Prognostic significance of DNA replication errors in young patients with colorectal cancer.
To determine the DNA replication error (RER) status in young patients with colorectal cancer (CRC), and to compare the clinical and pathologic characteristics of RER-positive and RER-negative cases. Recent studies suggest that patients with RER-positive CRC have an improved prognosis. Further data are required to confirm this observation in young CRC patients. All patients 40 years of age and younger with CRC admitted to the National Naval Medical Center between 1970 and 1992 were considered for inclusion in the study. After review, 36 patients for whom the original archived pathology specimen could be retrieved served as the study population. The RER status was determined using a previously described polymerase chain reaction-based assay. The clinical and pathologic features and survival data were compared to RER status. RER-positive tumors were found in 17 cases (47%). There was no significant difference in Dukes' stage or histologic grade at the time of diagnosis between patients with RER-positive tumors compared to RER-negative tumors. Patients with RER-positive tumors were found to have an improved prognosis: the 5-year survival probability for patients with RER-positive tumors was 68%, as compared to 32% for patients with RER-negative tumors (p < 0.05). RER-positive tumors are common in young patients with CRC, and patients with RER-positive tumors have a significantly improved prognosis. Because of their young age, survival data and prognosis play an important role in the overall treatment plan of young patients with CRC. Therefore, knowledge of RER status could affect initial therapy, postoperative chemotherapy, and follow-up.


Accuracy and influencing factors of maxillary and mandibular repositioning using pre-bent locking plates: a prospective study.
In-house repositioning methods based on three-dimensional (3D)-printing technology and the use of pre-bent plates has been gaining popularity in orthognathic surgery. However, there remains room for further improvement in methods and investigations on clinical factors that affect accuracy. This single-centre, prospective study included 34 patients and aimed to evaluate the accuracy and factors influencing maxillary and mandibular repositioning using pre-bent locking plates. The plates were manually pre-bent on the 3D-printed models of the planned position, and their hole positions were scanned and reproduced intraoperatively with osteotomy guides. The accuracy of repositioning and plate-hole positioning was calculated in three axes with the set landmarks. The following clinical factors that affect repositioning accuracy were also verified: deviation of the plate-hole positioning, amount of planned movement, and amount of simulated bony interference. The median deviations of the repositioning and hole positioning between the preoperative plan and postoperative results were 0.26 mm and 0.23 mm, respectively, in the maxilla, and 0.69 mm and 0.36 mm, respectively, in the mandible, suggesting that the method was highly accurate, and the repositioning concept based on the plate hole and form matching was more effective in the maxilla. Results of the correlation test suggest that large amounts of bony interference and plate-hole positioning errors in the up/down direction could reduce mandibular repositioning accuracy.


Determination of fipronil and bixafen pesticides residues using gas chromatography mass spectroscopy with matrix matching calibration strategy after binary dispersive liquid-liquid microextraction.
The objective of this study was to determine bixafen and fipronil residues in domestic and industrial wastewater, soil and mint samples by binary dispersive liquid-liquid microextraction method (BDLLME) prior to gas chromatography-mass spectrometry (GC-MS). Extraction efficiency for the selected analytes was improved by optimizing the parameters such as solvent type, ratio and volume, dispersive solvent type/volume, mixing type and duration to increase overall analytical performance. Under the optimum chromatographic and extraction conditions, limits of detection values for bixafen and fipronil were determined as 7.3 and 6.1 µg L-1, respectively. Spiking experiments were performed for domestic and industrial wastewater, soil and mint samples to evaluate applicability and accuracy of the proposed method. Recovery results for the samples were calculated in the range of 89.4%-112.6% via matrix matching calibration strategy. It was determined that the detection power of GC-MS system was improved 7.8 times for bixafen and 119 times for fipronil over LOD comparisons of conventional GC-MS and B-DLLME-GC-MS systems.


Perspective: agency and activism: rethinking health advocacy in the medical profession.
Health advocacy is being formalized as a professional activity for physicians across North America, but the accommodation of this activity into conceptions of daily practice has been controversial and confusing. There appears to be a lack of clarity around what a physician should do as a health advocate and how this should manifest in daily practice. In this article, the authors explore how the medical community has characterized the health advocate role and the roots of the debates regarding its place within training and practice, using the example of the CanMEDS Health Advocate Role. They argue that the confusion might be a result of subsuming two distinct activities, agency and activism, under the rubric of health advocacy. They propose that these activities and their associated skills are sufficiently distinct as to merit separate discussions. Agency involves advancing the health of individual patients ("working the system"), and activism involves advancing the health of communities and populations ("changing the system"). The authors suggest that distinguishing between agency and activism within health advocacy provides opportunities to explore their distinct goals and skill sets in a manner that will advance the debate about health advocacy, a conversation that remains critically important to the medical profession.


Comparison of the performance of the Panther Fusion respiratory virus panel to R-Gene and laboratory developed tests for diagnostic and hygiene screening specimens from the upper and lower respiratory tract.
Introduction. Diagnosis of acute respiratory infections (ARIs) can be facilitated by the Panther Fusion (PF) automatic, random access PCR system for the detection of influenzavirus A (Flu A) and B (Flu B), parainfluenzavirus (Paraflu), respiratory syncytial virus (RSV), human metapneumovirus (hMPV), rhinovirus (RV) and human adenovirus (AdV) in nasopharyngeal swabs.Aim. To evaluate the performance of PF in comparison with established methods, including subsets of (1) lower respiratory tract (LRT) specimens and (2) upper respiratory tract (URT) hygiene screening specimens of patients without ARI symptoms.Methodology. The performance characteristics of PF were compared with bioMérieux R-Gene and laboratory-developed PCR tests (LDTs). Overall, 1544 specimens with 6658 individual diagnostic requests were analysed.Results. The overall concordances of PF and LDTs for Flu A, Flu B and AdV were 98.4, 99.9 and 96.1%, respectively; by re-testing of discrepant specimens concordances increased to 99.4, 99.9 and 98.0%, respectively. Initial concordances of PF and R-Gene assays for RSV, Paraflu, hMPV and RV were 98.4, 96.3, 99.3 and 96.0%, respectively, and retest concordances were 99.7, 97.9, 99.9 and 98.9%, respectively. No differences to the overall performance were found for the subgroups of LRT and hygiene screening specimens. PCR cycle threshold (Ct) values correlated very well between methods, indicating that a semi-quantitative diagnostic approach using Ct values (e.g. highly vs. weakly positive) could augment the diagnostic information.Conclusion. PF performed similar to R-Gene and LDTs not only for its intended use but also for LRT and hygiene screening specimens with shorter hands-on and turnaround times.


A field comparison of inhalable and thoracic size selective sampling techniques.
We measured inhalable, thoracic, and so-called "total" wood dust exposure in British Columbia lumber mill workers. Particle-size selective sampling was conducted using the GSP and Seven hole inhalable samplers, the PEM thoracic sampler and the 37-mm closed-face cassette "total" sampler. All measurements were full-shift personal samples, obtained from randomly selected workers. We obtained intersampler comparison data for the following pairs of instruments: GSP and 37-mm sampler; GSP and seven-hole sampler (SHS); and PEM and 37-mm sampler. The intersampler measurement ratios were estimated as: GSP/37-mm sampler = 4.2; GSP/SHS = 1.7; and PEM/37-mm sampler = 1.6. The GSP/37-mm sampler ratio is consistent with previously reported findings, while PEM/37-mm sampler and GSP/SHS ratios were both larger than expected. We found that in all comparisons, the measurement ratio had significant variability that was greatest at low ambient dust concentrations. Although it was not possible to attribute the source of the variability to specific sampler types, we concluded that the GSP sampler might be susceptible to "projectile" particles not normally aspirated, and may be vulnerable to direct aspiration of dust from accidentally contacted surfaces. The PEM was designed for environmental monitoring, and it is possible that it is unsuited to the higher particulate concentrations found in some occupational settings. Disparities among inhalable sampling techniques such as that between GSP and SHS should be investigated further in light of the proposed adoption of the inhalable method as an industrial standard.


Pulmonary expansion manoeuvres compared to usual care on ventilatory mechanics, oxygenation, length of mechanical ventilation and hospital stay, extubation, atelectasis, and mortality of patients in mechanical ventilation: A randomized clinical trial.
Pulmonary expansion manoeuvres are therapeutic techniques used to prevent and reverse atelectasis; however, no randomized controlled trials have provided evidence supporting the use of this intervention among individuals on mechanical ventilation. To evaluate the effects of chest compression-decompression and chest block manoeuvres compared to usual care among patients on mechanical ventilation. The current study was a randomized clinical trial of adult subjects on mechanical ventilation for 12 to 48 hours. The control group received usual care (passive or active mobilization, manoeuvres for airway clearance and tracheal aspiration). The intervention group received usual care plus two lung expansion manoeuvres, i.e., chest decompression and chest block, while remaining on mechanical ventilation. Assessments were performed before and after usual care, immediately after the intervention and 30 minutes after the intervention. The primary outcome was static compliance. The secondary outcomes were the incidence of atelectasis, dynamic compliance, airway resistance, driving pressure, oxygenation, duration of mechanical ventilation, extubation success, length of hospital and ICU stay, and mortality. Fifty-one participants (67±15 years old, 53% men, 26 in the control group and 25 in the intervention group) were evaluated. No differences in static compliance were observed between groups (intervention minus control) before and after expansion manoeuvres [3.64 ml/cmH2O (95% CI: -0.36-7.65, p = 0.074)]. Peripheral oxygen saturation differed between groups before and after expansion manoeuvres, with more favourable outcome observed in the control group [-1.04% (95% CI: -1.94 --0.14), p = 0.027]. No differences were found in other outcomes. Chest compression-decompression and chest block manoeuvres did not improve ventilatory mechanics, the incidence of atelectasis, oxygenation, the duration of mechanical ventilation, the length of stay in the ICU and hospital, or mortality in individuals on mechanical ventilation. The findings of this study can be valuable for guiding evidence-based clinical practice and developing a therapeutic approach that provides real benefits for this population.


Plasma microRNA192 in combination with serum CA19-9 as non-invasive prognostic biomarker in periampullary carcinoma.
In this study, circulating microRNAs (miRNAs) are being investigated as non-invasive biomarkers for early diagnosis and prognosis of human cancers. Since the prognosis for pancreatobiliary subtype of periampullary carcinoma is poor, we assessed the prognostic relevance of miRNAs in combination with CA19-9 as noninvasive biomarker in periampullary carcinoma. Circulating miRNAs in plasma and serum CA19-9 were evaluated in periampullary carcinoma patients (n = 109) undergoing Whipple's pancreaticoduodenectomy and in healthy volunteers (n = 92). Tumour tissues were subjected to staging and subtyping prior to determining differentially expressed miRNAs in them by quantitative real-time polymerase chain reaction (qRT-PCR). Statistical analysis involved correlation, receiver operating characteristic, logistic regression, survival analyses and Cox-proportional regression. Of the three differentially expressed circulating miRNA, miRNA192 was significantly increased both in circulation and in tumour tissue and correlated with tumour stage and aggressiveness (r = 0.96, P < 0.0001). Area under the curve of circulating miRNA192 + CA19-9 combination was 0.877 (95% confidence interval, 0.72 to 0.96) for stage III and 0.92 (95% confidence interval, 0.77 to 0.88) for tumour aggressiveness. The combination was associated with poor survival (median: 22 months, P = 0.0008) in stage III patients. Cox-proportional regression analysis revealed prognostic importance of combination of circulating miR192 and CA19-9 (HR = 1.005, P = 0.0001) in periampullary carcinoma. In conclusion, combination of circulating miRNA192 with serum CA19-9 is a better prognostic biomarker than CA19-9 alone.


Laparoscopic management of ventriculoperitoneal and lumboperitoneal shunt complications.
Minimally invasive approaches for the initial placement of ventriculoperitoneal (VP) and lumboperitoneal (LP) shunts have been well described. A laparoscopic approach has multiple advantages over open techniques, including decreased morbidity, more rapid recovery, and ability to visually assess catheter function. However, few series have addressed the role of laparoscopy in the management of VP and LP shunt complications. We present here the largest published series of laparoscopic treatment of VP and LP shunt complications in adults, by retrospectively reviewing all cases performed in a 1-year interval by a single surgeon. Ten patients presented with complications of previous shunting; all were managed laparoscopically. Eighty percent of these patients had a successful single laparoscopic intervention. One patient developed a cerebrospinal fluid leak from the lumbar wound, and 2 patients required additional laparoscopic shunt revisions. We conclude that laparoscopy has great utility in the assessment of shunt function. Laparoscopic techniques should be considered not only for placement of peritoneal catheters, but also for the management of distal shunt malfunction and diagnosis of abdominal pain in these patients.


Gastric inhibitory polypeptide receptor methylation in newly diagnosed, drug-naïve patients with type 2 diabetes: a case-control study.
GIP action in type 2 diabetic (T2D) patients is altered. We hypothesized that methylation changes could be present in GIP receptor of T2D patients. This study aimed to assess the differences in DNA methylation profile of GIPR promoter between T2D patients and age- and Body Mass Index (BMI)-matched controls. We included 93 T2D patients (cases) that were uniquely on diet (without any anti-diabetic pharmacological treatment). We matched one control (with oral glucose tolerance test negative, non diabetic), by age and BMI, for every case. Cytokines and hormones were determined by ELISA. DNA was extracted from whole blood and DNA methylation was assessed using the Sequenom EpiTYPER system. Our results showed that T2D patients were more insulin resistant and had a poorer β cell function than their controls. Fasting adiponectin was lower in T2D patients as compared to controls (7.0±3.8 µgr/mL vs. 10.0±4.2 µgr/mL). Levels of IL 12 in serum were almost double in T2D patients (52.8±58.3 pg/mL vs. 29.7±37.4 pg/mL). We found that GIPR promoter was hypomethylated in T2D patients as compared to controls. In addition, HOMA-IR and fasting glucose correlated negatively with mean methylation of GIPR promoter, especially in T2D patients. This case-control study confirms that newly diagnosed, drug-naïve T2D patients are more insulin resistant and have worse β cell function than age- and BMI-matched controls, which is partly related to changes in the insulin-sensitizing metabolites (adiponectin), in the proinflammatory profile (IL12) and we suggest in the methylation pattern of GIPR. Our study provides novel findings on GIPR promoter methylation profile which may improve our ability to understand type 2 diabetes pathogenesis.


Inhibitors - genetic and environmental factors.
It is known that a large number of both genetic and environmental factors contribute to the risk of inhibitor development, but underlying pathogenetic mechanisms are still under investigation. The clinical research on inhibitors towards factor VIII (FVIII) is challenged by the fact that this is an infrequent event occurring in a rare disease. Therefore, it is widely accepted that complementary studies involving animal models can provide important insights into the pathogenesis and treatment of this complication. In this respect, mouse models have been studied for clues to FVIII immunogenicity, natural history of immunity and for different approaches to primary and secondary tolerance induction. In the clinical setting, the type of FVIII product used and the occurrence of product switching are considered important factors which may have an influence on inhibitor development. The evaluation of data currently available in the literature does not prove unequivocally that a difference in the immunogenicity exists between particular FVIII products (e.g. recombinant vs. plasma-derived, full length vs. B-domainless). In addition, national products switches have occurred and, in this context, switching was not associated with an enhanced inhibitor risk. In contrast with severe haemophilia A, patients with moderate and mild haemophilia A receive FVIII treatment infrequently for bleeds or surgery. In this condition the inhibitor risk is low but remains present lifelong, requiring continuous vigilance, particularly after intensive FVIII exposure.


Mechanistic implications for the formation of the diiron cluster in ribonucleotide reductase provided by quantitative EPR spectroscopy.
The small subunit of Escherichia coli ribonucleotide reductase (R2) is a homodimeric (betabeta) protein, in which each beta-peptide contains a diiron cluster composed of two inequivalent iron sites. R2 is capable of reductively activating O(2) to produce a stable tyrosine radical (Y122*), which is essential for production of deoxyribonucleotides on the larger R1 subunit. In this work, the paramagnetic Mn(II) ion is used as a spectroscopic probe to characterize the assembly of the R2 site with EPR spectroscopy. Upon titration of Mn(II) into samples of apoR2, we have been able to quantitatively follow three species (aquaMn(II), mononuclear Mn(II)R2, and dinuclear Mn(2)(II)R2) and fit each to a sequential two binding site model. As previously observed for Fe(II) binding within apoR2, one of the sites has a greater binding affinity relative to the other, K(1) = (5.5 +/- 1.1) x 10(5) M(-)(1) and K(2) = (3.9 +/- 0.6) x 10(4) M(-)(1), which are assigned to the B and A sites, respectively. In multiple titrations, only one dinuclear Mn(2)(II)R2 site was created per homodimer of R2, indicating that only one of the two beta-peptides of R2 is capable of binding Mn(II) following addition of Mn(II) to apoR2. Under anaerobic conditions, addition of only 2 equiv of Fe(II) to R2 (Fe(2)(II)R2) completely prevented the formation of any bound MnR2 species. Upon reaction of this sample with O(2) in the presence of Mn(II), both Y122* and Mn(2)(II)R2 were produced in equal amounts. Previous stopped-flow absorption spectroscopy studies have indicated that apoR2 undergoes a protein conformational change upon binding of metal (Tong et al. J. Am. Chem. Soc. 1996, 118, 2107-2108). On the basis of these observations, we propose a model for R2 metal incorporation that invokes an allosteric interaction between the two beta-peptides of R2. Upon binding the first equiv of metal to a beta-peptide (beta(I)), the aforementioned protein conformational change prevents metal binding in the adjacent beta-peptide (beta(II)) approximately 25 A away. Furthermore, we show that metal incorporation into beta(II) occurs only during the O(2) activation chemistry of the beta(I)-peptide. This is the first direct evidence of an allosteric interaction between the two beta-peptides of R2. Furthermore, this model can explain the generally observed low Fe occupancy of R2. We also demonstrate that metal uptake and this newly observed allosteric effect are buffer dependent. Higher levels of glycerol cause loss of the allosteric effect. Reductive cycling of samples in the presence of Mn(II) produced a novel mixed metal Fe(III)Mn(III)R2 species within the active site of R2. The magnitude of the exchange coupling (J) determined for both the Mn(2)(II)R2 and Fe(III)Mn(III)R2 species was determined to be -1.8 +/- 0.3 and -18 +/- 3 cm(-)(1), respectively. Quantitative spectral simulations for the Fe(III)Mn(III)R2 and mononuclear Mn(II)R2 species are provided. This work represents the first instance where both X- and Q-band simulations of perpendicular and parallel mode spectra were used to quantitatively predict the concentration of a protein bound mononuclear Mn(II) species.


Secondary use of data from hospital electronic prescribing and pharmacy systems to support the quality and safety of antimicrobial use: a systematic review.
Electronic prescribing (EP) and electronic hospital pharmacy (EHP) systems are increasingly common. A potential benefit is the extensive data in these systems that could be used to support antimicrobial stewardship, but there is little information on how such data are currently used to support the quality and safety of antimicrobial use. To summarize the literature on secondary use of data (SuD) from EP and EHP systems to support quality and safety of antimicrobial use, to describe any barriers to secondary use and to make recommendations for future work in this field. We conducted a systematic search within four databases; we included original research studies that were (1) based on SuD from hospital EP or EHP systems and (2) reported outcomes relating to quality and/or safety of antimicrobial use and/or qualitative findings relating to SuD in this context. Ninety-four full-text articles were obtained; 14 met our inclusion criteria. Only two described interventions based on SuD; seven described SuD to evaluate other antimicrobial stewardship interventions and five described descriptive or exploratory studies of potential applications of SuD. Types of data used were quantitative antibiotic usage data ( n = 9 studies), dose administration data ( n = 4) and user log data from an electronic dashboard ( n = 1). Barriers included data access, data accuracy and completeness, and complexity when using data from multiple systems or hospital sites. The literature suggests that SuD from EP and EHP systems is potentially useful to support or evaluate antimicrobial stewardship activities; greater system functionality would help to realize these benefits.


Role of an expanded inositol transporter repertoire in Cryptococcus neoformans sexual reproduction and virulence.
Cryptococcus neoformans and Cryptococcus gattii are globally distributed human fungal pathogens and the leading causes of fungal meningitis. Recent studies reveal that myo-inositol is an important factor for fungal sexual reproduction. That C. neoformans can utilize myo-inositol as a sole carbon source and the existence of abundant inositol in the human central nervous system suggest that inositol is important for Cryptococcus development and virulence. In accord with this central importance of inositol, an expanded myo-inositol transporter (ITR) gene family has been identified in Cryptococcus. This gene family contains two phylogenetically distinct groups, with a total of 10 or more members in C. neoformans and at least six members in the sibling species C. gattii. These inositol transporter genes are differentially expressed under inositol-inducing conditions based on quantitative real-time PCR analyses. Expression of ITR genes in a Saccharomyces cerevisiae itr1 itr2 mutant lacking inositol transport can complement the slow-growth phenotype of this strain, confirming that ITR genes are bona fide inositol transporters. Gene mutagenesis studies reveal that the Itr1 and Itr1A transporters are important for myo-inositol stimulation of mating and that functional redundancies among the myo-inositol transporters likely exist. Deletion of the inositol 1-phosphate synthase gene INO1 in an itr1 or itr1a mutant background compromised virulence in a murine inhalation model, indicating the importance of inositol sensing and acquisition for fungal infectivity. Our study provides a platform for further understanding the roles of inositol in fungal physiology and virulence.


Crystal structure and XANES investigation of petzite, Ag3AuTe2.
Petzite, Ag3AuTe2, crystallizes in the space group I4132, which is a Sohncke type of space group where chiral crystal structures can occur. The structure refinement of petzite reported long ago [Frueh (1959). Am. Mineral. 44, 693-701] did not provide any information about the absolute structure. A new single-crystal X-ray diffraction refinement has now been performed on a sample from Lake View Mine, Golden Mile, Kalgoorlie, Australia, which has resulted in a reliable absolute structure [a Flack parameter of 0.05 (3)], although this corresponds to the opposite enantiomorph reported previously. The minimum Te-Te distance is 3.767 (3) Å, slightly shorter than the van der Waals bonding distance, which suggests a weak interaction between the two chalcogens. XANES spectra near the Au and Te LIII edges suggest that the chemical-bonding character of Au in petzite is more metallic than in other gold minerals.


Evaluating the Impact of Prescription Fill Rates on Risk Stratification Model Performance.
Risk adjustment models are traditionally derived from administrative claims. Prescription fill rates-extracted by comparing electronic health record prescriptions and pharmacy claims fills-represent a novel measure of medication adherence and may improve the performance of risk adjustment models. We evaluated the impact of prescription fill rates on claims-based risk adjustment models in predicting both concurrent and prospective costs and utilization. We conducted a retrospective cohort study of 43,097 primary care patients from HealthPartners network between 2011 and 2012. Diagnosis and/or pharmacy claims of 2011 were used to build 3 base models using the Johns Hopkins ACG system, in addition to demographics. Model performances were compared before and after adding 3 types of prescription fill rates: primary 0-7 days, primary 0-30 days, and overall. Overall fill rates utilized all ordered prescriptions from electronic health record while primary fill rates excluded refill orders. The overall, primary 0-7, and 0-30 days fill rates were 72.30%, 59.82%, and 67.33%. The fill rates were similar between sexes but varied across different medication classifications, whereas the youngest had the highest rate. Adding fill rates modestly improved the performance of all models in explaining medical costs (improving concurrent R by 1.15% to 2.07%), followed by total costs (0.58% to 1.43%), and pharmacy costs (0.07% to 0.65%). The impact was greater for concurrent costs compared with prospective costs. Base models without diagnosis information showed the highest improvement using prescription fill rates. Prescription fill rates can modestly enhance claims-based risk prediction models; however, population-level improvements in predicting utilization are limited.


Selective externalization of an ATP-binding protein structurally related to the clathrin-uncoating ATPase/heat shock protein in vesicles containing terminal transferrin receptors during reticulocyte maturation.
Transferrin receptors are lost from reticulocytes in vesicles that are released during the final stage of erythrocyte maturation (Pan, B. T., and Johnstone, R. M. (1983) Cell 33, 967-977). Transferrin receptor-containing vesicles have a major protein component present in a 1:1 ratio with the receptor that migrates on sodium dodecyl sulfate gels as two polypeptides of Mr = 71,000 and 72,000. The Mr = 71,000/72,000 doublet is indistinguishable from the clathrin-uncoating ATPase/heat shock protein based on cross-reaction with affinity-purified antibody against the uncoating protein, by comparison of peptide maps of the Mr = 72,000 and 71,000 polypeptides and the uncoating protein, and by selective binding of these polypeptides to ATP-agarose. This finding suggests a possible activity of proteins related to the uncoating/heat shock protein family in the disposal of aged membrane proteins by a pathway independent of lysosomes.


[Early failure of restrained acetabuli in relapsing dislocation of total hip prostheses].
Dislocation is one of the major complications after a total hip replacement and several treatment alternatives have been described for it. One of them is the use of restrained acetabular implants, which led us to conducting an analysis of the failures of this type of components implanted in our center, focusing on the association between relapsing dislocation and the malpositioning of the locking ring. A total of 41 restrained components were implanted in 39 patients to treat the relapsing dislocation of total hip replacements. Mean postoperative follow-up was 24 months. The patients' demographics, the history of the causes for the dislocations and the treatment provided prior to the use of the restrained implant were all considered. The complications were also reviewed, with major ones being the revision rates and the re-dislocation of the prostheses and the acetabular insert locking ring, as well as the probable causes of failure of the antidislocation device. A total of 10 re-dislocations occurred during the follow-up; 70% of them in patients with malpositioning of the locking ring, with a statistically significant difference (p < 0.05). At the end of the follow-up period periacetabular radiolucent lines were seen in 9.09%. There are no papers in the literature that analyze the relation between malpositioning of the locking ring and repeated dislocations. The failure rate reported in our series was high. Restrained implants represent an alternative for relapsing dislocation, but they have to be very carefully placed.


Improving transition from paediatric to adult cystic fibrosis care: programme implementation and evaluation.
The paradigm of cystic fibrosis (CF) care has changed as effective therapies extend the lives of patients well into adulthood. Preparing for and maintaining high quality CF care into the adult healthcare setting is critical for prolonged survival. Unfortunately, this transfer process from the paediatric to the adult CF centre is met with a variety of challenges. The objective of this quality improvement (QI) project was to develop, implement and evaluate a theory-based programme for transition from paediatric to adult CF care. In a multi-phase process, the paediatric and adult programmes developed a transition curriculum, addressed care standards and standardised patient transfer protocols. We evaluated the impact of this process through staff surveys, review of field notes from QI meetings, tracking transfers and responses of patients to the Transition Readiness Assessment Questionnaire (TRAQ) at the start of the programme and 18 months after initiation. The collaboration between the paediatric and adult teams continued through quarterly meetings over the past 4 years. This has provided a forum that sustained our transition programme, harmonised care across CF centres and addressed other needs of our CF centre. Discussion of transition with families in the paediatric centre increased twofold (35% to 73% p<0.001), and resulted in a trend towards improved patient TRAQ self-advocacy scores and decreased in-hospital transfer. We successfully created a curriculum and process for transition from paediatric to adult CF care at our centres. This collaboration shapes the communication between our paediatric and adult CF care teams and enables ongoing feedback among patients, families and providers. The impact of our transition programme on long-term patient morbidity will require future evaluation.


[Axillary lymphadenectomy].
Axillary lymphadenectomy represents one of the historical hallmarks in the surgical strategy for breast cancer treatment. In recent years the role of axillary dissection is becoming matter for a renewed debate: up to date better therapeutic results together with new biologic acquisitions and progresses in early diagnosis have been influencing current indications and extension of axillary lymphadenectomy; also, cosmetic and financial considerations play a relevant role. This is the background of a large number of studies investigating the possibility of a more limited applications of axillary dissection provided that oncologic results remain unchanged. In this context sentinel node biopsy is one of the most promising lines of research. It can be stated that, as far as early stages are concerned, a trend towards more conservative resection techniques that started with the first quadrantectomies two decades ago, is now involving axillary dissection too. Possibly, in the next future, an elective axillary lymphadenectomy will not be performed any more if the presence of positive nodes will not be preliminarily ascertained. Nevertheless such conservative strategy can not be recommended to date, until the technique of sentinel node biopsy will be optimized and its results confirmed by randomized trials.


Growth factor superfamilies and mammalian embryogenesis.
With the availability of amino acid and nucleotide sequence information has come the realization that growth factors can be clustered in to superfamilies. Several of these superfamilies contain molecules that were not initially identified because of growth-promoting activities; rather they were discovered through their ability to regulate other processes. Certain members of these superfamilies are present during early mammalian embryogenesis. However, until recently, it has been difficult to manipulate the developing mammalian embryo to observe directly the effects of inappropriate, excessive, or reduced expression of these molecules. Despite this limitation, at least some of these molecules have been implicated in the control of differentiation and morphogenesis, two actions unpredicted from the cell biology of most of the growth factors. Moreover, these actions are reflected in nonmammalian species where homologues of the mammalian growth factors control crucial steps in the choice of developmental fate. This review describes five growth factor superfamilies and the role these molecules may have in controlling proliferation, differentiation, and morphogenesis during mammalian development.


Asymmetric cell division within the human hematopoietic stem and progenitor cell compartment: identification of asymmetrically segregating proteins.
The findings that many primitive human hematopoietic cells give rise to daughter cells that adopt different cell fates and/or show different proliferation kinetics suggest that hematopoietic stem cells (HSCs) and hematopoietic progenitor cells (HPCs) can divide asymmetrically. However, definitive experimental demonstration is lacking due to the current absence of asymmetrically segregating marker molecules within the primitive hematopoietic cell compartment. Thus, it remains an open question as to whether HSCs/HPCs have the capability to divide asymmetrically, or whether the differences that have been observed are established by extrinsic mechanisms that act on postmitotic progenitors. Here, we have identified 4 proteins (CD53, CD62L/L-selectin, CD63/lamp-3, and CD71/transferrin receptor) that segregate differentially in about 20% of primitive human hematopoietic cells that divide in stroma-free cultures. Therefore, this indicates for the first time that HSCs/HPCs have the capability to divide asymmetrically. Remarkably, these proteins, in combination with the surrogate stem-cell marker CD133, help to discriminate the more primitive human cultivated HSCs/HPCs. Since 3 of these proteins, the transferrin receptor and the tetraspanins CD53 and CD63, are endosomal-associated proteins, they may provide a link between the endosomal compartment and the process of asymmetric cell division within the HSC/HPC compartment.


Motor cognitive dual tasking: early detection of gait impairment, fall risk and cognitive decline.
Gait and cognition are closely associated. Older adults with gait deficits have an increased risk of developing cognitive deficits and cognitive deficits are associated with worsened gait. Both gait and cognitive impairments are risk factors for falls in older adults. The aims of this article are (1) to highlight the association between gait and cognition, particularly executive function, (2) to present motor cognitive dual tasking test paradigms and (3) to provide an algorithm for standardized mobility tests that can quickly and easily be performed in a private practice or on a hospital ward. A Pubmed review of current literature on the topic as well as the personal experience and recommendations of the authors are presented. Assessments summarized: clock drawing test, stops walking when talking test, normal walking speed, timed up and go test, regular, as a dual task and imagined. It is recommended that at least two of the presented assessments should be performed at each clinical visit in all patients age 65 years or older. If one of the assessments presented provides abnormal results, patients should be referred to a gait specialist for an in-depth quantitative gait analysis. Assessments of functional mobility, fall risk and cognition should be an integral part of every comprehensive geriatric assessment. Quantitative gait analysis allows not only the early detection of gait deficits and fall risk, but also of cognitive deficits. Early detection allows for timely implementation of targeted interventions to improve gait and/or cognition.


Pneumatic otoscopy in infants and children.
The pneumatic otoscope is indispensable in evaluating pressure within the middle ear of infants and children by measuring the tympanic membrane mobility. If the eardrum moves normally in response to positive and negative pressure, then a significant ear effusion, suppurative or nonsuppurative, is highly unlikely. Tympanic membrane color, luster, translucence, and light reflex should therefore be correlated with mobility before any decisions regarding therapy are made. In addition, the pneumatic otoscope with a veterinary otic speculum appears to be a simple and safe way to evaluate the vagina and rectum of infants and young children.


Magnetic fingerprint powder from a mineral indigenous to Thailand.
A study was conducted to investigate whether natural magnetite (Fe3O4), which is an abundant mineral in Thailand, could be used as a magnetic powder in the detection of latent fingerprints. Because of the presence of impurities, powdered magnetite is only weakly attracted by a magnet and cannot be used as a magnetic fingerprint powder by itself. Mixing a small amount of magnetite powder with nickel powder greatly enhances the magnetic attraction. A mixture of magnetite powder and nickel powder in a mass ratio of approximately 1:100 was found to be suitable for use as a magnetic fingerprint powder. Fingerprints developed using the magnetite/nickel mixture on nonporous surfaces were found to exhibit good adherence and clarity. Using an automated fingerprint identification system, the number of minutiae detected in fingerprints developed by using the prepared powder on nonporous surfaces was found to be comparable to those detected in fingerprints developed by using a commercial black magnetic powder. The cost is lowered by more than 60%.


Dynamics of Wastewater Effluent Contributions in Streams and Impacts on Drinking Water Supply via Riverbank Filtration in Germany-A National Reconnaissance.
The discharge of wastewater effluents to a stream that is subsequently used for drinking water abstraction has been previously referred to as de facto water reuse. Where the abstraction of surface water for drinking water production occurs via induced bank filtration or aquifer recharge, additional site-specific factors should be considered to assess the impact of wastewater effluents on bank-filtered water. This study represents the first national reconnaissance to quantify wastewater effluent contributions in streams across Germany and consequences for indirect drinking water abstraction from these streams. An automated assessment using ArcGIS was conducted for river basins considering minimum and mean average discharge conditions of streams as well as discharge from more than 7500 wastewater facilities. In urban areas, where the natural base discharge is low, wastewater effluent contributions greater than 30-50% were determined under mean minimum discharge conditions, which commonly prevail from May to September. A conceptual model was proposed to estimate critical bank filtrate shares resulting in exceedances of monitoring trigger levels for health-relevant chemicals as a universal qualitative assessment regarding the relevance of de facto reuse conditions in surface waters used for drinking water abstraction. This approach was validated using chemical monitoring data for three case study locations.


Human fetal mesenchymal stem cells differentiate into brown and white adipocytes: a role for ERRalpha in human UCP1 expression.
We investigated the ability of fetal mesenchymal stem cells (fMSCs) to differentiate into brown and white adipocytes and compared the expression of a number of marker genes and key regulatory factors. We showed that the expression of key adipocyte regulators and markers during differentiation is similar to that in other human and murine adipocyte models, including induction of PPARgamma2 and FABP4. Notably, we found that the preadipocyte marker, Pref-1, is induced early in differentiation and then declines markedly as the process continues, suggesting that fMSCs first acquire preadipocyte characteristics as they commit to the adipogenic lineage, prior to their differentiation into mature adipocytes. After adipogenic induction, some stem cell isolates differentiated into cells resembling brown adipocytes and others into white adipocytes. Detailed investigation of one isolate showed that the novel brown fat-determining factor PRDM16 is expressed both before and after differentiation. Importantly, these cells exhibited elevated basal UCP-1 expression, which was dependent on the activity of the orphan nuclear receptor ERRalpha, highlighting a novel role for ERRalpha in human brown fat. Thus fMSCs represent a useful in vitro model for human adipogenesis, and provide opportunities to study the stages prior to commitment to the adipocyte lineage. They also offer invaluable insights into the characteristics of human brown fat.


Nontrivial topology and topological phase transition in two-dimensional monolayer Tl.
Topological insulating material with dissipationless edge states is a rising star in spintronics. While most two-dimensional (2D) topological insulators belong to group-IV or -V elements in a honeycomb lattice, herein, we propose a new topological phase in the 2D hexagonal group-III crystal, h-Tl, based on a tight-binding model and density-functional theory calculation. Analysis of band dispersion reveals a Dirac nodal-ring near the Fermi level, which is attributed to px,y/pz band crossing. Upon inclusion of spin-orbit coupling (SOC), h-Tl turns into a quantum spin Hall insulator under 21% biaxial strain, confirmed by integrating spin Berry curvature in the Brillouin zone and spin-polarized edge states. A prominent feature of its electronic properties is that the effect of SOC plays two essential roles of both topological gap opening and band inversion between px,y/pz orbitals, which is the first observed phenomenon in 2D materials. This study extends the scope of 2D elemental topological insulators and presents a platform to design new 2D topotronics materials.


Molecular architecture and functional analysis of NetB, a pore-forming toxin from Clostridium perfringens.
NetB is a pore-forming toxin produced by Clostridium perfringens and has been reported to play a major role in the pathogenesis of avian necrotic enteritis, a disease that has emerged due to the removal of antibiotics in animal feedstuffs. Here we present the crystal structure of the pore form of NetB solved to 3.9 Å. The heptameric assembly shares structural homology to the staphylococcal α-hemolysin. However, the rim domain, a region that is thought to interact with the target cell membrane, shows sequence and structural divergence leading to the alteration of a phosphocholine binding pocket found in the staphylococcal toxins. Consistent with the structure we show that NetB does not bind phosphocholine efficiently but instead interacts directly with cholesterol leading to enhanced oligomerization and pore formation. Finally we have identified conserved and non-conserved amino acid positions within the rim loops that significantly affect binding and toxicity of NetB. These findings present new insights into the mode of action of these pore-forming toxins, enabling the design of more effective control measures against necrotic enteritis and providing potential new tools to the field of bionanotechnology.


Image analysis comparison of nerve staining with food dye, methylene blue or tissue marker.
Novel locoregional techniques use dye studies to confirm successful nerve targeting. The goal was to objectively quantify and compare nerve staining characteristics of dye mixtures commonly reported in the literature using image analysis software. Prospective, randomized cadaveric study. Thirty-six brachial plexus nerves from unpreserved pig cadavers were randomized into three groups of 12: FD (1:10 mixture of blue food dye and bupivacaine 0.5%), MB (methylene blue 1%) and TM (0.1:10 mixture of blue tissue marker and lidocaine 2%). Nerves were immersed in dye for 1, 15, 30 or 60 minutes (n = 3 each). Images of nerves before immersion (baseline) and at each time point with epineurium intact (superficial staining) and after longitudinal bisection (deep staining) were processed using image analysis software. Color saturation values were divided into quartiles (dark, medium-dark, medium-light or light). Percentage of stained nerve area in each quartile was calculated and compared using two-way anova. Superficially, at minute 1, dark saturation covered 40% of nerve area in FD versus 19% in MB (p = 0.04) and 0% in TM (p < 0.0001). In bisected nerves, dark and medium-dark saturations occurred only in FD; medium-light saturation comprised anywhere from 4% to 22.5% over time in FD versus <1% at any time in MB (p = 1.000; p = 0.343; p = 0.383; p = 0.262). Deep staining was not found in TM at any point. Food dye rapidly stains superficial and deep nerve layers. Based on these characteristics, investigators can choose the appropriate dye for their study.


Effect of managed care on children's relationships with their primary care physicians: differences by race.
Racial and ethnic disparities in the quality of the patient-provider relationship have been documented previously, but only among adults. Few studies have examined this aspect of primary care quality for children, and none has examined the role of managed care in mediating disparities. To explore variations in the associations among 3 managed care policies and the sustained patient-provider relationship for children by racial and ethnic group. Telephone survey of parents of a random sample of 413 children attending elementary school in a large school district spanning 3 cities in southern California. Self-reported managed care information was validated through contact with health plan representatives. Parent reporting of the strength of affiliation and interpersonal relationships among the child, the family, and the health care provider. Parents of Hispanic and black children reported significantly lower strength of affiliation than whites did (mean, 3.19 and 3.27 vs 3.57; P<.05 and P<.01, respectively). Asians and Hispanics reported significantly poorer interpersonal relationships with providers than whites did (mean, 3.35 and 3.38 vs 3.53; P<.05 for both). Managed care policies requiring patients to (1) sign up with a particular provider, (2) seek a referral for specialty care, and (3) stay within a network were associated with reductions in the interpersonal relationship. All minority groups, but not whites, reported significantly poorer relationships when managed care policies were present. Asian children were most sensitive to the managed care policies. Our data suggest that racial and ethnic minority children experience poorer patient-provider relationships compared with white children, even controlling for socioeconomic status and health system factors. Minority children are particularly sensitive to managed care policies that restrict patient freedom in choosing where to seek care. Implementing less restrictive policies may enhance the development of the patient-provider relationship.


The biopharmaceutics risk assessment roadmap for optimizing clinical drug product performance.
The biopharmaceutics risk assessment roadmap (BioRAM) optimizes drug product development and performance by using therapy-driven target drug delivery profiles as a framework to achieve the desired therapeutic outcome. Hence, clinical relevance is directly built into early formulation development. Biopharmaceutics tools are used to identify and address potential challenges to optimize the drug product for patient benefit. For illustration, BioRAM is applied to four relatively common therapy-driven drug delivery scenarios: rapid therapeutic onset, multiphasic delivery, delayed therapeutic onset, and maintenance of target exposure. BioRAM considers the therapeutic target with the drug substance characteristics and enables collection of critical knowledge for development of a dosage form that can perform consistently for meeting the patient's needs. Accordingly, the key factors are identified and in vitro, in vivo, and in silico modeling and simulation techniques are used to elucidate the optimal drug delivery rate and pattern. BioRAM enables (1) feasibility assessment for the dosage form, (2) development and conduct of appropriate "learning and confirming" studies, (3) transparency in decision-making, (4) assurance of drug product quality during lifecycle management, and (5) development of robust linkages between the desired clinical outcome and the necessary product quality attributes for inclusion in the quality target product profile.


Husband-wife communication and family planning practices among Malay married couples in Mukim Rusila, Terengganu.
The role of husband-wife communication in the practice of family planning was studied among the rural Malay couples in Mukim Rusila, Terengganu. It was a cross-sectional study in which a cluster sampling was used to select the study sample. A total of 193 (100.0%) wives and 74 (38.3%) husbands responded to a face-to-face interview. Visual Analogue Scale was used to measure the level of husband-wife communication. Husband-wife communication score was lower on family planning compared to other matters. There was a significant fair agreement on the scores between the husbands and the wives (p > 0.05) on family planning. There was a significant association between husband-wife communication on family planning and the current practice of family planning (p = 0.002). The 'likelihood' that couples who had had good husband-wife communication to practice family planning was about 2.8 times higher compared to those couples with poor husband-wife communication (95% C.I.: 1.4, 5.3). Further research need to be carried out, as the study was limited by certain constraints.


Attitude of Reproductive Healthcare Providers to Prenatal Diagnosis in a Low Resource Nigerian Setting.
Prenatal diagnosis comprises all diagnostic modalities aimed at gaining information about the embryo or fetal wellbeing. It enables antenatal care tailored to the individual need(s) of the fetus. To determine the knowledge, practice and prospect of prenatal diagnosis among reproductive health care providers in Abakaliki, Nigeria. This was a cross-sectional descriptive study in which completely filled self-administered semi-structured questionnaires were retrieved from 182 reproductive healthcare providers at Federal Teaching Hospital, Abakaliki (FETHA). The questionnaires contained 17 items covering the socio-demographic data, knowledge, practice and prospects of prenatal diagnosis among the respondents. A total of 179 respondents (98.4%) were aware of the prenatal diagnosis. One hundred and sixty four (90.1%) of the respondents agreed that, prenatal diagnostic services is offered in the study centre and 97% of these respondents cited ultrasound scan as the prenatal diagnostic investigation. While 133 respondents (73.1%) would allow parents to decide the next line of action after due counseling for the diagnosis of a condition not compatible with extrauterine life was made, 23(12.6%) of the respondents would offer termination of the pregnancy. Among the respondents, 173(95.1%) would encourage prenatal diagnosis at the study centre and 153(88.4%) of the 173 respondents would do so by educating the populace on the benefits of the procedure. However, 2(1.1%) of the respondent would not encourage the practice of prenatal diagnosis in FETHA citing adverse effects on the woman and her fetus. Reproductive healthcare providers in Abakaliki have a high level of awareness and favorable disposition to prenatal diagnosis. However, prenatal diagnosis is still rudimentary in this environment.


[Contractile function, glucose consumption and lactate release by the isolated rat heart during different regimens of alcohol administration and after discontinuation of ethanol].
Acute or chronic intoxication of rats with ethanol (intragastric administration at a dose of 8 g/kg or free-choice drinking of 10% ethanol for 3 months) produced no significant changes in contractile function, glycogen content, glucose uptake and lactate release in isolated hearts. Withdrawal syndrome simulated in rats following a short period of severe intoxication with ethanol at a dose of 4-5 g/kg twice daily has demonstrated a 15 and 28% decrease in peak systolic pressure and tension time index, respectively. In this case glucose uptake and lactate release were 2 times higher. Changes in glycogen level were observed three days after the last ethanol administration. The rats, survived after the abstinence period, revealed areas of perivascular myocardial necrosis. It is concluded that withdrawal syndrome plays an important role in pathogenesis of alcoholic cardiomyopathy.


Functional phenotyping of hepatic lymphocytes in murine MASH by mass cytometry.
Hepatic inflammation, driven by immune cells such as B and T lymphocytes, is a hallmark feature of metabolic dysfunction-associated steatohepatitis (MASH). Here, we detail a robust cytometry by time-of-flight (CyTOF) procedure to phenotype hepatic lymphocytes from mice with MASH. We employ custom metal conjugation of antibodies, isolation of hepatic lymphocytes, cell surface and intracellular staining, and data acquisition. This protocol overcomes the limitations of traditional flow cytometry by accommodating up to 40 markers for comprehensive immune phenotyping. For complete details on the use and execution of this protocol, please refer to Barrow et al.1.


Nuclear segmentation, condensation and bilateral symmetry in polymorphonuclear leukocytes reflect genomic order and favor immunologic function.
Segmentation, condensation and bilateral symmetry of the nuclei of polymorphonuclear leukocytes seem related to their function. Segmentation of the nuclei into two or more lobes and their condensation facilitate their passage (diapedesis) through the endothelial layer of blood vessels to the extravasal space and subsequent locomotion through the interstitial compartment of different tissues. Bilateral symmetry of these nuclei along with their association to the cytoskeletal fibers contribute to their efficiency in locomotion by alignment of the axis of nuclear symmetry to the axis of cellular polarity, which orients towards the direction of locomotion in response to cytokines and other stimuli. Observations of the cytogenetic facets of intranuclear order support these assumptions.


Investigation of the effect of astaxanthin on alveolar bone loss in experimental periodontitis.
Astaxanthin is a keto-carotenoid that has a strong antioxidant effect. The purpose of this study was to evaluate the effects of astaxanthin on alveolar bone loss and histopathological changes in ligature-induced periodontitis in rats. Wistar rats were divided into four experimental groups: non-ligated (C, n = 6); ligature only (L, n = 6); ligature and astaxanthin (1 mg/kg/day astaxanthin, AS1 group, n = 8); ligature and astaxanthin (5 mg/kg/day astaxanthin, AS5 group, n = 8). Silk ligatures were placed at the gingival margin of lower first molars of the mandibular quadrant. The study duration was 11 days and the animals were killed at the end of this period. Changes in alveolar bone levels were clinically measured and tissues were immunohistochemically examined, osteocalcin, bone morphogenic protein-2, inducible nitric oxide synthase, Bax and bcl-2 levels in alveolar bone and tartrate-resistant acid phosphatase-positive osteoclast cells, osteoblast and inflammatory cell counts were determined. Alveolar bone loss was highest in the L group and the differences among the L, AS1 and AS5 groups were also significant (P < .05). Both doses of astaxanthin decreased tartrate-resistant acid phosphatase-positive+ osteoclast cell and increased osteoblast cell counts (P < .05). The inflammation in the L group was also higher than those of the C and AS1 groups were (P < .05) indicating the anti-inflammatory effect of astaxanthin. Although inducible nitric oxide synthase, osteocalcin, bone morphogenic protein-2 and bax staining percentages were all highest in the AS5 group and bcl-2 staining percentage was highest in the AS1 group, values were close to each other (P > .05). Within the limits of this study, it can be suggested that astaxanthin administration may reduce alveolar bone loss by increasing osteoblastic activity and decrease osteoclastic activity in experimental periodontitis model.


The impact of employment impasse and its resolution in the allied health professions.
Although conflict within organizations is inevitable, any of its damaging consequences may be minimized or controlled if a consensual process for its rational resolution exists. Since collective bargaining is becoming an increasingly prevalent mechanism for governing the relationships between employees and employers in the allied health professions, knowledge about the various formal conflict resolution mechanisms available is vital. The concept of impasse and its resolution in negotiations and in administration of a labor contract are analyzed. The impact of impasse resolution is summarized, in order to emphasize the interdependent nature of the employment relationship.


The Development of Steady-State Activation Hubs between Adult LTi ILC3s and Primed Macrophages in Small Intestine.
Group 3 innate lymphoid cells (ILC3s) are important for intestinal health, particularly in controlling inflammation in response to epithelial dysregulation, but their role during homeostasis remains less well understood. We generated IL-22 reporter mice to assess production of this key cytokine by ILC3s in the small intestine during development and under basal conditions. Although IL-22 is produced by a variety of lymphocyte populations, constitutively high IL-22 expression was limited to lymphoid-tissue inducer (LTi) cells residing in lymph node-like structures in the gut called solitary intestinal lymphoid tissues (SILT). Constitutive IL-22 expression was dependent on the microbiota and MyD88 signaling, appeared upon weaning, and was present across the spectrum of SILT, including in cryptopatches. Activated SILT LTi cells colocalized with a rare subpopulation of activated macrophages constitutively positive for IL-12/23 p40 and capable of activating neonatal LTi cells in response to TLR stimulus. Thus, weaning leads to the organization of innate immune activation hubs at SILT that mature and are continuously sustained by signals from the microbiota. This functional and anatomic organization constitutes a significant portion of the steady-state IL-23/IL-22 axis.


A dual modality approach to quantitative quality control in emission tomography.
Routine quality control (QC) and optimization of image quality of reconstructed images in single photon emission computed tomography (SPECT) and positron emission tomography (PET) remains a relatively qualitative exercise. With the advent of combined SPECT/CT and PET/CT devices, and accurate post hoc co-registration algorithms, the potential exists to utilize high resolution structural information for QC evaluation in addition to their use for anatomical correlation in clinical studies. The aim of this work was to explore, in principle, the uses of x-ray CT data of QC phantoms used in SPECT and PET to develop more objective assessments of performance of the emission tomographic (ET) devices and reconstructed data. A CT reconstruction of a novel ET QC phantom was segmented into the various compartments it contained. Using software, the voxel values in the different compartments were then altered to correspond to the concentration of the radioactivity in the actual scan of the same phantom on the SPECT system. This produces a high resolution version of a 'perfect' ET scan. Image co-registration techniques were then used to spatially align the synthetic high resolution SPECT scan to the measured SPECT scan. Various parameters can then be objectively derived from the registered data, for example, image contrast, spatial resolution, spatial non-uniformity, etc. In this study, we have used this approach to estimate spatial resolution (full width at half maximum, FWHM) and recovered contrast in reconstructed images of a SPECT phantom. Two independent methods were used to measure spatial resolution, obtaining excellent agreement. In conclusion, the ability to produce high resolution synthetic phantoms in emission tomography QC affords an objective approach to assessing system performance and optimizing protocols which is readily automated and quantifiable.


Taming Electrowetting Using Highly Concentrated Aqueous Solutions.
Wetting of carbon surfaces is one of the most widespread, yet poorly understood, physical phenomena. Control over wetting properties underpins the operation of aqueous energy-storage devices and carbon-based filtration systems. Electrowetting, the variation in the contact angle with an applied potential, is the most straightforward way of introducing control over wetting. Here, we study electrowetting directly on graphitic surfaces with the use of aqueous electrolytes to show that reversible control of wetting can be achieved and quantitatively understood using models of the interfacial capacitance. We manifest that the use of highly concentrated aqueous electrolytes induces a fully symmetric and reversible wetting behavior without degradation of the substrate within the unprecedented potential window of 2.8 V. We demonstrate where the classical "Young-Lippmann" models apply, and break down, and discuss reasons for the latter, establishing relations among the applied bias, the electrolyte concentration, and the resultant contact angle. The approach is extended to electrowetting at the liquid|liquid interface, where a concentrated aqueous electrolyte drives reversibly the electrowetting response of an insulating organic phase with a significantly decreased potential threshold. In summary, this study highlights the beneficial effect of highly concentrated aqueous electrolytes on the electrowettability of carbon surfaces, being directly related to the performance of carbon-based aqueous energy-storage systems and electronic and microfluidic devices.


Castleman disease of plasma cell type accompanied with bronchiolitis obliterans: a case report and review of the literature.
Castleman disease, also known as giant lymph node hyperplasia or angiofollicular lymph node hyperplasia, is a highly heterogeneous clinicopathological entity that belongs to the family lymphoproliferative disorders. Castleman disease accompanied by bronchiolitis obliterans is uncommon and often poses a great diagnostic challenge, which is easily confused with respiratory diseases and impeding the correct diagnosis and treatment. The main aim in presenting such rare case studies is to raise awareness and expand the diagnostic horizon of clinicians for appropriate management. Here, we present a 69-year-old Chinese male who was admitted to our hospital due to right chest pain for 6 months, accompanied by cough, expectoration, and fever. Laboratory examinations revealed elevated immunoglobulin G and C-reactive protein, and normal serum levels of tumor markers and interleukin-6. Computed tomography scan detected diffuse bronchial wall thickening and patchy area of air trapping consistent with small airway disease. Pulmonary function test showed mild small airway obstructive ventilation dysfunction and moderate decrease in diffusion capacity. The pathological result of the right axillary lymph node was consistent with the plasma cell type Castleman disease. According to the above examinations, the patient was finally diagnosed with the plasma cell type Castleman disease accompanied with bronchiolitis obliterans. He received immunosuppressive medication after surgery and has been followed up for 11 months. Now the patient is currently in stable condition without recurrence. Castleman disease is a rare lymphoproliferative disorder with a variety of symptoms. At present, the treatment of Castleman disease accompanied with bronchiolitis obliterans is mostly based on experiences or previous case reports, and there is no standard treatment. Here, we report an uncommon case of Castleman disease accompanied with bronchiolitis obliterans in which the patient received immunosuppressive medication after surgery and has been followed up for 11 months without experiencing a recurrence, which may deepen and extend our understanding of this disease.


Differentiation-associated toxin receptor modulation, cytokine production, and sensitivity to Shiga-like toxins in human monocytes and monocytic cell lines.
Infections with Shiga toxin-producing Shigella dysenteriae type 1 or Shiga-like toxin (SLT)-producing Escherichia coli cause bloody diarrhea and are associated with an increased risk of acute renal failure and severe neurological complications. Histopathological examination of human and animal tissues suggests that the target cells for toxin action are vascular endothelial cells. Proinflammatory cytokines regulate endothelial cell membrane expression of the glycolipid globotriaosylceramide (Gb(3)) which serves as the toxin receptor, suggesting that the host response to the toxins or other bacterial products may contribute to pathogenesis by regulating target cell sensitivity to the toxins. We examined the effects of purified SLTs on human peripheral blood monocytes (PBMn) and two monocytic cell lines. Undifferentiated THP-1 cells were sensitive to SLTs. Treatment of the cells with a number of differentiation factors resulted in increased toxin resistance which was associated with decreased toxin receptor expression. U-937 cells, irrespective of maturation state, and PBMn were resistant to the toxins. U-937 cells expressed low levels of GB(3), and toxin receptor expression was not altered during differentiation. Treatment of monocytic cells with tumor necrosis factor alpha (TNF-alpha) did not markedly increase sensitivity or alter toxin receptor expression. Undifferentiated monocytic cells failed to synthesize TNF and interleukin 1beta when treated with sublethal concentrations of SLT type I (SLT-I), whereas cells treated with 12-0-tetradecanoylphorbol-13-acetate acquired the ability to produce cytokines when stimulated with SLT-I. When stimulated with SLT-I, U-937 cells produced lower levels of TNF than PBMn and THP-1 cells did.


Successful in vitro expansion and differentiation of cord blood derived CD34+ cells into early endothelial progenitor cells reveals highly differential gene expression.
Endothelial progenitor cells (EPCs) can be purified from peripheral blood, bone marrow or cord blood and are typically defined by a limited number of cell surface markers and a few functional tests. A detailed in vitro characterization is often restricted by the low cell numbers of circulating EPCs. Therefore in vitro culturing and expansion methods are applied, which allow at least distinguishing two different types of EPCs, early and late EPCs. Herein, we describe an in vitro culture technique with the aim to generate high numbers of phenotypically, functionally and genetically defined early EPCs from human cord blood. Characterization of EPCs was done by flow cytometry, immunofluorescence microscopy, colony forming unit (CFU) assay and endothelial tube formation assay. There was an average 48-fold increase in EPC numbers. EPCs expressed VEGFR-2, CD144, CD18, and CD61, and were positive for acetylated LDL uptake and ulex lectin binding. The cells stimulated endothelial tube formation only in co-cultures with mature endothelial cells and formed CFUs. Microarray analysis revealed highly up-regulated genes, including LL-37 (CAMP), PDK4, and alpha-2-macroglobulin. In addition, genes known to be associated with cardioprotective (GDF15) or pro-angiogenic (galectin-3) properties were also significantly up-regulated after a 72 h differentiation period on fibronectin. We present a novel method that allows to generate high numbers of phenotypically, functionally and genetically characterized early EPCs. Furthermore, we identified several genes newly linked to EPC differentiation, among them LL-37 (CAMP) was the most up-regulated gene.


Magneto-thermal properties and slow magnetic relaxation in Mn(II)Ln(III) complexes: influence of magnetic coupling on the magneto-caloric effect.
A family of Mn(II)Ln(III) dinuclear and tetranuclear complexes (Ln = Gd and Dy) has been prepared from the compartmental ligands N,N'-dimethyl-N,N'-bis(2-hydroxy-3-formyl-5-bromobenzyl)ethylenediamine (H2L1) and N,N',N''-trimethyl-N,N''-bis(2-hydroxy-3-methoxy-5-methylbenzyl)diethylenetriamine (H2L2). The Mn(II)Gd(III) complexes exhibit antiferromagnetic interactions between Mn(II) and Gd(III) ions in most cases, which are supported by Density Functional Theory (DFT) calculations. Experimental magneto-structural correlations carried out for the reported complexes and other related complexes found in bibliography show that the highest ferromagnetic coupling constants are observed in di-μ-phenoxido bridged complexes, which is due to the planarity of the Mn-(μ-O)2-Gd bridging fragment and to the high Mn-O-Gd angles. The effect of these angles has been studied by DFT calculations performed on a di-μ-phenoxido doubly bridged model. The magneto-thermal properties of the Mn(II)Gd(III) based complexes have also been measured, concluding that the magnitude of the Magneto-Caloric Effect (MCE) is due to the strength rather than to the nature of the magnetic coupling. Moreover, when two Mn(II)Gd(III) dinuclear units are connected by two carbonato-bridging ligands the MCE is enhanced, obtaining a maximum magnetic entropy change of 36.4 Jkg-1 K-1 at ΔB = 7 T and T = 2.2 K. On the other hand, one of the dinuclear Mn(II)Dy(III) complexes displays Single-Molecule Magnet (SMM) behaviour with an energy barrier of 14.8 K under an applied external field of 1000 Oe.


Exploring diazepam's effect on hemodynamic responses of mouse brain tissue by optical spectroscopic imaging.
In this study, a simple duel-optical spectroscopic imaging apparatus capable of simultaneously determining relative changes in brain oxy-and deoxy-hemoglobin concentrations was used following administration of the anxiolytic compound diazepam in mice with strong dominant (Dom) and submissive (Sub) behavioral traits. Three month old mice (n = 30) were anesthetized and after 10 min of baseline imaging, diazepam (1.5 mg/kg) was administered and measurements were taken for 80 min. The mouse head was illuminated by white light based LED's and diffused reflected light passing through different channels, consisting of a bandpass filter and a CCD camera, respectively, was collected and analyzed to measure the hemodynamic response. This work's major findings are threefold: first, Dom and Sub animals showed statistically significant differences in hemodynamic response to diazepam administration. Secondly, diazepam was found to more strongly affect the Sub group. Thirdly, different time-series profiles were observed post-injection, which can serve as a possible marker for the groups' differentiation. To the best of our knowledge, this is the first report on the effects of an anxiolytic drug on brain hemodynamic responses in mice using diffused light optical imaging.


Photoperiodic modulation of GnRH mRNA in the male Syrian hamster.
Male Syrian hamsters (Mesocricetus auratus) are seasonal breeders. They show marked testicular regression when exposed to short autumnal photoperiods, and then remain sexually quiescent for several months. By mid-winter, however, they show a loss in responsiveness to the inhibitory influence of short photoperiods and their testes begin to recrudesce. To shed light on the neuroendocrine mechanism responsible for mediating these reproductive changes, we examined the influence of photoperiod on the expression of GnRH mRNA in the hamster forebrain. Adult males were either exposed to short photoperiods (6L:18D) for 16 weeks or were maintained under long photoperiods (14L:10D); additional animals were exposed to short or long photoperiods for 22 weeks. As expected, exposure to short photoperiods for 12 weeks resulted in a marked decrease (P<0.01) in testicular mass and serum testosterone levels, but after 22 weeks these reproductive parameters were once again significantly elevated (P<0.01). In contrast, quantitative in situ hybridization histochemistry revealed no difference (P>0.05) between the GnRH mRNA levels of the short-photoperiod hamsters and their aged-matched long-photoperiod controls, although an age-related decrease (P<0.05) was evident in both photoperiod-treatment groups. These data emphasize that GnRH mRNA is highly expressed in hamsters even when their reproductive axis has been rendered sexually quiescent by exposure to short photoperiods, and that photoperiod-induced changes in GnRH secretion, rather than synthesis, are more likely to regulate the timing of the breeding season. On the other hand, the data indicate that GnRH mRNA levels show an aging-related decrease, regardless of photoperiod, suggesting that in the long term a decrease in GnRH gene expression may contribute to the reduced fertility of old hamsters.


S14-Phosphorylated RPN6 Mediates Proteasome Activation by PKA and Alleviates Proteinopathy.
A better understanding of the regulation of proteasome activities can facilitate the search for new therapeutic strategies. A cell culture study shows that PKA (cAMP-dependent protein kinase or protein kinase A) activates the 26S proteasome by pS14-Rpn6 (serine14-phosphorylated Rpn6), but this discovery and its physiological significance remain to be established in vivo. Male and female mice with Ser14 of Rpn6 (regulatory particle non-ATPase 6) mutated to Ala (S14A [Rpn6/Psmd11S14A]) or Asp (S14D) to respectively block or mimic pS14-Rpn6 were created and used along with cells derived from them. cAMP/PKA were manipulated pharmacologically. Ubiquitin-proteasome system functioning was evaluated with the GFPdgn (green fluorescence protein with carboxyl fusion of the CL1 degron) reporter mouse and proteasomal activity assays. Impact of S14A and S14D on proteotoxicity was tested in mice and cardiomyocytes overexpressing the misfolded protein R120G-CryAB (R120G [arginine120 to glycine missense mutant alpha B-crystallin]). PKA activation increased pS14-Rpn6 and 26S proteasome activities in wild-type but not S14A embryonic fibroblasts (mouse embryonic fibroblasts), adult cardiomyocytes, and mouse hearts. Basal 26S proteasome activities were significantly greater in S14D myocardium and adult mouse cardiomyocytes than in wild-type counterparts. S14D::GFPdgn mice displayed significantly lower myocardial GFPdgn protein but not mRNA levels than GFPdgn mice. In R120G mice, a classic model of cardiac proteotoxicity, basal myocardial pS14-Rpn6 was significantly lower compared with nontransgenic littermates, which was not always associated with reduction of other phosphorylated PKA substrates. Cultured S14D neonatal cardiomyocytes displayed significantly faster proteasomal degradation of R120G than wild-type neonatal cardiomyocytes. Compared with R120G mice, S14D/S14D::R120G mice showed significantly greater myocardial proteasome activities, lower levels of total and K48-linked ubiquitin conjugates, and of aberrant CryAB (alpha B-crystallin) protein aggregates, less fetal gene reactivation, and cardiac hypertrophy, and delays in cardiac malfunction. This study establishes in animals that pS14-Rpn6 mediates the activation of 26S proteasomes by PKA and that the reduced pS14-Rpn6 is a key pathogenic factor in cardiac proteinopathy, thereby identifying a new therapeutic target to reduce cardiac proteotoxicity.


Conditioning medicine for ischemic and hemorrhagic stroke.
Remote ischemic conditioning (RIC) is a promising safe, feasible, and inexpensive treatment for acute stroke, both ischemic and hemorrhagic. It is applied with a blood pressure cuff on the limbs and is ideal for the prehospital setting. RIC is a form of preconditioning with similarities to physical exercise. Its mechanisms of action are multiple and include improvement of collateral cerebral blood flow (CBF) and RIC acts as a "collateral therapeutic". The increased CBF is likely related to nitric oxide synthase 3 in the endothelium and more importantly in circulating blood cells like the red blood cell. The RESIST clinical trial is a 1500 subject multicenter, randomized, sham-controlled trial of RIC in the prehospital setting in Denmark and should address the questions of whether RIC is safe and effective in acute stroke and whether the effect is mediated by an effect on nitric oxide/nitrite metabolism.


Genetic characterization of the oral Actinomyces.
Actinomyces are difficult to identify using serological and biochemical methods but genotyping is an efficient and reliable means of bacterial characterization and can be used to determine clonal identity. The purpose here was to genotype 13 American type culture collection (ATCC) reference strains representing six different oral Actinomyces spp. by using chromosomal DNA fingerprinting (CDF), arbitrarily primed-polymerase chain reaction (AP-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In CDF analysis, BamHI, BstEII and SmaI yielded digestion patterns revealing characteristic differences among the known Actinomyces spp., with SmaI demonstrating optimal resolution. Amplicons generated by AP-PCR with primer OPB-07 displayed banding patterns that permitted discrimination of all Actinomyces strains tested. PCR-RFLP with MnlI digests generated fragment patterns that also characterized the reference strains. Collectively, genotypic profiles generated by CDF, AP-PCR and PCR-RFLP permitted differentiation of all 13 ATCC Actinomyces strains. SmaI CDF analysis of 18 clinical isolates of catalase-positive A. naeslundii genospecies 2 revealed extensive genetic diversity among these strains. These molecular approaches may be useful in determining genetic diversity within oral Actinomyces populations and fidelity of Actinomyces transmission between mother and child.


The functional role of the triceps surae muscle during human locomotion.
Despite numerous studies addressing the issue, it remains unclear whether the triceps surae muscle group generates forward propulsive force during gait, commonly identified as 'push-off'. In order to challenge the push-off postulate, one must probe the effect of varying the propulsive force while annulling the effect of the progression velocity. This can be obtained by adding a load to the subject while maintaining the same progression velocity. Ten healthy subjects initiated gait in both unloaded and loaded conditions (about 30% of body weight attached at abdominal level), for two walking velocities, spontaneous and fast. Ground reaction force and EMG activity of soleus and gastrocnemius medialis and lateralis muscles of the stance leg were recorded. Centre of mass velocity and position, centre of pressure position, and disequilibrium torque were calculated. At spontaneous velocity, adding the load increased disequilibrium torque and propulsive force. However, load had no effect on the vertical braking force or amplitude of triceps activity. At fast progression velocity, disequilibrium torque, vertical braking force and triceps EMG increased with respect to spontaneous velocity. Still, adding the load did not further increase braking force or EMG. Triceps surae is not responsible for the generation of propulsive force but is merely supporting the body during walking and restraining it from falling. By controlling the disequilibrium torque, however, triceps can affect the propulsive force through the exchange of potential into kinetic energy.


Trochlear and abducens nerve neurinomas accompanied by a cerebellopontine angle meningioma--case report.
A 66-year-old male presented with cerebellovestibular symptoms and hypesthesia of the V2 area of his face. Neuroimaging only detected a cerebellopontine angle (CPA) meningioma. The CPA meningioma was removed using the lateral suboccipital approach, which exposed small neurinomas arising from the trochlear and abducens nerves. Both neurinomas were removed intracapsularly. Postoperatively hypesthesia resolved, but other symptoms were unchanged. A karyotypic analysis of chromosome 22 and estrogen receptor analysis suggested absence of neurofibromatosis II, but the cause(s) of the genesis of the multiple diverse tumors was not determined. This extremely rare combination of neurinomas and meningioma was probably incidental, as there are no reports of any case which a combination of the trochlear and the abducens nerve neurinomas, much less one accompanied by a meningioma.


BL-P1654, ticarcillin, and carbenicillin: in vitro comparison alone and in combination with gentamicin against Pseudomonas aeruginosa.
Minimum inhibitory concentrations of carbenicillin, ticarcillin, and BL-P1654 were determined for 89 clinical isolates of Pseudomonas aeruginosa. Ticarcillin was generally twice as active and BL-P1654 eight to 16 times as active as carbenicillin. Usually carbenicillin and ticarcillin killed at the same concentration or twice the concentration needed to inhibit, whereas 400 mug of BL-P1654 per ml was not bactericidal for the majority of isolates tested. The inhibitory effect of all three drugs varied markedly with the size of bacterial inoculum. When therapeutically achievable concentrations were used, adding gentamicin enhanced the inhibitory and bactericidal activity of all three penicillin derivatives for the majority of isolates. However, inhibition of isolates highly resistant to gentamicin was not improved by combining the semisynthetic penicillins with gentamicin.


Metformin sensitizes endometrial cancer cells to chemotherapy through IDH1-induced Nrf2 expression via an epigenetic mechanism.
Chemoresistance is the major obstacle to cure endometrial cancer, whereas metformin has demonstrated sensitization to chemotherapy in endometrial cancer. A novel finding states that isocitrate dehydrogenase 1 (IDH1) involves in cancer chemoresistance. Recent studies have revealed that epigenetic modifications facilitate chemoresistance. However, whether IDH1 play a role in metformin-induced endometrial cancer chemosensitivity through epigenetic modification is incompletely understood. Immunohistochemistry and Elisa assays were used to evaluate the expression pattern of IDH1 in endometrial tissue and serum, respectively. Western blot was performed to determine changes in expression of key molecules in the IDH1-ɑ-KG-TET1-Nrf2 signaling pathway after various treatments. Dot blot assays were used to assess global hydroxymethylation levels after metformin administration or plasmid transfection. Antioxidant response element (ARE) activity in the IDH1 promoter region was monitored by luciferase assay. Cancer cell sensitivity to chemotherapy was detected by SRB assay. We found that activation of the IDH1 signaling pathway in endometrial cancer tissue resulting from aberrant expression of IDH1 and its downstream mediators conferred chemoresistance. We found that this effect was abated by metformin treatment. Dot blot and HMeDIP assays revealed that metformin blocked IDH1-ɑ-KG-TET1-mediated enhancement of Nrf2 hydroxymethylation levels, eliminating chemoresistance. Moreover, we observed that chemoresistance was enhanced via a regulatory loop in which Nrf2 activated IDH1-ɑ-KG-TET1-Nrf2 signaling via binding to the ARE sites in the IDH1 promoter region. Our findings highlight a critical role of IDH1-ɑ-KG-TET1-Nrf2 signaling in chemoresistance and suggest that rational combination therapy with metformin and chemotherapeutics has the potential to suppress chemoresistance.


[Percutaneous embolization of the kidney: indications and clinical results].
Renal embolisation offers a minimal invasive means of renal ablation under primary palliative indication. We evaluated our experience with this technique in a series of 40 patients. From 1996 to 2002 a total of 40 patients underwent total embolisation of the kidney. Our patients encompassed 15 females and 25-males (median age: 68 years, 46 - 89 years). In 15 patients indication for embolisation was to ablate a non-functioning kidney following obstruction due to advanced cancer, in 9 patients preoperatively with advanced renal cell carcinomas invading the vena cava, in 8 patients for control of hematuria and in 8 patients with non-operable advanced renal cancers. The embolisation was carried out in DAS-technique with a 7 Fr. selective catheter and central placement of macrocoils, from 2000 on we additionally performed a peripheral embolisation with polyvinylalcohol (PVA) particles. The intervention took place in epidural anesthesia. Perioperatively in all patients a complete occlusion of the renal artery as well as acessory arteries (n = 6) could be documented radiographically, perioperative complications were not observed. A post-embolisation syndrome occured in 32 patients. Reinterventions with repeat embolisation became necessary in 5 patients (persistent urine production-n = 4, persistent hematuria-n = 1). The combined central and peripheral embolisation of the kidney in epidural anesthesia is a valuable means in selected patients under palliative indication for defunctionalisation of the kidney as well as control of hematuria.


Crystal site location of iron and trace elements in a magnesium-iron olivine by a new crystallographic technique.
A new crysallographic technique has been developed, which has been applied to the problem of locating the cations in a natural olivine crystal with the composition (Mg(0.90)Fe(0.10)Ni(0.004)Mn(0.002))(2)SiO(4). The method uses the variation of characteristic x-ray emission with the direction of an exciting electron beam in an analytical transmission electron microscope. It may be applied to nanometer-sized areas and to concentrations as low as 0.1 atomic percent, is capable of distinguishing neighbors in the periodic table, and does not require external standards. The iron atoms in this crystal are evenly distributed between the two available crystal sites M1 and M2 (49.6 +/- 1 percent on M1), whereas the trace elements nickel and manganese occupy the M1 and M2 positions, respectively (97 +/- 5 percent nickel on M1 and 1 +/- 5 percent manganese on M1).


Comparison of laparoscopic performance in vivo with performance measured in a laparoscopic simulator.
Laparoscopic skill was measured objectively through a series of seven tasks in an inanimate laparoscopic simulator. Seven analogous skills were tested in an in vivo porcine model. These skills included transferring, cutting, clipping, placement of a ligating loop, mesh placement, and suturing with an intracorporeal and extracorporeal knot. Scoring of each task rewarded precision and speed. Twelve PGY3 residents were given a baseline evaluation in the simulator and in the animal model. They were then randomized to either five practice sessions in the simulator (group A) or no practice (group B). Each group was retested in the simulator and in the animal (final test). Scores in vivo were compared by t-test for baseline versus final evaluation for each group. Linear regression analysis was used to correlate in vivo and in vitro scores for each task and for the total score (sum of all scores). Group A showed significant improvement in performance in vivo for cutting, clipping, mesh placement, and suturing with an intracorporeal and extracorporeal knot, as well as in the total score (p < 0.05). Group B showed significant improvement in suturing with an intracorporeal and extracorporeal knot, and in the total score. The magnitude of improvement from baseline to final evaluation was significantly greater for group A (p < 0.05). There was significant correlation between in vitro and in vivo total scores and the score for each task (p < 0.05) except for placement of the ligating loop and mesh. Performance in an in vitro laparoscopic simulator correlated significantly with performance in an in vivo animal model. Practice in the simulator resulted in improved performance in vivo.


Genome-Wide Identification, Expression, and Molecular Characterization of the CONSTANS-like Gene Family in Seven Orchid Species.
The orchid is one of the most distinctive and highly valued flowering plants. Nevertheless, the CONSTANS-like (COL) gene family plays significant roles in the control of flowering, and its functions in Orchidaceae have been minimally explored. This research identified 68 potential COL genes within seven orchids' complete genome, divided into three groups (groups I, II, and III) via a phylogenetic tree. The modeled three-dimensional structure and the conserved domains exhibited a high degree of similarity among the orchid COL proteins. The selection pressure analysis showed that all orchid COLs suffered a strong purifying selection. Furthermore, the orchid COL genes exhibited functional and structural heterogeneity in terms of collinearity, gene structure, cis-acting elements within their promoters, and expression patterns. Moreover, we identified 50 genes in orchids with a homology to those involved in the COL transcriptional regulatory network in Arabidopsis. Additionally, the first overexpression of CsiCOL05 and CsiCOL09 in Cymbidium sinense protoplasts suggests that they may antagonize the regulation of flowering time and gynostemium development. Our study will undoubtedly provide new resources, ideas, and values for the modern breeding of orchids and other plants.


Outbreak of hepatitis B associated with acupuncture.
In the period February to May 1980, acute hepatitis B occurred in six patients who had received acupuncture at a chiropractic clinic. Of persons attending the clinic (October 1979 to April 1980), hepatitis B occurred more frequently among those who had received acupuncture (6 of 103) than among those who had not (0 of 408; P less than .0001). Hepatitis patients had received acupuncture during two exposure periods: November 27-28, 1979, and February 19-20, 1980. During these periods the clinic reused acupuncture needles after immersing them overnight in a 1:750 solution of benzalkonium chloride. The sequence of acupuncture sessions suggests that hepatitis B virus remained infectious on the acupuncture needles for 20 to 27 hours despite this procedure. It is recommended that only either disposable needles or reuseable needles that have been physically cleaned and autoclaved after each use be used in acupuncture procedures.


Factors associated with health-related quality of life in chronic leg ulceration.
Individuals with chronic leg ulceration may have significantly impaired health-related quality of life (HRQOL) due to pain, impaired mobility, poor sleep, depression, restricted work capacity, and social isolation. The study purpose was to examine the associations among sociodemographic and clinical factors and HRQOL in a large sample of community-dwelling adults being treated for leg ulcers. Data are from the cross-sectional baseline assessment of the Canadian Bandaging Trial, a multi-center, randomized controlled trial conducted to assess time to healing with two forms of high-compression bandaging. All participants received a comprehensive, standardized clinical assessment, and completed the 12-item Short Form (SF-12) and McGill Pain Questionnaire. SF-12 data were compared to age- and sex-adjusted norms, and multivariable logistic regression was used to identify factors associated with whether individuals were below, or at/above their normative values on the physical and mental component summary (PCS, MCS). Of 424 individuals enrolled over a 50-month period, 407 (96 %) completed the SF-12. Mean age was 65 ± 17 years, and 55 % were women. Mean PCS was 39.1 ± 9.9 with 91 (22.4 %) scoring at/above the mean value for their age and sex; equivalent values for the MCS were 51.4 ± 9.9 and 209 (51.4 %). Higher levels of pain, younger age, larger size and longer duration of ulcer, and limited mobility were associated with poorer HRQOL. Findings confirm the considerable burden of illness associated with leg ulcers. Given the chronic and recurring nature of the condition, strategies focused on improving HRQOL and healing are needed for this vulnerable population.


Effects of ammonium ions on endplate channels.
Miniature endplate currents, recorded from voltage-clamped toad sartorius muscle fibers in solutions containing ammonium ions substituted for sodium ions, were increased in amplitude and decayed exponentially with a slower time constant than in control (Na) solution. The peak conductance of miniature endplate currents was also greater in ammonium solutions. The acetylcholine null potential was -2.8 +/- 0.8 mV in control solution, and shifted to 0.9 +/- 1.6 mV in solutions in which NH4Cl replaced half the NaCl. In solutions containing NH4Cl substituted for all the NaCl, the null potential was 6.5 +/- 1.3 mV. Single channel conductance and average channel lifetime were both increased in solutions containing ammonium ions. The exponential relationship between the time constant of decay of miniature endplate currents or channel lifetime and membrane potential was unchanged in ammonium solutions. A slight but consistent increase in peak conductance during miniature endplate currents and single channel conductance was seen as membrane potential became more positive (depolarized) in both control and ammonium solutions. Net charge transfer was greater in ammonium solutions than in control solution, whether measured during a miniature endplate current or through a single channel. The results presented here are consistent with an endplate channel model containing high field strength, neutral sites.


Diplopia in Green Tobacco Sickness.
This paper examines the first reported case of Green Tobacco Sickness (GTS) in Poland. A 25-year-old man who worked in a tobacco field for 14 hours with no protective measures experienced symptoms of GTS, and additionally, diplopia, after leaving the field. Upon hospital admission, diplopia was no longer observed. Diplopia was most probably caused by disturbances to the cholinergic neuromuscular transmission, secondary to nicotine. These neurological disturbances, the first of their kind observed in the course of GTS, deserve special attention. The case shows a potential adverse health effect related to tobacco harvest, as most Polish tobacco plantations are not mechanized. Polish farmers should be obligated to protect their workers with protective clothing, shoes, gloves and masks. Recommendations for tobacco harvester health are put forth in the paper.


[Nursing activities in family medicine groups for patients with chronic pain].
Thousands of people treated in primary care are currently experiencing chronic pain (CP), for which management is often inadequate. In Quebec, nurses in family medicine groups (FMGs) play a key role in the management of chronic health problems. The present study aimed to describe the activities performed by FMG nurses in relation to CP management and to describe barriers to those activities. A descriptive correlational cross-sectional postal survey was used. The accessible population includes FMG nurses on the Ordre des infirmières et infirmiers du Québec list. All nurses on the list who provided consent to be contacted at home for research purposes were contacted. A self-administered postal questionnaire (Pain Management Activities Questionnaire) was completed by 53 FMG nurses. Three activities most often performed by nurses were to establish a therapeutic relationship with the client; discuss the effectiveness of therapeutic measures with the physician; and conduct personalized teaching for the patient. The average number of individuals seen by interviewed nurses that they believe suffer from CP was 2.68 per week. The lack of knowledge of possible interventions in pain management (71.7%) and the nonavailability of information on pain management (52.8%) are the main barriers perceived by FMG nurses. FMG nurses are currently performing few activities in CP management. The nonrecognition of CP may explain this situation.