High frame-rate multichannel beam-scanning microscopy based on Lissajous trajectories.
A simple beam-scanning optical design based on Lissajous trajectory imaging is described for achieving up to kHz frame-rate optical imaging on multiple simultaneous data acquisition channels. In brief, two fast-scan resonant mirrors direct the optical beam on a circuitous trajectory through the field of view, with the trajectory repeat-time given by the least common multiplier of the mirror periods. Dicing the raw time-domain data into sub-trajectories combined with model-based image reconstruction (MBIR) 3D in-painting algorithms allows for effective frame-rates much higher than the repeat time of the Lissajous trajectory. Since sub-trajectory and full-trajectory imaging are simply different methods of analyzing the same data, both high-frame rate images with relatively low resolution and low frame rate images with high resolution are simultaneously acquired. The optical hardware required to perform Lissajous imaging represents only a minor modification to established beam-scanning hardware, combined with additional control and data acquisition electronics. Preliminary studies based on laser transmittance imaging and polarization-dependent second harmonic generation microscopy support the viability of the approach both for detection of subtle changes in large signals and for trace-light detection of transient fluctuations.


Stability and Synthesis of Phospholipids during Desiccation and Rehydration of a Desiccation-Tolerant and a Desiccation-Intolerant Moss.
The fatty acid composition of the phospholipids from the desiccation-tolerant moss Tortula ruralis (Hedw.) Gaertn, Meyer and Scherb and the desiccation-intolerant moss Cratoneuron filicinum has been determined. No changes in composition occur in either moss as a consequence of rapid drying, but, after slow drying, there is a decline in some unsaturated fatty acids. Upon rehydration of T. ruralis after slow drying, these acids decline further; however, within 105 minutes, they regain the same levels as those in undesiccated controls. A smaller and more transient decline occurs after rapid desiccation. Most phospholipid unsaturated fatty acids decrease during rehydration of C. filicinum, and their levels are not recovered. After both rapid and slow drying of T. ruralis, acetate and glycerol are incorporated into the phospholipid fraction, although de novo synthesis, alone, might not account for the increase in unsaturated fatty acids upon rehydration. Very little acetate or glycerol is incorporated during rehydration of C. filicinum. Loss of unsaturated fatty acids from the phospholipids of T. ruralis does not appear to be associated with increased lipoxygenase activity. Furthermore, there is little correlation between the extent of peroxidation of fatty acids due to desiccation and changes in the phospholipid fraction.


COMPARISON OF ISOFLURANE GAS VERSUS A GUAIFENESIN, KETAMINE, AND MEDETOMIDINE CONSTANT-RATE INFUSION FOR MAINTENANCE ANESTHESIA IN AMERICAN BLACK BEARS (URSUS AMERICANUS).
Published anesthetic protocols for captive and free-ranging bears are limited to injectable inductions with maintenance via inhalants or additional injectable boluses. Though common in other species, intravenous (IV) continuous-rate infusions (CRI) using guaifenesin combinations have not been evaluated in ursids. This study evaluated the use of a CRI compared to an inhalant for maintenance anesthesia. Seven healthy American black bears (Ursus americanus) were anesthetized in a crossover design with two different anesthetic maintenance protocols. Bears were immobilized with ketamine (2.02 ± 0.14 mg/kg) and medetomidine (0.04 ± 0.003 mg/kg) for both protocols. The anesthetic maintenance control protocol consisted of isoflurane gas (ISO) started at 2% delivered by endotracheal tube; the experimental protocol consisted of guaifenesin, medetomidine, ketamine (GMK) IV CRI started at 50 mg/kg/hr guaifenesin, 0.01 mg/kg/hr medetomidine, and 1 mg/kg/hr ketamine. Induction and recovery parameters including time to first effect, recumbency, and hands on; duration of maintenance protocol; and time from reversals administered to head up, standing on all four feet, no ataxia, and to fully recovered were recorded and compared between protocols. Heart rate, respiratory rate, rectal temperature, blood pressure, end tidal carbon dioxide, and hemoglobin oxygen saturation were recorded at 5-min intervals and compared between protocols. Venous blood gases were obtained at the start, middle, and end of the maintenance anesthesia and compared between protocols. All bears exhibited hypertension with mild respiratory acidosis throughout procedures. Measured physiologic parameters did not differ significantly between the isoflurane and the GMK CRI maintenance protocols, with the exception of higher endpoint (ISO) pCO2 measurements. No adverse events were recorded with either protocol, and adequate depth of anesthesia was maintained with both protocols. GMK CRI provides a safe, effective, and more portable alternative to inhalant anesthetics for maintenance anesthesia in bears in captivity or in the field.


Genesis of self-identity as disother: life histories of people who stutter.
This paper explores the processes shaping self-identity formation as DisOther and the actions of participants who stutter. It illuminates the experiences of adults who stutter using a biographical, narrative, life history methodology. The participants were seven South African adults of diverse racial, social and economic backgrounds from KwaZulu Natal, South Africa. Five males and two females were invited to participate via purposive and convenience sampling processes. Their stories of living with stuttering in their life worlds over time were constructed via biographical interviews using personal, social and temporal lenses typical of life history methodology. The interviews were audio-recorded and transcribed. The data were analysed at two levels using a combination of strategies. The first level entailed a narrative analysis that was represented as research stories for each participant. The cross-case and thematic analysis of research stories constituted the second level analysis of narratives. The findings explain the complex and interrelated personal and social processes over time which contribute to the genesis of self-identity formation as DisOther Social inscriptions of difference occurred in immediate home, school and work contexts over time via multiple processes such as labelling, norming, judging and teasing. Personal processes included discoveries of difference via critical events, repeated reinforcement of difference, self-judgement and temporal burdening. Furthermore, the actions participants took in negotiating stuttering were examined. The implications of the findings and limitations of the study are presented.


Usefulness of the Snare Technique During Leadless Pacemaker Implantation for a Patient with a Severely Dilated Right Atrium.
We report here the case of a 92-year-old woman with atrial fibrillation bradycardia in which leadless pacemaker implantation was performed with a difficult delivery of the catheter sheath due to an extremely large right atrium. Using a snare technique with correction of the direction of the force on the catheter toward the right ventricle (RV) can result in successful delivery of the pacemaker catheter and stable placement of the pacemaker system in the RV septum. This specific snare technique has the potential to facilitate leadless pacemaker implantation safely in a severely dilated chamber of the heart, making this technique effective to use in clinical practice.


Oxidative stress and its possible relation to lower urinary tract functional pathology.
Oxidative stress is considered to reflect an imbalance between the systemic manifestation of reactive oxygen and nitrogen species (RONS) and a biological system's ability to readily detoxify the reactive intermediates or to repair the resulting damage. RONS are not only harmful agents that cause oxidative damage in pathologies; they also have important roles as regulatory agents in a range of biological phenomena. They are normally generated as by-products of oxygen metabolism; however, environmental stressors (i.e., ultraviolet radiation, ionizing radiations, pollutants, heavy metal, and xenobiotics) contribute to greatly increase RONS production. Several antioxidants have been exploited in recent years for their actual or supposed beneficial effect against oxidative stress, but to date, none has been approved for any indication because they have not met the criteria of efficacy for drug approval. The present review discusses the concept of oxidative stress, how to measure it, how to prevent it, and its occurrence in different organ systems with special reference to the lower urinary tract.


Impact of a Combination Antibiotic Bag on Compliance With Surviving Sepsis Campaign Goals in Emergency Department Patients With Severe Sepsis and Septic Shock.
Severe sepsis and septic shock represent common presentations in the emergency department (ED) and have high rates of mortality. Guideline-recommended goals of care have been shown to benefit these patients, but can be difficult to provide. To determine whether the use of a premixed bag consisting of 2 g cefepime and 1 g vancomycin in 1000 mL of normal saline increases the probability of patients receiving Surviving Sepsis Campaign (SSC) recommendations for the initiation of antimicrobials and fluid challenge. This was a 6-month retrospective analysis conducted to determine the impact of an intervention on time to antimicrobials and fluid administration in patients with severe sepsis and septic shock. Patients presenting to the ED who received a diagnosis of severe sepsis or septic shock and were administered 2 antibiotics were eligible for inclusion. The primary outcome assessed was compliance with SSC recommendations for antibiotic and fluid goals within 3 hours of ED arrival. A total of 160 patients were included. In the intervention group, 63.8% of patients met the primary outcome compared with 22.5% in the historical group (odds ratio = 2.32; 95% CI = 1.67-3.23). Time to administration of antibiotics was less with the combination antibiotic bag (CAB: median (IQR) = 72 (48-115) minutes; non-CAB: median (IQR) = 135 (102-244) minutes; P ≤ 0.001). This intervention significantly increased the proportion of patients provided with SSC goals of care. Such interventions have not been reported previously and could be meaningful in the management of severe sepsis and septic shock.


Radiofrequency and microwave energy sources in surgical ablation of atrial fibrillation: a comparative analysis.
Due to its complexity and risk of bleeding, the Maze III procedure has been largely replaced by surgical ablation for atrial fibrillation (AF) using alternative energy sources. Radiofrequency (RF) and microwave (MW) are the most commonly used energy forms. In this study, we sought to compare these energy modalities in terms of clinical outcomes. Two hundred five patients underwent surgical ablation of AF, from October 1999 to May 2004 at our institution via an endocardial approach. Patients were categorized into 2 groups: RF and MW. Baseline characteristics, operative details, and clinical outcomes were compared between the 2 groups. Rhythm success was defined as freedom from AF and atrial flutter as determined by postoperative electrocardiograms. One hundred twenty patients (58.5%) were ablated using RF, whereas 85 (41.5%) were ablated with MW. Most of the patients had persistent AF in both the RF and MW groups (85.7% versus 80.0%, respectively; P = .363). Intraoperative left atrial size was 6.4 +/- 1.7 cm for the RF group and 6.4 +/- 1.7 cm for the MW group (P = .820). Postoperative rhythm success at 6 and 12 months was 72.4% versus 71.4% (P +/- .611) and 75.0% versus 66.7% (P = .909) for the RF and MW groups, respectively. Hospital length of stay was comparable for both groups (15.4 +/- 14.0 versus 13.3 +/- 13.9 days; P = .307). Postoperative survival at 6 months, 1 year, and 3 years was 90.4%, 89.5%, and 86.1% for RF patients compared to 87.9%, 86.5%, and 84.4% for MW patients, respectively (log rank P = .490). RF and MW energy forms yield comparable postoperative rhythm success, hospital length of stay, and postoperative survival. Both sources are rapid, safe, and effective alternatives to "cut and sew" techniques for surgical treatment of AF.


[How to develop advanced practice nursing in complex health care systems?].
Over the last few decades, advanced nursing practitioners (ANP) have developed new roles in health care systems and this tendency is continuing to grow. Postgraduate trainedANP interact directly with the person, i.e. the individual and his/her family - in many fields of practice and in a context of collaboration withfellow nurses and other health care professionals. The potential benefits of ANP interventions have been demonstrated in many fields. In particular, ANP are public health actors, able to participate in the interdisciplinary response to supportive care ofpatients with chronic diseases. However, the development of advanced practice nursing (APN) in a complex health care system requires a systemic approach coordinated with the various levels of training of nursing practitioners and other health care professionals. This is an essential prerequisite to allow ANPs to develop new roles adapted to their capacities (legal qualification, high level, specialist training, modalities of collaboration, etc.). To achieve an added value for patients, for the health care system in terms ofresults and to ensure adequate nursing conditions, the authors emphasize the importance of structured development of APN and propose an awareness phase comprising adoption of a conceptual model of APN and the establishment of a structured list of existing nursing practices in order to prepare a methodical implementation strategy.


Comparison between persisting anti-lipopolysaccharide antibodies and culture at postmortem in salmonella-infected cattle herds.
Herds with recent clinical outbreaks of Salmonella dublin (7 herds) and S. typhimurium (4 herds) infections were followed serologically in O-antigen ELISAs over about one year, divided in four equal sampling phases. Animals found to be persistent high-reactors or seronegative at the end of the study were slaughtered and subsequently cultured for salmonella in a selected number of organ samples. Approximately 3% of all animals had high seroreactions up to 17 months after the outbreaks, and less than half of the seropositive animals in the S. dublin-infected herds were salmonella culture positive at slaughter (14/31). However, one persistently seronegative animal was also culture positive. Furthermore, as much as 70% of the male calves investigated at postmortem in the S. dublin-infected herds were high-reactors, among which approx. 56% were culture positive. Surprisingly, 2 of the 14 animals found culture positive turned out to be culture positive for S. typhimurium only. In the S. typhimurium study, none of the 17 animals investigated at postmortem were salmonella culture positive. All sera from these animals were negative in the O:9 blocking ELISA, and no serum sample was positive in the S. dublin ELISA, alone. In conclusion, although serology based on the O-antigens appears to be useful to identify salmonella-infected herds, it seems to be insufficient for identification of persistently infected animals.


Myocarditis following COVID-19 mRNA vaccination.
Messenger RNA vaccines are the main COVID-19 vaccines authorized for use in the United States. Side effects are typically minor and transient. We report a case series of four subjects with an acute myocarditis-like illness following mRNA COVID-19 vaccination who were hospitalized at our hospital in Lubbock, Texas. Three patients were young men who presented with acute chest pain after the second dose of the mRNA-1273 vaccine. Another patient was a 53-year-old white woman who presented with acute left arm pain 3 days after the first dose of the mRNA-1273 vaccine. She was later found to have acute decompensated heart failure, and endomyocardial biopsy revealed eosinophilic injury-mediated myocarditis.


Intraoperative Radiotherapy Versus Whole-Breast External Beam Radiotherapy in Early-Stage Breast Cancer: A Systematic Review and Meta-Analysis.
There has not been a clear answer about the efficacy of intraoperative radiotherapy (IORT) for women with early-stage breast cancer.The aim of this meta-analysis was to summarize the available evidence comparing the efficacy and safety of IORT with those of whole-breast external beam radiotherapy (EBRT) for women with early-stage breast cancer.MEDLINE, EMBASE, the Web of Science, and the Cochrane Library were searched up to October 2014. Two authors independently conducted the literature selection and data extraction.Studies that compared IORT with whole-breast EBRT were included in the systematic review. IORT was defined as a single dose of irradiation to the tumor bed during breast-conserving surgery rather than whole-breast irradiation.Qualities of RCTs were evaluated according to the PEDro scale. Qualities of non-RCTs were evaluated according to the Methodological Index for Non-Randomized Studies (MINORS). The risk ratios (RRs) of ipsilateral breast tumor recurrence, overall mortality, breast cancer mortality, non-breast cancer mortality, and distant metastasis were pooled using a random-effects model.Four studies with 5415 patients were included in this meta-analysis, including 2 randomized controlled trials (RCTs) and 2 non-RCTs. Ipsilateral breast tumor recurrence was significantly higher in patients with IORT compared to those with whole-breast EBRT (RR 2.83, 95% CI 1.23-6.51), but with significant heterogeneity (I = 58.5%, P = 0.065). Comparing IORT with whole-breast EBRT, the pooled RRs for overall mortality, breast cancer mortality, non-breast cancer mortality, and distant metastasis were 0.88 (95% CI: 0.66-1.17), 1.20 (95% CI: 0.77-1.86), 0.76 (95% CI: 0.44-1.31), and 0.95 (95% CI: 0.61-1.49), respectively.IORT had a significantly higher risk of ipsilateral breast tumor recurrence than whole-breast EBRT. Overall mortality did not differ significantly. IORT should be used in conjunction with the prudent selection of suitable patients. It is imperative to identify women with a low risk of local recurrence.


Afferent connections to the abducent nucleus in the cat.
The afferent connections to the abducent nucleus in the cat were studied by means of retrograde transport of WGA-HRP after implantations of the tracer in crystalline form. Retrogradely labelled cells were found bilaterally in the medial and descending vestibular nuclei, mainly in their ventral and medial portions, in the rostral part of the ipsilateral gigantocellular reticular nucleus, in the medial part of the contralateral caudal pontine reticular nucleus and bilaterally in the oculomotor nucleus, mainly in its dorsolateral division. Some labelled cells were also found bilaterally in the mesencephalic reticular formation, the periaqueductal grey and the nucleus of the trapezoid body.


Brain plasticity after peripheral nerve injury treatment with massage therapy based on resting-state functional magnetic resonance imaging.
Massage therapy is an alternative treatment for chronic pain that is potentially related to brain plasticity. However, the underlying mechanism remains unclear. We established a peripheral nerve injury model in rats by unilateral sciatic nerve transection and direct anastomosis. The experimental rats were treated over the gastrocnemius muscle of the affected hindlimb with a customized massage instrument (0.45 N, 120 times/min, 10 minutes daily, for 4 successive weeks). Resting-state functional magnetic resonance imaging revealed that compared with control rats, the amplitude of low-frequency fluctuations in the sensorimotor cortex contralateral to the affected limb was significantly lower after sciatic nerve transection. However, amplitudes were significantly higher in the massage group than in a sham-massage group. These findings suggest that massage therapy facilitated adaptive change in the somatosensory cortex that led to the recovery of peripheral nerve injury and repair. This study was approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine of China (approval No. 201701001) on January 12, 2017.


Interaction of cigarette smoke condensate and some of its components with chlorpromazine toxicity on Saccharomyces cerevisiae.
Chlorpromazine (CPZ) is an antipsychotic phenothiazine which is still commonly prescribed though it causes idiosyncratic toxicity such as cholestasis. CPZ toxicity mechanisms involve oxidative stress among others. Cigarette smoke (CS) causes deleterious effects through diverse mechanisms such as oxidative stress. CS alters drug metabolizing enzymes expression and drug transporters expression and activity in animal cell models as well as in Saccharomyces cerevisiae. CS therefore alters pharmacokinetic and pharmacodynamics of many drugs including CPZ and caffeine whose toxicity is promoted by CS condensate (CSC). CSC interaction with CPZ toxicity deserves investigation. In this study, CSC exerted mild toxicity on Saccharomyces cerevisiae which resisted to this chemical stress after several hours. CPZ toxicity on yeast was dose-dependent and the cells resisted to CPZ up to 40 µM after 24 h of treatment. Yeast cells treated simultaneously with CPZ and a nontoxic CSC dose were less sensitive to CPZ. CSC probably triggers cross-resistance to CPZ. Using Sod1 mutant strain, we showed that this gene is potentially involved in the potential cross-resistance. Other genes encoding stress-related transcription factors could be involved in this process. Nicotine and cadmium chloride, which caused a dose-dependent toxicity individually, acted with CPZ in an additive or synergistic manner in terms of toxicity. Although our results cannot be extrapolated to humans, they clearly show that CSC and its components interact with CPZ toxicity.


The Protein Translocation Defect of MCT8L291R Is Rescued by Sodium Phenylbutyrate.
The monocarboxylate transporter 8 (MCT8; SLC16A2) is a specific transporter for thyroid hormones. MCT8 deficiency, formerly known as the Allan-Herndon-Dudley syndrome, is a rare genetic disease that leads to neurological impairments and muscle weakness. Current experimental treatment options rely on thyromimetic agonists that do not depend on MCT8 for cellular uptake. Another approach comes from studies with the chemical chaperone sodium phenylbutyrate (NaPB), which was able to stabilize MCT8 mutants having protein folding defects in vitro. In addition, NaPB is known as a compound that assists with plasma membrane translocation. The pathogenic MCT8L291R leads to the same severe neurological impairments found for other MCT8-deficient patients but, unexpectedly, lacks alterations in plasma 3,3',5-triiodothyronine (T3) levels. Here we tried to unravel the underlying mechanism of MCT8 deficiency and tested whether the pathogenic MCT8L291R mutant responds to NaPB treatment. Therefore, we overexpressed the mutant in Madin-Darby canine kidney cells in the human choriocarcinoma cell line JEG1 and in COS7 cells of African green monkey origin. In our recent study we describe that the MCT8L291R mutation most likely leads to a translocation defect. The pathogenic mutant is not located at the plasma membrane, but shows overlapping expression with a marker protein of the lysosome. Mutation of the corresponding amino acid in murine Mct8 (Mct8L223R) displays a similar effect on cell surface expression and transport function as seen before for MCT8L291R. NaPB was able to correct the translocation defect of MCT8L291R/Mct8L223R and restored protein function by increasing T3 transport activity. Furthermore, we detected enhanced mRNA levels of wild-type and mutant MCT8/Mct8 after NaPB treatment. The increase in mRNA levels could be an explanation for the positive effect on protein expression and function detected for wild-type MCT8. NaPB is not only suitable for the treatment of mutations leading to misfolding and protein degradation, but also for a mutant wrongly sorted inside a cell which is otherwise functional.


PeSTZ1, a C2H2-type zinc finger transcription factor from Populus euphratica, enhances freezing tolerance through modulation of ROS scavenging by directly regulating PeAPX2.
In the present study, PeSTZ1, a cysteine-2/histidine-2-type zinc finger transcription factor, was isolated from the desert poplar, Populus euphratica, which serves as a model stress adaptation system for trees. PeSTZ1 was preferentially expressed in the young stems and was significantly up-regulated during chilling and freezing treatments. PeSTZ1 was localized to the nucleus and bound specifically to the PeAPX2 promoter. To examine the potential functions of PeSTZ1, we overexpressed it in poplar 84K hybrids (Populus alba × Populus glandulosa), which are known to be stress-sensitive. Upon exposure to freezing stress, transgenic poplars maintained higher photosynthetic activity and dissipated more excess light energy (in the form of heat) than wild-type poplars. Thus, PeSTZ1 functions as a transcription activator to enhance freezing tolerance without sacrificing growth. Under freezing stress, PeSTZ1 acts upstream of ASCORBATE PEROXIDASE2 (PeAPX2) and directly regulates its expression by binding to its promoter. Activated PeAPX2 promotes cytosolic APX that scavenges reactive oxygen species (ROS) under cold stress. PeSTZ1 may operate in parallel with C-REPEAT-BINDING FACTORS to regulate COLD-REGULATED gene expression. Moreover, PeSTZ1 up-regulation reduces malondialdehyde and ROS accumulation by activating the antioxidant system. Taken together, these results suggested that overexpressing PeSTZ1 in 84K poplar enhances freezing tolerance through the modulation of ROS scavenging via the direct regulation of PeAPX2 expression.


Contribution of histidine residues to oligomerization of theta-toxin (perfringolysin O), a cholesterol-binding cytolysin.
Theta-toxin (perfringolysin O) modified by diethyl pyrocarbonate, a histidine-specific reagent, lost its hemolytic activity. The modified toxin retains the activities of binding to and insertion into cholesterol-containing membranes but lacks the ability to form oligomers. These results suggest that histidine residues of theta-toxin contribute their share to cytolysis, especially the oligomerization process.


Internal micropulse structure of a storage-ring free-electron laser.
The longitudinal distribution of a free-electron laser (FEL) may present a complex internal structure. This phenomenon has been already observed in the case of LINAC based oscillators and self-amplified spontaneous emission devices (for which the presence of "spikes" in the temporal distribution is systematically observed). We investigate here the physical process responsible for the growth of complex substructures inside the micropulse of a storage-ring free-electron laser. This "hole-burning-like" process results from the localized character of the interaction between the ultrarelativistic electron beam circulating in the storage ring and the laser pulse. Experimental results concerning the case of the super-ACO FEL are presented and interpreted by means of a pass-to-pass tracking code containing all the relevant features of the system dynamics.


Objective and Perceived Weight: Associations with Risky Adolescent Sexual Behavior.
Studies have shown that obesity is associated with increased sexual risk-taking, particularly among adolescent females, but the relationships between obesity, perceived weight and sexual risk behaviors are poorly understood. Integrative data analysis was performed that combined baseline data from the 1994-1995 National Longitudinal Study of Adolescent Health (from 17,606 respondents in grades 7-12) and the 1997 National Longitudinal Survey of Youth (from 7,752 respondents aged 12-16). Using six sexual behaviors measured in both data sets (age at first intercourse, various measures of contraceptive use and number of partners), cluster analysis was conducted that identified five distinct behavior clusters. Multivariate ordinal logistic regression analysis examined associations between adolescents' weight status (categorized as underweight, normal-weight, overweight or obese) and weight perception and their cluster membership. Among males, being underweight, rather than normal-weight, was negatively associated with membership in increasingly risky clusters (odds ratio, 0.5), as was the perception of being overweight, as opposed to about the right weight (0.8). However, being overweight was positively associated with males' membership in increasingly risky clusters (1.3). Among females, being obese, rather than normal-weight, was negatively correlated with membership in increasingly risky clusters (0.8), while the perception of being overweight was positively correlated with such membership (1.1). Both objective and subjective assessments of weight are associated with the clustering of risky sexual behaviors among adolescents, and these behavioral patterns differ by gender.


Micro-capillary-based self-referencing surface plasmon resonance biosensor for determination of transferrin.
A novel self-referencing surface plasmon resonance (SPR) biosensor for detection of transferrin is demonstrated using a micro-capillary as the sensing element. The biosensor employs the SPR mode as a measuring signal and the Fabry-Perot (FP) mode as a referencing signal. The SPR mode is generated in the gold film that is coated on the outside of the capillary; instead, the FP mode is excited in the capillary, which is filled with de-ionized water. The FP mode is sensitive to temperature and insensitive to refractive index, which can be used as a referencing signal to compensate the effects caused by the temperature fluctuation. The sensor provides a high sensitivity of 1783.943 nm/RIU (refractive index unit) and a resolution of about 7.287×10-5 RIU. The self-referencing biosensor was applied to measurement of transferrin protein. It can monitor the interaction of transferrin protein with anti-transferrin in real time (0-5.228 μM). The simple and low-cost SPR sensor can be used for highly sensitive self-referencing biosensing for further investigations.


Caregiver Burden and Dementia: A Systematic Review of Self-Report Instruments.
As the demand for dementia care grows rapidly worldwide, heavy "caregiver burden" has been associated with stress and depression. Even so, standard metrics for interdisciplinary research of caregiver burden are limited. The objective of the present review is to recommend valid, reliable, and comprehensive self-report instruments of caregiver burden. A systematic review was performed using four databases, searched in April 2021. Articles that established or evaluated self-report instruments for dementia caregiver burden were included, while studies that involved non-dementia caregivers or did not clearly define caregiver burden were excluded. Established guidelines for reliability and agreement studies were used to assess quality and risk of bias. Assessments of self-report instruments were made based on reliability, validity, feasibility, and quality of psychometric evaluations, and comparative evaluations were presented in visual form using radar graphs. Search terms yielded 1,720 articles, and 40 were included in the systematic review after excluding those of low quality. Based on the results of these studies, we recommend the Zarit Burden Interview, Screen for Caregiver Burden, Caregiver Burden Interview, and Burden Scale for Family Caregivers, due to their validity, reliability, and inclusion of multiple subjective and objective dimensions of burden. Targeting specific sources of caregiver burden can help prevent negative outcomes for both dementia patients and caregivers. Future studies should apply self-report instruments to measure and address caregiver burden longitudinally.


β-Hairpin loop of eukaryotic initiation factor 1 (eIF1) mediates 40 S ribosome binding to regulate initiator tRNA(Met) recruitment and accuracy of AUG selection in vivo.
Recognition of the translation initiation codon is thought to require dissociation of eIF1 from the 40 S ribosomal subunit, enabling irreversible GTP hydrolysis (Pi release) by the eIF2·GTP·Met-tRNAi ternary complex (TC), rearrangement of the 40 S subunit to a closed conformation incompatible with scanning, and stable binding of Met-tRNAi to the P site. The crystal structure of a Tetrahymena 40 S·eIF1 complex revealed several basic amino acids in eIF1 contacting 18 S rRNA, and we tested the prediction that their counterparts in yeast eIF1 are required to prevent premature eIF1 dissociation from scanning ribosomes at non-AUG triplets. Supporting this idea, substituting Lys-60 in helix α1, or either Lys-37 or Arg-33 in β-hairpin loop-1, impairs binding of yeast eIF1 to 40 S·eIF1A complexes in vitro, and it confers increased initiation at UUG codons (Sui(-) phenotype) or lethality, in a manner suppressed by overexpressing the mutant proteins or by an eIF1A mutation (17-21) known to impede eIF1 dissociation in vitro. The eIF1 Sui(-) mutations also derepress translation of GCN4 mRNA, indicating impaired ternary complex loading, and this Gcd(-) phenotype is likewise suppressed by eIF1 overexpression or the 17-21 mutation. These findings indicate that direct contacts of eIF1 with 18 S rRNA seen in the Tetrahymena 40 S·eIF1 complex are crucial in yeast to stabilize the open conformation of the 40 S subunit and are required for rapid TC loading and ribosomal scanning and to impede rearrangement to the closed complex at non-AUG codons. Finally, we implicate the unstructured N-terminal tail of eIF1 in blocking rearrangement to the closed conformation in the scanning preinitiation complex.


Phylogeny of the Celastraceae inferred from 26S nuclear ribosomal DNA, phytochrome B, rbcL, atpB, and morphology.
Phylogenetic relationships within Celastraceae (spindle-tree family) were inferred from nucleotide sequence characters from the 5' end of 26S nuclear ribosomal DNA (including expansion segments D1-D3; 84 species sampled), phytochrome B (58 species), rbcL (31 species), atpB (23 species), and morphology (94 species). Among taxa of questionable affinity, Forsellesia is a member of Crossosomataceae, and Goupia is excluded from Celastraceae. However, Brexia, Canotia, Lepuropetalon, Parnassia, Siphonodon, and Stackhousiaceae are supported as members of Celastraceae. Gymnosporia and Tricerma are distinct from Maytenus, Cassine is supported as distinct from Elaeodendron, and Dicarpellum is distinct from Salacia. Catha, Maytenus, and Pristimera are not resolved as natural genera. Hippocrateaceae (including Plagiopteron and Lophopetalum) are a clade nested within a paraphyletic Celastraceae. These data also suggest that the Loesener's classification of Celastraceae sensu stricto and Hallé's classification of Hippocrateaceae are artificial. The diversification of the fruit and aril within Celastraceae appears to be complex, with multiple origins of most fruit and aril forms.


Effect of RNA secondary structure on the kinetics of DNA synthesis catalyzed by HIV-1 reverse transcriptase.
The effect of RNA secondary structure on the kinetics of DNA synthesis catalyzed by HIV-1 RT was determined using a 66 nucleotide RNA template containing a stable 12 base pair hairpin structure. Prior to reaching the hairpin structure, the primer elongation by RT was fast and the kinetics of polymerization was not affected by the presence of the secondary structure. Once within the regions of template secondary structure, polymerization was much slower and RT paused at five distinct sites [Suo, Z., & Johnson, K. A. (1997) Biochemistry (manuscript submitted for publication)]. Kinetic analysis of single nucleotide incorporation at the pause sites showed polymerization occurred by both a fast phase (54-76 s-1) and a slow phase (0.07-0.4 s-1) during a single binding event. The biphasic kinetics suggests that the DNA substrates are initially bound in both productive and nonproductive states at the polymerase site of RT. The nonproductively bound DNA is slowly converted into a productive state without dissociation from the enzyme. At the pause sites, the enzyme amplitudes of the fast phase are small (4.0-15%) while the amplitudes of the slow phase are large (11-40%). In contrast, only the reaction at the fast phase was observed at the nonpause sites and the enzyme amplitudes were large (63-66%) although the nucleotide incorporation rates (62-78 s-1) are similar to the fast phase rates at the pause sites. These indicate that DNA substrates were bound predominantly nonproductively at pause sites and productively at nonpause sites. However, the overall binding affinity of DNA substrates was measured by the nitrocellulose-DEAE double filter binding assay, binding affinity at both pause sites and nonpause sites was similar (9-38 nM). This indicates that substrates are bound tightly at the large binding cleft of HIV-1, although they may not be productively bound at the polymerase active site. These results and those reported elsewhere [Suo, Z., & Johnson, K. A. (1997) Biochemistry (manuscript submitted for publication)] are consistent with a model in which, at pause sites, HIV-1 RT remains bound to DNA substrates waiting for the melting of the next stem base pair of template secondary structure. Upon melting of the stem base pair, polymerization to fill the open template site is fast and largely irreversible, allowing RT to read through the stable hairpin structures.


Compliance with safety glasses wear in monocular children.
Many ophthalmologists prescribe polycarbonate "safety glasses" to protect the remaining eye of monocular patients. Others do not routinely do so when the remaining eye is emmetropic since they feel compliance is poor. To evaluate compliance with safety glasses wear in these children, we reviewed the charts of all children (< 21 years old) treated with enucleation and followed at the University of Iowa by the Pediatric Ophthalmology Service between 1962 and 1991. Safety glasses were defined as polycarbonate lenses in a frame suitable for protective wear with spherical equivalent < or = -0.75 or < or = +2.00 diopters. Fifty-six patients were followed after enucleation. Forty-six of these patients met the inclusion criteria and 33 patients were available for follow up. Eighty-five percent of patients wear safety glasses > or = 50% of their waking hours; 61% wear them > or = 80% of the time; 33% wear them 100% of their waking hours. Twenty-one of the 33 patients participate in sports. One hundred percent of the female participants and 80% to 93% of the male participants wear safety glasses, goggles, or face shields during sports. Fifteen of the 33 respondents reported at least one potentially serious accident in which the safety glasses had protected the good eye. Compliance with safety glasses wear appears to be good in this population and may prevent injury to the remaining eye.


Characterization of native and modified extensin monomers and oligomers by electron microscopy and gel filtration.
We isolated hydroxyproline-rich extensin precursors from suspension-cultured tomato, cucumber, and sycamore-maple by salt-elution of intact cells and cell wall preparations. Cation exchange chromatography and HPLC gel filtration resolved these precursors into monomeric and oligomeric fractions, confirmed by amino acid analysis, immunological cross-reactivity, and TEM visualization. After rotary shadowing monomers appeared as flexuous rods with a contour length of 70 to 100 nanometers and a ;persistence length' (maximum linear displacement) of 44 to 51 nanometers. Oligomers were larger branched assemblies with occasional pores. Native extensin monomers gave uniform gel filtration retention times (Rts), but the Rts of HF-deglycosylated monomers varied depending on concentration, implying ionic interaction between the highly basic deglycosylated monomers and a weakly cationic gel matrix. Succinylation of the deglycosylated monomers reversed the net charge, and restored the retention time to that of glycosylated monomers, confirming the ionic interaction. Succinylation enhanced visualization of the deglycosylated monomers, which previously were barely discernible flexuous rods. The persistence length:contour length ratios of succinylated deglycosylated monomers (tomato sdP2) and glycosylated monomers (sP2) were the same, implying a similar molecular flexibility for both glycosylated and deglycosylated monomers at room temperature. These molecular properties are consistent with suggestions that extensin monomers reptate into the wall as a transmural protein ;weft' which becomes progressively cross-linked forming a network penetrated by the cellulose ;warp.'


Multimodal Nonopioid Pain Protocol Provides Better or Equivalent Pain Control Compared to Opioid Analgesia Following Arthroscopic Rotator Cuff Surgery: A Prospective Randomized Controlled Trial.
To evaluate the efficacy of a multimodal nonopioid analgesic protocol in controlling postoperative pain compared to opioids following a primary arthroscopic rotator cuff repair. Seventy consecutive patients undergoing a primary rotator cuff repair were assessed for eligibility. An observer-blinded prospective randomized controlled trial was designed in accordance with the Consolidated Standards of Reporting Trials 2010 (CONSORT) statement. The two arms of the study included a multimodal nonopioid pain regimen for the experimental group, and a standard of care narcotics for the control group. The primary outcome was visual analog scale (VAS) pain scores for the first 10 postoperative days. Secondary outcomes included PROMIS-PI (Patient-Reported Outcomes Measurement Information System-Pain Interference) scale, patient satisfaction, and adverse drug events. Thirty patients declined to participate or were excluded, and 40 patients were included in the final analysis. A total of 23 patients were in the traditional group, and 17 patients were in the nonopioid group. Control patients on opioid pain management reported a significantly higher VAS pain score on postoperative day 1 (opioid: 5.7 ± 2, nonopioid: 3.7 ± 2.2; P = .011) and postoperative day 4 (opioid: 4.4 ± 2.7, nonopioid: 2.4 ± 2.2; P = .023). No significant difference was seen on any other postoperative day. When mixed measured models were used to control for confounding factors, the nonopioid group demonstrated significantly lower VAS and PROMIS-PI scores (P < .01) at every time point. Patients in the traditional analgesia group reported significantly more days with constipation (P = .003) and days with upset stomach (P = .020) than those in the nonopioid group. The present study found that a multimodal nonopioid pain protocol provided equivalent or better pain control compared to traditional opioid analgesics in patients undergoing primary arthroscopic rotator cuff repair. Minimal side effects were noted with some improvement in the multimodal nonopioid pain cohort. All patients reported satisfaction with their pain management. Level I, prospective randomized controlled trial.


Adaptors for clathrin-mediated traffic.
Clathrin-based systems are responsible for a large portion of vesicular traffic originating from the plasma membrane and the trans-Golgi network that reaches the endosomal compartment. The assembly of cytosolic clathrin forms the scaffold required for the local deformation of the membrane and for the formation of coated pits and vesicles. In this process, clathrin interacts in a coordinated fashion with a large number of protein partners. A subset designated clathrin adaptors links integral membrane proteins to the clathrin coat, a process that results in the recruitment of specific cargo proteins to the budding vesicle. This review focuses on the most recent advances dealing with the molecular basis for sorting by clathrin adaptors.


Geosocial Features and Loss of Biodiversity Underlie Variable Rates of Inflammatory Bowel Disease in a Large Developing Country: A Population-Based Study.
The epidemiology of inflammatory bowel disease (IBD) in developing countries may uncover etiopathogenic factors. We investigated IBD prevalence in Brazil by investigating its geographic, spatial, and temporal distribution, and attempted to identify factors associated with its recent increase. A drug prescription database was queried longitudinally to identify patients and verify population distribution and density, race, urbanicity, sanitation, and Human Development Index. Prevalence was calculated using the number of IBD patients and the population estimated during the same decade. Data were matched to indices using linear regression analyses. We identified 162 894 IBD patients, 59% with ulcerative colitis (UC) and 41% with Crohn's disease (CD). The overall prevalence of IBD was 80 per 100 000, with 46 per 100 000 for UC and 36 per 100 000 for CD. Estimated rates adjusted to total population showed that IBD more than triplicated from 2008 to 2017. The distribution of IBD demonstrated a South-to-North gradient that generally followed population apportionment. However, marked regional differences and disease clusters were identified that did not fit with conventionally accepted IBD epidemiological associations, revealing that the rise of IBD was variable. In some areas, loss of biodiversity was associated with high IBD prevalence. When distribution is considered in the context of IBD prevalence, marked regional differences become evident. Despite a background of Westernization, hotspots of IBD are recognized that are not explained by population density, urbanicity, sanitation, or other indices but apparently are explained by biodiversity loss. Thus, the rise of IBD in developing countries is not uniform, but rather is one that varies depending on yet unexplored factors like geoecological conditions.


A quantitative approach to radiography of the lower limb. Principles and applications.
A method is described which provides standardised reproducible radiographic images of the lower limb. Anteroposterior and lateral radiographs are digitised and processed by computer to provide graphic/numeric displays of angles and linear measurements, relating the centre points of the hip, knee, and ankle. Two cases illustrate how surgical planning is facilitated when standardised data are available. These data confirm the close relationship between postoperative limb alignment and positioning of prosthetic elements.


A reclassification of the millipede superfamily Trichopolydesmoidea, with descriptions of two new species from the Aegean region (Diplopoda, Polydesmida).
TWO NEW SPECIES ARE DESCRIBED FROM CAVES IN SEVERAL GREEK ISLANDS IN THE AEGEAN SEA: Galliocookia gracilis sp. n., a presumed troglobite from Rhodes, Dodecanese Islands, and Sphaeroparia simplex sp. n., likely a troglophile from Kithnos, Cyclades, and Chios, Eastern Sporades. These genera are assigned to the family Trichopolydesmidae Verhoeff, 1910. Because Sphaeroparia Attems, 1909, an Afrotropical genus, nicely bridges the gap, both morphologically and geographically, between the Euro-Mediterranean Trichopolydesmidae and the much more diverse, pantropical Fuhrmannodesmidae Brölemann, 1916, the latter family is considered as a new junior subjective synonym of the former, syn. n. Thus expanded, the family Trichopolydesmidae is rediagnosed and its position within the superfamily Trichopolydesmoidea refined. Because the families Mastigonodesmidae Attems, 1914, Macrosternodesmidae Brölemann, 1916 and Nearctodesmidae Chamberlin & Hoffman, 1950 are also formally synonymized with Trichopolydesmidae, syn. n., the Trichopolydesmoidea currently contains only two families, Trichopolydesmidae and Opisotretidae Hoffman, 1980.


Remote monitoring in nocturnal home hemodialysis 2003.
Over the past several years there has been growth in the number of dialysis units performing remote monitoring in nocturnal home hemodialysis, there has been growth in the technical options available for remote monitoring, and there have been more creative approaches to overcoming previous limitations. This article will review this growth and the diverse factors driving the innovations internationally.


Mapping quantitative trait loci with epistatic effects.
Epistatic variance can be an important source of variation for complex traits. However, detecting epistatic effects is difficult primarily due to insufficient sample sizes and lack of robust statistical methods. In this paper, we develop a Bayesian method to map multiple quantitative trait loci (QTLs) with epistatic effects. The method can map QTLs in complicated mating designs derived from the cross of two inbred lines. In addition to mapping QTLs for quantitative traits, the proposed method can even map genes underlying binary traits such as disease susceptibility using the threshold model. The parameters of interest are various QTL effects, including additive, dominance and epistatic effects of QTLs, the locations of identified QTLs and even the number of QTLs. When the number of QTLs is treated as an unknown parameter, the dimension of the model becomes a variable. This requires the reversible jump Markov chain Monte Carlo algorithm. The utility of the proposed method is demonstrated through analysis of simulation data.


[The CAPITOL study (Captopril Post Infarction Tolerance). A trial of progressive titration of captopril after myocardial infarct with left ventricular dysfunction].
The objective of CAPITOL (Captopril Post Infarction Tolerance) multicentre open trial was to study the tolerance of a protocol of titration of Captopril in patients with recent myocardial infarction complicated by left ventricular dysfunction. Five hundred and four patients, with a mean age of 62 +/- 12 years, were included during the hospital period in the 74 participating intensive care units, 9 +/- 6 days after myocardial infarction (ejection fraction 34 +/- 6%). After a 6.25 mg test dose of Captopril, the dosage was progressively increased to the target dose of 150 mg at the end of the first month. Of the 504 patients included, 343 finished the trial and 161 stopped the trial prematurely. At the end of the hospital period, 73% received 75 mg/day: at the first follow-up visit (27 +/- 16 days after inclusion), 59% had attained 150 mg/day, this proportion increasing to 71% at the end of the trial (79 +/- 33 days after inclusion). There was no significant change in blood pressure for the whole study population. However, the systolic blood pressure of the patients receiving 150 mg/day of Captopril at the end of the trial was slightly higher than that observed at the end of the hospital period (126 +/- 17 mmHg and 116 +/- 17 mmHg respectively, p = 0.006). Severe Intercurrent events were observed in 89 patients: 24 deaths, 7 recurrent infarctions, 58 hospital admissions (21 for cardiac failure, 15 for recurrence of angina, 11 aorto-coronary bypass operations, 7 coronary angioplasties, 2 cerebro-vascular accidents, 2 systemic emboli). Of the benign complications, hypotension was observed in 25% of patients, nearly half of which occurred during the hospital admission. The drugs prescribed in association with Captopril were Aspirin (78%), betablockers (57%), nitrate derivatives (42%) and diuretics (27%). Multivariate analysis showed 3 factors associated with good tolerance of the 150 mg dosage of Captopril: Killip Class I or II on admission, an ejection fraction > 30% and an initial systolic blood pressure > 100 mmHg. In conclusion, in this trial of dose titration, 3 out of 4 patients with myocardial infarction and left ventricular dysfunction, tolerated the 150 mg/day dosage of Captopril. Patients in the trial could also be treated with drugs recommended after myocardial infarction, in particular the betablockers. Arch Mal Coeur 1999: 92: 395-403.


How do family carers and care-home staff manage refusals when assisting a person with advanced dementia with their personal care?
Caregivers may encounter, or inadvertently cause, refusals of care by a care recipient. Managing refusals of care can be challenging and have potential negative consequences. We aimed to examine caregivers' (care-home staff and family carers) experiences of managing refusals of personal care in advanced dementia. One-to-one semi-structured interviews with 12 care assistants from six care homes and 20 family carers who were physically assisting a person with advanced dementia with their personal care in the UK. Interviews were audio recorded and transcribed verbatim, with data analysed using qualitative content analysis. Core to the caregiver experience of refusals of care was knowing the person. This underpinned five key themes identified as caregivers' strategies used in preventing or managing refusals of care: (1) finding the right moment to care; (2) using specific communication strategies; (3) being tactful: simplifying, leaving, or adapting care; (4) having confidence in care; and (5) seeking support from others when safety is at risk. Different caregiver relationships with the person with dementia influenced how they managed refusals of care. Refusals of care can place caregivers in tough situations with tensions between providing care when it is seemingly not wanted and leaving care incomplete. Both caregiver groups require support such as coaching, mentoring and/or advice from other health and social care practitioners to manage difficult personal care interactions before crisis points occur.


Reoperative surgery--indications, efficacy, and long-term follow-up.
The Roux-en-Y gastric bypass with fascial support (RYGBPF) has proved to be effective as a primary as well as a revision/conversion operation in the management of morbid obesity. Since May 1984, 361 primary and 100 revision RYGBPF operations have been reported with low morbidity and mortality although morbidity and mortality were higher in the revision than in the primary group. The most common late complications of gastric-reduction operations resulting in weight regain were staple line disruption and pouch or stoma enlargement. With the use of the RYGBPF as a primary as well as a revision operation, weight loss results at 5 y have been good and our revision operation rate has dropped to less than 1%.


An animal model of benign bile-duct stricture, sclerosing cholangitis and cholangiocarcinoma and the role of epidermal growth factor receptor in ductal proliferation.
To adapt an animal model of benign bile-duct stricture, sclerosing cholangitis and cholangiocarcinoma in order to determine if the expression of epidermal growth factor receptor (EGFr) could be used to differentiate these lesions. A prospective control study with blinded interpretation of liver biopsy histology and immunohistochemical staining as the criterion standards. A university-affiliated research centre. Male Syrian Golden hamsters (40 for benign duct stricture, 29 for sclerosing cholangitis and 27 for cholangiocarcinoma). Ligation of the common bile duct with 6-0 catgut for benign duct stricture; injection of the biliary tree with 0.15 mL of formalin for sclerosing cholangitis; and weekly subcutaneous injections of 500 mg/kg of di-isopropanolnitrosamine for 10 weeks followed by ligation of the common bile duct with 6-0 catgut for cholangiocarcinoma. Routine histologic preparation of liver biopsies obtained at autopsy 10 weeks postoperatively then immunohistochemical staining of specimens for EGFr. The development of benign or atypical biliary ductal proliferation, including adenoma and carcinoma formation. The presence or absence of immunohistochemical staining for EGFr. Benign ductal proliferation without atypia was seen in 15 of 21 animals in the bile-duct-stricture group that were sacrificed, in 15 of 24 animals in the sclerosing cholangitis group and in 17 of 18 animals in the cholangiocarcinoma group. Atypical proliferation was seen in 13 of 18 animals with cholangiocarcinoma but not in the other two groups. The differential occurrence of atypical ductal proliferation was statistically significant (p < 0.00001) for both groups. No evidence of EGFr expression was found in any group. Although the animal model was valid histologically for comparing benign and malignant biliary disease, EGFr does not play a role in biliary ductal proliferation and so cannot be used to differentiate between benign and malignant lesions.


Transient accumulation of heme O (cytochrome o) in the cytoplasmic membrane of semi-anaerobic Anacystis nidulans. Evidence for oxygenase-catalyzed heme O/A transformation.
Incubation of obligately photoautotrophic and aerobic cyanobacterium Anacystis nidulans (Synechococcus sp. PCC 6301) in the light in the presence of the photo-system II inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea and equilibrated with approximately 1% (v/v) O2 in N2 (10 microM O2 in solution) led to a decrease of the heme A content of isolated cytoplasmic membranes and to the appearance of heme O. The latter was not seen in membranes from fully aerated cells (> 210 microM dissolved O2). Non-covalently bound hemes extracted from the membranes were identified by reversed phase high performance liquid chromatography. Heme A and O contents of the membranes changed in a reversible fashion solely depending on the ambient oxygen regime. Both hemes A and O combine with the same apoprotein as suggested by immunoblotting. CO/reduced-minus-reduced optical difference spectra, photoaction spectra of CO-inhibited O2 uptake by the membranes, and pyridine hemochrome spectra pointed to either heme belonging to a functional form of the terminal oxidase. The NADH:O2 oxidoreductase reaction catalyzed by membranes from both high O2 and low O2 cells was strictly dependent on the addition of catalytic amounts of cytochrome c, fully inhibited by 1.2 microM KCN, and insensitive to 5 microM 2-n-heptyl-4-hydroxyquinoline-N-oxide. O2 uptake by the membranes was effectively catalyzed by N,N,N',N'-tetramethyl-p-phenylenediamine but not 2-methylnaphthoquinol or plastoquinol-1 as artificial substrates. Therefore we conclude that the cyanobacterial respiratory oxidase, irrespective of the type of heme in its O2-reducing center, is a cytochrome c rather than a quinol oxidase.


Sirolimus immunosuppression for graft-versus-host disease prophylaxis and therapy: an update.
Sirolimus is being used increasingly as an immunosuppressant in allogeneic hematopoietic stem cell transplantation. This article reviews recent results in sirolimus-based graft-versus-host disease (GVHD) prophylaxis, as well as outcomes using sirolimus for established acute and chronic GVHD. In uncontrolled experiments, sirolimus provides good control of steroid-resistant acute or chronic GVHD. Similarly, the addition of sirolimus to GVHD prophylaxis regimens appears to reduce the rate of acute GVHD in the myeloablative and reduced-intensity settings, when used in matched, related, and unrelated donor transplantation. The use of sirolimus in haploidentical transplantation is now being explored as well. Sirolimus should be considered for use in steroid-resistant acute and chronic GVHD. Sirolimus appears promising as primary prophylaxis for GVHD prevention. Randomized trials are currently being performed to determine whether sirolimus-based immunosuppression is superior to traditional GVHD prophylaxis.


Microbiome of pear psyllids: A tale about closely related species sharing their endosymbionts.
Psyllids are phloem-feeding insects that can transmit plant pathogens such as phytoplasmas, intracellular bacteria causing numerous plant diseases worldwide. Their microbiomes are essential for insect physiology and may also influence the capacity of vectors to transmit pathogens. Using 16S rRNA gene metabarcoding, we compared the microbiomes of three sympatric psyllid species associated with pear trees in Central Europe. All three species are able to transmit 'Candidatus Phytoplasma pyri', albeit with different efficiencies. Our results revealed potential relationships between insect biology and microbiome composition that varied during psyllid ontogeny and between generations in Cacopsylla pyri and C. pyricola, as well as between localities in C. pyri. In contrast, no variations related to psyllid life cycle and geography were detected in C. pyrisuga. In addition to the primary endosymbiont Carsonella ruddii, we detected another highly abundant endosymbiont (unclassified Enterobacteriaceae). C. pyri and C. pyricola shared the same taxon of Enterobacteriaceae which is related to endosymbionts harboured by other psyllid species from various families. In contrast, C. pyrisuga carried a different Enterobacteriaceae taxon related to the genus Sodalis. Our study provides new insights into host-symbiont interactions in psyllids and highlights the importance of host biology and geography in shaping microbiome structure.


Localized cytosolic domains of the erythropoietin receptor regulate growth signaling and down-modulate responsiveness to granulocyte-macrophage colony-stimulating factor.
Erythrocyte development in mammals depends in part upon the interaction of the glycopeptide hormone erythropoietin (EPO) with cell surface receptors on committed erythroid progenitor cells. Both this factor and an EPO receptor polypeptide previously have been cloned, yet little is presently understood concerning molecular mechanisms of receptor activation and signal transduction. To identify cytosolic receptor domains necessary for signaling, we have compared the activities of a series of deletionally mutated EPO receptor constructs by their expression in interleukin 3-dependent, myeloid FDC-P1 cells. EPO-induced growth was transduced efficiently in these cells by the full-length receptor (507 amino acids), and no measurable loss in activity resulted from the deletion of up to 111 carboxyl-terminal residues. In contrast, the deletion of 44 additional residues led to a dramatic loss (86.3% +/- 7.8%; mean +/- SD) in the ability of this receptor to mediate EPO-induced growth, thus indicating that residues between Gly-352 and Met-396 constitute a functionally critical cytosolic subdomain. Interestingly, the expression of full-length EPO receptors in FDC-P1 cells also led to a selective inhibition of normal proliferative responsiveness to the alternative hematopoietic factor granulocyte-macrophage colony-stimulating factor. Moreover, this inhibition was progressively reversed in forms of the EPO receptor in which distal cytosolic residues were sequentially deleted. These results suggest that EPO receptors normally may trans-modulate components in the pathway of granulocyte-macrophage colony-stimulating factor-induced proliferation and that this down-modulation, as exerted by intact EPO receptors, may play a role in promoting erythroid commitment during myeloid blood cell development.


Amelioration of cisplatin-induced acute kidney injury by recombinant neutrophil gelatinase-associated lipocalin.
We investigated the protective effect and mechanism of neutrophil gelatinase-associated lipocalin (NGAL) in a murine model of cisplatin-induced nephrotoxicity. Male Swiss-Webster mice were assigned to four groups (n = 10 in each group). Control mice received vehicle only. Mice in the experimental group were given a single intraperitoneal injection of cisplatin (20 mg/kg) to induce nephrotoxicity, and were divided into three groups. The first group received 100 μL of saline only via tail vein at the time of cisplatin administration. The second group was given biologically active recombinant NGAL via tail vein (250 μg/100 μL solution). The third group was injected with a 250 μg/100μL solution of inactivated NGAL. After 4 days, we measured serum creatinine and urinary N-acetyl-β-d-glucosaminidase (NAG), and performed histologic studies. Biologically active NGAL significantly blunted the rise in serum creatinine (NGAL plus cisplatin 1.33 ± 0.31 versus cisplatin alone 2.43 ± 0.31 mg/dL, p < .001) as well as the increase in urine NAG (NGAL plus cisplatin 60.7 ± 14.2 versus cisplatin alone 120.5 ± 22.5 units/gm creatinine, p < .005). In addition, NGAL conferred a marked reduction in tubule cell necrosis and apoptosis (NGAL plus cisplatin 6.9 ± 1.2 versus cisplatin alone 15.1 ± 3.4 TUNEL positive nuclei per 100 cells, p < .001). These beneficial effects were completely abolished when heat-inactivated NGAL was administered instead of the biologically active form. Since induction of NGAL in kidney tubules is a known physiologic response to cisplatin, the pharmacologic use of NGAL to prevent cisplatin nephrotoxicity is likely to be safe and effective.


Comparative epidemiology of CTX-M-14 and CTX-M-15 producing Escherichia coli: association with distinct demographic groups in the community in New Zealand.
Extended spectrum beta-lactamase producing E. coli (ESBL-EC) are an emerging public health issue. In New Zealand (NZ), bla (CTX-M-14) and bla (CTX-M-15) are the most common ESBL genes. Although many studies describe risk factors for ESBL-EC, few describe risk factors for specific ESBL genes. Between January 2006 and December 2007, we characterized 108 consecutive, non-duplicate isolates of ESBL-EC at the Auckland Hospital laboratory. Demographic and clinical data were recorded. Of the 108, 54.6% (59) were CTX-M-15-EC, 26.9% (29) were CTX-M-14-EC and 12.09% were CTX-M-9 (13). The remaining seven isolates carried CTX-M-3 (3; 2.7%), CTX-M-65 (2; 1.8%), CTX-M-27 (1; 0.9%) and CTX-M-57 (1; 0.9%). CTX-M-15-EC were more likely than CTX-M-14-EC to be fluoroquinolone-resistant (86.4% versus 32.4%; p=0.006) and to be non-susceptible to amoxicillin-clavulanate (84.7% versus 41.4%; p=0.0001). Patients with CTX-M-15-EC were more likely to be of Indian ethnicity (34.5% versus 0%; p=0.0012) and to have travelled recently (31.6% versus 4%; p=0.0088). Patients with CTX-M-14-EC were more likely to have Chinese or South-East Asian ethnicity (48.1% versus 5.2%; p<0.0001) and to have no history of either travel or prior hospital admission (44% versus 8.9%; p=0.0006). These data imply that CTX-M-15 and CTX-M-14 producing E. coli are associated with distinct demographic subgroups in NZ.


Characterization of molecular mechanisms driving Merkel cell polyomavirus oncogene transcription and tumorigenic potential.
Merkel cell polyomavirus (MCPyV) is associated with approximately 80% of cases of Merkel cell carcinoma (MCC), an aggressive type of skin cancer. The incidence of MCC has tripled over the past twenty years, but there are currently very few effective targeted treatments. A better understanding of the MCPyV life cycle and its oncogenic mechanisms is needed to unveil novel strategies for the prevention and treatment of MCC. MCPyV infection and oncogenesis are reliant on the expression of the early viral oncoproteins, which drive the viral life cycle and MCPyV+ MCC tumor cell growth. To date, the molecular mechanisms regulating the transcription of the MCPyV oncogenes remain largely uncharacterized. In this study, we investigated how MCPyV early transcription is regulated to support viral infection and MCC tumorigenesis. Our studies established the roles of multiple cellular factors in the control of MCPyV gene expression. Inhibitor screening experiments revealed that the histone acetyltransferases p300 and CBP positively regulate MCPyV transcription. Their regulation of viral gene expression occurs through coactivation of the transcription factor NF-κB, which binds to the viral genome to drive MCPyV oncogene expression in a manner that is tightly controlled through a negative feedback loop. Furthermore, we discovered that small molecule inhibitors specifically targeting p300/CBP histone acetyltransferase activity are effective at blocking MCPyV tumor antigen expression and MCPyV+ MCC cell proliferation. Together, our work establishes key cellular factors regulating MCPyV transcription, providing the basis for understanding the largely unknown mechanisms governing MCPyV transcription that defines its infectious host cell tropism, viral life cycle, and oncogenic potential. Our studies also identify a novel therapeutic strategy against MCPyV+ MCC through specific blockage of MCPyV oncogene expression and MCC tumor growth.


Insulin therapy in pediatric patients with type I diabetes: continuous subcutaneous insulin infusion versus multiple daily injections.
The goal of this article was to review studies comparing continuous subcutaneous insulin infusion (CSII) to multiple daily insulin injections (MDIs) in children and adolescents with type I diabetes on five parameters: glycemic control, adverse events, required insulin dosage, weight gain, and psychosocial implications. Two of the five studies (sample sizes 19-75 patients) showed that insulin pumps provided better glycemic control, fewer adverse events, and better psychosocial implications than MDIs. Three of the five showed a lower required insulin dosage in CSII and one of the five showed less weight gain with CSII. Studies 4 and 5, both done in preschoolers, showed no significant difference on any of the five parameters. The majority of the patients and families chose to continue with CSII after the completion of the studies, even in studies where insulin pumps showed no objective benefit. Insulin pumps appear to offer potential benefit over MDIs, but the efficacy, safety, and cost effectiveness of CSII versus MDIs as first-line therapy for type I diabetes in this population has not yet been clearly established. Until more data are available from large, randomized, controlled studies, MDIs may be used as first-line therapy and CSII should be considered in patients who have poor glycemic control or who are not compliant with multiple injections.


Summary of the published Indian data on chronic myeloid leukemia.
Chronic myelogenous leukemia (LML) was recognized as a distinct entity in the mid-1800s. Since Nowell and Hunagerford initiated their research on CML in1960 our understanding in CML has been increasing. Imatinib became the preferred treatment from 2000 onwards as a result of its unprecedented success. The lack of structured Indian data on CML led to the formation of a CML data cansortuim which invited CML data albiet retro spartive form around the country including major cancer service providers both government and private. We provide a summary of published Indian data on CML here.


CD1d1 intrinsic signaling in macrophages controls NLRP3 inflammasome expression during inflammation.
Dysregulation of immune responses in the gut often associates with inflammatory bowel diseases (IBD). Mouse CD1d1, an ortholog of human CD1d mainly participating in lipid-antigen presentation to NKT cells, is able to generate intrinsic signals upon stimulation. Mice with macrophage-specific CD1d1 deficiency (LymCD1d1-/- ) acquire resistance to dextran sodium sulfate (DSS)-induced colitis, attributing to the transcriptional inhibition of NLRP3 inflammasome components. The hyperactivation of NLRP3 inflammasome accounts for gut epithelial proliferation and intestine-blood barrier integrity. Mechanistically, occupancy by the natural ligand glycosphingolipid iGb3, CD1d1 responds with intracellular Ser330 dephosphorylation thus to reduce the Peroxiredoxin 1 (PRDX1)-associated AKT-STAT1 phosphorylation and subsequent NF-κB activation, eventually causing transcriptional down-regulation of Nlrp3 and its immediate substrates Il1b and Il18 in macrophages. Therefore, the counterbalancing role of CD1d1 in macrophages appears to determine severity of DSS-mediated colitis in mice. These findings propose new intervention strategies for treating IBD and other inflammatory disorders.


Flood and Contain: An Optimized Repeal-Based Flooding Algorithm for Wireless Ad Hoc and Sensor Networks.
Flooding is a simple yet reliable way of discovering resources in wireless ad hoc networks such as mobile ad hoc networks (MANETs), ad hoc sensors, and recently, IoT networks. However, its operation is resource-intensive, especially in densely populated networks. Several approaches can be found in the literature to reduce the impact of flooding. Many of these approaches follow a repeal-based operation, chasing and stopping further propagation of flooding packets once the target is found. However, repeal-based protocols might end up transmitting even more packets than the original flooding. This work characterizes a maximum repeal-flooding boundary beyond which it is counterproductive to chase the original flooding. We present the Flood and Contain (F&C) algorithm, a method that can quickly establish the maximum repeal-flooding boundary for each node while making no assumptions on the underlying network. F&C's packet overhead increases linearly with the hop count up to the maximum repeal-flooding boundary, in which case there is no attempt to chase the original flooding. In this latter case, F&C generates only as many packets as the original flooding. Simulations show that, on average, F&C reduces the total flooding overhead (compared to traditional flooding) up to 35 percent once considering all possible destinations, with only a slight increase in resource discovery latency, and it outperforms all other repeal-based protocols, particularly for longer routes.


The efficacy of different cleaning and disinfection procedures to reduce Salmonella and Enterobacteriaceae in the lairage environment of a pig abattoir.
This study investigated several cleaning and disinfection protocols for their ability to eliminate Salmonella and to reduce levels of Enterobacteriaceae, within the lairage pens of a commercial pig abattoir. Eight protocols were evaluated in each of 12 lairage pens at the end of the slaughtering day on 3 occasions (36 pens/protocol): (P1) high-pressure cold water wash (herein referred to as high-pressure wash); (P2) high-pressure wash followed by a quaternary ammonium compound (QAC)-based disinfectant without rinsing; (P3) high-pressure wash followed by a chlorocresol-based disinfectant without rinsing; (P4) high-pressure wash followed by a sodium hydroxide/sodium hypochlorite detergent with rinsing; (P5) P4 followed by P2; (P6) P4 followed by P3; (P7) P5 with drying for 24-48h; and (P8) P6 with drying for 24-48h. Two floor swabs and one wall swab were taken from each lairage pen before and after each protocol was applied, and examined for the presence of Salmonella and enumeration of Enterobacteriaceae. High-pressure washing alone (P1) did not reduce the prevalence of Salmonella in the lairage pens. When high-pressure washing, the probability of detecting Salmonella following application of the chlorocresol-based disinfectant (P3) was lower than with the QAC-based disinfectant, P2 (14.2% versus 34.0%, respectively; p<0.05). The probability of detecting Salmonella after the combined use of detergent and the chlorocresol-based disinfectant (P6) was also lower than application of detergent followed by the QAC-based disinfectant, P5 (2.2% versus 17.1%, respectively; p<0.05). Drying of pens (P7 and P8) greatly reduced the probability of detecting Salmonella. Only 3.8% of swabs were Salmonella-positive 48h after cleaning with detergent and the QAC-based disinfectant (P7); while an eradication of Salmonella was achieved 24h after cleaning with detergent and the chlorocresol-based disinfectant, P8. A reduction in Enterobacteriaceae counts to below the limit of detection (LOD; 10CFU/cm2) was achieved following cleaning with detergent and disinfection with the chlorocresol-based disinfectant, regardless of drying (p<0.05), whereas, applying detergent and the QAC-based disinfectant (P7) did not reduce Enterobacteriaceae counts to below the LOD. Therefore ensuring that lairage pens are allowed to dry after intensive cleaning with detergent and a chlorocresol-based disinfectant is recommended as the most effective hygiene routine to eliminate Salmonella and reduce Enterobacteriaceae counts.


Algorithmic approach to the management of the patient with lumbar spinal stenosis.
A 71-year-old generally healthy woman presents for her first visit in 3 years. She ambulates slowly from the waiting room, with a more stooped posture than previously. She reports a 2-year history of slowly worsening buttock and leg pain when she walks any distance. She has noticed that her symptoms are much less when she leans on a shopping cart in the grocery store. Her buttock/leg pain resolves within a few minutes when she sits down. The patient exhibits signs and symptoms suggestive of lumbar spinal stenosis.


Quantitative Effects of 2,4-Dichlorophenoxyacetic Acid on Growth of Suspension-cultured Acer pseudoplatanus Cells: II. Influence of 2,4-D Metabolism and Intracellular pH on the Control of Cell Division by Intracellular 2,4-D Concentration.
The utilization of 2,4-dichlorophenoxyacetic acid (2,4-D) molecules by Acer pseudoplatanus cells is governed mainly by a glucosylation process. Evidence that 2,4-D glucoside molecules are biologically inactive is presented. 2,3,5-Triiodobenzoic acid (TIBA), by inhibiting 2,4-D glucosylation, has a sparing effect on 2,4-D molecules; thus TIBA treatments increase growth yield (expressed as the ratio of the maximum number of cells produced to the initial concentration of 2,4-D in the culture medium).Significant amounts of intact 2,4-D molecules remain outside and inside the cells when cell division stops at the onset of the stationary phase. This result and the previous demonstration that, at the onset of the stationary phase, 2,4-D is the specific limiting factor of cell division (Leguay JJ, J Guern 1975 Plant Physiol 56: 356-359) suggest that a threshold concentration of auxin is needed for cell division to proceed.The distribution of 2,4-D molecules between the cells and the culture medium is dependent on the population density at the stationary phase. The extracellular 2,4-D concentration at that time is a linear function of the population density whereas intracellular amounts of 2,4-D and 2,4-D metabolites are constant. By using a modified 2-(14)C,-5,5-dimethyloxazolidine-2,4-dione technique, it has been shown that the intracellular pH is markedly lowered as the population density at the plateau is increased. This intracellular pH modification is likely to be responsible for a large modification of the ratio between intracellular and extracellular auxin concentrations.The intracellular auxin concentration reaches a constant value (about 3 x 10(-7)m), independent of population density when cell division stops at the onset of the stationary phase suggesting that it represents the threshold value of the control for cell division.


Overtly manifested diabetes mellitus after resection of insulinoma.
Insulinoma is the most common cause of endogenous hyperinsulinemic hypoglycemia in adults. However, the coincidence of insulinoma and diabetes is extremely uncommon. We describe a rare, but very interesting case of diabetes mellitus which was masked by insulinoma and was overtly manifest after the removal of the insulinoma.


Time to initiation of neo-adjuvant chemotherapy for breast cancer treatment does not influence patient survival: A study of US breast cancer patients.
Delays in the initiation of adjuvant chemotherapy or radiation therapy are associated with worse outcomes in patients undergoing treatment for breast cancer. However, the impact of the time to initiation of neo-adjuvant chemotherapy (NAC) on patient outcomes has not previously been studied. The purpose of this study was to determine whether delays in NAC initiation impact patient survival. The National Cancer Database was queried for women ≥ 18 years old who underwent NAC within 6 months of being diagnosed with stage I-III invasive breast cancer in 2010-2011. ER+ or PR+, Her2- cancers were excluded from analysis. Multivariable Cox proportional hazard modeling was used to evaluate the relationship between time to NAC, sociodemographic, diagnosis, and treatment factors with patient survival. The median age of the 12 806 women included in this study was 52 (range 21-90) with 62% presenting with cT2 disease and 62% with nodal involvement. Half of the women (50%) had triple negative, 30% ER/PR+Her2+ and 20% ER-PR-Her2+ cancers. The median time to starting NAC was 4 weeks (range 0-26) for all subtypes. Time to NAC initiation was not associated with a difference in survival for triple negative or Her2+ cancers. Delays from diagnosis to starting NAC are not associated with worse survival for patients with Her2+ or triple negative breast cancer. This study demonstrates that the majority of women in the modern era start NAC in a timely fashion and delays in starting NAC within 6 months of diagnosis do not impact long-term patient outcomes.


Kindness begins with yourself: The role of self-compassion in adolescent body satisfaction and eating pathology.
A wealth of evidence indicates that self-compassion is linked with positive psychological outcomes; however, little is known about the process through which self-compassion exerts its effect. The primary purpose of this research was to investigate the direct and indirect impact of self-compassion on body satisfaction and eating pathology in adolescents. Two hundred and thirty-eight students were recruited from three local high schools (Mage = 16.49, 43.7% boys). All participants completed the Self-Compassion Scale (SCS), Hopkins Symptom Checklist (SCL-5), Body Areas Satisfaction Scale (BASS), and Eating Disorder Examination Questionnaire - Adolescent Version (EDE-Q) at baseline. The SCL-5, BASS, and EDE-Q were completed 4 months later. Self-compassion was positively associated with body satisfaction and negatively associated with psychological distress and eating pathology in boys and girls at both time points. Longitudinal conditional process analyses revealed that self-compassion predicted changes in body satisfaction and eating pathology through changes in psychological distress for girls only. Notably, self-compassion was higher in boys than in girls. Results underscore how self-compassion may be an important factor to target in fostering a positive body image and preventing disordered eating in adolescents.


Attitudes toward domestic violence: a cultural perspective.
This study examines the effect of acculturation on the attitudes held by Ethiopian Jews in Israel toward domestic violence (DV). The study findings revealed the following: Ethiopians who immigrated to Israel (n = 31) held more lenient attitudes toward DV than Israeli born Jews (n = 62), which supported the hypothesis that culture influences attitudes toward DV; in addition, Ethiopians born in Israel (n = 29) held attitudes closer to those of Israeli-born Jews who were not from Ethiopian origin, thus supporting the hypothesis that integration into the host country results in changes in DV attitudes. These are important findings due to the extremely high number of DV episodes among immigrant populations in general and Ethiopian Jews living in Israel in particular. This study may provide optimism in that it is probable that the younger generation will prove to be less violent than the first-generation immigrants. Perhaps one conclusion that can be drawn is the importance of expediting the integration process of the second-generation Ethiopian Jews in Israel.


Scent marking in a territorial African antelope: I. The maintenance of borders between male oribi.
Scent marking is ubiquitous among the dwarf antelope and gazelles of Africa, but its function has been the subject of debate. This study examined preorbital gland scent marking in the oribi, Ourebia ourebi, a territorial African antelope. Several hypotheses for the function of scent marking by territorial antelope were tested with observational data. Of these, the hypotheses that scent marking is driven by intrasexual competition between neighbouring males, and that marks serve as an honest advertisement of a male's ability to defend his territory from rivals, were supported best. Thirty-three territorial male oribi on 23 territories marked most at borders shared with other territorial males, and territorial males marked more often at borders shared with multimale groups than at borders shared with a single male. This suggests that males perceived neighbouring male groups as a greater threat to territory ownership than neighbouring males that defended their territories without the aid of adult subordinates. Marking rate was unrelated to territory size or the number of females on adjacent territories, but males with many male neighbours marked at higher rates than those with fewer male neighbours. These results suggest that the presence of male neighbours has a greater effect on the scent marking behaviour of territorial antelope than has been considered previously. Copyright 1999 The Association for the Study of Animal Behaviour.


Water-Dispersible Phytosterol Nanoparticles: Preparation, Characterization, and in vitro Digestion.
For improving solubility and bioaccessibility of phytosterols (PS), phytosterol nanoparticles (PNPs) were prepared by emulsification-evaporation combined high-pressure homogenization method. The organic phase was formed with the dissolved PS and soybean lecithin (SL) in anhydrous ethanol, then mixed with soy protein isolate (SPI) solution, and homogenized into nanoparticles, followed by the evaporation of ethanol. The optimum fabrication conditions were determined as PS (1%, w/v): SL of 1:4, SPI content of 0.75% (w/v), and ethanol volume of 16 ml. PNPs were characterized to have average particle size 93.35 nm, polydispersity index (PDI) 0.179, zeta potential -29.3 mV, and encapsulation efficiency (EE) 97.3%. The impact of temperature, pH, and ionic strength on the stability of fabricated PNPs was determined. After 3-h in vitro digestion, the bioaccessibility of PS in nanoparticles reached 70.8%, significantly higher than the 18.2% of raw PS. Upon freeze-drying, the particle size of PNPs increased to 199.1 nm, resulting in a bimodal distribution. The solubility of PS in water could reach up to 2.122 mg/ml, ~155 times higher than that of raw PS. Therefore, this study contributes to the development of functional PS-food ingredients.


Amphipols can support the activity of a membrane enzyme.
Amphipathic polymers ("amphipols") were introduced several years ago (Tribet, C.; Audebert, R.; Popot, J.-L. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 15047-15050) as an alternative method for solubilizing integral membrane proteins in stable, nativelike conformations. However, direct maintenance of full membrane protein functionality in amphipol solutions has not previously been demonstrated in the absence of added lipid or detergent. In this contribution, the first zwitterionic amphipol "PMAL-B-100" is introduced. PMAL-B-100 not only maintains membrane protein structure and solubility, but also supports the full catalytic activity of an integral membrane enzyme, diacylglycerol kinase, in the complete absence of additional lipid or detergent. All of the roles which a lipid bilayer normally plays in maintaining diacylglycerol kinase's structure and in facilitating catalysis are satisfied by the environment and interactions supplied by PMAL-B-100.


Interferons at the placental interface.
The antiluteolytic factors secreted by sheep and cattle conceptuses are closely related structurally to alpha-interferons (IFN-alpha s). They are known as ovine and bovine trophoblast protein-1 (oTP-1 and bTP-1), respectively. The mRNAs for oTP-1 and bTP-1 are transcribed from multiple genes and are the major translatable messages of Day 13-17 sheep conceptuses and Day 15-20 cattle conceptuses. The proteins belong to the 172-amino acid IFN-alpha II (or IFN-omega) subfamily and have the typical antiviral and antiproliferative properties of the 166-residue IFN-alpha Is. These embryonic interferons also bind to the IFN-alpha receptor, which is present in uterine endometrium in high concentrations, and can influence the production of prostaglandin F-2 alpha and the pattern of protein secretion in that tissue. Through use of in-situ hybridization procedures on tissue sections and Northern and dot blot analyses of extracted conceptus RNA, ovine oTP-1 mRNA has been shown to increase markedly around Day 13 and to decrease after about Day 15 of pregnancy. The mRNA is confined entirely to cells of the trophectoderm. Significant induction of mRNA that hybridizes to an oTP-1 cDNA occurs in response to exposure to polyI:polyC in Day 11 sheep blastocysts which normally have low levels of oTP-1 expression. However, the basis for induction in the normal progression of embryonic development remains unclear. The fact that preimplantation conceptuses of other species, e.g. pig, release substances with antiviral activity suggests that IFNs may have an important role in pregnancy that extends beyond the domestic ruminants.


Validation of novel hub genes and molecular mechanisms in acute lung injury using an integrative bioinformatics approach.
Acute lung injury (ALI) is an inflammatory pulmonary disease that can easily develop into serious acute respiratory distress syndrome, which has high morbidity and mortality. However, the molecular mechanism of ALI remains unclear, and few molecular biomarkers for diagnosis and treatment have been identified. In this study, we aimed to identify novel molecular biomarkers using a bioinformatics approach. Gene expression data were obtained from the Gene Expression Omnibus database, co-expressed differentially expressed genes (CoDEGs) were identified using R software, and further functional enrichment analyses were conducted using the online tool Database for Annotation, Visualization, and Integrated Discovery. A protein-protein interaction network was established using the STRING database and Cytoscape software. Lipopolysaccharide (LPS)-induced ALI mouse model was constructed and verified. The hub genes were screened and validated in vivo. The transcription factors (TFs) and miRNAs associated with the hub genes were predicted using the NetworkAnalyst database. In total, 71 CoDEGs were screened and found to be mainly involved in the cytokine-cytokine receptor interactions, and the tumor necrosis factor and malaria signaling pathways. Animal experiments showed that the lung injury score, bronchoalveolar lavage fluid protein concentration, and wet-to-dry weight ratio were higher in the LPS group than those in the control group. Real-time polymerase chain reaction analysis indicated that most of the hub genes such as colony-stimulating factor 2 (Csf2) were overexpressed in the LPS group. A total of 20 TFs including nuclear respiratory factor 1 (NRF1) and two miRNAs were predicted to be regulators of the hub genes. In summary, Csf2 may serve as a novel diagnostic and therapeutic target for ALI. NRF1 and mmu-mir-122-5p may be key regulators in the development of ALI.


Obstetric analgesia, anaesthesia and the Apgar score.
This study examines the changing pattern of maternal analgesia administration during the decade 1970-1979. In addition, the relationship between analgesia, anaesthesia and the infant's Apgar score was documented. Approximately 40000 deliveries to South Glamorgan residents were included in the study. Over the two quinquennia 1970-1974 and 1975-1979 the administration of epidural block increased from 5.6% to 17.5%. The Apgar score of babies in the epidural block group was significantly better in the latter period. Respiratory depression following Caesarean section remains a problem. The group delivered by elective Caesarean section contained a substantial proportion of infants with a low Apgar score; 30% had an Apgar score less than 8 and 10% had a score of less than 4. Likely causative factors include undue sensitivity of the infants' respiratory centre and aortocaval compression during surgery.


Enzymuria of the rat: biorhythms and sex differences.
Alanine aminopeptidase, gamma-glutamyltransferase and N-acetyl-beta-D-glucosaminidase were measured daily over 65 days in 24-hour urine of male and female Wistar rats. The mathematical evaluation was based on the Fourier-analysis. The excretion of alanine aminopeptidase and gamma-glutamyltransferase was higher in male than in female rats. This sex-dependent difference was not observed for N-acetyl-beta-D-glucosaminidase. The excretion of the 3 enzymes followed a biorhythm with a dominant period of 7 days for gamma-glutamyltransferase and N-acetyl-beta-D-glucosaminidase and one of 9 days for alanine aminopeptidase. Biorhythms and sex differences of enzymuria should be considered in experimental designs.


Hypertriglyceridemia-associated acute pancreatitis: Response to continuous insulin infusion.
To assess the response of serum triglycerides (TG) to continuous insulin infusion (CII) in adults with hypertriglyceridemia-associated acute pancreatitis (HTGP). Retrospective analysis of TG response to standardized CII therapy in 77 adults admitted to intensive care with TG >1000 mg/dL and HTGP. Participants had initial TG 3869.0 [2713.5, 5443.5] mg/dL and were 39.3 ± 9.7 years old, 66.2% males, 58.4% Hispanic, BMI 30.2 [27.0, 34.8] kg/m2, 74.0% with diabetes mellitus (DM) and 50.6% with excess alcohol use. TG-goal, defined as ≤1,000 ± 100 mg/dL, was achieved in 95%. Among the 73 TG-goal achievers (responders), 53.4% reached TG-goal in <36 hours after CII initiation (rapid responders). When compared to slow responders taking≥36 hours, rapid responders had lower initial TG (2862.0 [1965.0, 4519.0] vs 4814.5 [3368.8, 6900.0] mg/dL), BMI (29.4 [25.9, 32.8] vs 31.9 [28.2, 38.3] kg/m2), DM prevalence (56.4 vs 94.1%), and reached TG-50% (half of respective initial TG) faster (12.0 [6.0, 17.0] vs 18.5 [13.0, 32.8] hours). Those with DM (n = 57) vs non-DM (n = 20) were obese (31.4 [28.0, 35.6] vs 27.8 [23.6, 30.3] kg/m2), took longer to reach TG-final (41.0 [25.0, 60.5] vs 14.5 [12.5, 25.5] hours) and used more daily insulin (1.7 [1.3, 2.1] vs 1.1 [0.5, 1.9] U/kg/day). Among those with DM, the rapid responders had higher daily use of insulin vs slow responders 1.9 [1.4, 2.3] vs 1.6 [1.1, 1.8] U/kg/day. All results significant. In multivariable analysis, predictors of faster TG response were absence of DM, lower BMI and initial TG. CII was effective in reaching TG-goal in 95% of patients with HTGP. Half achieved TG-goal within 36 hours. Presence of DM, higher BMI and initial TG slowed the time to reach TG-goal. These baseline parameters and rate of decline to TG-50% may be real-time indicators to initiate and adjust the CII for quicker response.


Stress-induced accumulation of wheat germ agglutinin and abscisic Acid in roots of wheat seedlings.
Wheat germ agglutinin (WGA) levels in roots of 2-day-old wheat seedlings increased up to three-fold when stressed by air-drying. Similar results were obtained when seedling roots were incubated either in 0.5 molar mannitol or 180 grams per liter polyethylene glycol 6000, with a peak level of WGA after 5 hours of stress. Longer periods of osmotic treatment resulted in a gradual decline of WGA in the roots. Since excised wheat roots incorporate more [(35)S]cysteine into WGA under stress conditions, the observed increase of lectin levels is due to de novo synthesis. Measurement of abscisic acid (ABA) levels in roots of control and stressed seedlings indicated a 10-fold increase upon air-drying. Similarly, a five- and seven-fold increase of ABA content of seedling roots was found after 2 hours of osmotic stress by polyethylene glycol 6000 and mannitol, respectively. Finally, the stress-induced increase of WGA in wheat roots could be inhibited by growing seedlings in the presence of fluridone, an inhibitor of ABA synthesis. These results indicate that roots of water-stressed wheat seedlings (a) contain more WGA as a result of an increased de novo synthesis of this lectin, and (b) exhibit higher ABA levels. The stress-induced increase of lectin accumulation seems to be under control of ABA.


Predictive Modeling of Tacrolimus Dose Requirement Based on High-Throughput Genetic Screening.
Any biochemical reaction underlying drug metabolism depends on individual gene-drug interactions and on groups of genes interacting together. Based on a high-throughput genetic approach, we sought to identify a set of covariant single-nucleotide polymorphisms predictive of interindividual tacrolimus (Tac) dose requirement variability. Tac blood concentrations (Tac C0 ) of 229 kidney transplant recipients were repeatedly monitored after transplantation over 3 mo. Given the high dimension of the genomic data in comparison to the low number of observations and the high multicolinearity among the variables (gene variants), we developed an original predictive approach that integrates an ensemble variable-selection strategy to reinforce the stability of the variable-selection process and multivariate modeling. Our predictive models explained up to 70% of total variability in Tac C0 per dose with a maximum of 44 gene variants (p-value <0.001 with a permutation test). These models included molecular networks of drug metabolism with oxidoreductase activities and the multidrug-resistant ABCC8 transporter, which was found in the most stringent model. Finally, we identified an intronic variant of the gene encoding SLC28A3, a drug transporter, as a key gene involved in Tac metabolism, and we confirmed it in an independent validation cohort.


High-performance liquid chromatographic assay for N-acetylcysteine in plasma and urine.
A sensitive high-performance liquid chromatographic (HPLC) assay for N-acetylcysteine in human plasma and urine has been developed. The method employs a prechromatographic stage to produce the dinitrophenyl derivative of N-acetylcysteine, which is chromatographed on a 5-microns C18 bonded reverse-phase column using an external standard method. The derivatized N-acetylcysteine molecule has a retention time of congruent to 13 min at a flow rate of 1.0 mL/min for a mobile phase of methanol-aqueous 0.05 M citrate-0.001 M EDTA buffer pH 7.0 (30:70). In a multistep extraction procedure, which is slightly modified for the N-acetylcysteine assay in urine, the limits of sensitivity for the plasma and urine assays were found to be congruent to 60 ng/mL and congruent to 200 micrograms/mL, respectively. Preliminary data from pilot studies in human subjects are presented.