Psilocin - The "real deal" or an extraction byproduct.
Most illicit drug casework samples at the Israel Police National Drug Laboratory are found to be mixtures of substances. Some are a mixture of an illicit drug with fillers, and others may contain more than one illicit drug. This study was triggered by a routine gas chromatography-mass spectrometry (GC-MS) analysis of an unusual casework sample. The sample chromatogram showed a mixture of two illicit drugs, 4-acetoxy-DMT and psilocin. Considering the two substances' similar skeletal structure, the authors wondered whether the sample was indeed a mixture of the two substances, or whether perhaps 4-acetoxy-DMT was hydrolyzed to psilocin during the analysis. This study hypothesized that indeed the base used in the pre-injection sample preparation hydrolyzed the ester group on the 4-acetoxy-DMT yielding a hydroxide group. This was tested using several concentrations of ammonium hydroxide and two additional bases - pyridine (a weak base) and sodium hydroxide (a strong base). Results showed that media with a higher pH (induced by the stronger base) yielded a higher psilocin to 4-acetoxy-DMT ratio which is compatible with degradation of 4-acetoxy-DMT. This study also explored the possibility that psilocin was a byproduct of thermal decomposition of 4-acetoxy-DMT and found it thermally stable in the temperature of the GC injection port (200°C). The 4-acetoxy-DMT case demonstrates how pre-injection laboratory procedures can inadvertently modify casework samples. Caution is clearly advisable in selecting reagents and processes in general, and specifically in the case of GC-MS pre-injection procedures conducted to analyze substances like the ones in the present study.


Detecting cell-free circulating hTERT mRNA in the plasma may identify a subset of nonsmall cell lung cancer patients.
Human telomerase reverse transcriptase (hTERT), the catalytic subunity of telomerase, a marker of cell immortalization, is upregulated in most tumors, including nonsmall cell lung cancer (NSCLC). However, little is known about the role of assessing cell-free plasma circulating hTERT mRNA for tracing these tumors. We investigated by RT-polymerase chain reaction (PCR) and real-time quantitative PCR the prevalence and functional implications of hTERT mRNA in both tumor tissue and paired plasma samples in 34 (27 males and 7 females) stages I-IIIB NSCLC patients (21 adenocarcinomas and 13 squamous-cell carcinomas) by using intron- and exon-spanning primers. Plasma samples of ten healthy volunteers and normal lung tissue were used as negative controls. We detected hTERT mRNA in the plasma of 4 out of 34 (12%) tumor patients, but none was detected in the ten plasma samples of healthy volunteers. Normal lung tissue was completely devoid of hTERT mRNA. No association was found between hTERT plasma mRNA and clinicopathologic variables of the patients' population. We conclude that cell-free circulating hTERT mRNA is detectable in a subset of patients, whereas it is consistently absent in healthy volunteers. It can be added to the panel of multiple genetic tracers to detect lung cancer in the plasma of patients, although, per se, it is not specific for this tumor.


Xenogeneic islet transplantation of microencapsulated porcine islets for therapy of type I diabetes: long-term normoglycemia in STZ-diabetic rats without immunosuppression.
Rejection and possible infection with porcine pathogens are obstacles in clinical xenogeneic transplantation of porcine pancreatic islets (PPI) to treat diabetic patients. A solution to this problem could be microencapsulation of the PPI. However, isolation and microencapsulation are highly demanding tasks with considerable risks of damaging the PPI. Thus, it is not surprising that the long-term function (>200 days) of microencapsulated PPI (mPPI), transplanted to diabetic rats, has been observed only in a few cases. Diabetes was induced in Wistar rats with streptozotozin (STZ 60 mg/kg body weight). Animals with consecutive blood glucose levels >300 mg/dl for more than 2 days were considered diabetic. PPI were isolated from brain-dead hybrid pigs (age 6-7 months or 2-3 years) using the Ricordi-technique and Liberase(PI). After in vitro culture PPI were microencapsulated with highly purified barium-alginate and 1,000 mPPI of 300-500 microm Ø were transplanted under the left kidney capsule and/or into the peritoneal cavity of STZ-diabetic rats (n = 15) without immunosuppression. Daily, later weekly, blood glucose level and body-weight were measured. mPPI showed normal glucose tolerance in vitro and also in vivo. Normoglycemia occurred between day 1 and 15 after transplantation. Four mPPI grafts functioned for more than 230 days, the longest now for >550 days. Three rats are currently normoglycemic for >40 days. Six rats lost xenograft function after 12-20 days, due to inflammatory reactions at the site of the grafts. Two xenografts failed to induce normoglycemia, because the capsules did not contain enough viable PPI. Microencapsulated xenogeneic islets can induce long term normoglycemia in rats without immunosuppression. However, very often the grafts fail to control the blood glucose level adequately. The reasons for these failures are currently under investigation. Nevertheless, our results are very promising and might lead the way towards preclinical trials in non-human primates.


Post-transplant monitoring of NK cell counts as a simple approach to predict the occurrence of opportunistic infection in liver transplant recipients.
Monitoring of peripheral blood lymphocyte subpopulation (PBLS) counts might be useful for estimating the risk of infection after liver transplantation (LT). We prospectively measured total lymphocyte and PBLS counts at baseline and post-transplant months 1 and 6 in 92 LT recipients. PBLS were enumerated by single-platform 6-color flow cytometry technology. Areas under receiver operating characteristic (ROC) curves were used to evaluate the accuracy of different PBLS for predicting cytomegalovirus (CMV) disease and overall opportunistic infection (OI). Adjusted hazard ratios (aHRs) for both outcomes were estimated by Cox regression. After a median follow-up of 730.0 days, 29 patients (31.5%) developed 38 episodes of OI (including 22 episodes of CMV disease). The counts of CD3(+) , CD4(+) , and CD8(+) T cells, and CD56(+) CD16(+) natural killer (NK) cells at month 1 were significantly lower in patients subsequently developing OI. The NK cell count was the best predictive parameter (area under ROC curve for predicting CMV disease: 0.78; P-value = 0.001). Patients with an NK cell count <0.050 × 10(3) cells/μL had higher cumulative incidences of CMV disease (P-value = 0.001) and overall OI (P-value <0.001). In the multivariate models, an NK cell count <0.050 × 10(3) cells/μL at month 1 post transplantation remained as an independent risk factor for CMV disease (aHR: 5.54; P-value = 0.003) and overall OI (aHR: 7.56; P-value <0.001). Post-transplant kinetics of NK cell counts may be used as a simple and affordable proxy to the cell-mediated immunity status in LT recipients and to their associated risk of OI.


Influenza virus glycoprotein-reactive human monoclonal antibodies.
Influenza continues to be a significant public health challenge. Two glycoproteins on the surface of influenza virus, hemagglutinin and neuraminidase, play a prominent role in the process of influenza virus infection and release. Monoclonal antibodies targeting glycoproteins can effectively prevent the spread of the virus. In this review, we summarized currently reported human monoclonal antibodies targeting glycoproteins of influenza A and B viruses.


R0 but not R1/R2 resection is associated with better survival than palliative photodynamic therapy in biliary tract cancer.
There is a need for better management strategies to improve the survival and quality of life in patients with biliary tract cancer (BTC). To assess prognostic factors for survival in a large, non-selective cohort of patients with BTC. We compared outcomes in 321 patients with a final diagnosis of BTC (cholangiocarcinoma n = 237, gallbladder cancer n = 84) seen in a tertiary referral cancer centre between 1998 and 2007. Survival according to disease stage and treatment category was compared using log-rank testing. Cox's regression analysis was used to determine independent prognostic factors. Eighty-nine (28%) patients underwent a surgical intervention with curative intent, of whom 38% had R0 resections. Among the 321 patients, 34% were given chemo- and/or radiotherapy, 14% were palliated with photodynamic therapy (PDT) and 37% with biliary drainage procedures alone. The overall median survival was 9 months (3-year survival, 14%). R0-resective surgery conferred the most favourable outcome (3-year survival, 57%). Although patients palliated with PDT had more advanced clinical T-stages, their survival was similar to those treated with attempted curative surgery but who had positive resection margins. On multivariable analysis, treatment modality, serum carbohydrate-associated antigen 19-9, distant metastases and vascular involvement were independent prognostic indicators of survival. In this large UK series of BTC, palliative PDT resulted in survival similar to those with curatively intended R1/R2 resections. Surgery conferred a survival advantage only in patients with R0 resection margins, emphasising the need for accurate pre-operative staging.


Design of band-pass filters in fluoroscopy by experimental and simulation methods for the 40 keV energy X-ray.
This study was carried out empirical computational and to design filters that while eliminating low-energy radiations according to the conventional methods attenuate high-energy beams that do not fundamentally affect images quality improvement and the absorbed patient dose reduction as well. In this regard, the impacts of thickness and filter material were examined on the contrast, resolution, absorbed patient dose, and image quality. We found that the use of filters increases the resolution, image quality and reduces the output dose intensity greatly, and the 0.1 mm thickness tin element was selected as the most suitable element for the filter.


Phospholipid dependence of calcium ion effects on electrophoretic mobilities of liposomes.
Electrophoretic light scattering (ELS) and depolarization of fluorescence have been used to determine the effect of membrane fluidity on the binding of Ca2+ to liposomes. ELS was used to measure the electrophoretic mobilities of the liposomes. Fluorescence depolarization was used to determine membrane fluidity. Zero to 30 mol% phosphatidylserine (PS) was incorporated into liposomes containing, as bulk phospholipids, one of the following: dimyristoyl-phosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), egg phosphatidylcholine (PC), or hydrogenated egg phosphatidylcholine (H egg PC). The binding of Ca2+ to the liposomes appears to be influenced by membrane fluidity. Liposomes containing bulk phospholipids whose phase transition temperature is higher than the experimental temperature exhibit enhanced binding of CA2+.


Evaluation of methods for measurement of regional skin temperature and heat flow in neonates.
A reliable method for measurement of regional heat loss is a prerequisite for studies of heat loss in the neonate with different clothing and under varying environmental conditions. In the present study we evaluated methods for measuring regional skin temperature and regional dry heat loss in neonates, and specifically the influence of contact sensors applied to the skin surface. A modified incubator with controlled climate conditions was used as the climate chamber. The operative temperature was measured continuously with a black globe thermometer. Skin temperature was recorded with small-mass contact sensors and their influence on skin temperature was verified by simultaneous measurement with an infrared camera. The correlation was 0.99 and 1 SD within 0.3 degree C from the line of identity. The influence of the regional heat flow meter on skin temperature was also checked with an infrared camera and was found to be neglible.


Differential scanning calorimetric studies on bovine serum albumin: I. Effects of pH and ionic strength.
Using defatted and SH-blocked bovine serum albumin (BSA), the measurement of differential scanning calorimetry (d.s.c.) was performed in the range pH 3-11 and ionic strength 0.001-1 M. The shape of the d.s.c. curve was classified into four regimes: (i) the curve with no peak, (ii) that with a peak, (iii) that with a peak having a shoulder, and (iv) that with two peaks. The presence of two peaks was interpreted by the concept of 'heat-induced transition'. The BSA molecule is composed of two domains, thermodynamically independent owing to the formation of a crevice in BSA in a particular range of pH and ionic strength; this gives two peaks in the d.s.c. curve. The enthalpy (delta H) from the d.s.c. curve was plotted against pH and against the NaCl concentration. The value of delta H increased with the increase in the ionic strength in the pH range 5.6-9.0. The temperature of thermal denaturation (the temperature of the peak maximum, Td) was raised with the increase in the ionic strength in the pH range 4.5-9.0, but was lowered in the pH range 3.5-4.0. BSA was stabilized in the neutral-alkaline pH range by the presence of NaCl, but was destabilized in the acidic pH range.


Swedish general practitioners' practice behaviour in hypertension care.
To examine care organization and practice behaviour in hypertension care among Swedish general practitioners. A postal questionnaire followed by a structured interview, in a two-step study design. Uppsala-Orebro region in mid-Sweden (population 1.9 mill.) General practitioners employed by counties, according to a register maintained by the Swedish Medical Association (N = 489). Practice behaviour score compared with a gold standard. 392 general practitioners returned the questionnaire, and the interview was accomplished with 222 of 232 who agreed to participate. Non-responder studies were carried out in both steps. Every fifth GP had access to a special hypertension clinic at their health centre, and 29% had specially trained nurses for hypertension care. 31% reported use of written guidelines. 47% declared they accepted higher BP-levels than recommended, although 78% agreed with official guidelines. The mean practice behaviour score was 8,77 (range 5-12, maximum score 12). Although a majority of GPs agree with official guidelines, there is a considerable variation in care organization and practice behaviour in hypertension care.


Modification of the atrophic effects of tenotomy on mouse soleus muscles by various hind limb nerve lesions and different levels of voluntary motor activity.
The response of mouse soleus muscles to transection of the proximal tendon was examined in a variety of circumstances which might be expected to modify the amount of activation of soleus motoneurons. (i) Denervation of ankle flexors antagonistic to the soleus produced slight atrophy of intact muscles but protected soleus to some extent from the effects of concurrent tenotomy. The onset of changes in tenotomized fibers was delayed several days. (ii) Tenotomy of the soleus soon after reinnervation of the muscle fibers (following tibial nerve transection) resulted in little loss of mass and no degenerative changes. (iii) Denervation during the first few days after tenotomy halted the progress of muscle shortening and alleviated the atrophic changes at the stage reached at the time of denervation. Anesthesia without surgical intervention at the same period after tenotomy also halted the atrophic changes, despite subsequent muscle shortening. (iv) Oral administration of diazepam (approximately 1 mg/kg/day) or chlorpromazine (approximately 12 mg/kg/day), which decreased motor activity during the early posttenotomy period, reduced the amount of muscle shortening and the degree of atrophy. The results suggest that both reflex and voluntary activation of soleus motoneurons contribute to the development of atrophy after tenotomy. The response in soleus muscle seems to be particularly sensitive to the pattern of motor activity during the first 2 to 3 days after tenotomy.


Breast Cancer Cryoablation: Assessment of the Impact of Fundamental Procedural Variables in an In Vitro Human Breast Cancer Model.
Breast cancer is the most prominent form of cancer and the second leading cause of death in women behind lung cancer. The primary modes of treatment today include surgical excision (lumpectomy, mastectomy), radiation, chemoablation, anti-HER2/neu therapy, and/or hormone therapy. The severe side effects associated with these therapies suggest a minimally invasive therapy with fewer quality of life issues would be advantageous for treatment of this pervasive disease. Cryoablation has been used in the treatment of other cancers, including prostate, skin, and cervical, for decades and has been shown to be a successful minimally invasive therapeutic option. To this end, the use of cryotherapy for the treatment of breast cancer has increased over the last several years. Although successful, one of the challenges in cryoablation is management of cancer destruction in the periphery of the ice ball as the tissue within this outer margin may not experience ablative temperatures. In breast cancer, this is of concern due to the lobular nature of the tumors. As such, in this study, we investigated the level of cell death at various temperatures associated with the margin of a cryogenic lesion as well as the impact of repetitive freezing and thawing methods on overall efficacy. Human breast cancer cells, MCF-7, were exposed to temperatures of -5°C, -10°C, -15°C, -20°C, or -25°C for 5-minute freeze intervals in a single or repeat freeze-thaw cycle. Samples were thawed with either passive or active warming for 5 or 10 minutes. Samples were assessed at 1, 2, and 3 days post-freeze to assess cell survival and recovery. In addition, the modes of cell death associated with freezing were assessed over the initial 24-hour post-thaw recovery period. Exposure of MCF-7 cells to -5°C and -10°C resulted in minimal cell death regardless of the freeze/thaw conditions. Freezing to a temperature of -25°C resulted in complete cell death 1 day post-thaw with no cell recovery in all freeze/thaw scenarios evaluated. Exposure to a single freeze event resulted in a gradual increase in cell death at -15°C and -20°C. Application of a repeat freeze-thaw cycle (dual 5-minute freeze) resulted in an increase in cell death with complete destruction at -20°C and near complete death at -15°C (day 1 survival: single -15°C freeze/thaw = 20%; repeated -15°C freeze/thaw = 4%). Analysis of thaw interval time (5 vs 10 minute) demonstrated that the shorter 5-minute thaw interval between freezes resulted in increased cell destruction. Furthermore, investigation of thaw rate (active vs passive thawing) demonstrated that active thawing resulted in increased cell survival thereby less effective ablation compared with passive thawing (eg, -15°C 5/10/5 procedure survival, passive thaw: 4% vs active thaw: 29%). In summary, these in vitro findings suggest that freezing to temperatures of 25°C results in a high degree of breast cancer cell destruction. Furthermore, the data demonstrate that the application of a repeat freeze procedure with a passive 5-minute or 10-minute thaw interval between freeze cycles increases the minimal lethal temperature to the -15°C to -20°C range. The data also demonstrate that the use of an active thawing procedure between freezes reduces ablation efficacy at temperatures associated with the iceball periphery. These findings may be important to improving future clinical applications of cryoablation for the treatment of breast cancer.


Stochastic approach for an unbiased estimation of the probability of a successful separation in conventional chromatography and sequential elution liquid chromatography.
A stochastic approach was utilized to estimate the probability of a successful isocratic or gradient separation in conventional chromatography for numbers of sample components, peak capacities, and saturation factors ranging from 2 to 30, 20-300, and 0.017-1, respectively. The stochastic probabilities were obtained under conditions of (i) constant peak width ("gradient" conditions) and (ii) peak width increasing linearly with time ("isocratic/constant N" conditions). The isocratic and gradient probabilities obtained stochastically were compared with the probabilities predicted by Martin et al. [Anal. Chem., 58 (1986) 2200-2207] and Davis and Stoll [J. Chromatogr. A, (2014) 128-142]; for a given number of components and peak capacity the same trend is always observed: probability obtained with the isocratic stochastic approach<probability obtained with the gradient stochastic approach≤probability predicted by Davis and Stoll << probability predicted by Martin et al. The differences are explained by the positive bias of the Martin equation and the lower average resolution observed for the isocratic simulations compared to the gradient simulations with the same peak capacity. When the stochastic results are applied to conventional HPLC and sequential elution liquid chromatography (SE-LC), the latter is shown to provide much greater probabilities of success for moderately complex samples (e.g., PHPLC=31.2% versus PSE-LC=69.1% for 12 components and the same analysis time). For a given number of components, the density of probability data provided over the range of peak capacities is sufficient to allow accurate interpolation of probabilities for peak capacities not reported, <1.5% error for saturation factors <0.20. Additional applications for the stochastic approach include isothermal and programmed-temperature gas chromatography.


Runt-related transcription factor 2 (RUNX2) inhibits p53-dependent apoptosis through the collaboration with HDAC6 in response to DNA damage.
Runt-related transcription factor 2 (RUNX2) is the best known as an essential protein for osteoblast differentiation. In this study, we have found for the first time that RUNX2 acts as a negative regulator for p53 in response to DNA damage. On DNA damage mediated by adriamycin (ADR) exposure, p53 as well as RUNX2 was induced at protein and mRNA level in human osteosarcoma-derived U2OS cells in association with a significant upregulation of various p53-target genes. Indirect immunostaining and co-immunoprecipitation experiments demonstrated that RUNX2 colocalizes with p53 in cell nucleus and forms a complex with p53 following ADR treatment. Chromatin immunoprecipitation assays revealed that RUNX2/p53 complex is efficiently recruited onto p53-target promoters in response to ADR, suggesting that RUNX2 might be involved in the regulation of transcriptional activation mediated by p53. Indeed, forced expression of RUNX2 resulted in a remarkable downregulation of p53-target genes. Consistent with these observations, knockdown of RUNX2 enhanced ADR-mediated apoptosis and also elevated p53-target gene expression in response to ADR. On the other hand, depletion of RUNX2 in p53-deficient human lung carcinoma-derived H1299 cells had an undetectable effect on p53-target gene expression regardless of ADR treatment, indicating that RUNX2-mediated downregulation of p53-target genes is dependent on p53. Furthermore, RUNX2/p53 complex included histone deacetylase 6 (HDAC6) and HDAC6 was also recruited onto p53-target promoters following ADR exposure. Of note, HDAC6-specific chemical inhibitor tubacin treatment enhanced ADR-mediated upregulation of p53-target gene expression, indicating that deacetylase activity of HDAC6 is required for RUNX2-mediated downregulation of p53-target gene. Taken together, our present findings strongly suggest that RUNX2 inhibits DNA damage-induced transcriptional as well as pro-apoptotic activity of p53 through the functional collaboration with HDAC6 and therefore might be an attractive therapeutic target for cancer treatment.


First-Trimester Ultrasound.
Since the 1980s, development of high-resolution transvaginal ultrasound transducers has significantly improved ultrasound evaluation at earlier gestational ages. Although many indications exist for first-trimester ultrasound in pregnancy, more emphasis has been placed on assessment of fetal anatomy recently. In turn, congenital diagnoses can also be made earlier in pregnancy, raising the question of whether anatomic assessment in the first trimester is one of choice or obligation. Combining transvaginal and transabdominal approach yields the highest detection rate overall. Some studies have shown that more than half of all anomalies and almost all severe anomalies can be detected on early scans.


5-{[4-(Di-methyl-amino)-phen-yl]ethyn-yl}pyrimidine-1,2,3,5-tetra-fluoro-4,6-di-iodo-benzene (1/2).
The treatment of 5-{[4-(di-methyl-amino)-phen-yl]ethyn-yl}pyrimidine with a threefold excess of 1,2,3,5-tetra-fluoro-4,6-di-iodo-benzene in di-chloro-methane solution led to the formation of the unexpected 1:2 title co-crystal, C14H13N3·2CF4I2. In the extended structure, two unique C-I⋯N halogen bonds from one of the 1,2,3,5-tetra-fluoro-4,6-di-iodo-benzene mol-ecules to the pyrimidine N atoms of the 5-{[4-(di-methyl-amino)-phen-yl]ethyn-yl}pyrimidine mol-ecule generate [110] chains and layers of these chains are π-stacked along the a-axis direction. The second 1,2,3,5-tetra-fluoro-4,6-di-iodo-benzene mol-ecule resides in channels formed parallel to the a-axis direction between stacks of 5-{[4-(di-methyl-amino)-phen-yl]ethyn-yl}pyrimidine mol-ecules and inter-acts with them via C-I⋯π(alkyne) contacts.


Revision of frailty assessment in kidney transplant recipients: Replacing unintentional weight loss with CT-assessed sarcopenia in the physical frailty phenotype.
Kidney transplantation (KT) experts did not support the use of subjective unintentional weight loss to measure shrinking in the physical frailty phenotype (PFP); a clinically feasible and predictive measure of shrinking is needed. To test whether unintentional weight loss could be replaced by an assessment of sarcopenia using existing CT scans, we performed a prospective cohort study of adult KT recipients with original PFP (oPFP) measured at admission (December 2008-February 2020). We ascertained sarcopenia by calculating skeletal muscle index from available, clinically obtained CTs within 1-year pre-KT (male < 50 cm2 /m2 ; female < 39 cm2 /m2 ) and combined it with the original four components to determine new PFP (nPFP) scores. Frailty was classified by frailty score: 0: non-frail; 1-2: pre-frail; ≥3: frail. Mortality and graft loss hazard ratios (HRs) were estimated using adjusted Cox proportional hazard models. Model discrimination was quantified using Harrell's C-statistic. Among 1113 recipients, 18.6% and 17.1% were frail by oPFP and nPFP, respectively. Compared to non-frail recipients, frail patients by either PFP had higher risks of mortality (oPFP HR = 1.67, 95% CI: 1.07-2.62, C = 0.710; nPFP HR = 1.68, 95% CI: 1.06-2.66, C = 0.710) and graft loss (oPFP HR = 1.67, 95% CI: 1.17-2.40, C = 0.631; nPFP HR = 1.66, 95% CI: 1.15-2.40, C = 0.634) with similar discriminations. oPFP and nPFP are equally useful in risk prediction for KT recipients; oPFP may aid in screening patients for pre-KT interventions, while nPFP may assist in nuanced clinical decision-making.


college women's preferred HIV prevention message mediums: mass media versus interpersonal relationships.
One quarter of HIV cases occur in women ages 15-44 years. We investigated preferential HIV prevention message mediums among college women (18-21 years of age) and their association with parent and partner communication. A nonexperimental cross-sectional survey assessed factors associated with parent and partner communication among 626 single female students who were sexually active in the previous 6 months and attending a 4-year public university in Florida. Women who perceived themselves to be at elevated risk of acquiring HIV were more likely to communicate with their parents (p < .05), but not their partners. In multivariable analysis, students were more likely to communicate about sexual risk behavior with their parents when mothers were younger and when less influenced by their peers. Reading items on the Internet about intravenous drug use and HIV were independently associated with higher parent and partner communication, respectively. Findings can guide future HIV prevention communication interventions.


A low-literacy skin care manual for spinal cord injury patients.
Most patient education materials are written at least two grade levels above the average grade achievement of health care consumers. This paper describes the development of a patient education manual for people with low literacy skills. The manual provides information about pressure ulcer prevention for patients with spinal cord injuries. Educational principles related to literacy skills were applied to produce a manual at the fifth-grade reading level.


Expert Perspectives On Clinical Challenges: Expert Perspectives: Challenges in Scleroderma.
You are consulted to evaluate a 56-year-old woman with known Raynaud's phenomenon, finger swelling of several; months' duration, and new hypertension with a blood pressure of 160/100 mm/Hg. She also reports progressive shortness of breath. Physical examination reveals telangiectasias, sclerodactyly, and proximal skin sclerosis (thick shiny skin on the chest and upper arms), and bibasilar crackles are found on chest examination. Laboratory tests reveal evidence of microangiopathic hemolytic anemia, thrombocytopenia, and elevation of the serum creatinine level (previously normal), and chest computed tomography shows evidence of ground-glass opacification in both lower lung fields.


Indoor air pollution in rural north-east India: Elemental compositions, changes in haematological indices, oxidative stress and health risks.
Chronic smoke exposure, emitted by biomass fuel burning leads to many diseases, which are originated due to oxidative stress. The present study investigated the levels of PM2.5, PM10 and PM2.5 bound trace metals released during cooking with fuelwood and subsequent changes in haematological parameters along with oxidative stress in rural tribal women of northeast India exposed to wood smoke. The levels of PM2.5, PM10 and trace metals associated with PM2.5 (nickel, cobalt, manganese, zinc, cadmium, lead and copper) were measured. In addition, blood samples were analyzed for concentrations of different blood related parameters (haemoglobin, platelet count, red blood cells and white blood cells) and levels of antioxidants (reduced glutathione, superoxide dismutase, and catalase). Plasma malondialdehyde (MDA) was measured as a biomarker of lipid peroxidation. Health risk assessment was done to assess the potential risk posed by inhalation of fine particles emitted from cooking with fuel wood. Levels of both PM2.5 and PM10 were higher in wood users compared to LPG users during cooking period (644.4 ± 368.3 µg/m³ vs 50 ± 23.8 µg/m³; 915 ± 441.3 µg/m³ vs 83.3 ± 33 µg/m³) and it exceeded the permissible limits of WHO. Levels of trace metals during the cooking period in fuel wood users were significantly higher than LPG users (p = 0.01). After controlling possible confounders, both platelet count and white blood cells (WBC) had a significant positive association with PM2.5and PM10. Similarly, haemoglobin had a negative association with both PM2.5 and PM10. Depleted levels of antioxidant enzymes and increase in lipid peroxidation (MDA) suggest a close association with pollutants released from wood smoke, indicating oxidative stress in women who used fuelwood for cooking. The total hazard quotient (HQ) of 0.11 was within the acceptable limit (i.e., 1.0). The total excess lifetime cancer risk (ELCR) was 5.4 × 10-6 which is five times higher than the acceptable limit of 1.0 × 10-6. Individual carcinogenic risk of Ni (2.3 × 10-6) and Cd (3.1 × 10-6) were also higher compared to acceptable limit. These results indicate that tribal women cooking with wood are at greater risk of developing cancer and also give support to the positive association between wood smoke and oxidative stress.


Influence of climate factors on pediatric alopecia areata flares in Philadelphia, Pennsylvania.
Patients with alopecia areata (AA) may experience episodic disease flares characterized by increasing hair loss that follow a seasonal pattern. However, no studies have examined whether specific climate factors contribute to the seasonal pattern of AA flares. Using Spearman rank correlation analyses, we assessed the association between climate variables and AA flare frequency per month in 336 children with AA in Philadelphia, Pennsylvania. Region-specific monthly values for average ambient temperature, air pressure, cloudiness, hours of sunlight, relative humidity, number of days with sun, number of days with rain, volume of precipitation, wind gust, wind speed, and UV index from January 2015 to December 2017 were obtained from World Weather Online. We found significant (P < 0.05) correlations between AA flare frequency and UV index (R = - 0.66), precipitation (R = - 0.66), number of days with rain (R = - 0.70), number of days with sun (R = 0.62), and air pressure (R = 0.80). Stratified analyses showed even stronger associations with UV index and precipitation in patients with an atopic comorbidity. New significant correlations appeared with temperature, wind speed, and UV index of the prior month. However, in patients who did not have atopic comorbidities, we generally observed weaker and non-significant correlations between climate and AA flare frequency. This study suggests that certain climate factors may mediate the seasonal pattern of AA flares and may contribute to AA pathogenesis. Atopic AA patients may be more susceptible to the influence of climate compared to those with no history of atopy.


Optimizing enrichment conditions for the isolation of Escherichia coli O157 in soils by immunomagnetic separation.
To compare a range of enrichment broths and enrichment temperatures for the isolation of Escherichia coli O157 by immunomagnetic separation (IMS) from sandy, loam and clay soils. Soils were spiked with cocktails of four atoxigenic strains of E. coli O157 and four strains of commensal E. coli. The organisms were stressed by subjecting soils to cycles of freeze/thawing, followed by drying at 20 degrees C for up to 4 days. Nine enrichment broths were trialled based on buffered peptone water, tryptone soya broths and EC broths supplemented with a range of selective additions. Enrichments were incubated for 6 h and assessed by target recovery after IMS on cefixime tellurite sorbitol MacConkey agar (CTSMAC) incubated at 37 degrees C for 24 h. A comparison of enrichment temperatures (37 and 42 degrees C) was also performed. Buffered peptone water (with or without vancomycin) and tryptone soya broth (with or without novobiocin) gave significant increases in recovery of E. coli O157 compared to others tested. In addition, broths incubated at 42 degrees C were superior to those at 37 degrees C for the recovery of E. coli O157. This study showed that sub-lethally damaged E. coli O157 surviving in soil can be sensitive to antimicrobial additions. The choice and concentration of these additions is vitally important to optimize target recovery. Some IMS protocols, established for the isolation of E. coli O157, may be unsuitable for the examination of soil samples.


Design and Synthesis of a Pyridine Based Chemosensor: Highly Selective Fluorescent Probe For Pb2+
A pyridine based imine-linked chemosensor has been synthesized and evaluated its binding affinity with library of transition metal ions. It has prominent selectivity towards Pb2+ among other metal ions in DMF/H2O (9:1, v/v) solvent system. The 1:1 stoichiometric was confirmed by job's plot and has a binding constant (Ka) = 5.142 × 103 M-1 on fluorescence. A B3LYP/6-31G and B3LYP/LanL2DZ basis sets were employed for optimization of 3 and 3.Pb2+.


Effect of long term androgen removal on androgen-induced proliferation of seminal vesicle cells in adult mice.
Male mice were castrated on day 60 after birth; daily injections of testosterone propionate (TP, 4 micrograms/g b.wt) were started 1.2 or 6 months after the castration. The incorporation of 5-[125I]iodo-2'-deoxyuridine [( 125I]IdUrd) into the whole seminal vesicles was determined on various days after starting the TP injections as an index for proliferation. Although the peak of [125I]IdUrd uptake was observed 3 days after starting the TP injections in both short (1-2 months) and long (6 months) term castrated mice, the peak was significantly lower and the period of proliferation was longer in the long term group than in the short term group; the weights of seminal vesicles before TP injections were 6 and 10 mg in the long and short term groups, respectively. Although TP injections induced the proliferation of only epithelial cells in the short term group, the same treatment induced the proliferation of both epithelial and fibromuscular cells in the long term group. The deficient responsiveness to androgen of the seminal vesicle cells found in the long term castrated mice was completely recovered by TP pretreatment for 2 weeks. The present findings suggest that so-called imprinted cells in the mouse seminal vesicle induced by neonatal and prepubertal testicular androgens are very slowly lost at least in part by androgen removal for long periods such as more than 6 months in adult mice and that the loss is at least in part due to the death of fibromuscular cells, which is recovered rather quickly by androgen pretreatment.


[Dexmedetomidine alleviates LPS/D-Gal-induced acute liver injury via up-regulation of LC3-II expression in mice].
The aim of the present study was to investigate the effects of dexmedetomidine (DEX) on acute liver injury induced by lipopolysaccharide (LPS)/D-galactosamine (D-Gal) and the underlying mechanism. Male BALB/c mice were intraperitoneally injected with LPS/D-Gal to induce acute liver injury model, and pretreated with DEX or in combination with the autophagy inhibitor, 3-methyladenine (3-MA) 30 min before injection. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity, as well as myeloperoxidase (MPO) activity in liver tissue were determined with the corresponding kits. Serum tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) levels were determined by ELISA. The protein expression levels of LC3-II and P62 in liver tissue were determined by Western blot. Liver histopathological changes were detected by HE staining. The results showed that, compared with control group, LPS/D-Gal enhanced ALT and AST activity, increased TNF-α and IL-6 levels, as well as MPO activity, up-regulated LC3-II and P62 protein expression levels, and significantly induced pathological damage in liver tissue. DEX reversed the above changes in the LPS/D-Gal group, whereas these protective effects of DEX were blocked by 3-MA. The above results suggest that DEX alleviates LPS/D-Gal-induced acute liver injury, which may be associated with the up-regulation of LC3-II protein expression and the activation of autophagy.


Presence of a Kirsten murine sarcoma virus ras oncogene in cells transformed by 3-methylcholanthrene.
Oncogenes have previously been reported in the DNAs of mouse fibroblast lines which had become transformed after in vitro exposure to the carcinogen 3-methylcholanthrene. These oncogenes are now shown to be versions of the cellular Kirsten ras gene and are therefore homologous to oncogenes detected in a variety of human tumor DNAs.


Leukaemic infiltration of the anus.
A case of biopsy-proven anal involvement in acute promyelocytic leukaemia is presented with management by local conservative measures and remission induction with chemotherapy. A brief review of the literature is given.


Studies of Athabasca asphaltene Langmuir films at air-water interface.
Asphaltenes are present in heavy oils and bitumen. They are a mixture of hydrocarbons having complex structures of polyaromatic rings and short side chains. In general, the high-molecular-weight asphaltene is the most aromatic fraction with the highest number of side chains and the low-molecular-weight asphaltene contains the lowest number of side chains, while the number of side chains of the whole asphaltene fraction lies in between. In this study, asphaltenes were extracted and/or fractionated from Athabasca oil sand bitumen. Subfractions of high and low molecular weight and the whole asphaltenes were characterized using a Langmuir trough and complementary techniques such as VPO, FTIR, AFM, and contact angle measurements. At an air-water interface, amphiphilic asphaltene molecules can form a monolayer. Various fractions (high, low, and whole) of the asphaltene molecules behave similarly at the air-water interface, characterized by close resemblance of their surface pressure-area, hysteresis, and relaxation isotherms. The high-molecular-weight asphaltene is the most expanded fraction, while the low-molecular-weight asphaltene fraction is the most condensed, with the whole asphaltene lying in between. At the air-water interface a monolayer of the low-molecular-weight asphaltene relaxes at a faster rate than one of the high-molecular-weight asphaltene.


Hemispheric asymmetry and somatotopy of afferent inhibition in healthy humans.
A conditioning electrical stimulus to a digital nerve can inhibit the motor-evoked potentials (MEPs) in adjacent hand muscles elicited by transcranial magnetic stimulation (TMS) to the contralateral primary motor cortex (M1) when given 25-50 ms before the TMS pulse. This is referred to as short-latency afferent inhibition (SAI). We studied inter-hemispheric differences (Experiment 1) and within-limb somatotopy (Experiment 2) of SAI in healthy right-handers. In Experiment 1, conditioning electrical pulses were applied to the right or left index finger (D2) and MEPs were recorded from relaxed first dorsal interosseus (FDI) and abductor digiti minimi (ADM) muscles ipsilateral to the conditioning stimulus. We found that SAI was more pronounced in right hand muscles. In Experiment 2, electrical stimulation was applied to the right D2 and MEPs were recorded from ipsilateral FDI, extensor digitorum communis (EDC) and biceps brachii (BB) muscles. The amount of SAI did not differ between FDI, EDC and BB muscles. These data demonstrate inter-hemispheric differences in the processing of cutaneous input from the hand, with stronger SAI in the dominant left hemisphere. We also found that SAI occurred not only in hand muscles adjacent to electrical digital stimulation, but also in distant hand and forearm and also proximal arm muscles. This suggests that SAI induced by electrical D2 stimulation is not focal and somatotopically specific, but a more widespread inhibitory phenomenon.


Proteinuria 1 year after renal transplantation is associated with impaired graft survival in children.
Proteinuria is a common manifestation of chronic kidney disease (CKD), and there is a high incidence of CDK and its complications following renal transplantation. However, little data are available on the association between proteinuria and graft/patient survival in the paediatric transplant population. The primary aim of this study was to investigate the associations between posttransplant proteinuria and graft/patient survival in children after renal transplantation. In this retrospective study, we screened all 91 children receiving renal allografts at a single institution between 1997 and 2007. The inclusion criteria were a functioning graft at 1 year posttransplant, data availability and no recurrence of focal-segmental glomerulosclerosis. The final cohort included 75 patients. Proteinuria was considered to be pathologic if the urinary protein/creatinine ratio was >30 mg/mmol. Donor and recipient characteristics, data on proteinuria, estimated glomerular filtration rate (eGFR) and rejection episodes were analysed. The most recent of the biopsies performed during the follow-up after 1 year posttransplant were analysed separately in the proteinuric group and the non-proteinuric group. Proteinuria at 1-year posttransplant was pathologic in 35 % of patients. The 5-year graft survival rate was significantly lower in the proteinuric group than in the non-proteinuric group (77 vs. 100 %; p < 0.001). Proteinuria at 1 year posttransplant was associated with reduced long-term graft survival independent of other risk factors, including decreased eGFR or episodes of acute corticosensitive and corticoresistant rejection. The most frequent histologic finding in the proteinuric group was chronic rejection. There was no significant difference in the 5-year patient survival rate between the proteinuric group and the non-proteinuric group. This study emphasizes the importance of proteinuria as a prognostic factor of renal allograft survival in children.


Overexpression of Cdk6 and Ccnd1 in chondrocytes inhibited chondrocyte maturation and caused p53-dependent apoptosis without enhancing proliferation.
Cell proliferation and differentiation are closely coupled. However, we previously showed that overexpression of cyclin-dependent kinase (Cdk6) blocks chondrocyte differentiation without affecting cell-cycle progression in vitro. To investigate whether Cdk6 inhibits chondrocyte differentiation in vivo, we generated chondrocyte-specific Cdk6 transgenic mice using Col2a1 promoter. Unexpectedly, differentiation and cell-cycle progression of chondrocytes in the Cdk6 transgenic mice were similar to those in wild-type mice. Then, we generated chondrocyte-specific Ccnd1 transgenic mice and Cdk6/Ccnd1 double transgenic mice to investigate the possibility that Cdk6 inhibits chondrocyte differentiation through E2f activation. Bromodeoxyuridine (BrdU)-positive chondrocytes and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive chondrocytes were increased in number, and chondrocyte maturation was inhibited only in Cdk6/Ccnd1 transgenic mice (K6(H)/D1(H) mice), which showed dwarfism. Retinoblastoma protein (pRb) was highly phosphorylated but p107 was upregulated, and the expression of E2f target genes was dysregulated as shown by upregulation of Cdc6 but downregulation of cyclin E, dihydrofolate reductase (dhfr), Cdc25a and B-Myb in chondrocytes of K6(H)/D1(H) mice. Similarly, overexpression of Cdk6/Ccnd1 in a chondrogenic cell line ATDC5 highly phosphorylated pRb, upregulated p107, induced apoptosis, upregulated Cdc6 and downregulated cyclin E, dhfr and B-Myb and p107 small interfering RNA reversed the expression of downregulated genes. Further, introduction of kinase-negative Cdk6 and cyclin D1 abolished all effects by Cdk6/cyclin D1 in ATDC5 cells, indicating the requirement of the kinase activity on these effects. p53 deletion partially restored the size of the skeleton and almost completely rescued chondrocyte apoptosis, but failed to enhance chondrocyte proliferation in K6(H)/D1(H) mice. These findings indicated that Cdk6/Ccnd1 overexpression inhibited chondrocyte maturation and enhanced G1/S cell-cycle transition by phosphorylating pRb, but the chondrocytes failed to accomplish the cell cycle, and underwent p53-dependent apoptosis probably due to the dysregulation of E2f target genes. Our findings also indicated that p53 deletion in addition to the inactivation of Rb was not sufficient to accelerate chondrocyte proliferation, suggesting the resistance of chondrocytes to sarcomagenesis.


Therapeutic consequences from changing voice ideals (clear to harsh, pleasant to jarring): summarizing report on a round-table discussion at the 5th World Voice Congress, Luxor, Egypt, 27-31 October 2012.
Starting out with an attempt to define an 'ideal voice', the discussion moved on to the hazards of pop singing, followed by the topics of harsh voices (from the desired effects in modern singing expression to devastating clinical cases), phoniatric targets (aesthetics and fitness), and, finally, the phoniatricians' attitudes comprising acceptance, neglect, and rejection. Therapy should aim at a resilient and effective voice regardless of the mere sound, unless the perfection of the vocal sound is the ultimate goal. In addition, the panellists agreed that voice specialists and all friends of healthy and efficient voices are obliged permanently to engage in voice care.


Immunologic reconstitution of foals with combined immunodeficiency.
Thirty-eight foals with combined immunodeficiency (CID) received transplanted fetal liver cells, fetal liver and thymus cells, histocompatible bone marrow cells, or equine lymphocyte antigen (ELA) haploidentical bone marrow cells in an attempt to reconstitute their deficient immune systems. Engraftment was infrequent, partial, and unpredictable when fetal cells were employed. Three of five CID foals receiving ELA haploidentical bone marrow cells demonstrated partial reconstitution, but engraftment was only temporary. Administration of histocompatible bone marrow cells resulted in rapid, full and sustained engraftment.


Informational landscapes in art, science, and evolution.
An informational landscape refers to an array of information related to a particular theme or function. The Internet is an example of an informational landscape designed by humans for purposes of communication. Once it exists, however, any informational landscape may be exploited to serve a new purpose. Listening Post is the name of a dynamic multimedia work of art that exploits the informational landscape of the Internet to produce a visual and auditory environment. Here, I use Listening Post as a prototypic example for considering the creative role of informational landscapes in the processes that beget evolution and science.


Ethylation of DNA and protamine by ethyl methanesulfonate in the germ cells of male mice and the relevancy of these molecular targets to the induction of dominant lethals.
The molecular dosimetry of ethyl methanesulfonate (EMS) in the germ cells of male mice has been investigated. The mice were injected i.p. with 200 mg/kg of [3H]EMS and the ethylations per sperm head, per deoxynucleotide, and per unit of protamine were then determined over a 2-week period. The ethylations per sperm head closely paralleled the dominant-lethal frequency curve for EMS, reaching a maximum of 5 to 6.5 million ethylations per vas sperm head at 8 to 10 days after treatment. Ethylation of sperm DNA was greatest at 4 h after treatment, with 5.7 ethylations/10(5) deoxynucleotides, and gradually decreased to 2.2 ethylations/10(5) deoxynucleotides at 15 days after treatment. The ethylation of sperm DNA did not increase in the germ-cell stages most sensitive to EMS, and was not correlated with the dominant-lethal frequency curve for EMS. However, ethylation of sperm protamine did increase in the germ-cell stages most sensitive to EMS, and showed an excellent correlation with the incidence of dominant lethals produced by EMS in the germ cells. A model is presented to explain, at a molecular level, how dominant lethals may be induced in mouse germ cells by EMS. Ethylation of cysteine sulfhydryl groups contained in mouse-sperm protamine could block normal disulfide-bond formation, preventing proper chromatin condensation in the sperm nucleus. Stresses in the chromatin structure could then eventually lead to chromosome breakage, with resultant dominant lethality.


Depletion of large dense-cored vesicles from parasympathetic nerve terminals in rat parotid glands after prolonged stimulation of the auriculotemporal nerve.
Large dense-cored vesicles (60-100 nm in diameter) have been assessed electron-microscopically in terminal parasympathetic axons at acinar neuro-effector sites in rat parotid glands. Their numbers in control unstimulated glands have been compared with those in the contralateral glands of the same animals after prolonged nerve stimulation. Bilateral postganglionic sympathectomy had been undertaken 4-6 weeks previously to remove adrenergic axons from the glands. Stimulation of the postganglionic parasympathetic nerve to the gland--the auriculotemporal nerve--for 80 min at 40 Hz caused a significant depletion of large dense-cored vesicles from the terminal axons. This depletion corresponded in time and magnitude to the depletion of vasoactive intestinal peptide and substance P from the glands that had been found previously to occur under identical conditions. This adds support to the belief that the neuropeptides are stored in such vesicles and that these vesicles release their contents at neuro-effector sites as a result of propagated impulse formation in the axons.


Human 25-hydroxyvitamin D 24-hydroxylase cytochrome P450 subunit maps to a different chromosomal location than that of pseudovitamin D-deficient rickets.
We have cloned part of the human 25-OHD 24-hydroxylase cytochrome P450 (P450cc24) cDNA. The characterized sequence consists of 776 bp of the coding and 720 bp of the 3'-untranslated region interrupted by an intron. In the coding region we found 79.8% similarity in DNA and 87.5% in deduced amino acid sequences between human and rat, with no similarity in the 3'-untranslated region. By Southern blot hybridization of DNA from human-hamster somatic cell hybrids and by in situ immunofluorescence hybridization, we mapped P450cc24 to human chromosome 20q13.1. This location of P450cc24 is different from that of pseudovitamin D-deficient rickets (PDDR), previously assigned to chromosome 12q14 by linkage analysis, thus excluding it as a target of the PDDR mutation. Since it is likely that PDDR is caused by a mutation in the 25-OHD 1 alpha-hydroxylase P450 subunit (P450cc1 alpha) our results do not support the hypothesis that the two cytochromes are encoded by a single gene.


Activation of interleukin-6/signal transducer and activator of transcription 3 by human papillomavirus early proteins 6 induces fibroblast senescence to promote cervical tumourigenesis through autocrine and paracrine pathways in tumour microenvironment.
Although it is reported that interleukin (IL)-6/signal transducer and activator of transcription 3 (STAT3) is activated by human papillomavirus (HPV) infection in cervical cancer cells, little is known about the role of IL-6/STAT3 in tumour microenvironment during development of the disease. In this study, we found that cancer-associated fibroblasts (CAF) but not normal fibroblasts (NF) secrete high level of IL-6 with activated STAT3 and appear senescent at early passages in culture or in cervical cancer tissues infected with high-risk HPV, and that treatment of NF with recombinant IL-6 or CAF conditioned medium (CM) induces activation of STAT3 and cellular senescence. IL-6 and STAT3 are either upregulated or activated in Siha and Hela cells infected with HPV 16 or 18, but not in C33A and ME180 cells without HPV 16 or 18 infection. Overexpression of HPV early proteins 6 (E6) activates STAT3, increases IL-6 expression and tumour burden in C33A and ME180 cells, while silencing of HPV E6 by specific shRNA reduces STAT3 activation, IL-6 expression, and tumour formation in Siha and HeLa cells, so does silencing of STAT3 by specific shRNA in HeLa and C33A/E6 cells. The tumour growth of cervical cancer cells reconstituted with CAF or NF is largely affected by inhibition of fibroblast senescence with STAT3 inhibitor or with IL-6 antibody treatment. Thus, we have uncovered a mechanism that fibroblast senescence promotes cervical cancer development through high-risk HPV E6-activated IL-6/STAT3 signalling in tumour microenvironment.


Cognitive and Motor Decline in Dementia with Lewy Bodies and Parkinson's Disease Dementia.
There is a need to better understand the rate of cognitive and motor decline of Dementia with Lewy bodies (DLB) and Parkinson's disease Dementia (PDD). To compare the rate of cognitive and motor decline in patients with DLB and PDD from the E-DLB Consortium and the Parkinson's Incidence Cohorts Collaboration (PICC) Cohorts. The annual change in MMSE and MDS-UPDRS part III was estimated using linear mixed regression models in patients with at least one follow-up (DLB n = 837 and PDD n = 157). When adjusting for confounders, we found no difference in the annual change in MMSE between DLB and PDD (-1.8 [95% CI -2.3, -1.3] vs. -1.9 [95% CI -2.6, -1.2] [P = 0.74]). MDS-UPDRS part III showed nearly identical annual changes (DLB 4.8 [95% CI 2.1, 7.5]) (PDD 4.8 [95% CI 2.7, 6.9], [P = 0.98]). DLB and PDD showed similar rates of cognitive and motor decline. This is relevant for future clinical trial designs.


Increased circulating levels of tumor necrosis factor-like cytokine 1A and decoy receptor 3 correlate with SYNTAX score in patients undergoing coronary surgery.
Chronic inflammation of the arteries is a critical mechanism responsible for coronary atherosclerosis. We aimed to determine if tumor necrosis factor (TNF)-like cytokine 1A (TL1A) and decoy receptor 3 (DcR3) were involved in promoting atherosclerosis. We compared plasma levels of TL1A and DcR3 in patients with coronary artery disease (CAD) undergoing coronary artery bypass grafting (n=40) and patients without CAD group (n=37, normal coronary artery angiogram) by enzyme-linked immunosorbent assay. We also analyzed the correlation between CAD and SYNTAX scores. Plasma levels of TL1A and DcR3 were significantly higher in the CAD compared with the no-CAD group. Multivariate analysis showed that TL1A and DcR3 were significantly correlated with the presence of CAD, and receiver operating characteristic curve analysis indicated that both TL1A and DcR3 showed high sensitivity and specificity for diagnosing CAD. Furthermore, TL1A was positively and significantly correlated with SYNTAX score in CAD patients. CAD patients requiring coronary artery bypass grafting have high circulating levels of both TL1A and DcR3, which may thus be useful biomarkers for diagnosing severe CAD. Furthermore, plasma levels of TL1A correlate with SYNTAX score, supporting its potential use as an indicator of the severity of CAD.


Depletion of the AP-1 repressor JDP2 induces cell death similar to apoptosis.
JDP2 is a ubiquitously expressed nuclear protein that efficiently represses the activity of the transcription factor AP-1. Thus far, all studies of JDP2 function have relied on the ectopic expression of the protein. In this study, we use a different approach: depletion of JDP2 from cells. Specific depletion of JDP2 resulted in p53-independent cell death that resembles apoptosis and was evident at 72 h. The death mechanism was caspase dependent as the cells could be rescued by treatment with caspase inhibitor zVAD. Our studies suggest that JDP2 functions as a general survival protein, not only following UV-irradiation, as reported earlier, but also under normal culture conditions. Thus, our data support that JDP2 is a cellular survival protein whose presence is necessary for normal cellular function.


Patterns of failure after use of (18)F-FDG PET/CT in integration of extended-field chemo-IMRT and 3D-brachytherapy plannings for advanced cervical cancers with extensive lymph node metastases.
The study is to evaluate the patterns of failure, toxicities and long-term outcomes of aggressive treatment using (18)F-FDG PET/CT-guided chemoradiation plannings for advanced cervical cancer with extensive nodal extent that has been regarded as a systemic disease. We retrospectively reviewed 72 consecutive patients with (18)F-FDG PET/CT-detected widespread pelvic, para-aortic and/or supraclavicular lymph nodes treated with curative-intent PET-guided cisplatin-based extended-field dose-escalating intensity-modulated radiotherapy (IMRT) and adaptive high-dose-rate intracavitary 3D-brachytherapy between 2002 and 2010. The failure sites were specifically localized by comparing recurrences on fusion of post-therapy recurrent (18)F-FDG PET/CT scans to the initial PET-guided radiation plannings for IMRT and brachytherapy. The median follow-up time for the 72 patients was 66 months (range, 3-142 months). The 5-year disease-free survival rate calculated by the Kaplan-Meier method for the patients with extensive N1 disease with the uppermost PET-positive pelvic-only nodes (26 patients), and the patients with M1 disease with the uppermost PET-positive para-aortic (31 patients) or supraclavicular (15 patients) nodes was 78.5 %, and 41.8-50 %, respectively (N1 vs. M1, p = 0.0465). Eight (11.1 %), 18 (25.0 %), and 3 (4.2 %) of the patients developed in-field recurrence, out-of-field and/or distant metastasis, and combined failure, respectively. The 6 (8.3 %) local failures around the uterine cervix were all at the junction between IMRT and brachytherapy in the parametrium. The rate of late grade 3/4 bladder and bowel toxicities was 4.2 and 9.7 %, respectively. When compared to conventional pelvic chemoradiation/2D-brachytherapy during 1990-2001, the adoption of (18)F-FDG PET-guided extended-field dose-escalating chemoradiation plannings in IMRT and 3D-brachytherapy after 2002 appeared to provide higher disease-free and overall survival rates with acceptable toxicities in advanced cervical cancer patients. For AJCC stage M1 cervical cancer with supraclavicular lymph node metastases, curability can be achieved in the era of PET and chemo-IMRT. However, the main pattern of failure is still out-of-field and/or distant metastasis. In addition to improving systemic treatment, how to optimize and integrate the junctional doses between IMRT and 3D-brachytherapy in PET-guided plannings to further decrease local recurrence warrants investigation.


Antimicrobial efficacy of two irrigation techniques in tapered and nontapered canal preparations: an in vitro study.
The aim of this in vitro study was to determine whether irrigation with apical negative pressure was more effective than traditional positive-pressure irrigation in eradicating Enterococcus faecalis from preshaped root canals. Fifty-four extracted mandibular molars were instrumented to produce either a non-tapered or tapered preparation, sterilized, inoculated with E. faecalis for 30 days, and then randomly assigned into the following groups: group 1--non-tapered preparation and negative-pressure irrigation, group 2--non-tapered preparation and positive-pressure irrigation, group 3--tapered preparation and positive-pressure irrigation, and group 4--tapered preparation and negative-pressure irrigation. Mesial canals were sampled before and after final irrigation and samples incubated aerobically for 48 hours at 37 degrees C. Scanning electron microscopic analysis confirmed dense bacterial colonies in the positive control, consistent with biofilm formation. A statistically significant difference was evident when comparing apical negative-pressure irrigation to positive-pressure irrigation (p=0.004). There was no statistically significant difference in colony-forming units (CFUs) between sizes #35 and #45, nor between tapered and non-tapered preparation. The results of this in vitro study showed that apical negative-pressure irrigation has the potential to achieve better microbial control than traditional irrigation delivery systems.


A comparison of the effects of IGF-I and insulin on glucose metabolism, fat metabolism and the cardiovascular system in normal human volunteers.
The metabolic and cardiovascular effects of recombinant human IGF-I were compared to insulin in six normal subjects. Subjects were studied twice and intravenously received an infusion of [6,6-2H2]glucose (0-480 min) and in random order either IGF-I 20 micrograms kg-1 h-1 (43.7 pmol kg-1 min-1 or insulin 0.5 mU kg-1 min-1 (3.4 pmol kg-1 min-1) with an euglycaemic clamp. One subject was withdrawn following a serious adverse event. During the IGF-I infusion glucose appearance rate (Ra) decreased from 1.79 +/- 0.13 at baseline (150-180 min) to 0.35 +/- 0.26 mg kg-1 min-1 (P < 0.01) at 360 min, and glucose utilization rate (Rd) increased from 1.79 +/- 0.28 to 4.17 +/- 0.84 mg kg-1 min-1 (P < 0.01). There was no change in free fatty acids (FFA) and an increase (percentage change from pre-infusion mean) in cardiac output +l37.3% +/- 9% (P < 0.01), heart rate +13% +/- 2% (P < 0.01) and stroke volume +21% +/- 7% (P < 0.05). During the insulin infusion glucose Ra decreased from 1.89 +/- 0.13 to 0.34 +/- 0.33 mg kg-1 min-1 (P < 0.01) and FFA from 0.546 mmol l-1 to 0.198 mmol l-1 (P < 0.01), glucose Rd increased from 1.89 +/- 0.18 to 5.41 +/- 1.47 mg kg-1 min-1 (P < 0.01) and there were no significant changes in the cardiovascular variables.


The Neuronal Migration Factor srGAP2 Achieves Specificity in Ligand Binding through a Two-Component Molecular Mechanism.
srGAP proteins regulate cell migration and morphogenesis by shaping the structure and dynamics of the cytoskeleton and membranes. First discovered as intracellular effectors for the Robo1 axon-guidance receptor, srGAPs were later identified as interacting with several other nuclear and cytoplasmic proteins. In all these cases, the srGAP SH3 domain mediates protein-protein interactions by recognizing a short proline-rich segment on the cognate-binding partner. However, as interactions between the isolated SH3 domain and a selected set of ligands show weak affinity and low specificity, it is not clear how srGAPs are precisely recruited to their signaling sites. Here, we report a two-component molecular mechanism that regulates ligand binding to srGAP2 by on the one hand dramatically tightening their association and on the other, moderately autoinhibiting and restricting binding. Our results allow the design of point mutations for better probing of srGAP2 activities, and may facilitate the identification of new srGAP2 ligands.


Interplay of structure and photophysics of individualized rod-shaped graphene quantum dots with up to 132 sp² carbon atoms.
Nanographene materials are promising building blocks for the growing field of low-dimensional materials for optics, electronics and biophotonics applications. In particular, bottom-up synthesized 0D graphene quantum dots show great potential as single quantum emitters. To fully exploit their exciting properties, the graphene quantum dots must be of high purity; the key parameter for efficient purification being the solubility of the starting materials. Here, we report the synthesis of a family of highly soluble and easily processable rod-shaped graphene quantum dots with fluorescence quantum yields up to 94%. This is uncommon for a red emission. The high solubility is directly related to the design of the structure, allowing for an accurate description of the photophysical properties of the graphene quantum dots both in solution and at the single molecule level. These photophysical properties were fully predicted by quantum-chemical calculations.


Changes in cardiac biomarkers in association with alterations in cardiac structure and function, and health status in heart failure with reduced ejection fraction: the EVALUATE-HF trial.
N-terminal pro-B-type natriuretic peptide (NT-proBNP), cardiac troponin T (cTnT) and soluble ST2 (sST2) provide complementary prognostic information in heart failure with reduced ejection fraction (HFrEF). We aimed to assess the association between changes in these markers with changes in cardiac structure, function and health status. Patients in the EVALUATE-HF trial (n = 464) were randomized to sacubitril/valsartan or enalapril for 12 weeks, followed by 12-week open-label sacubitril/valsartan. Cardiac biomarkers, echocardiography, and Kansas City Cardiomyopathy Questionnaires (KCCQ) were completed at baseline, and after 12 and 24 weeks. A total of 410 patients (88%) had serial biomarker measurements available (mean age 67 ± 9 years, 75% male and 75% white). After 24 weeks of treatment, NT-proBNP, sST2 and cTnT decreased by median (Q1, Q3) -31% (-55%, +6%), -6% (-19%, +8%) and - 3% (-13%, +8%), respectively (all p < 0.001). Decreases in NT-proBNP were associated with reductions in cardiac volumes and improvements in systolic and diastolic function and health status. Decreases in cTnT were associated with reductions in left ventricular mass, but not with changes in left ventricular function or KCCQ. Decreases in sST2 were consistently associated with improvements in health status, but not with measures of cardiac structure or function. There was no effect modification from treatment on the associations investigated (p for interaction >0.05) CONCLUSION: In HFrEF, serial changes in NT-proBNP correlate with changes in several key measures of cardiac structure and health status. cTnT changes correlate with changes in left ventricular mass and sST2 with changes in health status. These data highlight possible complementary pathophysiologic implications of changes in NT-proBNP, cTnT and sST2. ClinicalTrials.gov Identifier: NCT02874794.


Accelerated evolution at chaperone promoters among Antarctic notothenioid fishes.
Antarctic fishes of the Notothenioidei suborder constitutively upregulate multiple inducible chaperones, a highly derived adaptation that preserves proteostasis in extreme cold, and represent a system for studying the evolution of gene frontloading. We screened for Hsf1-binding sites, as Hsf1 is a master transcription factor of the heat shock response, and highly-conserved non-coding elements within proximal promoters of chaperone genes across 10 Antarctic notothens, 2 subpolar notothens, and 17 perciform fishes. We employed phylogenetic models of molecular evolution to determine whether (i) changes in motifs associated with Hsf1-binding and/or (ii) relaxed purifying selection or exaptation at ancestral cis-regulatory elements coincided with the evolution of chaperone frontloading in Antarctic notothens. Antarctic notothens exhibited significantly fewer Hsf1-binding sites per bp at chaperone promoters than subpolar notothens and Serranoidei, the most closely-related suborder to Notothenioidei included in this study. 90% of chaperone promoters exhibited accelerated substitution rates among Antarctic notothens relative to other perciformes. The proportion of bases undergoing accelerated evolution (i) was significantly greater in Antarctic notothens than in subpolar notothens and Perciformes in 70% of chaperone genes and (ii) increased among bases that were more conserved among perciformes. Lastly, we detected evidence of relaxed purifying selection and exaptation acting on ancestrally conserved cis-regulatory elements in the Antarctic notothen lineage and its major branches. A large degree of turnover has occurred in Notothenioidei at chaperone promoter regions that are conserved among perciform fishes following adaptation to the cooling of the Southern Ocean. Additionally, derived reductions in Hsf1-binding site frequency suggest cis-regulatory modifications to the classical heat shock response. Of note, turnover events within chaperone promoters were less frequent in the ancestral node of Antarctic notothens relative to younger Antarctic lineages. This suggests that cis-regulatory divergence at chaperone promoters may be greater between Antarctic notothen lineages than between subpolar and Antarctic clades. These findings demonstrate that strong selective forces have acted upon cis-regulatory elements of chaperone genes among Antarctic notothens.


White LED Lighting Increases the Root Productivity of Panax ginseng C. A. Meyer in a Hydroponic Cultivation System of a Plant Factory.
To identify effective light spectra for increasing the productivity of Panax ginseng, we conducted experiments in a controlled environment using a hydroponic cultivation system in a plant factory. We investigated the effect of single LEDs (red, blue, and yellow) and mixed LEDs (red + blue and red + blue + white). The relationships between four light spectra (red, blue, yellow, and white) and physiological responses (net photosynthetic rate, stomata conductance, transpiration rate, and intercellular CO2 partial pressure), as well as growth responses (shoot and root biomass), were analyzed using multivariate statistical analysis. Among the four physiological response variables, shoot biomass was not increased by any pathway, and root biomass was increased only by the intercellular CO2 partial pressure. Red and yellow light increased shoot biomass, whereas white light promoted an increase in the net photosynthetic rate and enhanced root biomass. In contrast, blue light was less effective than the other light spectra in increasing both shoot and root biomass. Therefore, red and yellow light are the most effective light spectra for increasing shoot biomass and white light is effective for increasing root biomass in a plant factory that uses artificial LED lighting. Furthermore, the intercellular CO2 partial pressure is an important physiological variable for increasing the root biomass of P. ginseng.


Switched Aβ43 generation in familial Alzheimer's disease with presenilin 1 mutation.
Presenilin (PS) with a genetic mutation generates abundant β-amyloid protein (Aβ) 43. Senile plaques are formed by Aβ43 in the cerebral parenchyma together with Aβ42 at middle ages. These brains cause the early onset of Alzheimer's disease (AD), which is known as familial Alzheimer's disease (FAD). Based on the stepwise processing model of Aβ generation by γ-secretase, we reassessed the levels of Aβs in the cerebrospinal fluid (CSF) of FAD participants. While low levels of Aβ38, Aβ40, and Aβ42 were generated in the CSF of FAD participants, the levels of Aβ43 were unchanged in some of them compared with other participants. We sought to investigate why the level of Aβ43 was unchanged in FAD participants. These characteristics of Aβ generation were observed in the γ-secretase assay in vitro using cells, which express FAD mutations in PS1. Aβ38 and Aβ40 generation from their precursors, Aβ42 and Aβ43, was decreased in PS1 mutants compared with wild-type (WT) PS1, as observed in the CSF. Both the ratios of Aβ38/Aβ42 and Aβ40/Aβ43 in PS1 mutants were lower than those in the WT. However, the ratio of Aβ43/amyloid precursor protein intracellular domain (AICD) increased in the PS1 mutants in an onset age dependency, while other Aβ/AICD ratios were decreased or unchanged. Importantly, liquid chromatography-mass spectrometry found that the generation of Aβ43 was stimulated from Aβ48 in PS1 mutants. This result indicates that PS1 mutants switched the Aβ43 generating line, which reflects the level of Aβ43 in the CSF and forming senile plaques.


Latent synaptic pathways revealed after tetanic stimulation in the hippocampus.
Synaptic plasticity may result from changes at existing synapses or from alterations in the number of functional synaptic connections. In the hippocampus excitatory synaptic strength is persistently enhanced after tetanic stimulation. Here we report that latent synaptic pathways may also become functional. Simultaneous intracellular recordings were made from pairs of CA3 pyramidal cells in slices from guinea pig hippocampus. After stimulating afferent fibres repetitively, polysynaptic excitatory pathways between previously unconnected cells became apparent. The efficacy of recurrent inhibitory circuits was also reduced. The loss of inhibitory control is of interest because latent excitatory pathways are revealed after pharmacological suppression of inhibition. This plasticity in local synaptic circuits leads to the emergence of synchronous firing in groups of CA3 cells. The formation of groups of associated cells and the ability of some cells to initiate synchronous firing in a larger cell group through recurrent pathways is reminiscent of several models of information storage and recall in the cortex.


Escitalopram reversibility of the impacts following chronic stress on central 5-HT profiles - Implications to depression and anxiety.
Stress is considered a crucial determinant influencing health capacity in modern society. Long-term stress makes individuals more susceptible to mental dysfunctions, among which depression and anxiety are two major mental disorders. The success of using selective serotonin reuptake inhibitors (SSRIs) to treat these two disorders highlights the involvement of the central serotonergic (5-HT) system. Later studies suggest both presynaptic and postsynaptic 5-HT profiles should be considered for the effects of SSRIs, making it difficult to interpret the etiological and therapeutic mechanisms underlying depression and anxiety. The present study aims to examine whether the intervention of escitalopram (Es, 5 mg/kg daily for 14 days) can reverse the behavioral phenotypes of both depression-like [by sucrose preference test (SPT) and forced swim test (FST)] and anxiety-like [by avoidance latency and escape latency in elevated-T maze (ETM)] behaviors, and the brain area-dependent neurochemical changes of 5-HT profiles of the terminal regions regarding both synaptic efflux and tissue levels in rats of chronic mild stress (CMS). Our results showed that: (i) Even mild stresses when presented in an unpredictable and long-term manner, can induce both depression-like and anxiety-like behaviors. (ii) Depressive profile indexed by SPT was more sensitive to reflect the Es effect than that of FST. (iii) Es did not significantly affect the CMS-induced anxiety-like symptoms indexed by ETM. (iv) Changes in the protein expression of 5-HT1A receptors in the prefrontal cortex and hippocampus were compatible with the treatment outcome. Our results contributed to the understanding of stress-induced mood dysfunction and the involvement of central 5-HT.


Preoperative Versus Perioperative Risk Factors for Delayed Pain and Opioid Cessation After Total Joint Arthroplasty: A Prospective Cohort Study.
The evolution of pre- versus postoperative risk factors remains unknown in the development of persistent postoperative pain and opioid use. We identified preoperative versus comprehensive perioperative models of delayed pain and opioid cessation after total joint arthroplasty including time-varying postoperative changes in emotional distress. We hypothesized that time-varying longitudinal measures of postoperative psychological distress, as well as pre- and postoperative use of opioids would be the most significant risk factors for both outcomes. A prospective cohort of 188 patients undergoing total hip or knee arthroplasty at Stanford Hospital completed baseline pain, opioid use, and emotional distress assessments. After surgery, a modified Brief Pain Inventory was assessed daily for 3 months, weekly thereafter up to 6 months, and monthly thereafter up to 1 year. Emotional distress and pain catastrophizing were assessed weekly to 6 months, then monthly thereafter. Stepwise multivariate time-varying Cox regression modeled preoperative variables alone, followed by all perioperative variables (before and after surgery) with time to postoperative opioid and pain cessation. The median time to opioid and pain cessation was 54 and 152 days, respectively. Preoperative total daily oral morphine equivalent use (hazard ratio-HR 0.97; 95% confidence interval-CI 0.96-0.98) was significantly associated with delayed postoperative opioid cessation in the perioperative model. In contrast, time-varying postoperative factors: elevated PROMIS (Patient-Reported Outcomes Measurement Information System) depression scores (HR 0.92; 95% CI 0.87-0.98), and higher Pain Catastrophizing Scale scores (HR 0.85; 95% CI 0.75-0.97) were independently associated with delayed postoperative pain resolution in the perioperative model. These findings highlight preoperative opioid use as a key determinant of delayed postoperative opioid cessation, while postoperative elevations in depressive symptoms and pain catastrophizing are associated with persistent pain after total joint arthroplasty providing the rationale for continued risk stratification before and after surgery to identify patients at highest risk for these distinct outcomes. Interventions targeting these perioperative risk factors may prevent prolonged postoperative pain and opioid use.


Qualitative factors shaping MS patients' experiences of infusible disease-modifying drugs: a critical incident technique analysis.
To explore factors shaping the experiences of patients with relapsing-remitting multiple sclerosis with infusible disease-modifying drugs in a hospital setting. The critical incident technique served as a framework for collecting and analysing patients' qualitative account practices involving infusible disease-modifying drugs. Data were collected through semistructured interviews and one single-case study. Participants were recruited from all five regions in Denmark. Inductive thematic analysis was used to identify and interpret factors shaping patients' infusion journey over time. Twenty-two patients with relapsing-remitting multiple sclerosis receiving infusion with disease-modifying drugs (natalizumab, alemtuzumab and ocrelizumab). Four time scenarios-preinfusion, day of infusion, long-term infusion and switch of infusion-associated with the infusion of disease-modifying drugs were analysed to reveal how different factors could both positively and negatively affect patient experience. Time taken to make the treatment decision was affected by participants' subjective perceptions of their disease activity; this may have set off a treatment dilemma in the event of a pressing need for treatment. Planning and routine made infusion practices manageable, but external and internal surroundings, including infusion room ambience and the quality of relationships with healthcare professionals and fellow patients, affected patients' cognitive state and well-being irrespective of the infusion regimen. Switching the infusion regimen can reactivate worries akin to the preinfusion scenario. This study provides novel insight into the positive and negative factors that shape patients' experience of infusion care practices. From a patient's perspective, an infusion practice is not a solitary event in time but includes planning and routine which become an integral part of their multiple sclerosis management. The quality of space and the ambience of the infusion room, combined with the relationship with healthcare professionals and fellow patients, can be a significant source of knowledge and support people with relapsing-remitting multiple sclerosis in their experience of agency in life.


Individualized Exercise Prescription for Athletes Using a Construct-Based Model.
Rehabilitation professionals prescribe exercise regularly with the goals of decreasing pain, increasing function, and returning athletes to competition. To maximize the effect of an exercise intervention, the program must be individualized and in context for the athlete considering biopsychosocial aspects of care. Current models of exercise prescription may not be ideal considering that less than 50% of injured athletes return to their pre-injury level. Advice on exercise prescription has been offered in the past, but the paradigms are either not user friendly or user friendly but linear, based on phases of recovery. As such, there is a need for a more flexible exercise prescription paradigm that should improve the individuality of exercise prescription. In this Current Opinion, we offer a user-friendly construct-oriented paradigm designed to facilitate the creation of individualized exercise programs for athletes.


Update on latest advances in time-of-flight PET.
This paper provides an update on time-of-flight (TOF) PET with a focus on latest hardware developments leading to current commercial PET/CT instruments. We describe advances in scintillator development, new photosensors and associated electronics readout, and detector designs for utilization in complete systems. Next, we introduce the latest commercial PET/CT scanners based on the aforementioned technologies, and discuss the detector design choices made that are relevant to differences in the system performance. Finally, we end with a discussion of the latest performance benchmarks for improved timing in PET detectors, challenges in scaling this performance to a complete system, and the outlook towards achieving a sub-50 ps coincidence timing resolution (CTR) in a PET detector.


Development and validation of a simple high performance thin layer chromatography method combined with direct 1,1-diphenyl-2-picrylhydrazyl assay to quantify free radical scavenging activity in wine.
The aim of this study was to: (a) develop a simple, high performance thin layer chromatographic (HPTLC) method combined with direct 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay to rapidly assess and compare free radical scavenging activity or anti-oxidant activity for major classes of polyphenolics present in wines; and (b) to investigate relationship between free radical scavenging activity to the total polyphenolic content (TPC) and total antioxidant capacity (TAC) in the wine samples. The most potent free radical scavengers that we tested for in the wine samples were found to be resveratrol (polyphenolic non-flavonoid) and rutin (flavonoid), while polyphenolic acids (caffeic acid and gallic acid) although present in all wine samples were found to be less potent free radical scavengers. Therefore, the total antioxidant capacity was mostly affected by the presence of resveratrol and rutin, while total polyphenolic content was mostly influenced by the presence of the less potent free radical scavengers gallic and caffeic acids.


Mothers' postpartum sleep disturbance is associated with the ability to sustain sensitivity toward infants.
Infancy is a period of rapid development when the quality of caregiving behavior may be particularly consequential for children's long-term functioning. During this critical period for caregiving behavior, parents experience changes in their sleep that may affect their ability to provide sensitive care. The current study investigated the association of mothers' sleep disturbance with both levels and trajectories of maternal sensitivity during interactions with their infants. At 18 weeks postpartum, mothers and their infants were observed during a home-based 10-minute "free play" interaction. Mothers' nighttime sleep was objectively measured using actigraphy and subjectively measured using sleep diaries. Maternal sensitivity was coded in two-minute intervals in order to characterize changes in sensitivity across the free play interaction. We used exploratory factor analysis to reduce the dimensionality of the objective and subjective measures of mothers' sleep, identifying a subjective sleep disturbance and an objective sleep continuity factor. Using multi-level modeling, we found that mothers with poorer objective sleep continuity evidenced decreasing sensitivity toward their infants across the interaction. Mothers' self-reports of sleep disturbance were not associated with maternal sensitivity. Although future research is necessary to identify the mechanisms that may explain the observed association between poor sleep continuity and the inability to sustain sensitivity toward infants, mothers' postpartum sleep continuity may be one factor to consider when designing interventions to improve the quality of caregiving. Clinicaltrials.gov, NCT01846585.


Cocaine abuse in pregnancy: effects on the fetus and newborn.
The outcome of infants born to cocaine-using drug dependent women was compared to that of infants of non-cocaine using drug dependent and non-drug dependent women. The study population included 150 pregnant women: 50 women used heroin and methadone plus cocaine, 50 used heroin and methadone minus cocaine, and 50 were non-drug dependent women. Significant differences were found between the cocaine and drug-free groups in infant birth weight, length, head circumference and Apgar scores, with the cocaine group having lower values for each variable. Average gestational age did not vary between the 3 groups. The cocaine group included 1 spontaneous abortion and 4 fetal deaths; non-cocaine drug dependent women had 2 fetal deaths, with none in the control group. Mean abstinence scores for 19 of the physiological and behavioral parameters were lower in the cocaine group than in the non-cocaine drug dependent women with the exception of vomiting and convulsions. These data suggest that: (1) infants born to drug dependent women have a poorer general outcome than those born to non-drug dependent women; (2) maternal cocaine use does not appear to increase the incidence of severe neonatal abstinence symptomatology; (3) pregnancies complicated by cocaine abuse have a greater chance for fetal loss resulting from both spontaneous abortions and fetal death; (4) infants born to cocaine abusing women had infants with decreased birth weight, head circumference, length and Apgar scores.


Two perspectives of proxy reporting of health-related quality of life using the Euroqol-5D, an investigation of agreement.
Proxy-reporting has been proposed as an alternative to self-report of health-related quality of life (HRQoL) for patients with poor cognition. There are 2 possible perspectives from which to complete a proxy-report, answer as the patient would (proxy-patient) or from the proxy's own perspective (proxy-proxy). Most research has not differentiated between perspectives. Agreement between patient and proxy-reports from either perspective has not been investigated using the Euroqol-5D (EQ-5D) among elderly hospital patients undergoing rehabilitation. Identify agreement levels between proxy-patient and patient self-report as well as proxy-proxy and patient self-report of the EQ-5D and investigate interaction effects of timing (admission vs. discharge) and basic cognition (intact vs. not intact). Repeated measures, inter-rater agreement investigation of clinician proxy-report, and patient self-report incorporating; proxy-patient reports (perspective A) and proxy-proxy reports (perspective B). Geriatric rehabilitation patients (n = 272) and their proxies (treating physiotherapists n = 29). EQ-5D for HRQoL and Mini Mental State Examination for cognition. One hundred fifty (89%) proxy-patient and 130 (98%) proxy-proxy datasets were complete, 51 perspective A and 52 perspective B patients did not have basic cognition intact. Proxy-patient assessments had strong agreement with self-report at discharge across all cognition levels (kappa = 0.76-0.95), but at admission had stronger agreement among patients with better cognition (kappa = 0.70-0.86) than patients with lower cognition (kappa = 0.47-0.76). At admission and discharge proxy-proxy assessments generally had moderate agreement with self-report among patients with poor cognition on most domains with proxies giving lower scores than patients (kappa = 0.23-0.81), this is in contrast to proxy-proxy assessments and patients with better cognition (kappa = 0.55-0.95). Clinician (physiotherapist) proxy-reports among this population generally had good agreement with patient self-report though this was affected by proxy perspective, patient cognition, and timing.


Effect of propranolol on accumulation of NEFA in the ischemic perfused rat heart.
The effect of propranolol on the accumulation of non-esterified fatty acids (NEFA) in the isolated, perfused working rat heart was investigated. Ischemia was induced by lowering the afterload pressure to 0 mm Hg for 20 min and for reperfusion, the pressure was raised to the pre-ischemic pressure (60 mm Hg) for 20 min. The heart was frozen for biochemical studies immediately after ischemia or reperfusion. Ischemia decreased mechanical function, increased the levels of palmitoleic, arachidonic and linoleic acids, left oleic, lauric, myristic, palmitic and stearic acids unchanged, decreased the levels of adenosine triphosphate (ATP), creatine phosphate (CrP) and the energy charge potential (ECP), and increased the level of lactate. Propranolol (10(-5) or 3 x 10(-5) M) restored mechanical function and inhibited the changes in NEFA, ATP and ECP caused by ischemia. It is suggested that propranolol inhibits the decrease in mechanical function and high energy phosphates caused by ischemia, and thereby inhibits the accumulation of NEFA, especially of the unsaturated fatty acids such as arachidonic acid, during ischemia.


The incidence of tuberculosis among hiv-positive individuals with high CD4 counts: implications for policy.
Intensified case finding (ICF) and earlier antiretroviral therapy (ART) initiation are strategies to reduce burden of HIV-associated tuberculosis (TB). We describe incidence of and associated factors for TB among HIV-positive individuals with CD4 counts > 350 cells/μl in South Africa. Prospective cohort study of individuals recruited for a TB vaccine trial. Eligible individuals without prevalent TB were followed up at 6 and 12 months after enrolment. Cox proportional hazards regression was used to determine factors associated with risk of incident TB. Six hundred thirty-four individuals were included in the analysis [80.9 % female, 57.9 % on ART, median CD4 count 562 cells/μl (IQR 466-694 cells/μl)]. TB incidence was 2.7 per 100 person-years (pyrs) (95 % CI 1.6-4.4 per 100 pyrs) and did not differ significantly between those on ART and those not on ART [HR 0.65 (95 % CI 0.24-1.81)]. Low body mass index (BMI <18.5 kg/m(2)) was associated with incident TB [HR 3.87 (95 % CI 1.09-13.73)]. Half of the cases occurred in the first 6 months of follow up and may have been prevalent or incubating cases at enrolment. TB incidence was high and associated with low BMI. Intensified case finding for TB should be strengthened for all HIV positive individuals regardless of their CD4 count or ART status.


An improved silver stain for the visualization of lipopolysaccharides on polyacrylamide gels.
A sensitive, brief, and user-friendly silver stain to meet the needs in high-efficiency detection of lipopolysaccharides (LPS) on polyacrylamide gels is described. In this study, the most commonly used formaldehyde-based LPS silver stain, which is potentially hazardous to the operator, is replaced by ascorbic acid (Vc) in alkaline sodium thiosulfate solution. It takes only about 35 min to complete all the protocol, with a detection limit of 4 ng of total LPS. The results indicate that this user-friendly method could be a good choice for LPS visualization on polyacrylamide gels.


Impaired activation of celiac ganglion neurons in vivo after damage to their sympathetic nerve terminals.
Because damage to sympathetic nerve terminals occurs in a variety of diseases, we tested the hypothesis that nerve terminal damage per se is sufficient to impair ganglionic neurotransmission in vivo. First, we measured the effect of nerve terminal damage produced by the sympathetic nerve terminal toxin 6-hydroxydopamine (6-OHDA) on ganglionic levels of several neurotrophins thought to promote neurotransmission. 6-OHDA-induced nerve terminal damage did not decrease the expression of neurotrophin-4 or brain-derived neurotrophic factor mRNA in the celiac ganglia but did decrease the ganglionic content of both nerve growth factor protein (nadir = -63%) and the mRNA of the alpha-3 subunit of the nicotinic cholinergic receptor (nadir = -49%), a subunit required for neurotransmission. Next, we tested whether this degree of receptor deficiency was sufficient to impair activation of celiac ganglia neurons. Impaired fos mRNA responses to nicotine administration in the celiac ganglia of 6-OHDA-pretreated rats correlated temporally with suppressed expression of functional nicotinic receptors. We verified by Fos protein immunohistochemistry that this ganglionic impairment was specific to principal ganglionic neurons. Last, we tested whether centrally initiated ganglionic neurotransmission is also impaired following nerve terminal damage. The principal neurons in rat celiac ganglia were reflexively activated by 2-deoxy-glucose-induced glucopenia, and the Fos response in the celiac ganglia was markedly inhibited by pretreatment with 6-OHDA. We conclude that sympathetic nerve terminal damage per se is sufficient to impair ganglionic neurotransmission in vivo and that decreased nicotinic receptor production is a likely mediator.


Persistent tubular conduction in vacuolated amphibian skeletal muscle following osmotic shock.
The transverse (T-)tubules primarily function in conducting the action potentials that initiate excitation contraction coupling in skeletal muscle but may additionally subserve longer-term roles in volume regulation, membrane fusion and other trafficking processes. Osmotic shock thus both electrically detaches the T-tubules from surface membrane ('detubulation') and produces tubular vacuolation. The present experiments separated these effects. An established, reference osmotic shock protocol that exposed muscles to Ca2+/Mg2+-Ringer and gradual cooling to 10 degrees C after 18 min in glycerol-Ringer accomplished significant detubulation (77.5+/-13.15%, mean +/- SEM; n = 4). In contrast, a test protocol conducted entirely at room temperature using Mg2+-rather than Ca2+/ Mg2+-Ringer yielded reduced (P < 0.05, post hoc Duncan's multiple range test) detubulation indices (1.67+/-1.67%, n = 6) statistically indistinguishable from findings in fibres spared osmotic shock. Yet both osmotic shocks caused a formation of closed vacuoles, demonstrated by Sulphorhodamine B trapping, that occupied statistically similar fractions of total fibre volume (reference procedure: 14.38+/-2.7%, n = 6; test procedure: 13.36+/-2.00%, n = 22) in turn higher than determinations in control fibres (P < 0.05). The findings reconcile reports associating detubulation with vacuolation in osmotically shocked muscle [S. Nik-Zainal et al. (1999) J Muscle Res Cell Motil 20: 45-53; K.N. Khan et al. (2000) J Muscle Res Cell Motil 21: 79-90] with the persistence of tubular electrical activity in extensively vacuolated amphibian fibres following fatigue [J. Lannergren and H. Westerblad (1987) Acta Physiol Scand 129: 311-318; J. Lannergren et al. (1999) J Muscle Res Cell Motil 20: 19-32]. Furthermore test protocols produced higher densities of open vacuoles (13.38+/-2.33%, n = 9) than did reference protocols (6.66+/-1.63%, n = 20) contrary to their possible involvement in the electrophysiological changes. Abolition of tubular electrophysiological activity thus either follows or is independent of tubular vacuolation whilst sharing some of its underlying osmotic mechanisms.