Modified heparin-Sepharose procedure for determination of plasma lipolytic activities of normolipidemic and hyperlipidemic subjects after injection of heparin.
A modified heparin-Sepharose affinity chromatography procedure (Boberg et al., J. Lipid Res. 18:544-547, 1977) was developed to determine two different triglyceride lipase activities in human post-heparin plasma: hepatic triglyceride lipase (I) and lipoprotein lipase (II). With this procedure, lipoproteins were separated from the eluted lipases. The total lipolytic activity of II was eluted from heparin-Sepharose by heparin. The use of heparin as eluting agent prevents the partial inhibition of II, in contrast to the procedure based on elution of II with a high concentration of NaCl. In a comparative study with the modified heparin-Sepharose affinity column chromatography, the immunochemical and protamine sulfate inhibition procedures, the results indicated that these three procedures are equally suitable for the determination of I and II from normolipidemic subjects. However, because of possible interference by plasma, the column-chromatographic procedure is the preferred method for measuring lipase concentrations in post-heparin plasma of hyperlipidemic patients. The II activity of post-heparin plasma from normolipidemic subjects was not significantly age-(20-39 and 40-60 years) or sex-related. I activity was also not significantly different with respect to age, but was significantly greater in men than in women.


Temperature and pressure effects on GFP mutants: explaining spectral changes by molecular dynamics simulations and TD-DFT calculations.
By combining spectroscopic measurements under high pressure with molecular dynamics simulations and quantum mechanics calculations we investigate how sub-angstrom structural perturbations are able to tune protein function. We monitored the variations in fluorescence output of two green fluorescent protein mutants (termed Mut2 and Mut2Y, the latter containing the key T203Y mutation) subjected to pressures up to 600 MPa, at various temperatures in the 280-320 K range. By performing 150 ns molecular dynamics simulations of the protein structures at various pressures, we evidenced subtle changes in conformation and dynamics around the light-absorbing chromophore. Such changes explain the measured spectral tuning in the case of the sizable 120 cm(-1) red-shift observed for pressurized Mut2Y, but absent in Mut2. Previous work [Barstow et al., Proc. Natl. Acad. Sci. U. S. A., 2008, 105, 13362] on pressure effects on GFP also involved a T203Y mutant. On the basis of cryocooling X-ray crystallography, the pressure-induced fluorescence blue shift at low temperature (77 K) was attributed to key changes in relative conformation of the chromophore and Tyr203 phenol ring. At room temperature, however, a red shift was observed at high pressure, analogous to the one we observe in Mut2Y. Our investigation of structural variations in compressed Mut2Y also explains their result, bridging the gap between low-temperature and room-temperature high-pressure effects.


Socio-demographic, economic and mental health problems were risk factors for suicidal ideation among Kenyan students aged 15 plus.
About a third of youth with suicidal ideation develop suicidal plans and about 60% of youth with suicidal plans make suicidal attempts. This study aimed to study different types of suicidal ideation and the risk factors in Kenyan youth. We studied a total of 9742 high school, college and university students using following self-administered instruments: - a researcher design socio-demographic questionnaire, Psychiatric Diagnostic Screening Questionnaire (PDSQ) to document psychiatric disorders and various types of suicidal ideas in previous two weeks, Washington Early Recognition Center Affectivity and Psychosis tool to assess stress, affectivity and psychosis, Wealth index questionnaire to document economic indicators based on household items for the families of the students. We used descriptive statistics, univariate analysis, bivariate logistic regression analysis and variables with a p-value of less than <.05 were entered into generalized linear models using logit links to identify independent predictors. The overall prevalence of different types of suicidal ideation was (22.6%), major depression was found in 20.0%, affectivity, psychosis and stress was found in 10.4%, 8.7% and 26.0% respectively. Female gender, major depression, stress, affectivity and psychosis and being in high school were significant (p < 0.05) predictors of suicidal ideation. This was a cross sectional study that focused only on suicidal ideas and associated economic factors and mental health disorders. It did not study suicidal behavior. Future studies are needed to study the progression from suicidal ideas to suicidal attempts and the factors associated with that progression.


Combined exposure to low doses of pesticides causes decreased birth weights in rats.
Decreased birth weight is a common effect of many pesticides in reproductive toxicity studies, but there are no empirical data on how pesticides act in combination on this endpoint. We hypothesized that a mixture of six pesticides (cyromazine, MCPB, pirimicarb, quinoclamine, thiram, and ziram) would decrease birth weight, and that these mixture effects could be predicted by the Dose Addition model. Data for the predictions were obtained from the Draft Assessment Reports of the individual pesticides. A mixture of equi-effective doses of these pesticides was tested in two studies in Wistar rats, showing mixture effects in good agreement with the additivity predictions. Significantly lower birth weights were observed when compounds were present at individual doses below their no-observed adverse effect levels (NOAELs). These results emphasize the need for cumulative risk assessment of pesticides to avoid potentially serious impact of mixed exposure on prenatal development and pregnancy in humans.


Anti-GBM disease with a mild relapsing course and low levels of anti-GBM autoantibodies.
Anti-glomerular basement membrane disease (anti-GBM) is usually characterized by rapidly progressive glomerulonephritis, and when autoantibody production has ceased, relapses are rare. Here, we report a 71-year-old women diagnosed at a stage of mild renal insufficiency. Over a period of 10 years, she experienced three mild relapses with return of anti-GBM antibodies, haematuria and slight elevations in serum creatinine level. All three relapses responded to immunosuppressive therapy, and all were preceded by peaks of myeloperoxidase-antineutrophil cytoplasm antibodies (MPO-ANCA). This case shows that long-term follow-up is warranted in patients treated for anti-GBM-mediated disease, but urinary dipsticks may be sufficient for early detection of relapses.


Synthesis, Herbicidal Activity, and QSAR of Novel N-Benzothiazolyl- pyrimidine-2,4-diones as Protoporphyrinogen Oxidase Inhibitors.
Protoporphyrinogen oxidase (PPO, E.C. 126.96.36.199) is known as a key action target for several structurally diverse herbicides. As a continuation of our research work on the development of new PPO-inhibiting herbicides, a series of novel 3-(2'-halo-5'-substituted-benzothiazol-1'-yl)-1-methyl-6-(trifluoromethyl)pyrimidine-2,4-diones 9 were designed and synthesized. The bioassay results indicated that a number of the newly synthesized compounds exhibited higher inhibition activity against tobacco PPO (mtPPO) than the controls, saflufenacil and sulfentrazone. Compound 9F-5 was identified as the most potent inhibitor with a Ki value of 0.0072 μM against mtPPO, showing about 4.2-fold and 1.4-fold higher potency than sulfentrazone (Ki = 0.03 μM) and saflufenacil (Ki = 0.01 μM), respectively. An additional green house assay demonstrated that compound 9F-6 (Ki = 0.012 μM) displayed the most promising postemergence herbicidal activity with a broad spectrum even at a concentration as low as 37.5 g of active ingredient (ai)/ha. Maize exhibits relative tolerance against compound 9F-6 at the dosage of 150 g ai/ha, but it is susceptible to saflufenacil even at 75 g ai/ha. Thus, compound 9F-6 exhibits the potential to be a new herbicide for weed control in maize fields.


Standard Setting in Simulation-based Training of Surgical Procedures: A Systematic Review.
This systematic review aims to examine the use of standard-setting methods in the context of simulation-based training of surgical procedures. Simulation-based training is increasingly used in surgical education. However, it is important to determine which level of competency trainees must reach during simulation-based training before operating on patients. Therefore, pass/fail standards must be established using systematic, transparent, and valid methods. Systematic literature search was done in 4 databases (Ovid MEDLINE, Embase, Web of Science, and Cochrane Library). Original studies investigating simulation-based assessment of surgical procedures with the application of a standard setting were included. Quality of evidence was appraised using GRADE. Of 24,299 studies identified by searches, 232 studies met the inclusion criteria. Publications using already established standard settings were excluded (N = 70), resulting in 162 original studies included in the final analyses. Most studies described how the standard setting was determined (N = 147, 91%) and most used the mean or median performance score of experienced surgeons (n = 65, 40%) for standard setting. We found considerable differences across most of the studies regarding study design, setup, and expert level classification. The studies were appraised as having low and moderate evidence. Surgical education is shifting toward competency-based education, and simulation-based training is increasingly used for acquiring skills and assessment. Most studies consider and describe how standard settings are established using more or less structured methods but for current and future educational programs, a critical approach is needed so that the learners receive a fair, valid, and reliable assessment.


No evidence of occult HBV infection in population born after mass vaccination.
Despite access to efficient hepatitis B virus (HBV) vaccine and universal immunization schedules, HBV infection remains a global health concern. HBV infection has decreased by this program. Nevertheless, breakthrough infections occur due to generation of occult HBV infection (OBI) and surface gene mutants in the immunized population. We aimed to determine the presence of OBI in a population born after initiation of nationwide HBV vaccination in Tehran, Iran. A HBV mass vaccination schedule was launched in Iran in 1993. For this study, we enrolled 1120 cases younger than 24 years. ELISA was applied to evaluate the presence of HBsAg, anti-HBs and anti-HBc. HBV-DNA presence was determined in all HBsAg-negative cases using nested polymerase chain reaction. The prevalence of HBsAg, anti-HBc and anti-HBs was 0.1, 0.54 and 39.9% respectively. Out of 6 anti-HBc-positive individuals, 4 cases also had anti-HBs. One case revealed HBsAg co-existence and the other one showed isolated anti-HBc. HBV-DNA was not detected in HBsAg-negative specimens. A very low prevalence of HBsAg and isolated anti-HBc was observed and no occult HBV infection was detected. It seems that evasion mutants are not a potential threat for HBV universal immunization efficacy in the vaccinated population.


Intrinsic differentiation potential of adolescent human tendon tissue: an in-vitro cell differentiation study.
Tendinosis lesions show an increase of glycosaminoglycan amount, calcifications, and lipid accumulation. Therefore, altered cellular differentiation might play a role in the etiology of tendinosis. This study investigates whether adolescent human tendon tissue contains a population of cells with intrinsic differentiation potential. Cells derived from adolescent non-degenerative hamstring tendons were characterized by immunohistochemistry and FACS-analysis. Cells were cultured for 21 days in osteogenic, adipogenic, and chondrogenic medium and phenotypical evaluation was carried out by immunohistochemical and qPCR analysis. The results were compared with the results of similar experiments on adult bone marrow-derived stromal cells (BMSCs). Tendon-derived cells stained D7-FIB (fibroblast-marker) positive, but alpha-SMA (marker for smooth muscle cells and pericytes) negative. Tendon-derived cells were 99% negative for CD34 (endothelial cell marker), and 73% positive for CD105 (mesenchymal progenitor-cell marker). In adipogenic medium, intracellular lipid vacuoles were visible and tendon-derived fibroblasts showed upregulation of adipogenic markers FABP4 (fatty-acid binding protein 4) and PPARG (peroxisome proliferative activated receptor gamma). In chondrogenic medium, some cells stained positive for collagen 2 and tendon-derived fibroblasts showed upregulation of collagen 2 and collagen 10. In osteogenic medium Von Kossa staining showed calcium deposition although osteogenic markers remained unaltered. Tendon-derived cells and BMCSs behaved largely comparable, although some distinct differences were present between the two cell populations. This study suggests that our population of explanted human tendon cells has an intrinsic differentiation potential. These results support the hypothesis that there might be a role for altered tendon-cell differentiation in the pathophysiology of tendinosis.


An Increase Incidence in Uric Acid Nephrolithiasis: Changing Patterns.
Nephrolithiasis is a complex disease affecting all age groups globally. As the causative factors for nephrolithiasis rises significantly, its incidence, prevalence and recurrence continues to baffle clinicians and patients. To study the prevalence of different types of renal stones extracted by Percutaneous Nephrolithotomy (PCNL) and open surgical procedures. Renal stones from 50 patients were retrieved by Percutaneous Nephrolithotomy (PCNL), Ureterorenoscopy (URS) and open surgical techniques for qualitative tests for detection of calcium, oxalate, uric acid, phosphate, ammonium ion, carbonate, cystine and xanthine. Three patients had stone removed by open surgery and rest had undergone PCNL. Nine of the stones were pure of calcium oxalate, 9 were of pure uric acid and 32 were mixed stones. Forty one stones had calcium. Among the mixed stones, oxalate was present in 25 samples (39 of total), uric acid was seen in 17 (25 of total stones), phosphate was present in 23 (23 of total) and carbonate was present in 4 stones (4 of total). Only 1 patient had triple phosphate stone. 12 were of staghorn appearance of which 6 were of struvite type, 6 were pure uric acid and remaining were mixed oxalate-phosphate stones. Our study, though in a small number of hospital based patients, found much higher prevalence of uric acid stones and mixed stones than reported by previous hospital based studies in north India (oxalate stones~90%, uric acid~1% and mixed stones~3%). Biochemical analysis of renal stones is warranted in all cases.


Breast Cancer Cells and PD-1/PD-L1 Blockade Upregulate the Expression of PD-1, CTLA-4, TIM-3 and LAG-3 Immune Checkpoints in CD4+ T Cells.
: Triple negative breast cancer (TNBC) is the most aggressive breast cancer subtype, and it exhibits resistance to common breast cancer therapies. Immune checkpoint inhibitors (ICIs) targeting programmed cell death 1 (PD-1) and its ligand, PD-L1, have been approved to treat various cancers. However, the therapeutic efficacy of targeting PD-1/PD-L1 axis in breast cancer is under clinical investigation. In addition, the mechanisms of action of drugs targeting PD-1 and PD-L1 have not been fully elucidated. In this study, we investigated the effect of human TNBC cell lines, MDA-MB-231 and MDA-MB-468, and the non-TNBC cell line, MCF-7, on the expression of immune checkpoints (ICs) on CD4+ T cell subsets, including regulatory T cells (Tregs), using a co-culture system. We also examined the effect of blocking PD-1 or PD-L1 separately and in combination on IC expression by CD4+ T cell subsets. We found that breast cancer cells upregulate the expression of ICs including PD-1, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) and lymphocyte activation gene-3 (LAG-3) in CD4+ T cell subsets. We also found that the co-blockade of PD-1 and PD-L1 further upregulates the co-expression of TIM-3 and LAG-3 on CD4+CD25+ T cells and CD4+CD25+FoxP3+Helios+ Tregs in the presence of TNBC cells, but not in non-TNBC cells. Our results indicate the emergence of compensatory inhibitory mechanisms, most likely mediated by Tregs and activated non-Tregs, which could lead to the development of TNBC resistance against PD-1/PD-L1 blockade.


Intracapsular hip fracture: increased cortical remodeling in the thinned and porous anterior region of the femoral neck.
It has been shown previously that the antero-inferior cortex is subjected to maximal tensile stress during a fall onto the greater trochanter. We have recently shown that in cases of femoral neck fracture, cortical thinning and porosity is greatest in the anterior and antero-inferior region of the femoral neck. To investigate whether this is due to increased remodeling, we have quantified surface-based parameters associated with Haversian remodeling in femoral neck biopsies from women with intracapsular hip fracture and post-mortem controls. Cryostat sections of chilled biopsies were reacted for either tartrate-resistant acid phosphatase (TRAP) or alkaline phosphatase (ALP) activity. Proportions of active canals were determined in each quadrant (inferior, anterior, superior, posterior) of the femoral neck. The biopsies were then embedded in methacrylate to permit histomorphometry using Goldner's and Solochrome sections. In the cases there was no significant increase in the proportion of canals undergoing remodeling in the cortex as a whole (p = 0.846), but the regional distribution of remodeling was markedly different from that in the controls. In the anterior cortex, the proportion of canals undergoing remodeling was increased by 56% (p = 0.0087); in contrast there was a relative decrease of 35% in the superior region (p = 0.0047). In the anterior cortex of cases there were 76% and 42% increases in the proportions of eroded (p = 0.019) and osteoid-bearing (p = 0.041) canals, respectively. In the superior region, the decrease in the proportion of remodeling sites was due to a marked decrease in canals with an osteoid surface (51%; p = 0.0031). Covariance analysis with cortical porosity as the dependent variable showed that porosity was significantly dependent on the regional distribution of eroded (p = 0.033) but not on the distribution of forming (p = 0.153) canals (R(2)adj = 0.51). Cellular levels of TRAP and ALP were significantly elevated in the anterior region of cases compared with the controls (TRAP 55%, p = 0.006; ALP 36%, p = 0.003). For the posterior and inferior regions there were no marked differences in cellular TRAP and ALP levels compared with control values. These data show that the increased cortical thinning and increased porosity we have previously observed in the anterior cortex in cases of hip fracture are associated with increased indices of Haversian remodeling. These findings are consistent with the hypothesis that, in cases of hip fracture, remodeling imbalance in the anterior cortex is a continuing process up to the time of fracture and is due to increased osteoclastic cellular activity associated with an osteoblastic response that is inadequate to prevent bone loss.


[alpha-Tocopherol in the combined treatment of vitiligo].
Photochemotherapy was combined with alpha-tocopherol in the treatment of 56 patients with vitiligo in order to shorten the treatment. alpha-tocopherol was used as antioxidant to reduce skin erythema and build up single UV doses. Due to 1.5-2-fold increase of UV exposure follicular mechanisms stimulation was enhanced in vitiligo foci, perifollicular pigmentation was achieved approximately 2 times sooner, and the length of treatment necessary to achieve manifest repigmentation (i.e. more than 50% of the maculae surface) was reduced twofold.


Microelectrode Arrays for Simultaneous Electrophysiology and Advanced Optical Microscopy.
Advanced optical imaging techniques address important biological questions in neuroscience, where structures such as synapses are below the resolution limit of a conventional microscope. At the same time, microelectrode arrays (MEAs) are indispensable in understanding the language of neurons. Here, the authors show transparent MEAs capable of recording action potentials from neurons and compatible with advanced microscopy. The electrodes are made of the conducting polymer poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS) and are patterned by optical lithography, ensuring scalable fabrication with good control over device parameters. A thickness of 380 nm ensures low enough impedance and >75% transparency throughout the visible part of the spectrum making them suitable for artefact-free recording in the presence of laser illumination. Using primary neuronal cells, the arrays record single units from multiple nearby sources with a signal-to-noise ratio of 7.7 (17.7 dB). Additionally, it is possible to perform calcium (Ca2+) imaging, a measure of neuronal activity, using the novel transparent electrodes. Different biomarkers are imaged through the electrodes using conventional and super-resolution microscopy (SRM), showing no qualitative differences compared to glass substrates. These transparent MEAs pave the way for harnessing the synergy between the superior temporal resolution of electrophysiology and the selectivity and high spatial resolution of optical imaging.


Impairment of ULK1 sulfhydration-mediated lipophagy by SREBF1/SREBP-1c in hepatic steatosis.
Nonalcoholic fatty liver disease (NAFLD) affects a quarter of the global population. However, its pathogenesis is not completely understood. In our recent study, we have demonstrated that in a high-fat diet-induced liver steatosis model, the activation of SREBF1/SREBP-1c (sterol regulatory element binding transcription factor 1) directly upregulates Mir216a transcription, which inhibits CTH/CSE (cystathionase (cystathionine gamma-lyase)) expression and its function in hydrogen sulfide (H2S) production. Reduced H2S production suppresses the sulfhydration of ULK1 (unc-51 like autophagy activating kinase 1), consequently inhibiting autophagic flux and lipid droplet turnover. A single substitution mutation (C951S) in ULK1 or the silencing of CTH impairs ULK1 sulfhydration-mediated lipophagy, thereby promoting hepatic steatosis in mice. Interestingly, the sulfhydration of ULK1 increases its intrinsic kinase activity to modulate autophagy at both initiation and progression stages of autophagic catabolic flux. This study reveals that SREBF1/SREBP-1c contributes to hepatic lipid accumulation through its combined effect of increased lipid synthesis coupled with decreased lipid degradation mediated by autophagic dysregulation.


Demographic, belief, and situational factors influencing the decision to utilize emergency medical services among chest pain patients. Rapid Early Action for Coronary Treatment (REACT) study.
Empirical evidence suggests that people value emergency medical services (EMS) but that they may not use the service when experiencing chest pain. This study evaluates this phenomenon and the factors associated with the failure to use EMS during a potential cardiac event. Baseline data were gathered from a randomized, controlled community trial (REACT) that was conducted in 20 US communities. A random-digit-dial survey documented bystander intentions to use EMS for cardiac symptoms in each community. An emergency department surveillance system documented the mode of transport among chest pain patients in each community and collected ancillary data, including situational factors surrounding the chest pain event. Logistic regression identified factors associated with failure to use EMS. A total of 962 community members responded to the phone survey, and data were collected on 875 chest pain emergency department arrivals. The mean proportion of community members intending to use EMS during a witnessed cardiac event was 89%; the mean proportion of patients observed using the service was 23%, with significant geographic differences (range, 10% to 48% use). After controlling for covariates, non-EMS users were more likely to try antacids/aspirin and call a doctor and were less likely to subscribe to (or participate in) an EMS prepayment plan. The results of this study indicate that indecision, self-treatment, physician contact, and financial concerns may undermine a chest pain patient's intention to use EMS.


Intracarotid cisplatin-based chemotherapy in patients with malignant melanoma and central nervous system (CNS) metastases.
Twenty-three patients with malignant melanoma metastatic to the central nervous system (CNS) were treated with intracarotid cisplatin-based chemotherapy. Twenty-two had failed prior cranial radiation therapy (one patient refused radiation therapy) and had progressive brain metastases documented by computerized tomography (CT). Intracarotid cisplatin 40-75 mg/m2 was administered alone (seven patients) with 1,3-bis(2-Chloroethyl)-1-nitrosourea (BCNU) (13 patients) or bleomycin (three patients). Courses were repeated monthly with CT scan. Seven patients (30%) had objective improvement in CT scans and three patients (13%) had stabilization of disease. The median time to tumor progression for responding patients was 20 weeks (range 8-34 weeks) while stable disease ranged from 9-12 weeks. In three patients the extracranial disease progressed while the brain metastases responded. Neurological and retinal toxicity were potential complications of therapy. Intracarotid cisplatin-based chemotherapy may be useful for palliation in selected patients with malignant melanoma and CNS metastases.


Baseline and acquired resistance to bedaquiline, linezolid and pretomanid, and impact on treatment outcomes in four tuberculosis clinical trials containing pretomanid.
Bedaquiline (B), pretomanid (Pa) and linezolid (L) are key components of new regimens for treating rifampicin-resistant tuberculosis (TB). However, there is limited information on the global prevalence of resistance to these drugs and the impact of resistance on treatment outcomes. Mycobacterium tuberculosis (MTB) phenotypic drug susceptibility and whole-genome sequence (WGS) data, as well as patient profiles from 4 pretomanid-containing trials-STAND, Nix-TB, ZeNix and SimpliciTB-were used to investigate the rates of baseline resistance (BR) and acquired resistance (AR) to BPaL drugs, as well as their genetic basis, risk factors and impact on treatment outcomes. Data from >1,000 TB patients enrolled from 2015 to 2020 in 12 countries was assessed. We identified 2 (0.3%) participants with linezolid BR. Pretomanid BR was also rare, with similar rates across TB drug resistance types (0-2.1%). In contrast, bedaquiline BR was more prevalent among participants with highly resistant TB or longer prior treatment histories than those with newly diagnosed disease (5.2-6.3% vs. 0-0.3%). Bedaquiline BR was a risk factor for bacteriological failure or relapse in Nix-TB/ZeNix; 3/12 (25%, 95% CI 5-57%) participants with vs. 6/185 (3.2%, 1.2-6.9%) without bedaquiline BR. Across trials, we observed no linezolid AR, and only 3 cases of bedaquiline AR, including 2 participants with poor adherence. Overall, pretomanid AR was also rare, except in ZeNix patients with bedaquiline BR. WGS analyses revealed novel mutations in canonical resistant genes and, in 7 MTB isolates, the genetic determinants could not be identified. The overall low rates of BR to linezolid and pretomanid, and to a lesser extent to bedaquiline, observed in the pretomanid trials are in support of the worldwide implementation of BPaL-based regimens. Similarly, the overall low AR rates observed suggest BPaL drugs are better protected in the regimens trialed here than in other regimens combining bedaquiline with more, but less effective drugs.


Relationship between bronchial airway responsiveness and clinical severity of asthma.
The minimum medication required to control symptoms was individually established in a systematic manner in 10 asthmatics. A scoring system for the severity of asthma was designed using the indices of initial airway calibre (a reflection of the degree of airway obstruction) and the minimum medication requirement; all the subjects were so scored. Bronchial airway responsiveness to histamine, methacholine and isocapnic hyperventilation of cold air was then measured in these subjects. The relationship between the level of bronchial responsiveness and the asthma severity score was examined. The mean airway responsiveness to histamine or methacholine for the subjects who required a combination of drugs was not significantly greater than that for those who required single medication intermittently or daily, while the airway responsiveness to cold air was significantly different between the subjects in the two treatment subgroups. Similarly, there was no correlation between the asthma severity score and airway responsiveness to methacholine and histamine (r = -0.38 and -0.48; P greater than 0.1) while a significant correlation was found with responsiveness to cold air (r = 0.72; P less than 0.02). The results suggest that there is a qualitative difference between the bronchoconstriction induced in asthmatic subjects by pharmacological constrictor substances and natural physical stimuli such as cold air.


Atypical infective endocarditis.
Although infective endocarditis has changed in the recent past as a result of microbiologic and risk factors, it continues to be clinically challenging. The disease is characterized by the formation of septic masses of platelets on the surfaces of heart valves. Several mechanisms can cause or contribute to the development of endocarditis. Although risk factors for infective endocarditis are well known, patients with atypical signs and symptoms continue to challenge us. We describe a case report of a patient admitted to our inpatient service with back pain and presumed pyelonephritis. A MEDLINE literature search was conducted, using the key words "endocarditis," "back pain," and "bacterial," for the years 1986 to the present. A 42-year-old woman with a history of intravenous drug abuse was admitted to the family practice service with back pain and pyelonephritis. She developed hypoxia and a new heart murmur and had continued fevers. Blood cultures drawn in the emergency department grew methicillin-resistant Staphylococcus aureus. A bone scan and magnetic resonance imaging led to the diagnosis of epidural abscess. What appeared to be a simple case of pyelonephritis with back pain became a case of infective endocarditis complicated by an epidural abscess.


Genetic Contributions to Attachment Stability Over Time: the Roles of CRHR1 Polymorphisms.
Corticotropin-releasing hormone receptor 1 (CRHR1), a hormone receptor essential to the activation of HPA axis and the subsequent release of cortisol, plays critical roles in emotional and behavioral responses relevant to attachment. However, the specific roles of CRHR1 polymorphisms in attachment remain unclear. To further clarify these genetic effects, this research conducted a three-wave study to investigate whether the CRHR1 polymorphisms (i.e., rs110402 and rs242924) are associated with the stability and variability of attachment by using a sample of freshmen (N = 604; Mage = 18.57 years, SD = 1.90; 68.8% girls). The results showed that rs110402 and rs242924 were associated with the stability of closeness-dependence. The G alleles of the both polymorphisms were found not to be related to lower attachment stability. However, these polymorphisms were not associated with the variability of attachment. Overall, these findings provide evidence for the contribution of CRHR1 to attachment stability.


Segmentectomy quality remains important in ground-glass-dominant stage I lung cancer.
Segmentectomy for early-stage lung cancer has benefits for survival and parenchymal preservation. However, segmentectomies are technically challenging, thereby resulting in considerable variability in the quality of resection. In this study, we aimed to review the quality of segmentectomies and analyze their clinical impact. This retrospective study reviewed patients diagnosed with stage I lung cancer after segmentectomies between 2013 and 2021. Segmentectomies were classified as anatomical or nonanatomical; anatomical resection included segmental bronchus and vessel (artery and/or vein) divisions; others were classified as nonanatomical. The primary outcome was recurrence-free and overall survival, and the secondary outcome was postoperative spirometry and lung plication, which is seen as a fibrotic line along the stapling site. Of the 132 segmental resections included in this study, 101 (76.5%) were anatomical segmentectomies. The median consolidation-tumor ratio was 0.40, and 83.3% (110/132) had ground-glass opacities (GGOs). Compared to nonanatomical resections, more N1 and total lymph node stations were retrieved after anatomical segmentectomies. Regarding clinical outcomes, recurrence-free survival was better after anatomical segmentectomy (p = 0.049); however, overall survival was not significantly different (p = 0.064). Furthermore, at 3-6 months postoperatively, thicker lung plication at the stapling site was observed in nonanatomical resections (p < 0.001). Subgroup analysis for complex segmentectomies revealed a larger decrease in forced-expiration volume in 1 s after nonanatomical resection. Anatomical segmentectomy resulted in better survival and a lower incidence of thick lung plication, even in GGO-dominant tumors. Therefore, further standardization and quality management of segmentectomy procedures will improve the clinical outcomes.


Interplay between proximal and distal promoter elements is required for squamous differentiation marker induction in the bronchial epithelium: role for ESE-1, Sp1, and AP-1 proteins.
Overexpression of SPRR1B in bronchial epithelial cells is a marker for early metaplastic changes induced by various toxicants/carcinogens. Previously, we have shown that the transcriptional stimulation of SPRR1B expression by phorbol 12-myristate 13-acetate (PMA) is mainly mediated by a -150/-94 bp enhancer harboring two critical 12-O-tetradecanoylphorbol-13-acetate-responsive elements (TREs) and by Jun.Fra-1 dimers. Here, we show that a region between -54 and -39 bp containing an ETS-binding site (EBS) and a GC box is essential for both basal and PMA-inducible SPRR1B transcription. In vivo footprinting demonstrated binding of transcription factors to these elements. However, unlike enhancer TREs, exposure of cells to PMA did not significantly alter the footprinting pattern at these elements. Mutations that crippled both the EBS and GC box suppressed both basal and PMA-inducible SPRR1B transcription. Consistent with this, overexpression of EBS-binding proteins ESE-1 and ESE-3 significantly stimulated SPRR1B promoter activity. Furthermore, preceding SPRR1B transcription, PMA up-regulated mRNA expression of ETS family members such as ESE-1 and ESE-3. Although ESE-1 synergistically activated c-Jun- and PMA-enhanced SPRR1B transcription, coexpression of Sp1 and ESE-1 showed no synergistic or additive effect on promoter activity, indicating an obligatory role for AP-1 proteins in such regulation. In support of this notion, deletion or mutation of two functional TREs inhibited ESE-1- and Sp1-enhanced promoter activation. Thus, the interaction between ESE-1 and Sp1, and AP-1 proteins that bind to the proximal and distal promoter regions, respectively, play a critical role in the induction of squamous differentiation marker expression in bronchial epithelial cells.


Cascade cyclization and intramolecular nitrone dipolar cycloaddition and formal synthesis of 19-hydroxyibogamine.
A cascade or domino sequence of condensation of hydroxylamine and an aldehyde to give an oxime, cyclization to a nitrone, and intramolecular 1,3-dipolar cycloaddition has been successfully employed where there is branching at C-4 as a route to the iboga alkaloids. Cyclization occurs with displacement of chloride as a leaving group and intramolecular cycloaddition occurs with an alkene as a dipolarophile. The reaction gives an azabicyclo[2.2.2]octane product containing a fused isoxazolidine as a single stereoisomer and this was converted to an isoquinuclidine that completed a formal synthesis of the alkaloid (±)-19-hydroxyibogamine.


The human Rh50 glycoprotein gene. Structural organization and associated splicing defect resulting in Rh(null) disease.
The Rh (Rhesus) protein family comprises Rh50 glycoprotein and Rh30 polypeptides, which form a complex essential for Rh antigen expression and erythrocyte membrane integrity. This article describes the structural organization of Rh50 gene and identification of its associated splicing defect causing Rhnull disease. The Rh50 gene, which maps at chromosome 6p11-21.1, has an exon/intron structure nearly identical to Rh30 genes, which map at 1p34-36. Of the 10 exons assigned, conservation of size and sequence is confined mainly to the region from exons 2 to 9, suggesting that RH50 and RH30 were formed as two separate genetic loci from a common ancestor via a transchromosomal insertion event. The available information on the structure of RH50 facilitated search for candidate mutations underlying the Rh deficiency syndrome, an autosomal recessive disorder characterized by mild to moderate chronic hemolytic anemia and spherostomatocytosis. In one patient with the Rhnull disease of regulator type, a shortened Rh50 transcript lacking the sequence of exon 7 was detected, while no abnormality was found in transcripts encoding Rh30 polypeptides and Rh-related CD47 glycoprotein. Amplification and sequencing of the genomic region spanning exon 7 revealed a G-->A transition in the invariant GT motif of the donor splice site in both Rh50 alleles. This splicing mutation caused not only a total skipping of exon 7 but also a frameshift and premature chain termination. Thus, the deduced translation product contained 351 instead of 409 amino acids, with an entirely different C-terminal sequence following Thr315. These results identify the donor splicing defect, for the first time, as a loss-of-function mutation at the RH50 locus and pinpoint the importance of the C-terminal region of Rh50 in Rh complex formation via protein-protein interactions.


The treatment of trigeminal neuralgia in patients with multiple sclerosis using percutaneous radiofrequency rhizotomy.
Trigeminal neuralgia (TN) has a higher incidence among patients with multiple sclerosis (MS) than in the general population. This cohort of MS patients with TN presents a series of management challenges including poor tolerance of antineuralgic medications and occasional bilateral presentation. We analyzed our surgical series of MS patients presenting with TN who were treated with percutaneous radiofrequency rhizotomy to estimate the success, failure and recurrence rate of this procedure for those patients. Surgical reports were retrospectively reviewed between the years 1996-2000. Patients with MS and TN who received a percutaneous rhizotomy during that time were included in the study and followed until the end of 2002. Data regarding age, sex, duration of MS and pain, response to medical treatment, pain distribution and surgical outcome were evaluated. There were thirteen patients with MS and medically refractory TN treated with percutaneous radiofrequency rhizotomy. The average age at diagnosis for MS was 41 with TN beginning an average of eight years later. Following rhizotomy, complete pain relief without the need for any medication was achieved in 81% of the patients. The addition of medications resulted in pain control in the remaining patients. During a mean follow-up period of 52 months, there was a 50% recurrence rate. There were no complications related to the procedure and the associated facial numbness was well-tolerated. Percutaneous radiofrequency rhizotomy is a safe and effective method for the treatment of TN in patients with MS. The unique susceptibility of this cohort to the side effects of antineuralgic medications may require early consideration of rhizotomy.


Spike chimera states and firing regularities in neuronal hypernetworks.
A complex spatiotemporal pattern with coexisting coherent and incoherent domains in a network of identically coupled oscillators is known as a chimera state. Here, we report the emergence and existence of a novel type of nonstationary chimera pattern in a network of identically coupled Hindmarsh-Rose neuronal oscillators in the presence of synaptic couplings. The development of brain function is mainly dependent on the interneuronal communications via bidirectional electrical gap junctions and unidirectional chemical synapses. In our study, we first consider a network of nonlocally coupled neurons where the interactions occur through chemical synapses. We uncover a new type of spatiotemporal pattern, which we call "spike chimera" induced by the desynchronized spikes of the coupled neurons with the coherent quiescent state. Thereafter, imperfect traveling chimera states emerge in a neuronal hypernetwork (which is characterized by the simultaneous presence of electrical and chemical synapses). Using suitable characterizations, such as local order parameter, strength of incoherence, and velocity profile, the existence of several dynamical states together with chimera states is identified in a wide range of parameter space. We also investigate the robustness of these nonstationary chimera states together with incoherent, coherent, and resting states with respect to initial conditions by using the basin stability measurement. Finally, we extend our study for the effect of firing regularity in the observed states. Interestingly, we find that the coherent motion of the neuronal network promotes the entire system to regular firing.


Bacillaemia in lepra reaction: its correlation with liver pathology.
Thirty patients of lepromatous leprosy in a state of reaction (ENL) were studied for bacillaemia and a correlation was made with liver pathology. Buffy coat smears were examined in all, eliminating as far as possible the chances of contamination of blood by the skin bacilli. Thirteen out of thirty patients showed presence of acid fast bacilli (AFB) in the buffy coat smear. Hepatic involvement in the form of leprous granuloma was observed in all, of which 93.3% showed presence of AFB. It is concluded that during reaction the possibility of heightened bacillaemia is less likely and that the systemic spread of infection is not accentuated during the bouts of reaction.


An Unusual Case of Tertiary Syphilis Behaving Like Tongue Squamous Cell Carcinoma.
Syphilis may present with a myriad of oral manifestations in the primary, secondary, and tertiary stages, and may be confused with malignancy. Despite a rise in the incidence of syphilis, tertiary syphilis is exceedingly rare. Tertiary syphilis gummas usually affect the hard palate, while tongue involvement is very rare. A 55-year-old male with extensive smoking and alcohol use was referred for malignancy evaluation with an ulcerative mass creating a tongue cleft, and a positron emission tomography scan suggestive for malignancy. Biopsy results demonstrated no carcinoma but histology demonstrated granulomatous inflammation. Further laboratory results demonstrated elevated rapid plasma reagin titers with Treponema pallidum immunoglobulin G antibodies present. The patient was diagnosed with tertiary syphilis, received appropriate antibiotic therapy, and had healing of the tongue with a persistent cleft. Syphilis may mimic many disease processes. As such, it is important to include this disease in the differential of an unusual tongue lesion. An oral lesion may be the first sign of infection.


Roles for RNA export factor, Nxt1, in ensuring muscle integrity and normal RNA expression in Drosophila.
The mRNA export pathway is responsible for the transport of mRNAs from the nucleus to the cytoplasm, and thus is essential for protein production and normal cellular functions. A partial loss of function allele of the mRNA export factor Nxt1 in Drosophila shows reduced viability and sterility. A previous study has shown that the male fertility defect is due to a defect in transcription and RNA stability, indicating the potential for this pathway to be implicated in processes beyond the known mRNA transport function. Here we investigate the reduced viability of Nxt1 partial loss of function mutants, and describe a defect in growth and maintenance of the larval muscles, leading to muscle degeneration. RNA-seq revealed reduced expression of a set of mRNAs, particularly from genes with long introns in Nxt1 mutant carcass. We detected differential expression of circRNA, and significantly fewer distinct circRNAs expressed in the mutants. Despite the widespread defects in gene expression, muscle degeneration was rescued by increased expression of the costamere component tn (abba) in muscles. This is the first report of a role for the RNA export pathway gene Nxt1 in the maintenance of muscle integrity. Our data also links the mRNA export pathway to a specific role in the expression of mRNA and circRNA from common precursor genes, in vivo.


Exploring the complex relationships between coping strategies, locus of control and self-esteem with psychopathology: structural equation modeling with a special focus on clinical high-risk of psychosis.
Coping strategies, competence, and locus of control (LOC) beliefs are important predictors of mental health (MH). However, research into their complex interactions has produced mixed results. Our study investigated them further in the previously unexplored context of clinical high-risk (CHR) of psychosis. We tested six alternative structural equation models in a community sample (N = 523), hypothesizing a mediating role of coping and treating CHR symptoms as (i) an additional mediator or (ii) a specific outcome. Our measurement model included two latent factors of MH: (1) psychopathology (PP), consisting of presence of mental disorders, global and psychosocial functioning, and (2) self-rated health (SRH) status. In the model with the best Akaike Information Criterion and the latent factors as outcome variables, maladaptive coping completely mediated the impact of maladaptive LOC on PP and SRH. Additionally, CHR symptoms partially mediated the effect of maladaptive coping on PP and SRH in the community sample, as long as sex was not entered into the model. In the clinical sample (N = 371), the model did not support a mediation by CHR symptoms, despite significant pathways with both coping and MH outcomes; further, competence beliefs directly impacted SRH. Coping strategies are an important intervention target for MH promotion, especially in the community. In clinical populations, interventions focusing on coping strategies may improve CHR symptoms, thus potentially supporting better MH, especially SRH. Additionally, due to their mostly cascading effects on MH, improving competence and LOC beliefs may also promote psychological well-being.


Rapid transcriptional down-regulation of c-myc expression during cyclic adenosine monophosphate-promoted differentiation of leukemic cells.
Pharmacologic elevation of cyclic AMP (cAMP) promotes growth arrest and differentiation in a variety of transformed mammalian cells, including the HL-60 human promyelocytic leukemia cell line. However, mechanisms underlying this phenomenon are poorly understood. Because cellular oncogenes play a pivotal role in regulating proliferation and differentiation, we examined whether cAMP-promoted differentiation of HL-60 was preceded by a decrease in the expression of c-myc, a cellular oncogene both amplified and constitutively expressed in HL-60. We find that cyclic AMP elevation in HL-60 caused by three different pharmacologic regimens is followed by an abrupt, greater than 90% decrease in steady state c-myc mRNA levels within 3 h, well before detectable changes in proliferation and differentiation. This decrease, which occurs despite protein synthetic blockade, is attributable to transcriptional down-regulation of c-myc and is accompanied by changes in chromatin structure near c-myc promoter sites. Our findings establish that cAMP, a ubiquitous intracellular regulatory messenger previously known only to enhance gene transcriptional activity in higher eukaryotic cells, can also suppress transcription of a cellular oncogene, thereby suggesting a potential mechanism for cAMP-promoted differentiation.


The Pedicled Sensate Osteocutaneous Groin Flap for Reconstruction of the Forearm and Hand.
The groin flap is axial pedicled and versatile. Until now, this flap has been used with many modifications for the reconstruction of forearm and hand defects. However, this flap has not been used in forearm, hand, and thumb reconstruction as a pedicled sensate osteocutaneous flap. In this study, a pedicled sensate osteocutaneous groin flap was used for the reconstruction of composite tissue defects on the forearm, hand, and thumb. A pedicled sensate osteocutaneous groin flap was used to reconstruction composite tissue defects on the forearm, hand, and thumb in 7 patients. The mean age of the patients was 42 years. The defects were located on the dorsal surface of the forearm and hand in 2 patients, the dorsal surface of the hand and finger in 2 patients, and the thumb in 3 patients. The dimensions of the flap skin paddle ranged from 7 × 11 cm to 8 × 23 cm, and the dimensions of the bone component ranged from 1 × 1.5 × 3.5 cm to 1 × 1.5 × 5 cm. The mean follow-up duration was 26 months. All the flaps survived. Flap debulking was performed using 3 flaps. Sensory recovery in the flaps was completed approximately 18 months after the first operation. When the results of static 2-point discrimination test and Semmes-Weinstein monofilament test were evaluated at 18 months postoperatively, it showed that protective sensation was obtained. Except for 1 patient, motion restriction did not develop in the wrist, elbow, or shoulder joints. An acceptable aesthetic result, minimal donor site deformity, and protective sensation were obtained in all patients. The pedicled sensate osteocutaneous groin flap can be safely used in the reconstruction of forearm and hand composite tissue defects that do not have available vascular structures for free flaps in the recipient area and in thumb reconstruction where toe transfer and pollicization cannot be performed.


Electroclinical characterization and classification of symptomatic epilepsies with very early onset by multiple correspondence analysis.
To characterize very early onset symptomatic epilepsies electroclinically, to identify meaningful factors to delineate epilepsy syndromes under this category, and to test validity of the current syndromic organization of these epilepsies by the International League Against Epilepsy (ILAE). Subjects were 38 epileptic patients with seizure onset before 3 months of age. We investigated the age of seizure onset, seizure types, myoclonus, EEG features, neuroimaging studies, outcome of seizures and development. We performed multiple correspondence analysis (MCA) to reveal associations among these characteristics and to find important characteristics to divide this epilepsy group into subgroups. Very early onset epilepsies were characterized by partial seizures and epileptic spasms, especially occurring in combination as a single ictal event (combined seizures), and poor outcome. Suppression-burst (SB), hypsarrhythmia, EEG asymmetry and structural brain abnormalities were common. MCA demonstrated that presence or absence of SB was the most meaningful factor to separate this group into subgroups. MCA on the subgroup with SB revealed a subset with early-onset spasms, partial seizures with single focus, no myoclonus, SB during wakefulness and sleep, no EEG asynchrony, hypsarrhythmia and combined seizures, and another subset with late-onset spasms, partial seizures with multiple foci, myoclonus, EEG asynchrony, no combined seizures, SB during sleep only, no EEG asymmetry, no structural abnormality and no hypsarrhythmia. These corresponded to Ohtahara syndrome and early myoclonic encephalopathy, respectively. Very early onset symptomatic epilepsies have the electroclinical characteristics distinct from those with later onset. This study supported the current ILAE organization on this epilepsy group from statistic approach.


Molecular profiles and clinical outcome of stage UICC II colon cancer patients.
Published multigene classifiers suggesting outcome prediction for patients with stage UICC II colon cancer have not been translated into a clinical application so far. Therefore, we aimed at validating own and published gene expression signatures employing methods which enable their reconstruction in routine diagnostic specimens. Immunohistochemistry was applied to 68 stage UICC II colon cancers to determine the protein expression of previously published prognostic classifier genes (CDH17, LAT, CA2, EMR3, and TNFRSF11A). RNA from macrodissected tumor samples from 53 of these 68 patients was profiled on Affymetrix GeneChips (HG-U133 Plus 2.0). Prognostic signatures were generated by "nearest shrunken centroids" with cross-validation. Previously published gene signatures were applied to our data set using "global tests" and leave-one-out cross-validation Correlation of protein expression with clinical outcome failed to separate patients with disease-free follow-up (group DF) and relapse (group R). Although gene expression profiling allowed the identification of differentially expressed genes ("DF" vs. "R"), a stable classification/prognosis signature was not discernable. Furthermore, the application of previously published gene signatures to our data was unable to predict clinical outcome (prediction rate 75.5% and 64.2%; n.s.). T-stage was the only independent prognostic factor for relapse with established clinical and pathological parameters including microsatellite status (multivariate analysis). Our protein and gene expression analyses do not support application of molecular classifiers for prediction of clinical outcome in current routine diagnostic as a basis for patient-orientated therapy in stage UICC II colon cancer. Further studies are needed to develop prognosis signatures applicable in patient care.


Heterogeneity in dendritic morphology of moth antennal lobe projection neurons.
A population of projection neurons (PNs) in the antennal lobe (AL) integrates sensory information from the antenna and is essential for processing odor information in the insect brain. We examined the anatomy of this neuronal population in the brain of the silkmoth Bombyx mori. Using intracellular dye injection, we labeled a total of 246 PNs and systematically analyzed their morphological features, including the soma position, antennocerebral tract, and number of innervating glomeruli. For example, we analyzed PNs that had somata in the different cell clusters, innervated overlapping but different groups of glomeruli, and ran through different pathways. We also identified glomeruli innervated by PNs using a previously established procedure that first classifies glomeruli into regional groups and then identifies individual glomeruli. We analyzed uniglomerular PNs (75.6% of the total) and found heterogeneity in the dendritic morphology of the PNs that was dependent on the regions and/or the innervating glomeruli. For example, most PNs innervating the macroglomerular complex did not have extraglomerular processes, whereas most PNs innervating ordinary glomeruli did. Moreover, PNs innervating the toroid glomerulus showed heterogeneity in their dendritic morphology. These PNs had dendritic arborization in different areas within the glomerulus. We found that, in some cases, the innervation pattern of the PN dendrite correlated with individual variation in the glomerular organization. These results indicate that PNs are not homogeneous populations, and in some cases morphological heterogeneity in PNs correlated with change in glomerular organization in the silkmoth AL.


Assessing bias in osteoarthritis trials included in Cochrane reviews: protocol for a meta-epidemiological study.
The validity of systematic reviews and meta-analysis depends on methodological quality and unbiased dissemination of trials. Our objective is to evaluate the association of estimates of treatment effects with different bias-related study characteristics in meta-analyses of interventions used for treating pain in osteoarthritis (OA). From the findings, we hope to consolidate guidance on interpreting OA trials in systematic reviews based on empirical evidence from Cochrane reviews. Only systematic reviews that compare experimental interventions with sham, placebo or no intervention control will be considered eligible. Bias will be assessed with the risk of bias tool, used according to the Cochrane Collaboration's recommendations. Furthermore, center status, trial size and funding will be assessed. The primary outcome (pain) will be abstracted from the first appearing forest plot for overall pain in the Cochrane review. Treatment effect sizes will be expressed as standardised mean differences (SMDs), where the difference in mean values available from the forest plots is divided by the pooled SD. To empirically assess the risk of bias in treatment benefits, we will perform stratified analyses of the trials from the included meta-analyses and assess the interaction between trial characteristics and treatment effect. A relevant study-level covariate is defined as one that decreases the between-study variance (τ(2), estimated as Tau-squared) as a consequence of inclusion in the mixed effects statistical model. Meta-analyses and randomised controlled trials provide the most reliable basis for treatment of patients with OA, but the actual impact of bias is unclear. This study will systematically examine the methodological quality in OA Cochrane reviews and explore the effect estimates behind possible bias. Since our study does not collect primary data, no formal ethical assessment and informed consent are required. PROSPERO (CRD42013006924).


How brain 'cleaners' fail: Mechanisms and therapeutic value of microglial phagocytosis in Alzheimer's disease.
Microglia are the resident phagocytes of the brain, where they primarily function in the clearance of dead cells and the removal of un- or misfolded proteins. The impaired activity of receptors or proteins involved in phagocytosis can result in enhanced inflammation and neurodegeneration. RNA-seq and genome-wide association studies have linked multiple phagocytosis-related genes to neurodegenerative diseases, while the knockout of such genes has been demonstrated to exert protective effects against neurodegeneration in animal models. The failure of microglial phagocytosis influences AD-linked pathologies, including amyloid β accumulation, tau propagation, neuroinflammation, and infection. However, a precise understanding of microglia-mediated phagocytosis in Alzheimer's disease (AD) is still lacking. In this review, we summarize current knowledge of the molecular mechanisms involved in microglial phagocytosis in AD across a wide range of pre-clinical, post-mortem, ex vivo, and clinical studies and review the current limitations regarding the detection of microglia phagocytosis in AD. Finally, we discuss the rationale of targeting microglial phagocytosis as a therapeutic strategy for preventing AD or slowing its progression.


Managing between the agendas: implementing health care reform policy in an acute care hospital.
This paper aims to assess administrative and clinical manager stances on health system reform. Understanding these stances will help to identify cultural differences and competing agendas between these two key health service stakeholders and contribute to developing strategies to improve organisational performance. A qualitative methodology was used comprising in-depth open-ended interviews conducted in 2007 with 26 administrative and clinical managers who managed clinical units. This paper provides empirical insights into the ways that administrative and clinical mangers conceive of their managerial roles in relation to health care reform and performance improvement in health services. The findings suggest that developing a hybrid clinical manager culture as a means to bridge the gap between administrative and clinical manager stances on reform objectives, while possible, is not yet being realised. The research has relevance for health services that are experiencing organisational transformation. However, its location in one health service limits the generalisability of findings to other sites. Further research is needed to assess the opportunities for a hybrid culture to emerge as well as its effect. While attention is predominantly directed to clinician groups as a key stakeholder in implementing health reform policies, this paper has implications for how administrative managers also structure their roles and responsibilities to create an organisational climate conducive to change. This will include strategies to support clinical managers to make the transition from a predominantly clinical, to a clinical managerial, orientation. This paper addresses a significant problem in health service governance, namely the divide between the value stances of dual hierarchies. This problem is only now gaining prominence as a significant barrier to health reform.


Non-uniform refinement: adaptive regularization improves single-particle cryo-EM reconstruction.
Cryogenic electron microscopy (cryo-EM) is widely used to study biological macromolecules that comprise regions with disorder, flexibility or partial occupancy. For example, membrane proteins are often kept in solution with detergent micelles and lipid nanodiscs that are locally disordered. Such spatial variability negatively impacts computational three-dimensional (3D) reconstruction with existing iterative refinement algorithms that assume rigidity. We introduce non-uniform refinement, an algorithm based on cross-validation optimization, which automatically regularizes 3D density maps during refinement to account for spatial variability. Unlike common shift-invariant regularizers, non-uniform refinement systematically removes noise from disordered regions, while retaining signal useful for aligning particle images, yielding dramatically improved resolution and 3D map quality in many cases. We obtain high-resolution reconstructions for multiple membrane proteins as small as 100 kDa, demonstrating increased effectiveness of cryo-EM for this class of targets critical in structural biology and drug discovery. Non-uniform refinement is implemented in the cryoSPARC software package.


[Effect of slow rates of freezing on the functional state of mitochondria from various types of tissue preparations of kidney cortex].
Changes in functional activity of mitochondria, in concentration of respiration enzymes and the state of internal membrane after slow freezing down to -5 degrees, -12 degrees, -21 degrees and -30 degrees of various tissue preparations of renal cortex (homogenates, slices, etc) were studied. The following phenomena were found: an increase in the rate of mitochondrial respiration at the second metabolic state, inhibition of electron transport most distinct during mitochondrial oxidation of alpha-ketoglutarate, a decrease in concentration of cytochromes, an increase in the internal membrane permeability. Considerable damage of mitochondria was observed at temperatures between -5 degrees and -12 degrees but the most pronounced impairment of the organelles occurred at -10 degrees -20 degrees, i. e. at the freezing point of free water in the suspension and at eutectic temperatures.


Obesity and thermogenesis related to the consumption of caffeine, ephedrine, capsaicin, and green tea.
The global prevalence of obesity has increased considerably in the last decade. Tools for obesity management, including caffeine, ephedrine, capsaicin, and green tea have been proposed as strategies for weight loss and weight maintenance, since they may increase energy expenditure and have been proposed to counteract the decrease in metabolic rate that is present during weight loss. A combination of caffeine and ephedrine has shown to be effective in long-term weight management, likely due to different mechanisms that may operate synergistically, e.g., respectively inhibiting the phosphodiesterase-induced degradation of cAMP and enhancing the sympathetic release of catecholamines. However, adverse effects of ephedrine prevent the feasibility of this approach. Capsaicin has been shown to be effective, yet when it is used clinically it requires a strong compliance to a certain dosage, that has not been shown to be feasible yet. Also positive effects on body-weight management have been shown using green tea mixtures. Green tea, by containing both tea catechins and caffeine, may act through inhibition of catechol O-methyl-transferase, and inhibition of phosphodiesterase. Here, the mechanisms may also operate synergistically. In addition, tea catechins have antiangiogenic properties that may prevent development of overweight and obesity. Furthermore, the sympathetic nervous system is involved in the regulation of lipolysis, and the sympathetic innervation of white adipose tissue may play an important role in the regulation of total body fat in general.


[Tics in children and adolescents: a retrospective analysis of 78 cases].
Tics are the most frequent abnormal movement in children. A familial history of tics and a personal and familial history of neurobehavioral disturbances are common in children with this abnormality. Tics may seriously compromise daily activities in affected individuals. To identify the characteristics of tics in children and adolescents followed-up in the Neuropediatric Unit of the Hospital Geral de Santo António. We performed a retrospective analysis of patients with tics based on information collected from medical records. The diagnostic criteria of the DSM IV-TR 2000 of the American Psychiatric Association were used. The medical records of 78 children were analyzed, 84.6 % of whom were boys. More than one third of the patients were aged 4 to 8 years old. In 5.1 % of the patients tics developed before the age of 2 years. A familial history of tics, depression and obsessive disorder traits was found in approximately 30 % of patients. The most frequent comorbidity was attention deficit hyperactivity disorder (67.9 %). The occurrence of pediatric autoimmune neuropsychiatric disorders associated with streptococcus infection (PANDAS) was suggested in five patients. In all patients, motor tics occurred before vocal tics. In more than two thirds of the patients, tics were simple. In 59.0 % of the patients, tics were chronic, and in 45.7 % of these met the criteria for Tourette's syndrome. A total of 43.1 % of the patients with chronic tics received pharmacotherapy, risperidone being the most frequently used drug. In general the results of the present study are in agreement with those of previous studies, underlining the need to consider a diagnosis of tics in young children and highlighting the importance of identification and appropriate treatment of comorbidities.


Apigenin Ameliorates the Obesity-Induced Skeletal Muscle Atrophy by Attenuating Mitochondrial Dysfunction in the Muscle of Obese Mice.
It was investigated whether apigenin (AP) protected against skeletal muscle atrophy induced by obesity. Mice were fed a high-fat diet (HFD) for 9 weeks to induce obesity, and then were assigned to two groups; the HFD group received a high-fat diet, and the HFD+AP group received a 0.1% AP-containing HFD. After additional feeding of the experimental diet for 8 weeks, mice in the HFD group were highly obese compared with the mice in the standard diet fed mice group. The mice in the AP-treated group showed less fat pad accumulation and less inflammatory cytokines without body weight reduction. The weight of skeletal muscle in the AP group tended to increase as compared with that of the HFD group. Furthermore, AP reduced the expression of atrophic genes, including MuRF1 and Atrogin-1, but increased the exercise capacity. The mitochondrial function and mitochondrial biogenesis were enhanced by AP. In cultured C2C12 cells, AP also suppressed palmitic acid-induced muscle atrophy and mitochondrial dysfunction. In addition, AP activated AMP-activated protein kinase (AMPK) in the C2C12 and the muscle of HFD-induced obese mice. The results suggested that AP ameliorated the obesity-induced skeletal muscle atrophy by attenuating mitochondrial dysfunction.


On physiological edema in man's lower extremity.
To examine whether the so-called musculovenous pump counteracts the development of interstitial edema in the lower extremities of man in the upright position, the volume changes in the calf which occurred during twenty minutes of rhythmic muscular exercise were measured in twenty-three subjects by impedance-plethysmography. The results were compared with the volume increase found during quiet relaxed standing for the same length of time. Contrary to the hypothesis, and edema-protective effect of the musculovenous pump could only be shown in about half the number of the subjects. In the others, muscular exercise led to increases in calf volume which were higher than those measured in the normal upright position. These results show that the calf muscle pump does not generally have a edema-protective effect but rather that muscle contractions also activate mechanisms which stimulate the extravasation of fluid. In a second test-series with twenty subjects, changes in calf volume were measured during the course of the day. In nearly all cases, the calf volume was greater in the evening than in the morning. It could be shown that the volume increases in the evening are caused by an increase in extravascular fluid. Compared to the increase in extravascular volume occurring during twenty minutes, in a normal upright position, the accumulation of extravascular fluid during the day is, however, remarkably low. Although it is still unknown how interstitial edema in man's lower extremities is prevented during the day, these findings lead to the hypothesis that the edema-preventing mechanisms, for instance the muscle-lymphpump, do not become maximally effective until a certain volume has accumulated in the interstitial space.


A major outer-membrane protein functions as a porin in Haemophilus influenzae.
Porins are pore-forming outer-membrane proteins which serve as a non-specific pathway for the entry of hydrophilic molecules into Gram-negative bacteria. We studied four strains of Haemophilus influenzae that had decreased permeability to chloramphenicol associated with diminished quantities of a 40 kDa major outer-membrane protein. Isogenic pairs of organisms containing and lacking this protein were compared. The latter strains grew more slowly and were less permeable to sucrose and raffinose. They were also more resistant to multiple hydrophilic antibiotics than an isogenic strain containing the 40 kDa protein and were less permeable to penicillin G and chloramphenicol. We conclude that the 40 kDa outer-membrane protein functions as a porin in H. influenzae.


Altered Expression of Hippo Signaling Pathway Molecules in Intrahepatic Cholangiocarcinoma.
MOB1, a core component of the Hippo signaling pathway, suppresses cell proliferation, and MOB1 liver conditional knockout mice develop intrahepatic cholangiocarcinoma (ICC). However, its clinical significance in human ICC has not been established. The aim of this study was to characterize protein levels and the role of Hippo and TGF pathways in ICCs. The protein levels of yes-associated protein 1 (YAP1), MOB1, Smad2, and TGFβ2 in 88 ICC cases were analyzed. Protein level was graded by a scoring system; then, the clinicopathological factors, including prognosis, were analyzed based on protein level. Nuclear overexpression of YAP1 was seen in 28 cases (31.8%), and it was significantly associated with a poor overall survival rate (p = 0.01). MOB1 expression decreased in 42 cases (47.7%) and was associated with a poor overall survival rate (p = 0.02). SMAD2 nuclear localization was significantly correlated with a high YAP1 level independent of TGFβ2. Multivariate analysis revealed that a high YAP1 level, a low MOB1 level, and lymphatic permeation were independent risk factors for overall survival. These results showed that key components of the Hippo signaling pathway are aberrantly expressed and associated with the malignant potential of human ICC.


A Paracoccidioides brasiliensis polysaccharide having granuloma-inducing, toxic and macrophage-stimulating activity.
The occurrence of a polysaccharide fraction of Paracoccidioides brasiliensis cell wall with toxic, granuloma-inducing and macrophage-stimulating activities was demonstrated. After fractionation of the lipid-extracted wall with 1 M-NaOH, three fractions were obtained: (1) an alkali-insoluble fraction; (2) an alkali-soluble, acid-insoluble fraction and (3) an alkali-soluble, acid-soluble fraction. When the three fractions were injected into mice only fraction (1) was able to induce chronic lung inflammation, causing a marked loss in body weight and death at a dose of 6 mg per animal. Analysis of the stimulation of peritoneal macrophages of mice (measured by cell spreading on glass) after intraperitoneal injection of fraction 1 showed that 75% of the cells were able to spread even 20 d after inoculation.


A periplasmic cupredoxin with a green CuT1.5 center is involved in bacterial copper tolerance.
The importance of copper resistance pathways in pathogenic bacteria is now well recognized, since macrophages use copper to fight bacterial infections. Additionally, considering the increase of antibiotic resistance, growing attention is given to the antimicrobial properties of copper. It is of primary importance to understand how bacteria deal with copper. The Cu-resistant cuproprotein CopI is present in many human bacterial pathogens and environmental bacteria and crucial under microaerobiosis (conditions for most pathogens to thrive within their host). Hence, understanding its mechanism of function is essential. CopI proteins share conserved histidine, cysteine, and methionine residues that could be ligands for different copper binding sites, among which the cupredoxin center could be involved in the protein function. Here, we demonstrated that Vibrio cholerae and Pseudomonas aeruginosa CopI restore the Cu-resistant phenotype in the Rubrivivax gelatinosus ΔcopI mutant. We identified that Cys125 (ligand in the cupredoxin center) and conserved histidines and methionines are essential for R. gelatinosus CopI (RgCopI) function. We also performed spectroscopic analyses of the purified RgCopI protein and showed that it is a green cupredoxin able to bind a maximum of three Cu(II) ions: (i) a green Cu site (CuT1.5), (ii) a type 2 Cu binding site (T2) located in the N-terminal region, and (iii) a third site with a yet unidentified location. CopI is therefore one member of the poorly described CuT1.5 center cupredoxin family. It is unique, since it is a single-domain cupredoxin with more than one Cu site involved in Cu resistance.


The Fibromyalgia Impact Questionnaire: a validated Spanish version to assess the health status in women with fibromyalgia.
To translate, adapt, validate and assess the sensitivity to change of a Spanish version of the Fibromyalgia Impact Questionnaire (FIQ-S). The FIQ-S was adapted following the translation and back-translation methodology. Female patients with fibromyalgia (FM) were invited to participate. Reliability was analyzed by the Spearman correlation coefficient between test and retest. Internal consistency was checked by the Cronbach's alpha coefficient. Construct validity was analyzed comparing FIQ-S with: HAQ, FHAQ, SF-36, SCL90-R, and the visual analogue scale for pain. Sensitivity to change was assessed in an 8-week randomized trial of exercise therapy. Feasibility was analyzed by the time taken in completing the FIQ-S and the proportion of patients able to complete the questionnaire. Translation was concordant. Adaptation affected at 4 sub-items of physical function. One-hundred and two FM patients completed the protocol. Mean age was 48.7 years with a mean of 9.2 years of evolution. Test-retest correlations were between 0.61-0.85 (p < 0.0001). Internal consistency showed alpha = 0.82 for all items and alpha = 0.86 for the sub-items of physical function. Significant correlations (p < 0.0001) were found between the FIQ-S items and HAQ, FHAQ, SF-36 and SCL90-R. For patients treated with the exercise program, the pre-treatment FIQ-S score was 52.0 +/- 11.5 and the post-treatment score was 40.8 +/- 13.7 (p < 0.003). Mean time for completing FIQ-S was 3.3 minutes. In 4% of the patients external help was needed. The FIQ-S is a reliable, valid and responsive to changes questionnaire for measuring health status and physical function in Spanish speaking FM patients.


The influence of subject and test design on dynamometric measurements of extremity muscles.
In the context of broader discussions of clinical dynamometry, earlier reviews have raised concerns about the potential effects of variations in subject factors and test procedures on measurements. None, however, have dealt exclusively with these effects. We therefore reviewed more than 200 articles to evaluate in detail the effects of variations in subject factors and test procedures on measurements. Factors relating to subjects that affected measurements were age, gender, weight, athletic background, disability, and limb dominance. Test conditions that led to variations in measurements were range of movement in which values were obtained, type of contraction or movement (concentric, eccentric, isokinetic, isometric, isotonic), pretest procedures (warm-up and gravity-correction procedures, starting position, stabilization, axes alignment, lever arm length, preload, damp/ramp settings), test conditions (speed, test sequence, rest intervals, feedback), and type of data analysis (the data selected and how they are manipulated). In the majority of the publications, the authors failed to provide sufficient detail for accurate replication of test procedures or for comparison with other studies. We advocate that the factors identified in this review be included whenever measurements obtained with a dynamometer are reported. Effective development of normative data, formation of ratios, comparison of measurements across studies, and relating measurements with other performance criteria (eg, measurements of functional performance) all require descriptions of variables relating to subjects and testing. Similarly, meaningful use of these measurements in clinical practice requires consideration and documentation of these variables.


Impact of heat on metabolic and hemodynamic changes in transcranial infrared laser stimulation measured by broadband near-infrared spectroscopy.
Transcranial infrared laser stimulation (TILS) has shown effectiveness in improving human cognition and was investigated using broadband near-infrared spectroscopy (bb-NIRS) in our previous study, but the effect of laser heating on the actual bb-NIRS measurements was not investigated. To address this potential confounding factor, 11 human participants were studied. First, we measured time-dependent temperature increases on forehead skin using clinical-grade thermometers following the TILS experimental protocol used in our previous study. Second, a subject-averaged, time-dependent temperature alteration curve was obtained, based on which a heat generator was controlled to induce the same temperature increase at the same forehead location that TILS was delivered on each participant. Third, the same bb-NIRS system was employed to monitor hemodynamic and metabolic changes of forehead tissue near the thermal stimulation site before, during, and after the heat stimulation. The results showed that cytochrome-c-oxidase of forehead tissue was not significantly modified by this heat stimulation. Significant differences in oxyhemoglobin, total hemoglobin, and differential hemoglobin concentrations were observed during the heat stimulation period versus the laser stimulation. The study demonstrated a transient hemodynamic effect of heat-based stimulation distinct to that of TILS. We concluded that the observed effects of TILS on cerebral hemodynamics and metabolism are not induced by heating the skin.


[Histological, histochemical, and ultrastructural findings in malignant hyperthermia (author's transl)].
Malignant hyperthermia is a rare but severe complication of modern anesthesia, induced by halothane and succinylcholine. The syndrome is characterized by a rapid sustained and extreme rise in body temperature associated with muscular rigidity, tachycardia, tachypnoea and cyanosis. The lethality is about 60%. The present paper describes the histological, histochemical and electron microscopical findings performed on muscle biopsies of 3 patients with malignant hyperthermia (1 patient died) and a so called risk patient. In all patients morphological findings consistent with a pre-existent myopathy were found. Histologically there were acute necrotic muscular fibers as well as in types I and II, variations in the fiber diameter and centralization of the nuclei. In two cases even fibers that had a normal aspect in HE slides, showed a pathologic pattern after phosphorylase reaction. In addition to acute rhabdomyolysis, electron-microscopic investigations revealed cystic expansion of the cisterns of the sarcoplasmic reticulum with a peculiar proliferation of the sarcolemma. In a degenerating mitochondrium, a crystalline inclusion was identified. These findings support the pathogenetic concept of Britt and coworkers of a functional defect in the calcium release or binding mechanism of sarcoplasmic reticulum. Since it is known that malignant hyperthermia has a familial predilection, it seems very important that clinical, biochemical, and morphological investigations be performed such as CPK estimations and muscular biopsies not only of the patients but also of the relatives in order to rule out this type of latent myopathy.


Accuracy of ECG-based screening for sleep-disordered breathing: a survey of all male workers in a transport company.
Sleep-disordered breathing (SDB) is associated with increased risk for cardiovascular morbidity and mortality and for sleepiness-related accidents, but >75 % of the patients remain undiagnosed. We sought to determine the diagnostic accuracy of ECG-based detection of SDB when used for population-based screening. All male workers, mostly truck drivers, of a transport company (n = 165; age, 43 ± 12 years) underwent standard attended overnight polysomnography. Cyclic variation of heart rate (CVHR), a characteristic pattern of heart rate associated with SDB, was detected from single-lead ECG signals during the polysomnography by a newly developed automated algorithm of autocorrelated wave detection with adaptive threshold (ACAT). Among 165 subjects, the apnea-hypopnea index (AHI) was ≥5 in 62 (38 %), ≥15 in 26 (16 %), and ≥30 in 16 (10 %). The number of CVHR per hour (CVHR index) closely correlated with AHI [r = 0.868 (95 % CI, 0.825-0.901)]. The areas under the receiver operating characteristic curves for detecting subjects with AHI ≥5, ≥15, and ≥30 were 0.796 (95 % CI, 0.727-0.855), 0.974 (0.937-0.993), and 0.997 (0.971-0.999), respectively. With a predetermined criterion of CVHR index ≥15, subjects with AHI ≥15 were identified with 88 % sensitivity and 97 % specificity (likelihood ratios for positive and negative test, 30.7 and 0.12). The classification performance was retained in subgroups of subjects with obesity, hypertension, diabetes mellitus, dyslipidemia, and decreased autonomic function. The CVHR obtained by the ACAT algorithm may provide a useful marker for screening for moderate-to-severe SDB among apparently healthy male workers.


Monoaminergic involvement in the behavioral effects of ergot derivatives.
Monoaminergic neuronal systems have been implicated in the mechanisms of action of most ergot derivatives (including those that are clinically useful and those that are "hallucinogenic"). Among the various assays that have been useful in determining the extent of such involvement is drug discrimination. In this procedure, animals are trained to respond in one way (e.g., press the right lever) following saline and to respond differently (e.g., press the left lever) after either a particular ergot or a neurotransmitter agonist, and are subsequently tested with other ergots, transmitter agonists or antagonists (alone or in combination with the training drug). The results to date indicate that: 1) There are similarities in the discriminable effects of lergotrile, lisuride and bromocriptine, which probably involve catecholaminergic neuronal systems (DA or NE). 2) These effects are clearly separable from those of ergonovine and LSD, which may be relatively more serotonergic. More precise delineation of the mechanisms underlying these cues (e.g., D1, D2-selective agonism, antagonism, etc.) awaits further testing with highly specific pharmacological interventions.


A mechanism of alkali metal carbonates catalysing the synthesis of β-hydroxyethyl sulfide with mercaptan and ethylene carbonate.
The reaction of β-hydroxyethylation is essential to the current practice of organic chemistry. Here, we proposed a new and green route to synthesize 2-hydroxyethyl n-alkyl sulfide with n-alkyl mercaptan and ethylene carbonate (EC) in the presence of alkali carbonates as catalysts and revealed the mechanism by experiments and theoretical calculations. The reaction reported proceeds rapidly with high yields when it is performed at 120 °C and the catalytic loading is ∼1 mol%. This protocol is applicable to other mercaptans to synthesize the corresponding β-hydroxyethyl sulfide. Density functional theory-based calculations show the energy profile for the reaction pathway. The rate-determining step is the ring-opening of EC. A negatively charged O atom of alkali carbonates approaches the S atom of -SH under the influence of hydrogen bonds. An activated S atom that carries more negative charge serves as a nucleophilic reagent and assists in the ring-opening of EC by reducing the Mayer bond orders of the C1-O1 bond in EC. Alkali cations also contribute to the C1-O1 bond cleavage. The energy barrier for the ring-opening of EC decreases with the decrease of electronegativity of alkali cations. Subsequent transference of a H atom leads to the formation of β-hydroxyethyl sulfide, the dissociation of CO2 and the reduction of K2CO3.


The extracellular deposition of mast cell products is increased in hypertrophic airways smooth muscles in allergic asthma but not in nonallergic asthma.
Bronchial asthma is characterized by airways smooth muscle hypertrophy and infiltration of mast cells in the bronchial mucosa. The aim of this investigation was to study the distribution of mast cells in different compartments in the bronchial mucosa of allergic and nonallergic asthma in relation to airways remodeling. Bronchial biopsies were obtained from 29 subjects with allergic and nonallergic asthma and healthy controls. The biopsies were stained for mast cells by means of the tryptase specific antibody AA1. Extracellular deposition of mast cell products were judged on a semi-quantitative scale. Mast cells per mm(2) were counted in epithelium, lamina propria and the smooth muscle compartment. Smooth muscle was visualized by actin antibodies and the proportion of staining of the biopsy estimated. Laminin and tenascin layers were visualized by their respective antibodies. Airways smooth muscle thickness was greater in allergic vs nonallergic asthma (P < 0.001). Mast cells were increased in all three compartments in both allergic and nonallergic asthma, with significantly higher numbers in smooth muscles in allergic asthma (P < 0.03). The extracellular deposition of mast cell products was more common in allergic than nonallergic asthma in lamina propria and smooth muscles (P = 0.025; P = 0.002, respectively). In patients with allergic asthma the numbers of mast cells with extracellular deposition of mast cell products were significantly correlated to the thickness of the laminin and tenascin layers. Our results suggest that there are large differences between allergic and nonallergic asthmatics as to mast cell activation and airways smooth muscle thickness. Our data implies that mast cells are causally involved in structural alterations in allergic asthma.


Psychological flexibility may reduce insomnia in persons with chronic pain: a preliminary retrospective study.
Sleep disturbance is a common complaint in people with chronic pain, and is associated with a range of adverse outcomes including reports of greater pain and disability. Research into insomnia with chronic pain sufferers has not yet examined the role of psychological flexibility, a process from acceptance and commitment therapy. We examined this role. Participants in this study were 159 adult patients attending an assessment at a specialist pain center. They were mostly women, 63%, and the mean age was 43.8 years. In preliminary analyses 79% of participants met the criteria for significant insomnia based on their self-reported symptoms. As predicted, significant positive correlations were found between components of psychological flexibility, particularly acceptance of pain and values-based action, and all measures of sleep quality. In regression analyses, the components of psychological flexibility considered together accounted for between 11% and 19% of variance across a range of measures of sleep quality. These results suggest the need to further develop treatment services for people with chronic pain and insomnia.


Perioperative hypertension management.
Perioperative hypertension is commonly encountered in patients that undergo surgery. While attempts have been made to standardize the method to characterize the intraoperative hemodynamics, these methods still vary widely. In addition, there is a lack of consensus concerning treatment thresholds and appropriate therapeutic targets, making absolute recommendations about treatment difficult. Nevertheless, perioperative hypertension requires careful management. When treatment is necessary, therapy should be individualized for the patient. This paper reviews the pharmacologic agents and strategies commonly used in the management of perioperative hypertension.


Improved methodology for identification of protists and microalgae from plankton samples preserved in Lugol's iodine solution: combining microscopic analysis with single-cell PCR.
Here we introduce a method for quantitative analysis of planktonic protists and microalgae from preserved field samples combining morphological and small-subunit (SSU) rRNA gene sequence analysis. We linked a microscopic screening with PCR of single cells using field samples preserved with Lugol's iodine solution. Cells possessing a rigid cell wall were incubated with Viscozyme and subsequently with proteinase K for cell disruption; this was unnecessary for fragile cells. The addition of sodium thiosulfate to the PCR tube considerably decreased the inhibiting effect of the fixative (iodine) on the PCR and thus allowed for successful single-cell PCR even of long DNA fragments (up to as many as 3,000 base pairs). We further applied the protocol to investigate the dominant SSU rRNA genotypes in distinct flagellate morphospecies originating from different samples. We hypothesized that despite the morphological similarity, protist morphospecies in different habitats or sampled during different seasons are represented by different genotypes. Our results indicate species-specific differences: the two species Ochromonas sp. and Dinobryon divergens were represented by several different genotypes each, and for the latter species, the dominating genotype differed with habitat. In contrast, Dinobryon pediforme, Dinobryon bavaricum, and Synura sphagnicola were exclusively represented by a single genotype each, and the respective genotype was the same in different samples. In summary, our results highlight the significance of molecular variation within protist morphospecies.


Courtship rate signals fertility in an externally fertilizing fish.
Sperm limitation is widespread across many animal species. Several mechanisms of sperm allocation have been proposed, including optimal allocation according to clutch size and equal allocation across females. However, considerably less effort has been directed at investigating the behavioural signals associated with sperm limitation in males, which may include mating rate and the intensity of courtship. We investigated whether multiple successive spawnings affect individual male fertilization success, mating rates and courtship rates in Japanese medaka (Oryzias latipes). Across an average of 17 spawning events per male, fertilization success decreased from 83.7 per cent for the first spawning to 40 per cent for the last spawning while courtship rate decreased from 3.4 to 1.5 min⁻¹. Females appeared to respond to male sperm depletion by reducing clutch size. Our results suggest that male Japanese medaka are sperm-limited, and that courtship rate may be an honest indication of fertilization ability.


Impact of clinical registries on quality of patient care and health outcomes: protocol for a systematic review.
Many developed countries have regional and national clinical registries aimed at improving health outcomes of patients diagnosed with particular diseases or cared for in particular healthcare settings. Clinical quality registries (CQRs) are clinical registries established with the purpose of monitoring quality of care and providing feedback to improve health outcomes. The aim of this systematic review is to understand the impact of CQRs on (1) mortality/survival; (2) measures of outcome that reflect a process or outcome of healthcare; (3) healthcare utilisation and (4) costs. The PRISMA-P methodology, checklist and standard strategy using predefined inclusion and exclusion criteria and structured data abstraction tools will be followed. A search of the electronic databases MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials (CENTRAL) and CINAHL will be undertaken, in addition to Google Scholar and grey literature, to identify studies in English covering the period January 1980 to December 2014. Case-control, cohort, randomised controlled trials and controlled clinical trials which describe the registry as an intervention will be eligible for inclusion. Narrative synthesis of study findings will be conducted, guided by a conceptual framework developed to analyse the outcome measure of the registry using defined criteria. If sufficient studies are identified with a similar outcome of interest and measure using the same comparator and time of interval, results will be pooled for random-effects meta-analysis. Test for heterogeneity and sensitivity analysis will be conducted. To identify reporting bias, forest plots and funnel plots will be created and, if required, Egger's test will be conducted. Ethical approval is not required as primary data will not be collected. Review results will be published as a part of thesis, peer-reviewed journal and conferences. CRD42015017319.


Plasma catestatin level in patients with acute myocardial infarction and its correlation with ventricular remodelling.
The evaluation of ventricular remodelling and functional recovery is essential in predicting the prognosis of patients with acute myocardial infarction (AMI). To determine the plasma catestatin level in patients with AMI, and investigate the association between plasma catestatin and heart function, and with left ventricular remodelling (LVR). Fifty-eight consecutive patients who were admitted within 12 h of the onset of their ST-segment elevation myocardial infarction symptoms between 1 October 2009 and 30 June 2011 were prospectively recruited. Circulating catestatin was measured by ELISA. All patients underwent an echocardiography examination during the first week; 31 patients had a second echocardiography examination 3 months after the myocardial infarction. Plasma catestatin at the time of admission was significantly higher in patients than in normal controls. The level increased further in the first week after AMI. Three months after AMI, the plasma catestatin level of patients was comparable to that of normal controls. The plasma level of catestatin correlated with anterior AMI and left ventricular ejection fraction (LVEF) in the acute stage. Compared with patients without LVR, those with LVR had significantly higher level of plasma brain natriuretic peptide on day 7 and a significantly higher level of plasma catestatin on admission and on days 3 and 7 (p=0.033, p=0.001, p=0.006, p=0.021, respectively). Plasma catestatin levels were raised after AMI. An early increase of catestatin correlated with anterior AMI and LVEF. Plasma catestatin after the onset of AMI might be associated with the magnitude of progressive ventricular remodelling 3 months after AMI.