Analysis of the factors affecting the loss of correction effect in patients with congenital scoliosis after one stage posterior hemivertebrae resection and orthosis fusion.
To analyze the factors affecting the loss of correction effect in patients with congenital scoliosis after one stage posterior hemivertebra resection, orthosis, fusion and internal fixation. Thirty-nine patients with congenital scoliosis (CS) who underwent one-stage posterior hemivertebra resection, orthosis, fusion and internal fixation were retrospectively included in Hebei Children's Hospital General demographic information of patients was collected. Preoperative and postoperative imaging indicators were compared, Including cobb Angle of the main curvature of the spine, segmental Cobb Angle, compensatory cephalic curve, compensatory curve on the caudal side, segmental kyphosis, coronal balance, sagittal balance, thoracic kyphosis, lumbar lordosis, and apical vertebra translation. Correlation analysis is used to evaluate the factors affecting the loss of judgment and correction effect, and the correlation indicators are included in the multi-factor Logistics regression. In terms of radiographic indicators in the coronal plane, compared to preoperative values, significant improvements were observed in postoperative Cobb Angle of main curve (8.00°±4.62° vs. 33.30°±9.86°), Segmental Cobb angle (11.87°±6.55° vs. 31.29°±10.03°), Compensatory cephalic curve (6.22°±6.33° vs. 14.75°±12.50°), Compensatory curve on the caudal side (5.58°±3.43° vs. 12.61°±8.72°), coronal balance (10.95 mm ± 8.65 mm vs. 13.52 mm ± 11.03 mm), and apical vertebra translation (5.96 mm ± 5.07 mm vs. 16.55 mm ± 8.39 mm) (all P < 0.05). In the sagittal plane, significant improvements were observed in Segmental kyposis Angle (7.60°±9.36° vs. 21.89°±14.62°, P < 0.05) as compared to preoperative values. At the last follow-up, Segmental kyphosis Angle (6.09°±9.75° vs. 21.89°±14.62°, P < 0.05), Thoracic kyphosis (26.57°±7.68° vs. 24.06°±10.49°, P < 0.05) and Lumbar lordosis (32.12°±13.15° vs. 27.84°±16.68°, P < 0.05) had statistical significance compared with the preoperative department. The correlation analysis showed that the correction effect of the main curve Cobb angle was correlated with fixed segment length (rs=-0.318, P = 0.048), postoperative segment Cobb angle (rs=-0.600, P < 0.001), preoperative apical vertebra translation (rs = 0.440, P = 0.005), and spinal cord malformation (rs=-0.437, P = 0.005). The correction effect of segmental kyphosis was correlated with age (rs = 0.388, P = 0.037). The results of the multivariate logistic regression analysis revealed that postoperative segmental Cobb angle > 10° (OR = 0.011, 95%CI:0.001-0.234, P = 0.004), associated spinal cord anomalies (OR = 24.369, 95%CI:1.057-561.793, P = 0.046), and preoperative apical translation > 10 mm (OR = 0.012, 95%CI:0.000-0.438, P = 0.016) were influential factors in the progression of the main curve Cobb angle. The one-stage posterior hemivertebra resection and short-segment corrective fusion with internal fixation are effective means to treat congenital scoliosis. However, attention should be paid to the loss of correction and curve progression during follow-up. Patients with spinal cord malformation and a large preoperative apical vertebra translation have a greater risk of losing the correction after surgery.


Is there a role for liquid biopsy in the surveillance of parathyroid carcinoma?
Parathyroid carcinoma is a rare malignancy with high recurrence rates. Liquid biopsy is a stratifying tool in disease recurrence/progression in other malignant processes. This study sought to assess the feasibility and application of liquid biopsy in parathyroid carcinoma and its impact on surveillance. Retrospective review of a prospectively maintained database of adults treated for parathyroid carcinoma at a tertiary care center (2017-2023). Demographics, clinical characteristics, and laboratory variables were collected. Circulating cell-free deoxyribonucleic acid enrichment and circulating tumor cell enumeration were obtained from serial blood samples. A total of 25 patients were identified-64% were male patients, with a median age of 56 years (interquartile range 45-63). Fifty blood samples were collected postoperatively. At first, circulating tumor cell enumeration, 56% (14/25) of patients had no evidence of disease, and 32% (8/25) had distant metastasis. Median follow-up was 53 months (interquartile range 23-107). At the last follow-up, 40% (10/25) of patients were found to have distant metastasis. Serial circulating tumor cell enumeration was performed in 52% of patients, median highest circulating tumor cell was (interquartile range 1-22). Circulating cell-free deoxyribonucleic acid was assessed in 64% of patients (16/25). There was no difference in circulating tumor cells or circulating cell-free deoxyribonucleic acid between those with distant metastasis and those without distant metastasis. The most common mutation identified was TP53, present in 88% of circulating cell-free deoxyribonucleic acid samples with a mutation. Circulating cell-free deoxyribonucleic acid and parathyroid hormone levels were not found to have any association (r = -0.27, P = .39), but parathyroid hormone and circulating tumor cell had a linear relationship (r = 0.76, P < .001). Liquid biopsy appears to be a feasible tool in parathyroid carcinoma surveillance. The relationship between circulating cell-free deoxyribonucleic acid and parathyroid hormone levels remains unclear, and the association between circulating tumor cell enumeration and parathyroid hormone levels may be impactful. The finding that TP53 mutation is more prevalent in patients with distant metastasis may impact further management.


Cytochrome P450 CYP1A1: wider roles in cancer progression and prevention.
CYP1A1 is one of the main cytochrome P450 enzymes, examined extensively for its capacity to activate compounds with carcinogenic properties. Continuous exposure to inhalation chemicals and environmental carcinogens is thought to increase the level of CYP1A1 expression in extrahepatic tissues, through the aryl hydrocarbon receptor (AhR). Although the latter has long been recognized as a ligand-induced transcription factor, which is responsible for the xenobiotic activating pathway of several phase I and phase II metabolizing enzymes, recent evidence suggests that the AhR is involved in various cell signaling pathways critical to cell cycle regulation and normal homeostasis. Disregulation of these pathways is implicated in tumor progression. In addition, it is becoming increasingly evident that CYP1A1 plays an important role in the detoxication of environmental carcinogens, as well as in the metabolic activation of dietary compounds with cancer preventative activity. Ultimately the contribution of CYP1A1 to cancer progression or prevention may depend on the balance of procarcinogen activation/detoxication and dietary natural product extrahepatic metabolism.


List-mode likelihood.
As photon-counting imaging systems become more complex, there is a trend toward measuring more attributes of each individual event. In various imaging systems the attributes can include several position variables, time variables, and energies. If more than about four attributes are measured for each event, it is not practical to record the data in an image matrix. Instead it is more efficient to use a simple list where every attribute is stored for every event. It is the purpose of this paper to discuss the concept of likelihood for such list-mode data. We present expressions for list-mode likelihood with an arbitrary number of attributes per photon and for both preset counts and preset time. Maximization of this likelihood can lead to a practical reconstruction algorithm with list-mode data, but that aspect is covered in a separate paper [IEEE Trans. Med. Imaging (to be published)]. An expression for lesion detectability for list-mode data is also derived and compared with the corresponding expression for conventional binned data.


Sex-specific Evaluation of Ventricular Ejection Fraction and End-Systolic Volume Applied to Cardiac Resynchronization Therapy.
Cardiac resynchronization therapy (CRT) can decrease the risk of heart failure (HF) events in relatively asymptomatic patients with a reduced ejection fraction (EF) and wide QRS complex. However, individual response to this type of therapy varies widely. Often based on either EF increase or end-systolic volume (ESV) decrease as criterion, a subgroup of super-responders has been described. Therefore, it is important to determine factors that can predict a favorable response and identify those patients who may benefit from CRT. With this goal in mind we explored the possible role of ESV.To improve insight in ventricular pump function we previously introduced the volume regulation graph (VRG), relating ESV to end-diastolic volume (EDV). An individual patient is uniquely defined by the prevailing working point in the volume domain. The traditional metric EF can be graphically derived for each working point. The nonlinear association between EF and ESV is given by EF = 1 + γ {ESV / (δ - ESV)}, with empirical constants γ and δ. The impact of CRT super-responders on EF can be evaluated, taking into account sex-specific ESV values. Based on available regression equations we modeled the impact on EF (as percent points) resulting from CRT-induced fractional ESV changes expressed as % of baseline ESV. Our analysis confirms clinical findings, indicating that CRT super-responders are likely to be women, and clarify why a specific reduction of ESV cannot be directly translated into EF improvement. We propose that the EF as CRT criterion should be abandoned and replaced by sex-specific ESV evaluations.Clinical Relevance- Response to CRT should be evaluated in a sex-specific manner. The smaller heart size in women has implications for the interpretation of percentwise reductions of ESV and their translation into an associated increase of EF.


Structure, function and gene expression of epithelial mucins.
In this review the main characteristics, i.e., structure, function and gene expression, of the different mucins are discussed. Mucin-type molecules consist of a core protein moiety (apomucin) where a number of carbohydrate chains are attached to serines and threonines by glycosidic bonds. O-linked carbohydrates form up to 80% of the molecule and the length of the glucidic side chains varies from one to more than 20 residues. At least eight mucin-like genes have been isolated so far, and the main characteristic is the presence of a central domain composed of a variable number of "tandem repeats". The sequence homology of the central domain among the different members of the mucin-type family is limited, indicating that this internal domain is unique for each mucin. Thanks to the integrated results of genetic, immunological and biochemical studies, it is now possible to identify eight apomucin genes, namely MUC1, MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC6 and MUC7. MUC1 is the best characterized mucin and it is expressed on the apical surface of most polarized epithelial cells. The MUC1 gene has been cloned and sequenced. The MUC2 gene encodes a typical secretory gel-forming mucin which represents the predominant form in human intestinal and colon tissues. Another intestinal mucin is MUC3. The MUC4, MUC5AC and MUC5B genes have been isolated from a bronchial tissue cDNA library. The MUC4 and MUC5AC genes are mainly expressed in the respiratory tract, in gastric and reproductive mucosa, while MUC5B is highly detectable only in the bronchial glands. The MUC6 gene is expressed by gastric tissue and, recently, MUC7 has been cloned and sequenced using a salivary cDNA library.


Preclinical and dose-ranging assessment of hESC-derived dopaminergic progenitors for a clinical trial on Parkinson's disease.
Human embryonic stem cell (hESC)-derived midbrain dopaminergic (mDA) cell transplantation is a promising therapeutic strategy for Parkinson's disease (PD). Here, we present the derivation of high-purity mDA progenitors from clinical-grade hESCs on a large scale under rigorous good manufacturing practice (GMP) conditions. We also assessed the toxicity, biodistribution, and tumorigenicity of these cells in immunodeficient rats in good laboratory practice (GLP)-compliant facilities. Various doses of mDA progenitors were transplanted into hemi-parkinsonian rats, and a significant dose-dependent behavioral improvement was observed with a minimal effective dose range of 5,000-10,000 mDA progenitor cells. These results provided insights into determining a low cell dosage (3.15 million cells) for human clinical trials. Based on these results, approval for a phase 1/2a clinical trial for PD cell therapy was obtained from the Ministry of Food and Drug Safety in Korea, and a clinical trial for treating patients with PD has commenced.


High performance organic field-effect transistors based on single-crystal microribbons and microsheets of solution-processed dithieno[3,2-b:2',3'-d]thiophene derivatives.
We present π-conjugated dithieno[3,2-b:2',3'-d]thiophene derivatives that act as high-performance p-type organic semiconductors. These molecules self-organize into single-crystal microribbons or microsheets. High carrier mobilities of up to 10.2 cm(2) V(-1) s(-1) and high on/off ratios of ~10(7) have been achieved in organic single-crystal field-effect transistors.


Sport Specialization Behaviors Are Associated With History of Reported Injury in Youth Basketball.
There is significant emerging evidence that early sport specialization is a potential risk factor for injury in youth sports. Despite basketball being the most popular youth team sport in the United States, sport specialization research, specifically in youth basketball players, has been limited. The purpose of this paper was to examine the association of sport specialization behaviors with injury history by surveying a nationally representative sample of parents of youth basketball athletes. We hypothesized that athletes who specialized in basketball, participated on multiple teams at the same time, and traveled regularly for basketball competitions would be more likely to report a basketball-related injury in the previous year. A nationally representative sample of 805 parents of 805 youth basketball players (female N=241, 29.9%; age: 12.9±2.5 y old) completed an online questionnaire that had 3 sections: (1) parent/child demographics, (2) child basketball participation information for the previous year, and (3) child basketball injury history in the previous year. Multivariate logistic regression examined the associations between variables of interest and injury history, adjusting for covariates. Odds ratios (ORs) with 95% confidence intervals (95% CI) were calculated for the variables of interest from the logistic regression model. Highly specialized athletes were more likely than low specialization athletes to report history of basketball injury in the previous year [OR (95% CI): 2.47 (1.25-4.88), P=0.009]. The odds of reporting an injury in the previous year were twice as great among athletes who played on a basketball team at the same time as another sport team compared with those who played basketball only [OR (95% CI): 1.98 (1.30-3.01), P=0.001]. The odds of reporting an injury in the previous year were 3 times greater among athletes who received private coaching compared with those who did not receive private coaching [OR (95% CI): 2.91 (1.97-4.31), P<0.001]. Specialization in basketball, along with several other behaviors that have become typical of modern youth sport participation, were associated with reported injury history. Further prospective research is necessary to determine whether sport specialization behaviors increase the risk of injury in youth basketball. Level III-cross-sectional study.


Proneural clusters: equivalence groups in the epithelium of Drosophila.
The segregation of neural precursors from epidermal cells during development of the nervous system of Drosophila relies on interactions between cells that are thought to be initially equivalent. During development of the adult peripheral nervous system, failure of the cellular interactions leads to the differentiation of a tuft of sensory bristles at the site where usually only one develops. It is thus thought that a group of cells at that site (a proneural cluster) has the potential to make a bristle but that in normal development only one cell will do so. The question addressed here is do these cells constitute an equivalence group (Kimble, J., Sulston, J. and White, J. (1979). In Cell Lineage, Stem Cells and Cell Determination (ed. N. Le Douarin). Inserm Symposium No. 10 pp. 59-68, Elsevier, Amsterdam)? Within clusters mutant for shaggy, where several cells of a cluster follow the neural fate and differentiate bristles, it is shown that these display identical neuronal specificity: stimulation of the bristles evoke the same leg cleaning response and backfilling of single neurons reveal similar axonal projections in the central nervous system. This provides direct experimental evidence that the cells of a proneural cluster are developmentally equivalent.


A polymerase chain reaction-based methodology to detect gene doping.
The non-therapeutic use of genes to enhance athletic performance (gene doping) is a novel threat to the world of sports. Skeletal muscle is a prime target of gene therapy and we asked whether we can develop a test system to produce and detect gene doping. Towards this end, we introduced a plasmid (pCMV-FAK, 3.8 kb, 50 μg) for constitutive expression of the chicken homologue for the regulator of muscle growth, focal adhesion kinase (FAK), via gene electro transfer in the anti-gravitational muscle, m. soleus, or gastrocnemius medialis of rats. Activation of hypertrophy signalling was monitored by assessing the ribosomal kinase p70S6K and muscle fibre cross section. Detectability of the introduced plasmid was monitored with polymerase chain reaction in deoxyribonucleic acids (DNA) from transfected muscle and serum. Muscle transfection with pCMV-FAK elevated FAK expression 7- and 73-fold, respectively, and increased mean cross section by 52 and 16% in targeted muscle fibres of soleus and gastrocnemius muscle 7 days after gene electro transfer. Concomitantly p70S6K content was increased in transfected soleus muscle (+110%). Detection of the exogenous plasmid sequence was possible in DNA and cDNA of muscle until 7 days after transfection, but not in serum except close to the site of plasmid deposition, 1 h after injection and surgery. The findings suggest that the reliable detection of gene doping in the immoral athlete is not possible unless a change in the current practice of tissue sampling is applied involving the collection of muscle biopsy close to the site of gene injection.


Multicenter evaluation of the Glucometer Elite XL meter, an instrument specifically designed for use with neonates.
To evaluate the clinical performance of the Glucometer Elite XL Diabetes Care System in neonatal settings using a multicenter study A total of 388 blood specimens from 333 neonates were included in the study. A capillary or arterial sample was analyzed for determination of glucose with the Glucometer Elite XL system by an attending trained nurse. Through the same sampling site, a specimen was collected and sent to the laboratory for measurement of plasma glucose, bilirubin, and hematocrit. The regression analysis between the results of the Glucometer Elite XL system and comparative methods resulted in the following: Glucometer Elite XL meter = 1.01 x laboratory method + 0.02 mmol/l (n = 388). For the 1.1-4.0 mmol/l plasma glucose range, the regression was Glucometer Elite XL meter = 1.07 x laboratory method + 0.12 mmol/l (n = 150). A difference plot indicated a mean bias of 0.04 mmol/l (95% CI -0.01 to 0.10). No relationship was found between meter glucose biases and hematocrit levels (r = 0.10, P = 0.14). Although a statistically significant correlation existed between bilirubin levels and the glucose meter biases (r = 0.14, P = 0.005), the predicted mean biases were of little clinical significance. The Glucometer Elite XL system showed a good performance when used in neonatal settings.


Immunological response and overall survival in a subset of advanced renal cell carcinoma patients from a randomized phase 2/3 study of naptumomab estafenatox plus IFN-α versus IFN-α.
Naptumomab estafenatox/ABR-217620/ANYARA (Nap) has been evaluated in clinical phase 1 and 2/3 studies. RCC patients in the phase 2/3 trial were randomized 1:1 in an open label study to receive Nap+IFN-α or IFN-α. In this study, we analyzed the UK patients for their immunological response in relation to prolonged overall survival (OS). We found that Nap-specific T cells were reduced after 3 treatment days in patients' peripheral blood. Levels of both Nap-specific CD4+ and CD8+ T cells were significantly higher 8 days after the first treatment. Patients with such pattern of reduction and expansion of Nap-binding T cells also showed increased levels of IL-2 and IFN-γ in plasma 3 hours after the first Nap treatment. In addition, Nap caused an increase of IL-6, IL-10 and TNF-α. The patients in the UK subset showed a tendency of OS benefit after Nap treatment. Most Nap treated patients with long OS had low baseline IL-6 and normal levels of anti-SEA/E-120 antibodies. Furthermore, patients with pronounced Nap induced IL-2 and T cell expansion had long OS. In conclusion, patients with low baseline IL-6 and normal anti-SEA/E-120 may respond well to Nap by T cell activation and expansion paving the way for anti-tumour effects.


Altered retrobulbar hemodynamics in patients who have transient monocular blindness without carotid stenosis.
This study was to evaluate the retrobulbar hemodynamics in patients who have transient monocular blindness (TMB) without carotid stenosis. Fifty-nine patients who have TMB without carotid stenosis were studied along with 59 age- and sex-matched controls. Color Doppler-imaging was used to study the retrobulbar hemodynamic by measuring the flow velocities (peak-systolic velocity, and end-diastolic velocity), vascular resistance indices (pulsatility index, and resistance index) in central retinal arteries, short posterior ciliary arteries, and ophthalmic arteries. The patients were divided into 2 groups according to the attack frequency: group 1 (occasional TMB, 2 or fewer attacks, 26 patients) and group 2 (frequent TMB, 3 or more attacks, 33 patients). The risk factors for atherosclerosis were similar between the cases and controls. The means of end-diastolic velocity were significantly lower in central retinal arteries and ophthalmic arteries, and the pulsatility index and resistance index were significantly higher in all the 3 retrobulbar vessels in TMB patients than for the controls. The differences between patients and controls were greater for the group-2 patients. Patients who have TMB without carotid stenosis had altered retrobulbar hemodynamics with a generalized increase in vascular resistance in the retrobulbar arteries. The role of venous hypertension as an etiology needs further study.


Shedding of Epstein-Barr virus and cytomegalovirus from the genital tract of women in a periurban community in Andhra Pradesh, India.
We found a large number of false-positive readings by visual inspection with acetic acid (VIA) in a study of cervical cancer screening strategies (VIA, human papillomavirus HPV DNA testing, and Pap cytology) in a periurban community in Andhra Pradesh, India. We evaluated whether these false-positive readings might be occurring as a result of infections with Epstein-Barr virus (EBV) or cytomegalovirus (CMV), prevalent latent herpesviruses known to be shed from the female genital tract. While we found that there was no association between VIA results and the presence of EBV or CMV in the cervix, we did find a high prevalence of both viruses: 20% for EBV and 26% for CMV. In multivariate analyses, CMV prevalence was associated with younger age, lack of running water in the home, and visually apparent cervical inflammation. EBV prevalence was associated with older age and a diagnosis of cervical intraepithelial neoplasia grade 1 or greater. The biological and clinical implications of these viruses at the cervix remain to be determined. The strong association between the presence of EBV and cervical disease warrants future exploration to determine whether EBV plays a causal role in disease development or if it is merely a bystander in the process.


Laparoscopic relief of obstructing folded muscular cuff after transanal pull-through for aganglionosis.
Transanal endorectal resection and colonic pull-through (TERPT) is a good technique for the management of Hirschsprungs disease. This procedure is feasible in the vast majority of patients and is associated with excellent results, early postoperative recovery, and no visible scars. We report the case of a patient who developed early postoperative severe constipation after TERPT due to unusual folding of the muscular cuff rim, which tightly narrowed the pulled-through colon. This complication was diagnosed and treated by laparoscopy. To prevent it, we recommend splitting of the aganglionic muscular cuff during TERPT.


Localization of Burkholderia cepacia complex bacteria in cystic fibrosis lungs and interactions with Pseudomonas aeruginosa in hypoxic mucus.
The localization of Burkholderia cepacia complex (Bcc) bacteria in cystic fibrosis (CF) lungs, alone or during coinfection with Pseudomonas aeruginosa, is poorly understood. We performed immunohistochemistry for Bcc and P. aeruginosa bacteria on 21 coinfected or singly infected CF lungs obtained at transplantation or autopsy. Parallel in vitro experiments examined the growth of two Bcc species, Burkholderia cenocepacia and Burkholderia multivorans, in environments similar to those occupied by P. aeruginosa in the CF lung. Bcc bacteria were predominantly identified in the CF lung as single cells or small clusters within phagocytes and mucus but not as "biofilm-like structures." In contrast, P. aeruginosa was identified in biofilm-like masses, but densities appeared to be reduced during coinfection with Bcc bacteria. Based on chemical analyses of CF and non-CF respiratory secretions, a test medium was defined to study Bcc growth and interactions with P. aeruginosa in an environment mimicking the CF lung. When test medium was supplemented with alternative electron acceptors under anaerobic conditions, B. cenocepacia and B. multivorans used fermentation rather than anaerobic respiration to gain energy, consistent with the identification of fermentation products by high-performance liquid chromatography (HPLC). Both Bcc species also expressed mucinases that produced carbon sources from mucins for growth. In the presence of P. aeruginosa in vitro, both Bcc species grew anaerobically but not aerobically. We propose that Bcc bacteria (i) invade a P. aeruginosa-infected CF lung when the airway lumen is anaerobic, (ii) inhibit P. aeruginosa biofilm-like growth, and (iii) expand the host bacterial niche from mucus to also include macrophages.


Brain stem auditory-evoked responses in the dog.
Brain stem auditory-evoked responses (BAER) were recorded from 58 dogs that did not have a known history of hearing problems. The BAER wave forms had an overall mean amplitude approximately 3.0 microV and typically consisted of a series of 4 to 5 vertex-positive peaks (peaks I through V). When acoustic clicks having intensities of 60-dB hearing level (decibels relative to the subjective hearing threshold) were used as stimuli, peak I had a latency of 1.49 +/- 0.13 ms; peak II, 2.32 +/- 0.14 ms; peak III, 3.01 +/- 0.25 ms; peak IV, 4.22 +/- 0.27 ms; and peak V, 5.55 +/- 0.37 ms. Latency values were influenced by a number of nonpathologic factors, including stimulus intensity and the body temperature of the dog. As stimulus intensity was decreased, there was a lengthening of the latency of each peak coupled with a decrease in the overall amplitude of BAER. Decreases in rectal temperature caused a similar lengthening of peak latencies. Age may have an influence on BAER, but under the conditions of the present study, the effect was not significant.


Extraction of Microbial and Host DNA, RNA, and Proteins from Oak Bark Tissue.
The application of high-throughput nucleic acid and protein sequencing technologies is transforming our understanding of plant microbiomes and their interactions with their hosts in health and disease. However, progress in studying host-microbiome interactions in above-ground compartments of the tree (the phyllosphere) has been hampered due to high concentrations of phenolic compounds, lignin, and other compounds in tree bark that severely limit the success of DNA, RNA, and protein extraction. Here we present modified sample-preparation and kit-based protocols for the extraction of host and microbiome DNA and RNA from oak (Quercus robus and Quercus petraea) bark tissue for subsequent high-throughput sequencing. In addition, reducing the quantity of bark tissue used for an established protein extraction protocol yielded high quality protein for parallel analysis of the oak-microbiota metaproteome. These procedures demonstrate the successful extraction of nucleic acids and proteins from oak tissue using as little as 50 mg of sample input, producing sufficient quantities for nucleic acid sequencing and protein mass spectrometry of tree stem tissues and their associated microbiota.


Prediction of smallpox outbreak and evaluation of control-measure policy in Japan, using a mathematical model.
Since the September 1 terrorist attacks and moreover, since the anthrax exposure events in 2001 in the United States, bioterrorism attacks seem to be a real threat. Of course, the public health authorities in Japan have started to prepare control measures for such events. We report here our attempts, using a mathematical model, to estimate outbreak size and to examine the most effective measures; comparing ring vaccination (contact tracing, isolation, and vaccination among contacts) and mass vaccination of the susceptible population in the area. The basic framework of the mathematical model follows a model used in previous research. The initial susceptible population is assumed to be 30 million persons. Concerning the important parameters, such as the number of initial-exposure cases, R0 (infectious power, or natural history) and, the starting day of intervention after the initial exposure, we checked the robustness of our conclusions by sensitivity analysis. We found that mass vaccination is preferable to ring vaccination when the values for the initial-exposure cases and R0 are high and when the start of intervention by public health authorities is delayed. In the base-case situation, the mass vaccination strategy needs almost 30 million vaccine doses. On the other hand, though ring vaccination needs fewer doses, it needs fewer than 50,000 doses in the worst-case scenario, that with larger first exposure, higher R0, or later start of public health authority intervention. This mathematical model can measure the prevalence of an infectious disease and can evaluate control measures for it before an outbreak. Especially, it is useful for the planning of the outbreaks of emerging diseases such as severe acute respiratory syndrome (SARS) or for bioterrorism attacks involving such diseases as smallpox. In further research, we will have to take into account the population people vaccinated of for smallpox, who account for about 70% of the total population in Japan.


Impact of extended maceration and regulated deficit irrigation (RDI) in Cabernet Sauvignon wines: characterization of proanthocyanidin distribution, anthocyanin extraction, and chromatic properties.
The impact of extended maceration (EM) was studied in Cabernet Sauvignon grapes sourced from a vineyard subjected to four regulated deficit irrigation (RDI) treatments: (I) 100% replenishment of crop evapotranspiration (100% ETc), (II) 70% ETc, (III) 25% ETc until véraison, followed by 100% ETc until harvest, and IV) 25% ETc. Each vineyard replicate was made into wine with two replicates designated as controls (10-day skin contact) and two as extended maceration (EM, 30-day skin contact). The mean degree of polymerization (mDP), size distribution, concentration, and composition of wine proanthocyanidins (PAs) and monomeric flavan-3-ols of 90 fractions were characterized by preparative and analytical HPLC techniques. The maceration length imparted a larger effect on most chemical parameters. The RDI treatment had no effect on the extraction patterns of anthocyanins, PAs, and/or on the origin of the PAs extracted into the wines. Conversely, EM led to anthocyanin losses and increased PA extraction during maceration, with ~73% of seed-derived PAs. Accordingly, the concentration of monomeric flavan-3-ols, oligomeric (2 ≤ mDP < 5) and polymeric PAs (mDP ≥ 5) was higher in EM wines. The size distribution of the wines' PAs revealed two major peaks as a function of concentration at mDP 2 (22-27% of total PAs mass) and at mDP 6-7 (12-17% of total PAs mass) and was found to follow a non-normal Rayleigh-type distribution.


Genetic structure of populations of the threatened eastern massasauga rattlesnake, Sistrurus c. catenatus: evidence from microsatellite DNA markers.
Throughout its distribution in North America, the threatened eastern massasauga rattlesnake (Sistrurus c. catenatus) persists in a series of habitat-isolated disjunct populations of varying size. Here, we use six microsatellite DNA loci to generate information on the degree of genetic differentiation between, and the levels of inbreeding within populations to understand how evolutionary processes operate in these populations and aid the development of conservation plans for this species. Samples were collected from 199 individuals from five populations in Ontario, New York and Ohio. Our results show that all sampled populations: (i) differ significantly in allele frequencies even though some populations are < 50 km apart, and may contain genetically distinct subpopulations < 2 km apart; (ii) have an average of 23% of alleles that are population specific; and (iii) have significant FIS values (mean overall FIS = 0.194) probably due to a combination of Wahlund effects resulting from fine-scale genetic differentiation within populations and the presence of null alleles. Our results imply that massasauga populations may be genetically structured on an extremely fine scale even within continuous populations, possibly due to limited dispersal. Additional information is needed to determine if dispersal and mating behaviour within populations can account for this structure and whether the observed differentiation is due to random processes such as drift or to local adaptation. From a conservation perspective, our results imply that these massasauga populations should be managed as demographically independent units and that each has high conservation value in terms of containing unique genetic variation.


Nose-To-Brain Delivery of PLGA-Diazepam Nanoparticles.
The objective of the present investigation was to optimize diazepam (Dzp)-loaded poly(lactic-co-glycolic acid) nanoparticles (NP) to achieve delivery in the brain through intranasal administration. Dzp nanoparticles (DNP) were formulated by nanoprecipitation and optimized using Box-Behnken design. The influence of various independent process variables (polymer, surfactant, aqueous to organic (w/o) phase ratio, and drug) on resulting properties of DNP (z-average and drug entrapment) was investigated. Developed DNP showed z-average 148-337 d.nm, polydispersity index 0.04-0.45, drug entrapment 69-92%, and zeta potential in the range of -15 to -29.24 mV. Optimized DNP were further analyzed by differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), ex-vivo drug release, and in-vitro cytotoxicity. Ex-vivo drug release study via sheep nasal mucosa from DNP showed a controlled release of 64.4% for 24 h. 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay performed on Vero cell line showed less toxicity for DNP as compared to Dzp suspension (DS). Gamma scintigraphy and biodistribution study of DNP and DS was performed on Sprague-Dawley rats using technetium-99m-labeled ((99m)Tc) Dzp formulations to investigate the nose-to-brain drug delivery pathway. Brain/blood uptake ratios, drug targeting efficiency, and direct nose-to-brain transport were found to be 1.23-1.45, 258, and 61% for (99m)Tc-DNP (i.n) compared to (99m)Tc-DS (i.n) (0.38-1.06, 125, and 1%). Scintigraphy images showed uptake of Dzp from nose-to-brain, and this observation was in agreement with the biodistribution results. These results suggest that the developed poly(D,L-lactide-co-glycolide) (PLGA) NP could serve as a potential carrier of Dzp for nose-to-brain delivery in outpatient management of status epilepticus.


The effect of diffusible acids on potassium ion uptake by yeast.
1. When yeast oxidizes ethanol at different pH values the uptake of K(+) corresponds closely to the amount of acetate accumulated at each pH value. 2. The addition of semicarbazide to the suspension buffered at pH4.75 inhibited both the K(+) uptake and the acetate accumulation by about 50%. 3. The addition of either acetate or propionate to the suspensions markedly increased the K(+) uptake. 4. The addition of acetate to the suspensions lowered the intracellular pH of the yeast from a resting value of pH5.80 to 5.56. 5. The ratio of the initial rate of K(+) uptake to O(2) consumption was 0.77. This ratio was increased to 1.77 in the presence of 10mmol of propionate/l.


The time-course of broiler intestinal microbiota development after administration of cecal contents to incubating eggs.
The microbial populations that inhabit the gastrointestinal tract (GIT) are known to influence the health and growth performance of the host. Clean hatcheries and machine-based incubation practices in the commercial poultry industry can lead to the acquisition of aberrant microbiota in the GIT of chickens and a very high level of bird-to-bird variation. The lack of microbial profile flock uniformity presents challenges for harnessing and manipulating intestinal bacteria to better serve the host. Cecal contents from high or low performing chickens were used to inoculate the surface of eggs prior to hatching and then the initial gut colonisation was monitored and subsequent changes in gut microbiota composition were followed over time. Two different cecal treatment groups were compared to an untreated control group (n = 32). Bacterial communities were characterised using high-throughput 16S rRNA gene sequencing techniques. Cecal microbiota transfer via egg surface application did not transfer the performance profile of the donors to the recipient birds. One of the cecal inoculations provided a more uniform gut microbiota, but this was not reproduced in the second group with a different inoculum. Development of the intestinal community was reproducible in all three groups with some genera like Lactobacillus showing no change, others like Faecalibacterium increased in abundance slowly and steadily over time and others like Enterobacter were abundant only in the first days of life. The cecal treatment reduced bird-to-bird variation in microbiota composition. Although the high FCR performance of donor birds was not transferred with the cecal microbiota, all three groups, including the control, performed better than standard for the breed. The pattern of microbiota development was similar in all three flocks, indicating that the normal processes of microbiota acquisition largely swamped any effect of the cecal material applied to eggs.


Biochemical indicators for the bioavailability of organic carbon in ground water.
The bioavailability of total organic carbon (TOC) was examined in ground water from two hydrologically distinct aquifers using biochemical indicators widely employed in chemical oceanography. Concentrations of total hydrolyzable neutral sugars (THNS), total hydrolyzable amino acids (THAA), and carbon-normalized percentages of TOC present as THNS and THAA (referred to as "yields") were assessed as indicators of bioavailability. A shallow coastal plain aquifer in Kings Bay, Georgia, was characterized by relatively high concentrations (425 to 1492 microM; 5.1 to 17.9 mg/L) of TOC but relatively low THNS and THAA yields (approximately 0.2%-1.0%). These low yields are consistent with the highly biodegraded nature of TOC mobilized from relatively ancient (Pleistocene) sediments overlying the aquifer. In contrast, a shallow fractured rock aquifer in West Trenton, New Jersey, exhibited lower TOC concentrations (47 to 325 microM; 0.6 to 3.9 mg/L) but higher THNS and THAA yields (approximately 1% to 4%). These higher yields were consistent with the younger, and thus more bioavailable, TOC being mobilized from modern soils overlying the aquifer. Consistent with these apparent differences in TOC bioavailability, no significant correlation between TOC and dissolved inorganic carbon (DIC), a product of organic carbon mineralization, was observed at Kings Bay, whereas a strong correlation was observed at West Trenton. In contrast to TOC, THNS and THAA concentrations were observed to correlate with DIC at the Kings Bay site. These observations suggest that biochemical indicators such as THNS and THAA may provide information concerning the bioavailability of organic carbon present in ground water that is not available from TOC measurements alone.


Antiplasticization and local elastic constants in trehalose and glycerol mixtures.
We have performed molecular dynamics simulations of glassy trehalose with various amounts of glycerol in order to explore the tendency for glycerol to antiplasticize the glass. We find that below a temperature of 300 K, the average density of the system containing 5%(wt) glycerol is larger than that of the pure trehalose system; the glass transition temperature is decreased, and the elastic constants are essentially unchanged. Taken together, these phenomena are indicative of mild antiplasticization, a type of behavior generally observed in polymeric systems. We have calculated the local elastic constants in our glassy materials and, consistent with previous simulations on a coarse-grained polymer, we find evidence of domains having negative elastic moduli. We have explored the ability of various measures of the Debye-Waller factor u(2) to predict the stiffness of our systems in terms of their elastic constants. We find that u(2) is indeed correlated with the behavior of the bulk elastic constants. On a local level, a correlation exists between the local moduli and u(2); however, that correlation is not strong enough to arrive at conclusive statements about the local elastic properties.


Contribution of extracranial lymphatics and arachnoid villi to the clearance of a CSF tracer in the rat.
The objective of this study was to determine the relative roles of arachnoid villi and cervical lymphatics in the clearance of a cerebrospinal fluid (CSF) tracer in rats. 125I-labeled human serum albumin (125I-HSA; 100 micrograms) was injected into one lateral ventricle, and an Evans blue dye-rat protein complex was injected intravenously. Arterial blood was sampled for 3 h. Immediately after this, multiple cervical vessels were ligated in the same animals, and plasma recoveries were monitored for a further 3 h after the intracerebroventricular injection of 100 micrograms 131I-HSA. Tracer recovery in plasma at 3 h averaged (%injected dose) 0.697 +/- 0.042 before lymphatic ligation and dropped significantly to 0.357 +/- 0. 060 after ligation. Estimates of the rate constant associated with the transport of the CSF tracer to plasma were also significantly lower after obstruction of cervical lymphatics (from 0.584 +/- 0. 072/h to 0.217 +/- 0.056/h). No significant changes were observed in sham-operated animals. Assuming that the movement of the CSF tracer to plasma in lymph-ligated animals was a result of arachnoid villi clearance, we conclude that arachnoid villi and extracranial lymphatic pathways contributed equally to the clearance of the CSF tracer from the cranial vault.


A new combination chemotherapy with cis-diammine-glycolatoplatinum (Nedaplatin) and 5-fluorouracil for advanced esophageal cancers.
The efficacy of a new chemotherapeutic combination consisting of Cis-diammineglycolatoplatinum (Nedaplatin), a derivative of cisplatin (CDDP), and 5-fluorouracil (5FU) was evaluated in patients with advanced esophageal carcinomas. Nedaplatin was administered at a dose of 80 or 100 mg/m2 with 500 ml of saline by slow drip infusion for 120 minutes on day 1. 5FU at a dose of 350 or 500 mg/m2 was mixed with 1,000 ml of saline and administered by continuous infusion for 24 hours on days 1 to 5. This combination chemotherapy was tried in 17 patients with metastatic, recurrent, or bulky unresectable esophageal cancers. Of these, 15 evaluable patients received at least two courses of chemotherapy. The response rates in assessable and all patients were 60% and 52.9%, respectively. Cases with lymph node and liver metastases, as well as primary lesions, showed excellent response to the therapy with positive response rates of 54.5% (6/11), 100% (5/5) and 58.4% (7/12), respectively. The median response duration was 7 (range 3 to 37+) months for patients who achieved a partial response. Adverse drug reactions were limited to three cases of grade 3 toxicity, including allergy, and decreased hemoglobin and platelets, which were well tolerated by the patients. The present study thus indicated the combination chemotherapy of Nedaplatin and 5FU to be safe and efficacious for advanced esophageal cancer. Further investigations are clearly warranted.


Ultracompact and broadband silicon-based TE-pass 1 × 2 power splitter using subwavelength grating couplers and hybrid plasmonic gratings.
An ultracompact TE-pass 1 × 2 power splitter, using subwavelength grating (SWG) couplers and hybrid plasmonic gratings (HPGs), is proposed and analyzed in detail. As the input strip waveguide is tapered to a narrow wire, where TE mode is cutoff, the launched TE-mode can be evenly divided with high efficiency with the help of the SWG couplers, which are placed between the central silicon wire and two nearby output branches. The injected TM-mode will be perfectly reflected by the carefully designed HPG, whose periodically varied metal layer is located above the bottom strip and SWG waveguides. Consequently, a single device combining both the functions of polarization selection and power division can be realized. This is valuable for highly dense integrated circuits. Results show that, with a period number of 4 in HPG, the present device is only ~6.2-μm-long with an extinction ratio (ER) and an insertion loss (IL) of 25.4 and 0.53 dB at 1.55 μm, respectively, and its bandwidth of ER > 20 dB is ~180 nm with the IL < 0.8 dB, showing a broadband property. Besides, fabrication tolerances to the key dimensions are analyzed and modal field evolution through the device is also presented.


Microreactors-A Powerful Tool to Synthesize Peroxycarboxylic Esters.
The synthesis of peroxycarboxylic esters, as one subgroup of organic peroxides, is characterized by a high thermal hazard potential regarding process safety. In case of failure in the production process, e.g., if the heat of reaction cannot be removed sufficiently fast, decomposition reactions can be triggered, and as a result, remarkable amounts of heat and gas can be released and can cause a high extent of damage. Multifarious technical and organizational measures are necessary to ensure the safe industrial production of peroxides. With the introduction of microreaction technology plenty of possibilities have been opened to carry out highly exothermic reactions in smaller volumes and with more efficient heat removal. In this paper we report the application of three different microstructured reactors, representing different mixing strategies, to synthesize two peroxymonocarboxylic esters, namely tert-butyl peroxypivalate and tert-butyl peroxy-2-ethylhexanoate. The following reactor types were considered: an orifice microreactor, a split and recombine microreactor and a capillary tube reactor in combination with ultrasonication. The efficiency of the two phase liquid/liquid reaction is expressed in comparison of conversion and selectivity. With microreaction technology a remarkable increase in space-time-yield, ranging from 12,500 kg·m(-3)·h(-1) to 414,000 kg·m(-3)·h(-1), is achieved.


Association of single nucleotide variants in VEGFA and KDR with the risk and angiogenic features of diffuse large B-cell lymphoma.
Diffuse large B-cell lymphoma (DLBCL) is the most common lymphoma subtype and dependent on angiogenesis (AG), whose main effectors are VEGFA and VEGFR2. Functional single nucleotide variants (SNVs) are described in VEGFA and KDR genes. However, it still unknown whether VEGFA - 2578C/A, -2489C/T, -1154G/A, -634G/C, -460C/T and KDR-604T/C, -271G/A, +1192G/A and +1719A/T SNVs act on DLBCL risk and angiogenic features. Genomic DNA from 168 DLBCL patients and 205 controls was used for SNV genotyping. Angiogenesis was immunohistochemically assessed in tumor biopsies, with reactions for VEGFA, VEGFR2, and CD34. VEGFA -1154GG genotype were associated with 1.6-fold higher DLBCL risk. KDR + 1192GG plus KDR + 1719 TT and KDR + 1192GG plus VEGFA - 2578CC combined genotypes are associated with 2.19- and 2.04-fold higher risks of DLBCL, respectively. VEGFA - 634GG or GC genotypes are associated with increased microvessel density and VEGFA levels. No relationship was observed between SNVs and cell-of-origin classification of DLBCL, but higher VEGFA and VEGFR2 were seen in non-germinal center tumors.


Observation of a large-scale anisotropy in the arrival directions of cosmic rays above 8 × 1018 eV.
Cosmic rays are atomic nuclei arriving from outer space that reach the highest energies observed in nature. Clues to their origin come from studying the distribution of their arrival directions. Using 3 × 104 cosmic rays with energies above 8 × 1018 electron volts, recorded with the Pierre Auger Observatory from a total exposure of 76,800 km2 sr year, we determined the existence of anisotropy in arrival directions. The anisotropy, detected at more than a 5.2σ level of significance, can be described by a dipole with an amplitude of [Formula: see text] percent toward right ascension αd = 100 ± 10 degrees and declination δd = [Formula: see text] degrees. That direction indicates an extragalactic origin for these ultrahigh-energy particles.


The cell polarity proteins Boi1p and Boi2p stimulate vesicle fusion at the plasma membrane of yeast cells.
Eukaryotic cells can direct secretion to defined regions of their plasma membrane. These regions are distinguished by an elaborate architecture of proteins and lipids that are specialized to capture and fuse post-Golgi vesicles. Here, we show that the proteins Boi1p and Boi2p are important elements of this area of active exocytosis at the tip of growing yeast cells. Cells lacking Boi1p and Boi2p accumulate secretory vesicles in their buds. The essential PH domains of Boi1p and Boi2p interact with Sec1p, a protein required for SNARE complex formation and vesicle fusion. Sec1p loses its tip localization in cells depleted of Boi1p and Boi2p but overexpression of Sec1p can partially compensate for their loss. The capacity to simultaneously bind phospholipids, Sec1p, multiple subunits of the exocyst, Cdc42p and the module for generating active Cdc42p identify Boi1p and Boi2p as essential mediators between exocytosis and polar growth.


Mammary analog secretory carcinoma, low-grade salivary duct carcinoma, and mimickers: a comparative study.
Mammary analog secretory carcinoma (MASC) is a recently recognized low-grade salivary carcinoma characterized by a specific ETV6 rearrangement. We describe 14 new MASCs and examine their immunophenotypic and genetic profiles in the context of look-alikes, namely, low-and high-grade salivary duct carcinoma and acinic cell carcinoma. ETV6 rearrangement, and robust expression of mammaglobin and S100, were demonstrated in 11/11, 14/14, and 12/14 MASCs, respectively. All low-grade salivary duct carcinomas coexpressed S100/mammaglobin (6/6); none harbored ETV6 rearrangements (0/5). Given that S100/mammaglobin coexpression and absence of zymogen granules are features of both MASC and low-grade salivary duct carcinoma, these two are best distinguished histologically. The former is predominantly an extraductal neoplasm with bubbly pink cytoplasm, whereas the latter is a distinct intraductal micropapillary and cribriform process. Querying ETV6 gene status may be necessary for difficult cases. No acinic cell carcinoma expressed mammaglobin (0/13) or harbored an ETV6 rearrangement (0/7); only 1/13 acinic cell carcinomas weakly expressed S100. DOG1 expression was limited or absent among all tumor types, except acinic cell carcinoma which expressed DOG1 diffusely in a canalicular pattern. Therefore, histology and immunohistochemistry (mammaglobin, S100, DOG1) suffices in distinguishing acinic cell carcinoma from both MASC and low-grade salivary duct carcinoma. HER2 (ERBB2) amplification was detected in only 1/10 acinic cell carcinomas, but none of the MASCs or low-grade salivary duct carcinomas tested. High-grade salivary duct carcinomas frequently expressed mammaglobin (11/18) and harbored HER2 amplifications (13/15); none harbored ETV6 rearrangements (0/12). High-grade salivary duct carcinomas can easily be distinguished from these other entities by histology and HER2 amplification.


CD8+ cellular immunity mediates rAd5 vaccine protection against Ebola virus infection of nonhuman primates.
Vaccine-induced immunity to Ebola virus infection in nonhuman primates (NHPs) is marked by potent antigen-specific cellular and humoral immune responses; however, the immune mechanism of protection remains unknown. Here we define the immune basis of protection conferred by a highly protective recombinant adenovirus virus serotype 5 (rAd5) encoding Ebola virus glycoprotein (GP) in NHPs. Passive transfer of high-titer polyclonal antibodies from vaccinated Ebola virus-immune cynomolgus macaques to naive macaques failed to confer protection against disease, suggesting a limited role of humoral immunity. In contrast, depletion of CD3(+) T cells in vivo after vaccination and immediately before challenge eliminated immunity in two vaccinated macaques, indicating a crucial requirement for T cells in this setting. The protective effect was mediated largely by CD8(+) cells, as depletion of CD8(+) cells in vivo using the cM-T807 monoclonal antibody (mAb), which does not affect CD4(+) T cell or humoral immune responses, abrogated protection in four out of five subjects. These findings indicate that CD8(+) cells have a major role in rAd5-GP-induced immune protection against Ebola virus infection in NHPs. Understanding the immunologic mechanism of Ebola virus protection will facilitate the development of vaccines for Ebola and related hemorrhagic fever viruses in humans.


Evidence-Based Guidelines for Prehospital Pain Management: Recommendations.
This project sought to develop evidence-based guidelines for the administration of analgesics for moderate to severe pain by Emergency Medical Services (EMS) clinicians based on a separate, previously published, systematic review of the comparative effectiveness of analgesics in the prehospital setting prepared by the University of Connecticut Evidence-Based Practice Center for the Agency for Healthcare Research and Quality (AHRQ). A technical expert panel (TEP) was assembled consisting of subject matter experts in prehospital and emergency care, and the development of evidence-based guidelines and patient care guidelines. A series of nine "patient/population-intervention-comparison-outcome" (PICO) questions were developed based on the Key Questions identified in the AHRQ systematic review, and an additional PICO question was developed to specifically address analgesia in pediatric patients. The panel made a strong recommendation for the use of intranasal fentanyl over intravenous (IV) opioids for pediatric patients without intravenous access given the supporting evidence, its effectiveness, ease of administration, and acceptance by patients and providers. The panel made a conditional recommendation for the use of IV non-steroidal anti-inflammatory drugs (NSAIDs) over IV acetaminophen (APAP). The panel made conditional recommendations for the use of either IV ketamine or IV opioids; for either IV NSAIDs or IV opioids; for either IV fentanyl or IV morphine; and for either IV ketamine or IV NSAIDs. A conditional recommendation was made for IV APAP over IV opioids. The panel made a conditional recommendation against the use of weight-based IV ketamine in combination with weight-based IV opioids versus weight-based IV opioids alone. The panel considered the use of oral analgesics and a conditional recommendation was made for either oral APAP or oral NSAIDs when the oral route of administration was preferred. Given the lack of a supporting evidence base, the panel was unable to make recommendations for the use of nitrous oxide versus IV opioids, or for IV ketamine in combination with IV opioids versus IV ketamine alone. Taken together, the recommendations emphasize that EMS medical directors and EMS clinicians have a variety of effective options for the management of moderate to severe pain in addition to opioids when designing patient care guidelines and caring for patients suffering from acute pain.


V-RISK-10: validation of a screen for risk of violence after discharge from acute psychiatry.
Current violence risk assessment instruments are time-consuming and mainly developed for forensic psychiatry. A paucity of violence screens for acute psychiatry instigated the development and validation of the V-RISK-10. The aim of this prospective naturalistic study was to test the predictive validity of the V-RISK-10 as a screen of violence risk after discharge from two acute psychiatric wards. Patients were screened with V-RISK-10 before discharge, and incidents of violence were recorded 3, 6, 9 and 12 months after discharge. A total of 381 of the 1017 patients that were screened completed the follow up. The ROC-AUC values for any violent behaviour were 0.80 and 0.75 (p<0.001) for the 3 and 12 months follow-up periods, respectively, and significant for both genders. The most accurate risk estimates were obtained for severe violence. For persons without a known history of violence prior to the screening, AUCs were 0.74 (p=0.004) and 0.68 (p=0.002). Results indicate that the V-RISK-10 is a valid and clinically useful screen for violence risk after discharge from acute psychiatry, and even significant for patients without a known previous history of violence.


Hydrogen alleviates mitochondrial dysfunction and organ damage via autophagy‑mediated NLRP3 inflammasome inactivation in sepsis.
Sepsis is a highly heterogeneous syndrome that is caused by a dysregulated host response to infection. The disproportionate inflammatory response to invasive infection is a triggering event inducing sepsis. The activation of inflammasomes in sepsis can amplify inflammatory responses. It has been reported that damaged mitochondria contribute to NACHT, LRR and PYD domains‑containing protein 3 (NLRP3) inflammasome‑related sepsis. Our previous study revealed that hydrogen (H2) exerts anti‑inflammatory effects in sepsis but the detailed mechanism remains to be elucidated. In the present study, septic mice induced by cecal ligation and puncture (CLP) and macrophages induced by lipopolysaccharide (LPS) were used as models of sepsis in vivo and in vitro, respectively. An inducer and inhibitor of autophagy and the NLRP3 inflammasome were administered to investigate the detailed mechanism of action of H2 treatment in sepsis. The results demonstrated that LPS and ATP led to NLRP3 inflammasome pathway activation, excessive cytokine release, mitochondrial dysfunction and the activation of autophagy. CLP induced organ injury and NLRP3 pathway activation. H2 treatment ameliorated vital organ damage, the inflammatory response, mitochondrial dysfunction and NLRP3 pathway activation, and promoted autophagy in macrophages induced by LPS and in CLP mice. However, the inhibitor of autophagy and the inducer of NLRP3 reversed the protective effect of H2 against organ damage, the inflammatory response and mitochondrial dysfunction in vivo and in vitro. Collectively, the results demonstrated that H2 alleviated mitochondrial dysfunction and cytokine release via autophagy‑mediated NLRP3 inflammasome inactivation.


Phage-displayed mimotopes elicit monoclonal antibodies specific for a malaria vaccine candidate.
The phage-displayed peptide CGRVCLRC (C15) has been isolated from a random library by affinity screening with the D14-3 monoclonal antibody, which was raised to the 42 kDa C-terminal fragment of the major merozoite surface protein 1 of Plasmodium vivax (Pv42). In order to investigate the use of such mimotopes as possible vaccine components, we studied the antibody response in Biozzi mice immunized with C15. High titers of antibodies cross-reacting with Pv42 were generated and the IC50 of all immune sera were in the 5 x 10(-9) M range. Two monoclonal antibodies that specifically bind the Pv42 fragment were isolated. Although these mAbs had a lower affinity for Pv42 when compared to D14-3, they reproduced the cross-reactivity of D14-3 with the equivalent protein in P. cynomolgi, a close relative of P. vivax. DNA sequence analysis showed similarities between the germline genes and the canonical CDR conformations of all three antibodies, but molecular modeling failed to reveal common structural features of their paratopes that could account for their cross-reacting patterns. These data demonstrate that mimotopes selected from random repertoires do not necessarily represent structural equivalents of the original antigen but provide functional images that could replace it for vaccine development.


Will the SAFE strategy be sufficient to eliminate trachoma by 2020? Puzzlements and possible solutions.
Since the inception of (the Global Elimination of Blinding Trachoma) GET 2020 in 1997 and the implementation of the SAFE strategy a year later, much progress has been made toward lowering the prevalence of trachoma worldwide with elimination of the disease in some countries. However, high recurrence of trichiasis after surgery, difficulty in controlling the reemergence of infection after mass distribution of azithromycin in some communities, the incomplete understanding of environment in relation to the disease, and the difficulty in establishing the prevalence of the disease in low endemic areas are some of the issues still facing completion of the GET 2020 goals. In this narrative review, literature was searched from 1998 to January 2013 in PubMed for original studies and reviews. Reasons for these ongoing problems are discussed, and several suggestions are made as avenues for exploration in relation to improving the SAFE strategy with emphasis on improving surgical quality and management of the mass treatment with antibiotics. In addition, more research needs to be done to better understand the approach to improve sanitation, hygiene, and environment. The main conclusion of this review is that scale-up is needed for all SAFE components, and more research should be generated from communities outside of Africa and Asia.


Somatostatins and their role in thyroid cancer.
Somatostatins are neuropeptides that exert a downregulatory effect on various physiological processes. Somatostatin analogues are used in the imaging and management of various tumour types. Their role in thyroid cancer has not as yet been fully elucidated. A systematic review of the literature using the keywords thyroid, cancer and somatostatin revealed 263 references. This paper summarizes the current knowledge of the role of somatostatins in thyroid cancer and assesses their future potential.


Importance of a nanoscience approach in the understanding of major aqueous contamination scenarios: case study from a recent coal ash spill.
A coal ash spill that occurred from an ash impoundment pond into the Dan River, North Carolina, provided a unique opportunity to study the significance and role of naturally occurring and incidental nanomaterials associated with contaminant distribution from a large-scale, acute aquatic contamination event. Besides traditional measurements of bulk watercolumn and sediment metal concentrations, the nanoparticle (NP) analyses are based on cross-flow ultrafiltration (CFUF) and advanced transmission electron microscopy (TEM) techniques. A drain pipe fed by coal ash impoundment seepage showed a high level of arsenic, with concentrations many times over the EPA limit. The majority of the arsenic was found sorbed to large aggregates dominated by incidental iron oxyhydroxide (ferrihydrite) NPs, while the remainder of the arsenic was truly dissolved. These ferrihydrites were probably formed in situ where Fe(II) was leached through subsurface flowpaths into an aerobic environment, and further act as a significant contributor to the elevated As concentrations in downstream sediments after the spill. In addition, we discovered and describe a photocatalytic nano-TiO2 phase (anatase) present in the coal ash impacted river water that was also carrying/transporting transition metals (Cu, Fe), which may also have environmental consequences.


Ecological divergence combined with ancient allopatry in lizard populations from a small volcanic island.
Population divergence and speciation are often explained by geographical isolation, but may also be possible under high gene flow due to strong ecology-related differences in selection pressures. This study combines coalescent analyses of genetic data (11 microsatellite loci and 1 Kbp of mtDNA) and ecological modelling to examine the relative contributions of isolation and ecology to incipient speciation in the scincid lizard Chalcides sexlineatus within the volcanic island of Gran Canaria. Bayesian multispecies coalescent dating of within-island genetic divergence of northern and southern populations showed correspondence with the timing of volcanic activity in the north of the island 1.5-3.0 Ma ago. Coalescent estimates of demographic changes reveal historical size increases in northern populations, consistent with expansions from a volcanic refuge. Nevertheless, ecological divergence is also supported. First, the two morphs showed non-equivalence of ecological niches and species distribution modelling associated the northern morph with mesic habitat types and the southern morph with xeric habitat types. It seems likely that the colour morphs are associated with different antipredator strategies in the different habitats. Second, coalescent estimation of gene copy migration (based on microsatellites and mtDNA) suggest high rates from northern to southern morphs demonstrating the strength of ecology-mediated selection pressures that maintain the divergent southern morph. Together, these findings underline the complexity of the speciation process by providing evidence for the combined effects of ecological divergence and ancient divergence in allopatry.


Metastatic breast cancer masquerading as gastrointestinal primary.
Seven patients with metastatic breast cancer presenting as gastrointestinal primary are described. These included six gastric and one colonic lesions. None of the patients had known systemic metastases at the time of diagnosis. The mean age at presentation was 66.7 yr (range 55-78). Median interval between breast cancer and gastrointestinal metastasis diagnosis was 6 yr (range 0.25-12.5). Original breast cancer histology included infiltrating lobular cancer (n = 4), infiltrating ductal cancer (n = 1), and a mixed type (n = 2). All patients with gastric involvement presented with epigastric pain and early satiety; the patient with colonic involvement had heme-positive stool. In three cases of gastric tumor and the one case of colonic tumor presentation, a definitive diagnosis of metastatic breast cancer was only confirmed after surgical resection of a presumed primary gastric or colonic malignancy. In the other three cases, pathological diagnostic confirmation was obtained through endoscopic biopsies and comparison to breast biopsy material, and operative treatment was avoided in favor of systemic cytotoxic therapy. The diagnosis was confirmed through similarities between mammary and gastric histopathology with regard to growth pattern, hormone receptor status, or gross cystic disease fluid protein. A high level of suspicion for metastatic breast cancer and a detailed pathological analysis will help avoid unnecessary surgical treatment in patients with a history of mammary carcinoma presenting with a newly diagnosed gastrointestinal neoplasm.


Pathologic studies in colorectal cancer. A guide to the surgical pathology examination of colorectal specimens and review of features of prognostic significance.
The results of a careful and systematic examination of surgical specimens from patients with tumors of the colorectum play an important role in patient care and the assessment of prognosis. An accurate and detailed gross examination of the specimen is important. Photographs and line drawings are very useful. Proper handling of the specimen and fixation for 24 to 48 hours are essential before satisfactory blocks can be taken for sectioning. Lymph nodes adjacent to the lesion and at the point of ligation of the vascular pedicle should be removed for sectioning. Multiple blocks of the tumor and adjacent tissues should be taken for histologic study. All blocks should be appropriately labeled and properly identified. I have found the use of large sections of the entire lesion a very satisfactory method of studying cancers and polyps; in such preparations the anatomic relationships remain undisturbed. The pathology report should include information on the site or sites of tumor, the size, configuration, and circumference of the bowel wall involved, obstruction, distance of the resected margin from the tumor, depth of infiltration, tumor grade, tumor margin, local inflammatory reaction, lymph node involvement and location, and venous and perineural invasions. There is evidence in the literature to suggest that all of the above pathologic features are either essential or desirable for pathology reports and have varying degrees of prognostic value. The literature on the prognostic value of immunomorphologic features in nontumorous regional lymph nodes is contradictory. Cortical, paracortical, and sinus hyperplasia have been reported to be both associated with or to have no relation to prognosis. However, careful documentation of the changes in lymph nodes may in time shed light on their value, if any, with regard to survival. We have found a standardized pathology reporting system very useful in compiling data and evaluating patient prognosis. Moreover, this system of reporting can be used as a method of quality control in establishing minimum standards for data collection, as originally suggested by Buckwalter and Kent. In a regional multiinstitutional study such as ours, differences attributable to subjective variations among different observers are likely to occur, and may have some bearing when different pathologic features are eventually correlated with survival.


Management of methicillin-resistant Staphylococcus aureus bacteremia.
Staphylococcus aureus bacteremia and endocarditis are serious infections that demand prompt clinical attention to ensure good outcomes. Of foremost importance is identifying and managing the source of infection and any associated complications. Evaluation for the presence of cardiac involvement is essential because inadequately managed S. aureus endocarditis is life threatening. Thus, physicians must aggressively negotiate treatment paths, considering whether the S. aureus bacteremia is complicated, whether foreign sources of infection should be removed or replaced, and whether surgical intervention is necessary. Selection of an antibiotic treatment is also an essential factor for optimal management. The increasing prevalence of methicillin-resistant S. aureus (MRSA) infections has created a tremendous demand for effective and safe antimicrobial agents other than the historic anti-MRSA agent vancomycin.


Repeated range expansions and inter-/postglacial recolonization routes of Sargentodoxa cuneata (Oliv.) Rehd. et Wils. (Lardizabalaceae) in subtropical China revealed by chloroplast phylogeography.
Most plant phylogeographic studies in subtropical China have stressed the importance of multiple refugia and limited admixture among refugia. Little attention has been paid to range expansion and recolonization routes in this region. In this study, we implemented a phylogeographic survey on Sargentodoxa cuneata, a widespread woody deciduous climber in subtropical China to determine if it conforms to the expansion-contraction (EC) model during the Pleistocene. Sequence variation of two chloroplast intergenic spacers (IGSs) in 369 individuals from 54 populations of S. cuneata was examined. Twenty-six chloroplast haplotypes were recovered. One of these (H5) occurred across the range of S. cuneata and was absent from only 13 populations. Sixteen of the 26 haplotypes were connected to H5 by one mutation and displayed a star-like pattern in the haplotype network. All chloroplast haplotypes clustered into two lineages (A and B) in a Bayesian tree, and most haplotypes (18 out of 26) originated during the mid-Pleistocene (0.63-1.07Ma). Demographic analyses detected a recent range expansion that occurred at 95.98ka (CI: 61.7-112.53ka) for Lineage A. The genetic signature of an ancient range expansion after the Middle Pleistocene Transition (MPT) was also evident. Three recolonization routes were identified in subtropical China. The results suggest that temperate plants in subtropical China may conform to the EC model to some extent. However, the genetic signature from multiple historical processes may complicate the phylogeographic patterns of organisms in the region due to the mild Pleistocene climate. This study provides a new perspective for understanding the evolutionary history of temperate plants in subtropical China.


A reaction model on the self-assembly process of octahedron-shaped coordination capsules.
Herein, the self-assembly process of an octahedron-shaped coordination capsule was analyzed based on a master equation approach using a reaction network model. This model was found to adequately reproduce the overall experimentally observed time evolutions and enabled us to trace the real-time evolution of transient intermediates, ranging from milli-second to hours. The time evolution of the distribution of individual intermediates species was obtained; a few linear-oligomers located near the reactant were produced at first, followed by an explosive increase in several types of intermediates. All of them were then consolidated into a few species just before the formation of the final product. A long-lived [Pd618Py]12+ is a key compound, which acts as a kinetic trap in the reaction dynamics.


Standardization of a model to study revaccination against Marek's disease under laboratory conditions.
Revaccination, the practice of administering Marek's disease (MD) vaccine a second time, has been used in commercial poultry flocks for many years. The rationale is largely anecdotal as the few published reports have failed to provide support for the value of the practice. In the present work, we have standardized a model to study MD revaccination under laboratory conditions. Nine bird experiments were conducted to evaluate homologous revaccination (same vaccine administered twice) and heterologous revaccination (administration of two different vaccines) with various challenge models. Our results demonstrated that heterologous revaccination (with a second vaccine more protective than the first vaccine) but not homologous revaccination provided a beneficial increase in protection. Administration of the first vaccine at 18 days of embryonation followed by a more protective second vaccine at hatch reproduced systematically the benefits of revaccination. In addition, our results show that revaccination protocols might aid in solving major drawbacks associated with various highly protective experimental MD vaccines; that is, lymphoid organ atrophy and residual virulence. Strain RM1 is one of the most protective vaccines against early challenge with highly virulent MD virus but it induces severe lymphoid atrophy in chickens lacking maternal antibodies against MD virus. In this study, strain RM1 did not induce lymphoid organ atrophy when administered as second vaccine in a revaccination protocol. Similarly, strain 648A100/BP5 maintains residual virulence in chickens lacking maternal antibodies against MD virus but did not induce any lesions when used as a second vaccine. Until now, arbitrary revaccination protocols have been occasionally proven useful to the poultry industry. The model developed in this study will allow for a better understanding of this phenomenon and its optimization. A more rational use of this practice will be of great help to control MD outbreaks until better vaccines are available.


Social phobia: epidemiology and health care.
This paper gives an overview on the epidemiology of social phobia. About 4.5% of the adult general populations suffer from social phobia, i.e. it is the most frequent of all anxiety disorders. Social phobia is clearly more frequent among women than among men. About the half of all individuals with social phobia suffer from any comorbid mental disorders. Reviews show a large variability between single studies, probably due to methodological differences. Several population surveys indicate that a marked proportion of those with social phobia do not receive adequate treatment.


'We never expected this to happen': narratives of ageing with HIV among gay men living in London, UK.
The proportion of HIV-positive people over the age of 50 is rapidly increasing in the UK. This reflects the use of antiretroviral therapies and the transformation of HIV from life-threatening disease to chronic treatable illness. In this study a biographical narrative approach was used to explore the lived experience of ageing in 10 HIV-positive gay men aged between 50 and 78. While some participants regarded ageing as an opportunity to continue progressing towards valued life goals, others were more ambivalent about their future prospects. The findings suggest that these differences were particularly influenced by an individual's biographic relationship to the history of the HIV epidemic rather than chronological age. Those with long histories of involvement with HIV were more likely to be disadvantaged by careers interrupted by illness, to be dependent on state benefits and to have social networks damaged by multiple AIDS-related bereavements. The research identifies a cohort of older gay men likely to require additional support in adapting to the challenge of growing older with HIV. The article also explores the construction of moral identities in relation to discourses of 'successful ageing' and the possibility of building supportive communities that are sensitive to the needs of older gay men.


Efficacy of US-guided transversus abdominis plane block and rectus sheath block with ropivacaine and dexmedetomidine in elderly high-risk patients.
This study was designed to evaluate the effectiveness and safety of ultrasound-guided transversus abdominis plane (TAP) and rectus sheath (RS) blocks with ropivacaine and dexmedetomidine in elderly, high-risk patients undergoing emergency abdominal surgery. Sixty elderly patients undergoing emergency abdominal surgery receiving both bilateral ultrasound-guided TAP and RS blocks were randomly divided into two groups: R+D (10 mL of 0.25% ropivacaine+0.5μg/kg dexmedetomidine) and R (10 mL of 0.25% ropivacaine). Pain scores (at rest and with movement) and overall analgesia satisfaction were assessed. The total amount of sufentanil administered postoperatively during the first 24 h, duration of sensory blockade, first time and total number of patient-controlled intravenous analgesia (PCIA) pump activations on the first postoperative day were recorded. The duration of sensory blockade and the first time to PCIA pump activation in the R+D group were longer than that of the R group (P<0.05). The total number of PCIA pump activations on the first postoperative day as well as the total amount of sufentanil administered during the first 24 h in R+D group were less than in the R group (P<0.05). VAS scores at rest and during activity in the R+D group were lower than those in the R group at 2, 6, and 12 h after surgery (P<0.05). Ultrasound-guided TAP and RS blocks with ropivacaine and dexmedetomidine are more effective in promoting block effect, prolonging the duration of analgesia, reducing postoperative pain in elderly high-risk patients undergoing emergency abdominal surgery.


[Clinical case of the month. An unusual presentation of superior vena cava syndrome].
We report the case of a 48-year-old man who was admitted to our emergency department because of a superior vena caval syndrome from which the symptoms occurred only during exercise. His past history included an episode of lower limb deep venous thrombosis. Because an atrio-ventricular partial block was fortuitously evidenced, the patient was submitted to a pacemaker insertion. A few years later, the patient suffered from rapid dyspnea on exercise, associated with facial cyanosis and systemic hypotension. The exploration by phlebography evidenced that the inferior vena cava was completely obstructed. Therefore the venous return from the lower part of the body to right heart was obtained through hypertrophy of the vena azygos. In addition, the presence of cardiac pacing electrodes induced a stenosis at the level of the superior vena cava, just before its entrance into the right atrium. Such a venous network accounted for the occurrence of a chronic superior vena caval syndrome associated with peripheral arterial hypotension during exercise. Treatment consisted of a superior vena caval percutaneous transluminal balloon angioplasty followed by a Wallstent insertion. This procedure led to a correction of all hemodynamic abnormalities responsible for reported pathophysiological limitations to stress.


Confinement Dynamics of Nanodroplets between Two Surfaces: Effects of Wettability and Electric Field.
The electrowetting effect and related applications of tiny droplets have aroused widespread research interest. In this work, we report molecular dynamics simulations of confinement dynamics of nanodroplets under different droplet-surface interactions and surface distances under an external electric field. So far, the effect of the surface-droplet interactions on electric field-induced dynamics behaviors of droplets in confined spaces has not been extensively studied. Our results show that in the absence of electric field there is a critical value of surface wettability for the shape transition of droplets. Above this value, the droplet is divided into small droplets adhered on the bottom and top surfaces; below this value, the droplets are detached from the surfaces. When an external electric field is applied parallel to the surfaces, the droplet spreads on the surface along the direction of the electric field. It was found that the surface separation significantly influences the transition of the droplet shape. The steady morphology of the droplets under the electric field depends on the surface-droplet interaction and surface separation. We explore the underlying mechanism causing the morphological transition through analyzing the molecular interactions, the number of interracial molecules and the interaction force between the droplets and surfaces. These results provide basic insights into the molecular interactions of nanodroplets under different confined environments, and clues for applications of confined nanodroplets under the control of electric field.


Myocardial blood flow and flow reserve after coronary reimplantation in patients after arterial switch and ross operation.
Coronary reimplantation is used in therapy for congenital heart disease, such as in the arterial switch (ASO) and Ross operations. The adequacy of myocardial perfusion may remain a matter of concern. The aim of the present study was to stratify the effect of coronary reimplantation on myocardial perfusion and to highlight the clinical relevance of any attenuation in myocardial perfusion. A total of 21 children with transposition of the great arteries at a mean interval of 11.2+/-2.9 years after ASO and 9 adolescents at a mean interval of 4.2+/-2.1 years after the Ross procedure were investigated. All patients were asymptomatic and had a normal exercise capacity. On stress echocardiography, 2 of the ASO patients had dyskinetic areas within the left ventricular myocardium, and 5 had adenosine-induced perfusion defects on positron emission tomography. No coronary obstruction was detected on coronary angiography in any patient, but a common finding was right coronary dominance and a small caliber of the distal part of the left anterior descending artery. Coronary flow reserve (CFR) was significantly reduced in all patients after ASO when compared with 10 normal healthy volunteers (age, 25.6+/-5.3 years). CFR was normal in the 9 patients who had the Ross operation (age, 19.2+/-7.6 years); exercise-induced perfusion defects were not detected in the Ross patients. Children after ASO are asymptomatic, without clinical signs of coronary dysfunction. In contrast to patients who had the Ross operation, stress-induced perfusion defects and an attenuated CFR were documented. The prognostic implications of these findings and the clinical consequences are unclear; nevertheless, close clinical follow-up of ASO patients is mandatory.


Effect on sport hemolysis of cold water leg immersion in athletes after training sessions.
The principal source of increased turnover of erythrocytes in athletes is sport hemolysis, the intravascular hemolysis that characteristically occurs with athletic performance in sport. The use of the parameter mean sphered cell volume (MSCV), automatically measured by means of the Coulter LH750, could be useful for diagnosing the presence of sport hemolysis. We studied the behavior of MSCV and mean corpuscular volume (MCV) in 30 top-level rugby players who underwent a heavy training session followed by 3 different recovery methods, administered to 3 subgroups of 10 athletes. We tested the use of active recovery consisting of cold water (5 degrees C) immersion of legs for 10 minutes either before (n = 10) or after (n = 10) cycling at 180 W for 10 minutes. In the whole group of athletes, measurements performed at rest and after training session and recovery showed no differences in MCV and MSCV values. The difference between MCV and MSCV was significant in the whole group and in the subgroup performing passive recovery, whereas the difference was not significant in the subgroups performing active recovery. This finding indicates that the use of active recovery in the top-level rugby players prevented the modifications of erythrocyte volume and shape. We outline that the values of the difference between MCV and MSCV was significantly modified in the whole group but the variations were not significant in the active recovery subgroups. The use of an index of erythrocyte shape modification (MCV - MSCV) can be very useful for evaluating sport hemolysis.


New class of dynamics in concentrated polymer gels.
We show that concentrated poly(methyl methacrylate) solution exhibits a new class of coupled dynamics, which can be regarded as an intermediate between the collective diffusion of solutions and the structural relaxations of glasses. This class of dynamics have a relaxation rate that is directly proportional to the wave vector. The transition from diffusive to coupled collective dynamics occurs at smaller length scales with increasing polymer concentration and decreasing temperature. The experimental observations can be understood by considering the contributions from physical cross-links interconnected by stiff polymer segments.


Repetitive sequence-based PCR versus pulsed-field gel electrophoresis for typing of Enterococcus faecalis at the subspecies level.
Repetitive sequence-based PCR was compared to pulsed-field gel electrophoresis (PFGE) for the ability to discriminate Enterococcus faecalis isolates at the subspecies level. The BOXA2R primer, derived from repetitive sequences in Streptococcus pneumoniae, was applied to 41 isolates of E. faecalis collected from various sources. The REP1R-Dt and REP2-Dt primers, derived from the gram-negative repetitive extragenic palindromic element, were also applied to 18 selected isolates. Of the 41 isolates examined, 7 were beta-lactamase producing and 8 were vancomycin resistant. By PFGE, 17 isolates had distinct patterns; the other 24 were classified into eight different clonal groups. By PCR using the BOXA2R primer, 16 isolates generated distinct patterns; the other 25 were classified into nine different clonal groups. There were only minor differences in the PCR results obtained by using the BOXA2R primer and the REP1R-Dt and REP2-Dt primers. Two isolates among vancomycin-resistant enterococci from the greater Houston, Tex., area were related by PFGE, distinct by PCR with the BOXA2R primer, and related by PCR with the REP1R-Dt and REP2-Dt primers. Clonal relationships among the remaining 39 isolates were similar by both PFGE and PCR. PCR reliably discriminated all epidemiologically unrelated isolates. Although PCR is less time consuming than PFGE, PCR results were more difficult to interpret than PFGE results, perhaps because fewer bands were generated by PCR than by PFGE and some PCR products were inconsistently seen.


Modern fluid management in thoracic surgery.
To provide an approach to perioperative fluid management for lung resection patients that incorporates the entire patient pathway in the context of international guidelines on enhanced recovery after surgery (ERAS). The concern with intraoperative fluid management is that giving too little or too much fluid is associated with worse outcomes after lung resection. However, it has not emerged as a key care element in thoracic ERAS programs probably due to the influence of other ERAS elements. Carbohydrate loading 2 h before surgery and the allowance of water until just prior to induction ensures the patient is both well hydrated and metabolically normal when they enter the operating room. Consequently, maintaining a euvolemic state during anesthesia can be achieved without goal-directed fluid therapy despite the recommendations of some guidelines. Intravenous fluids can be safely stopped in the immediate postoperative period. The goal of perioperative euvolemia can be achieved with the ongoing evolution and application of ERAS principles. A focus on the pre and postoperative phases of fluid management and a pragmatic approach to intraoperative fluid management negates the need for goal-directed fluid therapy in most cases.


Development and initial feedback about a human papillomavirus (HPV) vaccine comic book for adolescents.
Human papillomavirus (HPV) vaccination rates do not meet the Healthy People 2020 objective of 80% coverage among adolescent females. We describe the development and initial feedback about an HPV vaccine comic book for young adolescents. The comic book is one component of a multilevel intervention to improve HPV vaccination rates among adolescents. Parents suggested and provided input into the development of a HPV vaccine comic book. Following the development of the comic book, we conducted a pilot study to obtain initial feedback about the comic book among parents (n = 20) and their adolescents ages 9 to 14 (n = 17) recruited from a community-based organization. Parents completed a pre-post test including items addressing HPV knowledge, HPV vaccine attitudes, and about the content of the comic book. Adolescents completed a brief interview after reading the comic book. After reading the comic book, HPV knowledge improved (2.7 to 4.6 correct answers on a 0-5 scale; p < 0.01) and more positive attitudes toward HPV vaccination (p < 0.05) were reported among parents. Parents confirmed that the comic book's content was acceptable and adolescents liked the story, found it easy to read, and thought the comic book was a good way to learn about being healthy. Parents provided valuable information in the development of a theoretically-based comic book and the comic book appears to be an acceptable format for providing HPV vaccine information to adolescents. Future research will include the comic book in an intervention study to improve HPV vaccination rates.


Moisture content of tea dhool for the scenting process affects the aroma quality and volatile compounds of osmanthus black tea.
To improve the quality of osmanthus black tea, samples produced with different scenting methods were prepared. The sensory quality was assessed and the characteristic aromatic components were explored using solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry. According to the results, osmanthus black tea obtained by adding osmanthus scenting in the fermentation process had the strongest floral aroma. The major contributors to the aroma of osmanthus black tea were identified as β-ionone, dihydro-β-ionone, benzeneacetaldehyde, citral, geraniol, and linalool by calculating their relative odor activity values. An analysis of the causes revealed that the moisture content of tea dhool significantly affected the adsorption of fresh flower aroma by tea. The experimental results showed that osmanthus black tea produced using tea dhool containing 30% moisture content had the highest content of crucial aroma components, suggesting the tea dhool under this condition had the strongest adsorption capacity for osmanthus aroma.


A truly ecological epigenetics study.
Until a few years ago, epigenetics was a field of research that had nothing to do with ecology and that virtually no ecologist had ever heard of. This is now changing, as more and more ecologists learn about epigenetic processes and their potential ecological and evolutionary relevance, and a new research field of ecological epigenetics is beginning to take shape. One question that is particularly intriguing ecologists is to what extent epigenetic variation is an additional, and hitherto overlooked, source of natural variation in ecologically important traits. In this issue of Molecular Ecology, Herrera & Bazaga (2011) provide one of the first attempts to truly address this question in an ecological setting. They study variation of DNA methylation in a wild population of the rare, long-lived violet Viola cazorlensis, and they use these data to explore interrelations between environmental, genetic and epigenetic variation, and in particular the extent to which these factors are related to long-term differences in herbivore damage among plants. They find substantial epigenetic variation among plant individuals. Interestingly, this epigenetic variation is significantly correlated with long-term differences in herbivory, but only weakly with herbivory-related DNA sequence variation, which suggests that besides habitat, substrate and genetic variation, epigenetic variation may be an additional, and at least partly independent, factor influencing plant–herbivore interactions in the field. Although the study by Herrera & Bazaga (2011) raises at least as many new questions as it answers, it is a pioneering example of how epigenetics can be incorporated into ecological field studies, and it illustrates the value and potential novel insights to be gained from such efforts.


Copper binding dynamics and olfactory impairment in fathead minnows (Pimephales promelas).
When fish are exposed to sublethal, environmentally relevant Cu concentrations, olfactory acuity is impaired. The goals of the present study were to investigate the binding dynamics of waterborne Cu in the olfactory epithelium (OE), to examine the influence of calcium (Ca(2+)) on Cu binding, and to link Cu-OE binding to changes in olfactory acuity. Using short-term in vivo waterborne exposures to (64)Cu, we found that Cu accumulates rapidly in the OE, reaching a plateau by 3 h. The binding affinity (log K(Cu-OE)) and binding capacity (B(max)) of (64)Cu in the OE were 6.7 and 10.0 nmol Cu g(-1), respectively. As waterborne Ca(2+) was increased from 50 to 1000 microM L(-1), the B(max) of Cu decreased by approximately 50% while the log K(Cu-OE) remained constant, indicative of noncompetitive inhibition. Using electro-olfactograms (EOG), short-term exposures to 160 and 240 nmol Cu L(-1) were found to reduce olfactory responses to 10(-5) M l-arginine by 72 and 79%, respectively. Short-term exposure to 160 nmol Cu L(-1) also caused a 15-fold reduction in behavioral responses to a food stimulus. Interestingly, increasing waterborne Ca(2+) did not reduce the effects of Cu on EOG or behavioral responses. These results demonstrate that short-term, environmentally realistic concentrations of Cu not only bind to the OE of fathead minnows but also impair their olfactory sensitivity and behavioral responses to olfactory stimuli. Waterborne Ca(2+) reduces Cu-OE binding but does not protect against olfactory impairment.