle-aged (18 months) and five old (29 months) Fischer 344xBrown Norway rats received saline infusion; five old (29 months) rats received IGF-1 infusion for 28 days preceding sacrifice. Pyramidal neurons, total synaptic profiles as well as synaptic profiles in multiple spine bouton (MSB) complexes in CA1 were quantified stereologically with the physical disector technique and the postsynaptic density (PSD) length was determined as well. The results indicated a decrease of total synapses between middle and old age but a maintenance of PSD length and MSB synapses throughout life. IGF-1 infusion in old rats did not reverse the aging-related decline in total synapses but did increase PSD length and the number of MSB synapses. These changes in synaptic configurations are morphological correlates of enhanced synaptic efficacy. Thus, aging and IGF-1 affect different, but complementary, aspects of synapses in hippocampal CA1.


Self-reported pain after orthodontic treatments: a randomized controlled study on the effects of two follow-up procedures.
To assess the effects of a follow-up text message and a telephone call after bonding on participants' self-reported level of pain. Eighty-four participants were randomly assigned to one of three trial arms. Randomization was performed by the Department of Epidemiology and Biostatistics of IRCCS G.Gaslini. Participants were enrolled from patients with a permanent dentition who were beginning fixed no extraction treatment at the Orthodontic Department, Gaslini Hospital. Participants completed baseline questionnaires to assess their levels of pain prior to treatment. After the initial appointment, participants were completed a pain questionnaire at the same time, daily, for 7 days. The first group, served as control, did not receive any post-procedure communication; the second group received a structured text message; and the third group received a structured telephone call. Participants were blinded to group assignment. A larger sample size should have been considered in order to increase the ability to generalize this study's results. Participants in both the telephone call group and the text message group reported lower level of pain than participants in the control group with a larger and more consistent effect for the telephone call group. Most participants reported a higher level of pain during the first 48 hours post-bonding. The analgesic's consumption significantly correlated with the level of pain during the previous 24 hours. Female participants appeared to be more sensitive to pain than male participants. A telephone follow-up after orthodontic treatment may be an effective procedure to reduce participants' level of pain. The research protocol was approved by the Italian Comitato Etico Regionale della Liguria-sezione 3^ c/o IRCCS- Istituto G.Gaslini 845/2014. 182 Reg 2014, 16/09/2014 Comitato Etico Regione Liguria, Sez.3.


The impact of diabetes on productivity in China.
Diabetes increases the risk of premature death and reduces work productivity. We estimated the impact of diabetes in China in terms of mortality, years of life lost, and productivity-adjusted life years (PALYs) lost in the Chinese population. Life table modelling was used with simulated follow-up of those with diabetes in the Chinese population of working age (20-49 years in women and 20-59 years in men) until retirement age (50 years for women and 60 years for men). Data regarding the prevalence of diabetes, as well as excess mortality, labour force dropout and productivity loss attributable to diabetes, were taken from published sources. Models were constructed for the cohort with diabetes and repeated for the same cohort assuming that they had no diabetes. The differences in number of deaths, years of life lived and PALYs lived between the two models reflected the impact of diabetes. The WHO standard 3% annual discount rate was applied to years of life and PALYs lived. In 2017, an estimated 56.4 million people of working age in China (7.1%) had diabetes. With simulated follow-up until retirement, those with diabetes were predicted to experience an estimated 4.1 million more deaths, the loss of an additional 22.7 million years of life (3.7%) and the loss of an additional 75.8 million PALYs (15.1%). This was equivalent to an average of 1.3 PALYs lost per person with diabetes. Based on gross domestic product (GDP) per full-time worker in 2017, the loss in PALYs equated to a total of Chinese ¥17.4 trillion (US$2.6 trillion) in lost GDP owing to reduced productivity, with an average of ¥307,925 (US$45,959) lost per person with diabetes. Our study demonstrates the significant cumulative impact of diabetes on productivity across the working lifetime in the Chinese population, highlighting the potential economic benefits of diabetes prevention in the longer term.


Biomechanical analysis of cancerous and normal cells based on bulge generation in a microfluidic device.
This paper presents a new biomechanical analysis method for discrimination between cancerous and normal cells through compression by poly(dimethylsiloxane) (PDMS) membrane deflection in a microfluidic device. When a cell is compressed, cellular membrane will expand and then small bulges will appear on the peripheral cell membrane beyond the allowable strain. It is well known that the amount of F-actin in cancer cells is less than that of normal cells and bulges occur at the sites where cytoskeleton becomes detached from the membrane bilayer. Accordingly, we have demonstrated the difference of the bulge generation between breast cancer cells (MCF7) and normal cells (MCF10A). After excessive deformation, the bulges generated in MCF7 cells were not evenly distributed on the cell periphery. Contrary to this, the bulges of MCF10A cells showed an even distribution. In addition, the morphologies of bulges of MCF7 and MCF10A cells looked swollen protrusion and tubular protrusion, respectively. Peripheral strains at the moment of the bulge generation were also 72% in MCF7 and 46% in MCF10A. The results show that the bulge generation can be correlated with the cytoskeleton quantity inside the cell, providing the first step of a new biomechanical approach.


Endoscopic ultrasound characteristics of mucinous cystic neoplasms of the pancreas.
Mucinous cystic neoplasms of the pancreas have a more favorable prognosis than ductal adenocarcinoma. Management of a subgroup, intraductal papillary-mucinous neoplasms, is controversial. Endoscopic ultrasound (EUS) with fine-needle aspiration biopsy may emerge as the imaging modality of choice. There are few studies describing the EUS features of these tumors. A total of 35 consecutive cases of cystic tumors of the pancreas with an established pathological diagnosis were analyzed for characteristic EUS features. Mucinous cystadenocarcinomas (n = 14) were more likely to be characterized by hypoechoic cystic/solid mass or complex cyst and were frequently associated with a dilated main pancreatic duct. Benign mucinous duct ectasia (n = 6) were characterized by a dilated main pancreatic duct in conjunction with hyperechoic thickening of the duct wall. The two cases of intraductal mucinous hyperplasia additionally showed a hypoechoic mass. Intraductal papillary carcinoma (n = 11) had features in common with mucinous cystadenocarcinoma but also had echogenic foci in the mass and intraductal hyperechoic lesions. The two cases of microcystic cystadenoma showed either a mixed hypoechoic solid/cystic mass or a complex cyst without the additional features seen in mucinous cystadenocarcinoma. EUS features seem to exist that may help to differentiate cystic neoplasms from adenocarcinoma of the pancreas and, thus, to establish the preoperative diagnosis of cystic tumors of the pancreas.


Low- and standard-dose peginterferon alfa-2a for chronic hepatitis C, genotype 2 or 3: efficacy, tolerability, viral kinetics and cytokine response.
Chronic infection with hepatitis C, genotype 2/3, responds better than other genotypes to peginterferon and ribavirin treatment. We hypothesized that a lower dose of peginterferon would be as effective, but less toxic than standard doses. To test the hypothesis that a lower dose of peginterferon would be as effective as, but less toxic than, standard doses. A total of 30 patients were treated with low-dose peginterferon alfa-2a (90 microg/week) and 27 patients with standard doses (180 microg/week) for 24 weeks in combination with 800 mg/day of ribavirin. Patients who failed treatment were offered 48 weeks of standard-dose treatment. Viral and serum inducible protein 10 (IP-10) levels were measured and early viral kinetic parameters were calculated. Sustained virological response was achieved in 68% of the low-dose and 87% of the standard-dose patients (per protocol, P = 0.79 for non-inferiority). Re-treatment was successful in all patients who tolerated full dose and duration. The standard-dose group had greater first-phase declines of viral levels and faster time to negativity. The second-phase slope was not dose-dependent. IP-10 induction was significantly greater with the standard dose. Although fatigue and general feeling during treatment were worse for standard dose, haematological toxicity and depression did not differ between groups. A lower dose of peginterferon is associated with some symptomatic benefit, but the response is not equivalent to standard dosing.


Magnitude and Determinants of the Ratio between Prevalence of Low Vision and Blindness in Rapid Assessment of Avoidable Blindness Surveys.
To determine the magnitude and determinants of the ratio between prevalence of low vision and prevalence of blindness in rapid assessment of avoidable blindness (RAAB) surveys globally. Standard RAAB reports were downloaded from the repository or requested from principal investigators. Potential predictor variables included prevalence of uncorrected refractive error (URE) as well as gross domestic product (GDP) per capita, health expenditure per capita of the country across World Bank regions. Univariate and multivariate linear regression were used to investigate the correlation between potential predictor variables and the ratio. The results of 94 surveys from 43 countries showed that the ratio ranged from 1.35 in Mozambique to 11.03 in India with a median value of 3.90 (Interquartile range 3.06;5.38). Univariate regression analysis showed that prevalence of URE (p = 0.04), logarithm of GDP per capita (p = 0.01) and logarithm of health expenditure per capita (p = 0.03) were significantly associated with the higher ratio. However, only prevalence of URE was found to be significant in multivariate regression analysis (p = 0.03). There is a wide variation in the ratio of the prevalence of low vision to the prevalence of blindness. Eye care service utilization indicators such as the prevalence of URE may explain some of the variation across the regions.


Could Japonica Rice Be an Alternative Variety for Increased Global Food Security and Climate Change Mitigation?
The growing importance of rice globally over the past three decades is evident in its strategic place in many countries' food security planning policies. Still, its cultivation emits substantial greenhouse gases (GHGs). The Indica and Japonica sub-species of Oryza sativa L. are mainly grown, with Indica holding the largest market share. The awareness, economics, and acceptability of Japonica rice in a food-insecure Indica rice-consuming population were surveyed. The impact of parboiling on Japonica rice was studied and the factors which most impacted stickiness were investigated through sensory and statistical analyses. A comparison of the growing climate and greenhouse gas emissions of Japonica and Indica rice was carried out by reviewing previous studies. Survey results indicated that non-adhesiveness and pleasant aroma were the most preferred properties. Parboiling treatment altered Japonica rice's physical and chemical properties, introducing gelatinization of starch and reducing adhesiveness while retaining micronutrient concentrations. Regions with high food insecurity and high consumption of Indica rice were found to have suitable climatic conditions for growing Japonica rice. Adopting the higher-yielding, nutritious Japonica rice whose cultivation emits less GHG in these regions could help strengthen food security while reducing GHGs in global rice cultivation.


Platelet binding properties of monoclonal lupus autoantibodies produced by human hybridomas.
The platelet binding properties of human monoclonal lupus autoantibodies have been studied. These IgM autoantibodies, produced by human X human hybridomas derived from lymphocytes of patients with systemic lupus erythematosus, are known to bind to single-stranded DNA. Four anti-DNA antibodies that express the dominant 16/6 idiotype--HF2-1/17, HF2-18/2, HF2-1/13b, and HF3-16/6--bound to glutaraldehyde-fixed platelets. In contrast, HF6-21/28, HF9-11/3, and polyclonal IgM bound poorly to platelets. [35S]Methionine was incorporated into HF2-1/17, and the interaction of the intrinsically radiolabeled HF2-1/17 with fixed platelets was evaluated in a solution phase radioimmunoassay. [35S]Methionine HF2-1/17 bound to fixed platelets and could be displaced by equivalent amounts of HF2-1/17, HF2-18/2, HF2-1/13b, and HF3-16/6. HF2-1/17 bound with greater affinity to fresh platelets and to thrombin-activated platelets than to glutaraldehyde-fixed platelets. Single-stranded DNA competed with platelets for the HF2-1/17 combining site. Treatment of fresh platelets with nuclease I, trypsin, chymotrypsin, and neuraminidase did not alter the binding of antibody to the platelet surface. No binding of antibody to phospholipid micelles was observed. Purified IgM autoantibodies did not inhibit platelet aggregation induced with ADP, thrombin, or ristocetin in platelet-rich plasma. These results indicate that the human IgM monoclonal anti-DNA autoantibodies that express the dominant 16/6 idiotype are polyspecific, bind to platelets, and interact with a platelet epitope that does not appear to involve DNA, protein, or sialic acid. These antibodies interact with platelets through the same sites responsible for antibody-DNA binding.


Development of Fully Degradable Phosphonium-Functionalized Amphiphilic Diblock Copolymers for Nucleic Acids Delivery.
To expand the range of functional polymer materials to include fully hydrolytically degradable systems that bear bioinspired phosphorus-containing linkages both along the backbone and as cationic side chain moieties for packaging and delivery of nucleic acids, phosphonium-functionalized polyphosphoester- block-poly(l-lactide) copolymers of various compositions were synthesized, fully characterized, and their self-assembly into nanoparticles were studied. First, an alkyne-functionalized polyphosphoester- block-poly(l-lactide) copolymer was synthesized via a one pot sequential ring opening polymerization of an alkyne-functionalized phospholane monomer, followed by the addition of l-lactide to grow the second block. Second, the alkynyl side groups of the polyphosphoester block were functionalized via photoinitiated thiol-yne radical addition of a phosphonium-functionalized free thiol. The polymers of varying phosphonium substitution degrees were self-assembled in aqueous buffers to afford formation of well-defined core-shell assemblies with an average size ranging between 30 and 50 nm, as determined by dynamic light scattering. Intracellular delivery of the nanoparticles and their effects on cell viability and capability at enhancing transfection efficiency of nucleic acids (e.g., siRNA) were investigated. Cell viability assays demonstrated limited toxicity of the assembly to RAW 264.7 mouse macrophages, except at high polymer concentrations, where the polymer of high degree of phosphonium functionalization induced relatively higher cytotoxicity. Transfection efficiency was strongly affected by the phosphonium-to-phosphate (P+/P-) ratios of the polymers and siRNA, respectively. The AllStars Hs Cell Death siRNA complexed to the various copolymers at a P+/P- ratio of 10:1 induced comparable cell death to Lipofectamine. These fully degradable nanoparticles might provide biocompatible nanocarriers for therapeutic nucleic acid delivery.


Daily oral 25-hydroxycholecalciferol supplementation for vitamin D deficiency in haemodialysis patients: effects on mineral metabolism and bone markers.
Vitamin D deficiency is frequently observed in end-stage renal disease (ESRD) patients; however, the effects of vitamin D supplementation have rarely been reported. We aimed to assess the effects of daily 25(OH)D(3) supplementation on mineral metabolism, bone markers and Kidney Disease Outcomes Quality Initiative (KDOQI) targets in haemodialysis (HD) patients for a period of 6 months. HD patients were included in this study if their serum 25(OH)D level was <75 mmol/L. Oral 25(OH)D(3) was administered daily at 10-30 microg/day based on the severity of the deficiency. Characteristics of the patients were compared from the baseline to 6 months on the basis of their response to 25(OH)D(3) administration and the patients were divided into three groups. Patients who showed partial response [serum 25(OH)D <75 nmol/L] were placed in group 1, those who showed normal response [serum 25(OH)D ranging from 75 to 150 nmol/L] were placed in group 2 and those who showed excessive response [serum 25(OH)D >150 nmol/L] were placed in group 3. Of the 253 HD patients, 225 (89%) showed vitamin D insufficiency or deficiency, 172 were included in the study and 149 patients completed the study. After 6 months of treatment [mean daily 25(OH)D(3): 16 +/- 5 microg/day], the serum 25(OH)D level increased (30 +/- 19 to 126 +/- 46 nmol/ L, P < 0.001), with 13% of patients in group 1, 57% in group 2 and 30% in group 3. The serum intact parathyroid hormone (iPTH) level decreased (235 +/- 186 to 189 +/- 137 pg/mL, P = 0.05), except in group 1. Bone alkaline phosphatase (BALP) showed a tendency to normalize (23 +/- 16 to 18.3 +/- 11 microg/L, P < 0.05), leading to a decrease in alfacalcidol administration from 66% to 43% (P < 0.05), except in group 1. The KDOQI targets achieved increased significantly for serum calcium (76% to 85%) and phosphate levels (66% to 77%) in all patients. The serum albumin level increased in all groups (34.6 +/- 4 to 36.8 +/- 4 g/L, P < 0.05), without any significant improvement in normalized protein catabolic rate (nPCR) or C-reactive proteins (CRP). With a daily dose ranging from 10 to 30 microg, daily oral 25(OH)D(3) supplementation corrects most vitamin D deficiencies or insufficiencies in HD patients, without any evident toxicity. The main effects observed included correction of excessive bone turnover, despite less alfacalcidol administration, increase in serum albumin level and increase in the percentage of patients with serum calcium and phosphorus levels within the recommendation of the KDOQI guidelines.


Associations between absolute neutrophil count and lymphocyte-predominant breast cancer.
Tumor-infiltrating lymphocytes (TILs) might be associated with host-cell mediated immunity, which could be partly reflected by peripheral blood cell counts. In addition, lymphocyte-predominant breast cancer (LPBC), which was defined as tumors having high TIL levels, showed a favorable prognosis among triple-negative breast cancer or HER2-positive breast cancer. We aimed to investigate whether peripheral blood cell counts are associated with LPBC. We evaluated the percentage of stromal TILs in breast cancer patients who underwent primary surgery, using the standardized methodology proposed by the international TIL Working Group. Lymphocyte-predominant breast cancer (LPBC) was defined as tumors having high TIL levels (≥50%). Peripheral blood cell counts including absolute neutrophil counts (ANC), absolute lymphocyte counts (ALC) and neutrophil-to-lymphocyte ratio (NLR) was obtained from pretreatment laboratory data. Of the 810 patients, 132 (16.3%) had LPBC, and 678 (83.7%) had non-LBPC. In a comparison of 3 markers of peripheral blood counts, LPBC had a significantly lower mean ANC than non-LPBC (3,304 vs. 3,564; P = 0.023), but the other means were not different. In multivariable analysis, each 1K increment in ANC corresponded to an odds ratio of 0.790 (95% CI, 0.642 to 0.971) for LPBC. In the ER-negative and high-Ki67 subgroups identified by interaction tests, significant inverse correlations between continuous ANC and TILs were noted. Low peripheral ANC could be linked with LPBC, supporting the hypothesis that systemic immune cell counts might be associated with the tumor-immune microenvironment.


Significance of interspecies matches when evolutionary rate varies.
We develop techniques to estimate the statistical significance of gap-free alignments between two genomic DNA sequences, using human-mouse alignments as an example. The sequences are assumed to be sufficiently similar that some but not all of the neutrally evolving regions (i.e., those under no evolutionary constraint) can be reliably aligned. Our goal is to model the situation in which the neutral rate of evolution, and hence the extent of the aligning intervals, varies across the genome. In some cases, this permits the weaker of two matches to be judged as less likely to have arisen by chance, provided it lies in a genomic interval with a high level of background divergence. We employ a hidden Markov model to capture variations in divergence rates and assign probability values to gap-free alignments using techniques of Dembo and Karlin, which are related to those used for the same purpose by BLAST. Our methods are illustrated in detail using a 1.49 Mb genomic region. Results obtained from the analysis of human chromosome 22 using these techniques are also provided.


invMap: a sensitive mapping tool for long noisy reads with inversion structural variants.
Longer reads produced by PacBio or Oxford Nanopore sequencers could more frequently span the breakpoints of structural variants (SVs) than shorter reads. Therefore, existing long-read mapping methods often generate wrong alignments and variant calls. Compared to deletions and insertions, inversion events are more difficult to be detected since the anchors in inversion regions are nonlinear to those in SV-free regions. To address this issue, this study presents a novel long-read mapping algorithm (named as invMap). For each long noisy read, invMap first locates the aligned region with a specifically designed scoring method for chaining, then checks the remaining anchors in the aligned region to discover potential inversions. We benchmark invMap on simulated datasets across different genomes and sequencing coverages, experimental results demonstrate that invMap is more accurate to locate aligned regions and call SVs for inversions than the competing methods. The real human genome sequencing dataset of NA12878 illustrates that invMap can effectively find more candidate variant calls for inversions than the competing methods. The invMap software is available at https://github.com/zhang134/invMap.git. Supplementary data are available at Bioinformatics online.


Human papillomavirus cervical infection and associated risk factors in a region of Argentina with a high incidence of cervical carcinoma.
To assess the prevalence and potential risk factors associated with human papillomavirus (HPV) cervical infection among women residing in a region of northeastern Argentina with a high incidence of cervical cancer. A case-control study of 330 women participating in a cervical cytological screening program conducted in Posadas city, Misiones, Argentina, from February 1997 to November 1998 was carried out. Standardized questionnaires were administered, and clinical examination including colposcopy was performed. Fresh endocervical specimens for HPV DNA detection by generic polymerase chain reaction were collected and the products typed by dot-blot hybridization. Human papillomavirus DNA was found in 61% of samples analyzed (185/301). Samples with normal cytology had a 43% infection rate (85/199), while those classified as low-grade squamous intraepithelial lesion, high-grade squamous intraepithelial lesion, and invasive cervical carcinoma had an infection rate of 96% (53/55), 100% (29/29), and 100% (18/18), respectively. Human papillomavirus typing showed a 64% (118/185) prevalence of type 16 among all the infected population analyzed; type 16 was detected among 49% (42/85) of infected samples with normal cytology and in an average of 74% (74/100) with abnormal cytology. Sexual behavior, residence in southern Paraguay, and history of a previous sexually transmitted diseases were the main risk factors associated with high-grade cervical lesions. An elevated prevalence of HPV infection was detected in this population, which also has a high incidence of cervical cancer. The broad distribution of high-risk HPV type 16 in women with normal cytology and colposcopy suggests that viral infection is an important determinant of regional cancer incidence.


Does hospital type affect pyloromyotomy outcomes? Analysis of the Kids' Inpatient Database.
Pyloromyotomy is a common operative procedure performed on infants. The purpose of this study was to determine if hospital type affects lengths of stay (LOS), charges, and morbidity. Patients undergoing pyloromyotomy were identified in the Kids' Inpatients Database from 2000, 2003, and 2006. Freestanding children's hospitals (CH) were compared with children's units within general hospitals (CUGH) and general/nonchildren's hospitals (GH). Of the 10,969 patients, 25% received care at 30 CH, 35% received care at 94 CUGH, and 40% received care at 662 GH. Adjusted LOS were 2.41 days for CH, 2.75 days for CUGH, and 2.82 days for GH (P < .01). Adjusted mean charges were $11,160 for CH, $12,284 for CUGH, and $10,197 for GH (P = .01). CH had the lowest unadjusted complication rate at 1.2% compared with 1.6% at CUGH and 2.2% at GH (P < .01). GH were more likely to have patients with prolonged LOS (> or =4 days) compared with CH after adjusting for patient and hospital factors (odds ratio [OR], 1.7; 95% confidence interval [CI], 1.2-2.5). After accounting for LOS, CUGH were more likely to have higher charges (> or =$11,057) compared with CH (OR, 3.4; 95% CI, 1.03-11.18). The adjusted mean charges rose from $7,733 in 2000 to $11,335 in 2003 and to $14,572 in 2006 (P < .01). CH had the shortest LOS and lowest complication rates. Despite a higher complication rate and longer LOS, GH had the lowest charges. There is an opportunity to identify best practices, to improve quality, and to lower costs for pyloromyotomy in the United States, regardless of hospital type.


The effect of surgery and grade on outcome of gastrointestinal stromal tumors.
Gastrointestinal stromal tumors (GIST) are aggressive, rare, and difficult-to-cure gastrointestinal tumors. We believe that the clinical behavior of these tumors can be predicted by reproducible prognostic factors. A retrospective review of all patients (N = 70) with GIST treated at a tertiary care center from 1973 to 1998. Adequate data for evaluation were available for 69 patients. Male-female distribution was 40:29. Median age was 60 years. Median follow-up duration was 38 months. Tumor grade, stage, and histologic subtype at presentation; effect of grade, surgery and adjuvant therapy on recurrence, salvage, and survival. Tumor distribution included 61% in the upper, 23% in the middle, and 16% in the lower digestive tract, with a median tumor size of 7.9 cm (range, 1.8-25 cm). Tumors with more than 1 mitosis per 10 high-power fields constituted 57% of neoplasia in the series. Distant disease at initial visit occurred in 49% of patients. Complete gross resection occurred in 59% of patients. After complete resection, the 5-year survival rate was 42%, compared with 9% after incomplete resection (hazard ratio = 0.27, P<.001). Neither radiation nor chemotherapy demonstrated any significant benefit. Among 39 patients who were disease free after complete resection, 2% developed lymph node recurrence, 25% developed local recurrence, and 33% developed distant recurrences (54% liver, 20% peritoneum). By multivariate analysis the risk of local and/or distant metastases was significantly increased for tumors with more than 1 mitosis and size larger than 5 cm (P<.05). Multivariate analysis in all 69 patients revealed that incomplete resection, age greater than 50 years, non-smooth muscle histological feature, tumor with more than 1 mitosis, and tumor size larger than 5 cm significantly decreased survival. Complete gross surgical resection is presently the only means of cure for GIST. Tumors with more than 1 mitosis and a size larger than 5 cm have an especially poor prognosis, with decreased survival, and increased local and/or distant recurrence.


Pattern changes of EEG oscillations and BOLD signals associated with temporal lobe epilepsy as revealed by a working memory task.
It is known that the abnormal neural activity in epilepsy may be associated to the reorganization of neural circuits and brain plasticity in various ways. On that basis, we hypothesized that changes in neuronal circuitry due to epilepsy could lead to measurable variations in patterns of both EEG and BOLD signals in patients performing some cognitive task as compared to what would be obtained in normal condition. Thus, the aim of this study was to compare the cerebral areas involved in EEG oscillations versus fMRI signal patterns during a working memory (WM) task in normal controls and patients with refractory mesial temporal lobe epilepsy (MTLE) associated with hippocampal sclerosis (HS). The study included six patients with left MTLE-HS (left-HS group) and seven normal controls (control group) matched to the patients by age and educational level, both groups undergoing a blocked design paradigm based on Sternberg test during separated EEG and fMRI sessions. This test consisted of encoding and maintenance of a variable number of consonant letters on WM. EEG analysis for the encoding period revealed the presence of theta and alpha oscillations in the frontal and parietal areas, respectively. Likewise, fMRI showed the co-occurrence of positive and negative BOLD signals in both brain regions. As for the maintenance period, whereas EEG analysis revealed disappearance of theta oscillation, fMRI showed decrease of positive BOLD in frontal area and increase of negative BOLD in the posterior part of the brain. Generally speaking, these patterns of electrophysiological and hemodynamic signals were observed for both control and left-HS groups. However, the data also revealed remarkable differences between these groups that are consistent with the hypothesis of reorganization of brain circuitry associated with epilepsy.


The pharmacological profile of the acetylcholine response of a crustacean muscle.
A pharmacological analysis was made of the depolarizing acetylcholine (ACh) response found on the gastric mill I muscles of the crabs Cancer pagurus, Cancer irroratus and Cancer borealis. Acetylcholine, carbamylcholine, trimethylammonium, nicotine, and dimethyl-4-phenyl-piperazinium were effective in producing contractures and depolarizations in these muscles. No response to decamethonium, suberyldicholine, acetyl-beta-methylcholine, carbamyl-beta-methylcholine, pilocarpine and oxotremorine could be detected. High concentrations of muscarinic agonists (10(-4) to 10(-3) M) potentiated and prolonged the ACh iontophoretic response. When the acetylcholinesterase activity was inhibited with neostigmine, or when the response was elicited with carbamylcholine, muscarinic agonists partially inhibited the response. ACh responses were most effectively blocked by vertebrate nicotinic ganglionic antagonists, including dihydro-beta-erythroidine, pempidine, and mecamylamine. alpha-Bungarotoxin was without effect on the ACh response.


Changes in broiler chick tissue concentrations of lipid-soluble antioxidants immediately post-hatch.
The antioxidant protection of the chicken (Gallus gallus) embryo during incubation and early postnatal development plays an important role in chick viability. To assess the antioxidant capacity of the newly hatched chick, we determined the concentrations of vitamin A, vitamin E, carotenoids and coenzyme Q₁₀ in the major tissues of chicks which had been held in an incubator for up to 36 h post-hatch. Concentrations of total carotenoids and free retinol and retinol esters in the tissues did not differ significantly over the 36 h period post-hatch (p>0.05). In contrast concentrations of vitamin E (α-tocopherol, γ-tocopherol, and α-tocotrienol and γ-tocotrienol) in various tissues (liver, heart, brain and leg muscle) decreased significantly in chicks that had been held in the incubator for 36 h when compared to younger chicks that were held for up to 18 h. Comparatively high concentrations of coenzyme Q₁₀ were detected in the yolk sac membrane, liver and heart, the concentrations being dependent on age of chicks, the highest value being recorded 18 h post-hatch. In most of the tissues studied, coenzyme Q₁₀ concentrations decreased substantially between 18 and 36 h post-hatch. This study demonstrated that there are tissue-specific changes in the concentrations of the major antioxidants (vitamin E and coenzyme Q₁₀) during the 36 h post-hatch.


Influence of garlic (Allium sativum) on the pharmacokinetics of docetaxel.
The herbal supplement garlic (Allium sativum) is commonly used by cancer patients. Preclinical studies have shown that allicin, a major component of garlic, may affect cytochrome P450 3A4 (CYP3A4) activity. This study examines the influence of garlic supplementation on the pharmacokinetics of docetaxel, a CYP3A4 substrate. Women with metastatic breast cancer were treated with docetaxel (30 mg/m(2)) given weekly for 3 of 4 weeks. Three days after the initial dose of docetaxel, patients received 600 mg of garlic twice daily for 12 consecutive days. Docetaxel pharmacokinetics were assessed during the first three administrations. In 10 evaluable patients, the mean baseline clearance of docetaxel was 30.8 L/h/m(2) [95% confidence intervals (95% CI), 16.7-44.9]. Coadministration of garlic reduced mean clearance of docetaxel to 23.7 L/h/m(2) (95% CI, 15.5-31.8) and 20.0 L/h/m(2) (95% CI, 13.3-26.7) on days 8 and 15, respectively (P = 0.17). Additional pharmacokinetic variables of docetaxel, including peak concentration (P = 0.79), area under the curve (P = 0.36), volume of distribution (P = 0.84), and half-life (P = 0.36), were also not statistically significantly different. The mean area under the curve ratio between day 15 and day 1 was 3.74 in three individuals with the CYP3A5*1A/*1A genotype (all African American) compared with 1.02 in six individuals with the CYP3A5*3C/*3C genotype (all Caucasian). This study indicates that garlic does not significantly affect the disposition of docetaxel. However, it cannot be excluded that garlic decreases the clearance of docetaxel in patients carrying a CYP3A5*1A allele.


Characterization of gut microbiomes of household pets in the United States using a direct-to-consumer approach.
The role of gut microbiomes as important regulators of mammalian health is increasingly recognized, although feline and canine gut microbiomes remain poorly characterized. In this proof-of-concept study, we assessed the utility of a direct-to-consumer approach to executing pet microbiome studies. We characterized the gut microbiomes of 238 pets (46 cats and 192 dogs) by generating ~11 million merged reads that were mapped to the V4 region of 16S ribosomal RNA gene at a sequencing depth of 45,806 (±22,325) reads per sample. Analyses of these reads revealed that both feline and canine gut microbiomes are dominated by three major phyla, namely Firmicutes, Proteobacteria, and Bacteroides and that alpha diversity is higher in the feline gut. In addition to interspecies differences between the feline and canine gut, we also detected appreciable intraspecies bacterial variation within the canine population. While the dogs in this dataset could be assigned to three distinct clusters based on their gut microbiome, no clustering was observed within the feline population. Integration of additional data obtained from survey questionnaires revealed that geography and body weight may be associated with canine gut microbiome composition. Furthermore, we found that both the inter and intraspecies differences are more pronounced at finer taxonomic levels, indicating that strain-level investigations may be necessary in the future. This study demonstrates that the direct-to-consumer approach overcomes existing limitations in pet microbiome research, for example, it allows collection of large numbers of pet samples. The direct-to-consumer approach has proven successful in human genomics as well as human microbiomics and this study demonstrates that by building partnerships with an engaged general public this approach can also propel the field of pet microbiomics forward.


Primary Repair of Upper Extremity Peripheral Nerve Injuries: An NSQIP Analysis From 2010 to 2016.
To identify the rate of 30-day complications after primary repair of upper extremity peripheral nerve injuries, associated diagnoses, and postoperative complication rate. The American College of Surgeons National Surgical Quality Improvement Program database was reviewed from 2010 to 2016. Current Procedural Terminology codes consistent with primary nerve repair of the upper extremity were identified and included in the analysis. Patient demographics, comorbidities, type of procedure (elective/emergent), wound class, operative time, and 30-day complications were recorded. Patients with isolated upper extremity nerve injuries (isolated) were compared with those with peripheral nerve injuries in addition to bone, tendon, or soft tissue injuries (multiple). In all, 785 patients were identified as having upper extremity nerve repairs (0.16%). Of them, 64% were men and 36% were women; the average patient age was 40 years. The most common indication for surgery was injury to the digits (54% of cases). Thirty-day adverse events occurred in 3% of all cases. Isolated nerve injury occurred in 43% of patients, whereas 57% had additional injuries. The multiple injury group had a significantly higher complication rate compared with the isolated group (1% vs 4.5%) (P = .007). Repair of tendon at forearm or wrist was the most common concurrent procedure performed. Thirty-day complications among upper extremity peripheral nerve injuries are low, accounting for 3% of cases. Return to the operating room accounted for nearly half of all complications. Patients in the multiple injury group accounted for more than half of these and had a significantly higher complication rate compared with patients with isolated nerve injuries.


Use of nuclear receptor luciferase-based bioassays to detect endocrine active chemicals in a biosolids-biochar amended soil.
Biosolids are a potentially valuable source of carbon and nutrients for agricultural soils; however, potential unintended impacts on human health and the environment must be considered. Virtually all biosolids contain trace amounts endocrine-disrupting chemicals derived from human use of pharmaceuticals and personal care products (PPCPs). One potential way to reduce the bioavailability of PPCPs is to co-apply biosolids with biochar to soil, because biochar's chemical (e.g., aromaticity) and physical properties (e.g., surface area) give it a high affinity to bind many organic chemicals in the environment. We developed a soil-specific extraction method and utilized a luciferase-based bioassay (CALUX) to detect endocrine active chemicals in a biosolids-biochar co-amendment soil greenhouse study. Both biochar (walnut shell, 900 °C) and biosolids had positive impacts on carrot and lettuce biomass accumulation over our study period. However, the walnut shell biochar stimulated aryl hydrocarbon receptor activity, suggesting the presence of potential endocrine active chemicals in the biochar. Since the biochar rate tested (100 t ha-1) is above the average agronomic rate (10-20 t ha-1), endocrine effects would not be expected in most environmental applications. The effect of high temperature biochars on endocrine system pathways must be explored further, using both quantitative analytical tools to identify potential endocrine active chemicals and highly sensitive bioanalytical assays such as CALUX to measure the resulting biological activity of such compounds.


[Detection of toxin-producing pathogenic bacterial strains by polymerase chain reaction].
Polymerase chain reaction (PCR) was used for detection of pathogenic Clostridium botulinum, Clostridium perfringens, Clostridium difficile, and Escherichia coli. With this aim in view, primers to botulinic toxins types A, B, C1, D, E, F, and G, perfringens enterotoxin, difficile toxin, and types 1 and 2 Shigella-like toxins were chosen and synthesized. Optimal amplification conditions were selected for each pair of primers, with DNA and the respective agent as the reaction mixture matrices. PCR was highly specific and sensitive in all cases. Its sensitivity was 10-100 cells/sample. Among the tested C. botulinum, C. perfringens, C. difficile, and E. coli strains, specific amplification products of expected size were observed only in the strains containing the respective toxin genes. These findings recommend the use of these methods in clinical microbiology. Strains containing type 2 Shigella-like toxin gene were detected among E. coli strains isolated from patients with the hemolytic uremic syndrome, which for the first time indicates that the problem with E. coli epidemic strain O157 is valid for Russia. As a result of our studies, test systems for detection of types A, B, C, D, E, F, and G C. botulinum strains, C. perfringens and C. difficile, and E. coli O157 strains are now available.