Diagnostic performance of multislice CT coronary angiography in the assessment of significant coronary artery disease.
The use of noninvasive assessment tools such as multi-slice CT coronary angiography (MSCT-CA-CA) is recently considered mainly because it offers safety, patient convenience, and faster performance. The aim of the present study was to determine the ability of MSCT-CA-CA for the detection of significant stenoses in the coronary arteries, in comparison to conventional invasive coronary angiography (ICA). A total of 58 consecutive patients who were candidate for coronary angiography, with the diagnosis of acute coronary syndrome, from September 2006 to March 2006 were entered into the study. They underwent both coronary MSCT-CA-CA and ICA. The findings of each coronary segment were compared to MSCT-CA-CA in comparison with ICA. Based on artery analysis, sensitivity and specificity of MSCT-CA for the detection of involvement in RCA were 90.0% and 92.8%, in LAD were 71.8% and 92.9% and in LCX were 67.9% and 92.6%, respectively. On a per-segment basis, the sensitivity of MSCT-CA in the detection of injured segments ranged between 33.3% (for segment 11) and 100% (for segments 1, 2 and 12). Also, specificity ranged from 63.6% (for segment 15) and 98.1% (for segment 6). The presence of hypertension, hyperlipidemia, and smoking led to the reduction of the specificity and accuracy of MSCT-CA, whereas history of diabetes mellitus could increase the specificity and accuracy of this tool. MSCT-CA has high diagnostic performance in the assessment of significant coronary artery disease. Risk factors for coronary artery disease may influence this performance.


Characterization of promoter elements mediating ethanol regulation of hsc70 gene transcription.
Chronic exposure to ethanol increases transcription of the molecular chaperone Hsc70 in NG108-15 neuroblastoma X glioma cells. This and other ethanol-induced changes in gene expression may contribute to central nervous system tolerance and dependence in alcoholics. Here, we characterized sequences in the hsc70 promoter that are required for ethanol-induced transcriptional regulation. Deletion analysis of the hsc70 promoter showed that the 74-base pair region proximal to the transcription start site was sufficient for ethanol responsiveness. Point mutation or deletion of a consensus Spl-binding site at -67/-61 base pairs greatly reduced the induction by ethanol. Hsc70 promoter constructs with diminished ethanol responsiveness in NG108-15 cells similarly had decreased transcriptional activation by exogenous Sp1 in Drosophila SL2 cells. Some artificial promoter constructs containing multiple Sp1 sites were highly responsive to ethanol, but others were not, suggesting that the organization of the proximal promoter region was an additional factor that affected the ethanol response. Gel mobility shift analysis confirmed that an Sp1-like protein bound to the -67/-61 consensus Sp1 site. However ethanol exposure did not alter Sp1 DNA-binding activity. Together, our findings show that ethanol induction of Hsc70 requires a functional Sp1-binding site. Additional proximal promoter elements may also play a role in determining whether an Sp1-containing promoter will respond to ethanol.


A method for estimating urban background concentrations in support of hybrid air pollution modeling for environmental health studies.
Exposure studies rely on detailed characterization of air quality, either from sparsely located routine ambient monitors or from central monitoring sites that may lack spatial representativeness. Alternatively, some studies use models of various complexities to characterize local-scale air quality, but often with poor representation of background concentrations. A hybrid approach that addresses this drawback combines a regional-scale model to provide background concentrations and a local-scale model to assess impacts of local sources. However, this approach may double-count sources in the study regions. To address these limitations, we carefully define the background concentration as the concentration that would be measured if local sources were not present, and to estimate these background concentrations we developed a novel technique that combines space-time ordinary kriging (STOK) of observations with outputs from a detailed chemistry-transport model with local sources zeroed out. We applied this technique to support an exposure study in Detroit, Michigan, for several pollutants (including NOx and PM2.5), and evaluated the estimated hybrid concentrations (calculated by combining the background estimates that addresses this issue of double counting with local-scale dispersion model estimates) using observations. Our results demonstrate the strength of this approach specifically by eliminating the problem of double-counting reported in previous hybrid modeling approaches leading to improved estimates of background concentrations, and further highlight the relative importance of NOx vs. PM2.5 in their relative contributions to total concentrations. While a key limitation of this approach is the requirement for another detailed model simulation to avoid double-counting, STOK improves the overall characterization of background concentrations at very fine spatial scales.


Autoresuscitation: A Case and Discussion of the Lazarus Phenomenon.
Lazarus phenomenon or autoresuscitation is a very rare condition defined as delayed unassisted return of spontaneous circulation after cessation of cardiopulmonary resuscitation. Based on a case with a 67-year-old male who came back to life after discontinuation of cardiopulmonary resuscitation, we discuss the background and possible countermeasures related to the Lazarus phenomenon.


In vitro activity of azlocillin, carbenicillin, mezlocillin and piperacillin against Pseudomonas aeruginosa.
50% inhibitory concentration (IC50) of azlocillin, carbenicillin, mezlocillin and piperacillin against 157 strains of P. aeruginosa were determined by means of the agar plate-dilution method. The 3 new semisynthetic penicillins were significantly more active against P. aeruginosa than carbenicillin; azlocillin and piperacillin were 8-fold, and mezlocillin was 2-4-fold more active than carbenicillin. More than 95% of the P. aeruginosa strains were inhibited by 16 micrograms/ml of azlocillin or piperacillin whereas 64 micrograms/ml of mezlocillin and 256 micrograms/ml of carbenicillin, respectively, were necessary to inhibit 95% of the strains.


Prevention of adhesions in surgery of the flexor tendons of the hand: what is the evidence?
Despite advances in knowledge and refinements of technique, the management of flexor tendon injuries within the digital sheath continues to present a formidable challenge. This in turn has led to a massive expansion in search of modified surgical therapies and various adjuvant therapies, which could prevent adhesion formation without compromising digital function. A search of PubMed, Medline, CINAHL and Embase databases was performed using the keywords 'tendon adhesion prevention', 'tendon healing', 'adhesion prevention in tendons' and 'adjuvants for adhesion prevention'. Studies detailing the use of surgical, pharmacological and non-pharmacological agents for adhesion prevention in digital flexor tendons were identified, and their bibliographies were thoroughly reviewed to identify further related articles. This search identified 41 studies, which investigated the use of various pharmacological agents in adhesion prevention in digital tendons. There is a need to develop and utilize an optimal method for the prevention of adhesions in the flexor tendons of the hand, due to post-surgical complications. Even though there have been significant advances in the prevention of adhesions in flexor tendons, it remains to be proved which, if any, of the current methods are the most beneficial. The only thing that appears clinically justified in adhesion prevention is the need for early post-operative mobilization of digits after tendon injury or repair but the best method of mobilization remains controversial. Suggested changes in surgical techniques and various proposed pharmacological and non-pharmacological modalities need to withstand the test of adequately powered human trials, before their justification for potential benefit in clinical practice is accepted.


Effects of gradual increases in o(2) concentration on nodule activity in soybean.
The objectives of this study were to determine whether attached nodules of soybean (Glycine max L. Merr.) could adjust to gradual increases in rhizosphere pO(2) without nitrogenase inhibition and to determine whether the nitrogenase activity of the nodules is limited by pO(2) under ambient conditions. A computer-controlled gas blending apparatus was used to produce linear increases (ramps) in pO(2) around attached nodulated roots of soybean plants in an open gas exchange system. Nitrogenase activity (H(2) production in N(2):O(2) and Ar:O(2)) and respiration (CO(2) evolution) were monitored continuously as pO(2) was ramped from 20 to 30 kilopascals over periods of 0, 5, 10, 15, and 30 minutes. The 0, 5, and 10 minute ramps caused inhibitions of nitrogenase and respiration rates followed by recoveries of these rates to their initial values within 30 minutes. Distinct oscillations in nitrogenase activity and respiration were observed during the recovery period, and the possible basis for these oscillations is discussed. The 15 and 30 minute ramps did not inhibit nitrogenase activity, suggesting that such inhibition is not a factor in the regulation of nodule diffusion resistance. During the 30 minute ramp, a stimulation of nitrogenase activity was observed, indicating that an O(2)-based limitation to nitrogenase activity occurs in soybean nodules under ambient conditions.


Climate change and community disassembly: impacts of warming on tropical and temperate montane community structure.
Both tropical and temperate species are responding to global warming through range shifts, but our understanding of the consequences of these shifts for whole communities is limited. Here, we use current elevational range data for six taxonomic groups spanning 90° in latitude to examine the potential impacts of climate-driven range shifts on community change, or 'disassembly', across latitude. Elevational ranges are smaller at low latitudes for most groups and, as a consequence, tropical communities appear to be more sensitive to temperature increases compared with temperate communities. Under site-specific temperature projections, we generally found greater community disassembly in tropical compared with temperate communities, although this varied by dispersal assumptions. Mountain height can impact the amount of community disassembly, with greater change occurring on smaller mountains. Finally, projected community disassembly was higher for ectotherms than endotherms, although the variation among ectotherms was greater than the variation separating endotherms and ectotherms.


Transcranial Doppler evaluation of cerebrovascular reactivity to acetazolamide in normal subjects.
Acetazolamide (AZ), the selective inhibitor of carbonic anhydrase, was proved by intravenous Xenon 133 technique to increase cerebral blood flow (CBF). In this study cerebrovascular reactivity to AZ was evaluated in 10 normal subjects by transcranial Doppler (TCD) within the middle cerebral artery (MCA), since several reports have demonstrated that velocity of cerebral blood flow is well correlated to CBF. After 1 gr AZ injection blood flow velocity markedly increased in all subjects, with a peak in both systolic and diastolic velocity 20 min after drug administration. At that time systolic velocity increased by 35% and diastolic velocity by 50% in comparison to basal values. In contrast with Xenon 133 technique which gives one measurement only for each investigation, TCD allows a continuous monitoring of haemodynamic change following AZ infusion. MCA diastolic velocity at rest was inversely related to age (r = -.804, p < 0.01); baseline diastolic velocity was inversely related to the maximum percentage increment (r = -.745 p < .05). No change in blood pressure and heart rate was observed under experimental conditions. These results, confirm the usefulness of the AZ test in the evaluation of cerebrovascular reactivity and strongly support the use of TCD technique applied to AZ in order to investigate cerebrovascular haemodynamics in normal healthy subjects and in patients at risk of cerebrovascular insufficiency.


The Usefulness of Ultrasound Surveillance for Axillary Recurrence in Women With Personal History of Breast Cancer.
To evaluate the axillary recurrence rate and usefulness of axillary ultrasound (AUS) during supplementary whole-breast ultrasound (US) screening in women with a personal history of breast cancer (PHBC). A retrospective database search identified consecutive asymptomatic women who underwent postoperative supplemental whole-breast US screening, including that of the bilateral axillae, after negative findings on mammography between January and June 2017. Using the pathologic data or at least 1-year follow-up data as reference standards, the axillary recurrence rate, cancer detection rate (CDR), interval axillary recurrence rate per 1,000 screenings, sensitivity, specificity, and abnormal interpretation rate (AIR) were estimated. From the data of 4,430 women (mean age, 55.0 ± 10.1 years) analyzed in this study, there were five axillary recurrence cases (1.1/1,000) in the median follow-up period of 57.2 months. AUS showed a CDR of 0.2 (1/4,430; 95% confidence interval [CI], 0.01-1.3) and an interval axillary recurrence rate of 0.9 (4/4,402; 95% CI, 0.2-2.3) per 1,000 examinations. The sensitivity and specificity were 20.0% (1/5; 95% CI, 0.5-71.6), and 99.4% (4,398/4,425; 95% CI, 99.1-99.6), respectively, while the AIR was 0.6% (28/4,430; 95% CI, 0.4-0.9%). In asymptomatic women with a PHBC and negative findings on mammography, axillary recurrence after breast cancer and axillary treatment was uncommon, and the supplemental AUS screening yielded 0.2 cancers per 1,000 examinations.


Racial variation in spontaneous fetal deaths at 20 weeks or older in upstate New York, 1980-86.
The distribution of spontaneous fetal deaths (at age 20 weeks or more) by maternal race has received considerably less study than other adverse pregnancy outcomes. The purpose of this study was twofold--(a) to describe spontaneous fetal deaths among white, black, and American Indian women and (b) to determine if there was any variation by International Classification of Diseases, Ninth Revision (ICD-9) cause of death, gestational age at death, or maternal age at loss among these groups of mothers. Using the fetal death certificate registry maintained by the New York State Department of Health, 8,592 spontaneous fetal deaths at age 20 weeks or more were identified among upstate (exclusive of New York City) mothers between 1980 and 1986. By race it was 7,300 for white women, 1,257 for black women, and 27 for American Indian women. Spontaneous fetal death rates varied by maternal race as listed on vital records--black, 13.5 per 1,000 total births, white, 8.3, and American Indian, 8.1. The three leading causes of death (ICD-9,779, 762, and 761) did not vary by maternal race. Gestational age at death, imputed from last menstrual period, did vary by maternal race. Fetal deaths to white and black mothers were observed to occur most often between 24 weeks of pregnancy (39 percent) and 32 weeks (43 percent), while American Indian fetal deaths generally occurred later (more than 33 weeks) in pregnancy (41 percent). Most spontaneous fetal deaths occurred to mothers ages 20-29 regardless of race. Black teenage mothers, however, experienced the largest proportion of losses(23 percent) compared with white (10 percent) and American Indian (I I percent) teenage mothers.


Surgical valvotomy versus balloon dilatation for children with severe aortic valve stenosis: a systematic review.
Aim: To evaluate outcomes of interventions for severe aortic valve stenosis (AS), whether it is done by surgical aortic valvotomy (SAV) or balloon aortic dilatation (BAD). Results: Eleven studies with total number of 1733 patients; 743 patients had SAV, while 990 patients received BAD. There was no significant difference in early mortality (odds ratio [OR]: 0.96, p = 0.86), late mortality (OR: 1.28, p = 0.25), total mortality (OR: 1.10, p = 0.56), and freedom from aortic valve replacement (OR: 1.00, p = 1.00). Reduction of aortic systolic gradient was significantly higher in the SAV group (OR: 2.24, p = 0.00001), and postprocedural AR rate was lower in SAV group (OR: 0.21, p = 0.00001). Conclusion: SAV is associated with better reduction of aortic systolic gradient and lesser post procedural AR which reduce when compared with BAD.


The relationship of child neglect and physical maltreatment to placement outcomes and behavioral adjustment in children in foster care: a Canadian perspective.
Dramatic increases in child welfare rates in Canada over recent years have been largely driven by an increased reporting of neglect cases (Trocmé, Fallon, MacLaurin, & Neves, 2005). To a large extent, exploring the importance of neglect separate from physical maltreatment has been ignored in the child maltreatment literature. This study examined the differential effects of foster care in the child welfare system with children who presented as either experiencing physical maltreatment or neglect prior to their admission to care. Findings from this study are important to child welfare decision making about the differential needs of these two groups of children. The files of a sample of 110 children (79 neglected children and 31 physically maltreated children) were examined for differences in their adjustment while in foster care and on discharge. Some distinct differences in presentation were noted between the children experiencing the two types of maltreatment. Children experiencing neglect were younger, were more likely to have caregivers diagnosed with a substance abuse disorder, and had higher rates of exposure to spousal violence than maltreated children. Physically maltreated children displayed greater difficulty during their foster care adjustment. Once discharged from care, neglected children were more likely to be returned to the care of the agency. This study draws attention to the differential needs of children who experience neglect prior to their admission to a child welfare agency. Longer-term outcome studies are necessary to more completely understand how these two types of maltreatment influence the outcomes of children who are provided care within the child welfare system.


Cell metabolism: an essential link between cell growth and apoptosis.
Growth factor-stimulated or cancerous cells require sufficient nutrients to meet the metabolic demands of cell growth and division. If nutrients are insufficient, metabolic checkpoints are triggered that lead to cell cycle arrest and the activation of the intrinsic apoptotic cascade through a process dependent on the Bcl-2 family of proteins. Given the connections between metabolism and apoptosis, the notion of targeting metabolism to induce cell death in cancer cells has recently garnered much attention. However, the signaling pathways by which metabolic stresses induce apoptosis have not as of yet been fully elucidated. Thus, the best approach to this promising therapeutic avenue remains unclear. This review will discuss the intricate links between metabolism, growth, and intrinsic apoptosis and will consider ways in which manipulation of metabolism might be exploited to promote apoptotic cell death in cancer cells. This article is part of a Special Issue entitled Mitochondria: the deadly organelle.


The Static Stiffness Index: an important parameter to characterise compression therapy in vivo.
The compression pressure, which corresponds to the dosage of compression therapy, has been widely neglected up to now, not only concerning scientific literature, but also in clinical practice. It is evident that compression pressures in the upright position and during walking are clinically more relevant than just the resting pressure. The Static Stiffness Index (SSI), which is the difference between standing and resting pressure, is a valuable parameter characterising the efficacy of a specific compression product to narrow/occlude the venous lumen. This is a prerequisite for reducing venous reflux and exerting a massaging effect necessary to improve the venous pumping function during movement. This article provides an overview of the recent literature on the SSI, which supports the recommendations of the International Compression Club. In addition, it aims to provide an insight on the importance of the SSI in daily practice, as an educational tool as well as in defining the properties of applied compression therapy in clinical research.


Biodegradable gelatin-based films with nisin and EDTA that inhibit Escherichia coli.
In this study, we developed gelatin-based films for active packaging with the ability to inhibit E. coli. We created these novel biodegradable gelatin-based films with a nisin-EDTA mix. FT-IR, TGA, and SEM analysis showed that nisin interacted with the gelatin by modifying its thermal stability and morphology. The use of nisin (2,500 IU/mL) with concentrations of Na-EDTA (1.052 M stock solution) distributed in the polymer matrix generated a significant decrease in the growth of E. coli when compared to the control. In freshly made films (t0), the growth of E. coli ATCC 25922 was reduced by approximately 3 logarithmic cycles. Two weeks after the films were made, a reduction in antimicrobial activity was observed in approximately 1, 1 and 3 logarithmic cycles of the films with 5%, 10% and 20% of the compound (nisin/Na-EDTA) distributed in the polymer matrix, respectively. This evidences an antimicrobial effect over time. Also, biodegradation tests showed that the films were completely degraded after 10 days. With all these results, an active and biodegradable packaging was successfully obtained to be potentially applied in perishable foods. These biodegradable, gelatin-based films are a versatile active packaging option. Further research on the barrier properties of these films is needed.


Oxidative stress and inflammatory response biomarkers in dogs with mammary carcinoma.
Mammary carcinoma is the most common cancer that affects dogs, and in many cases it leads to death. Thus, given the importance of this disease, to clarify its pathogenesis is an important measure. In this sense, the aim of this study was to investigate the levels of cytokines and nitric oxide (NO), oxidative and antioxidant status, as well as the activity of adenosine deaminase (ADA) and butyrylcholinesterase (BChE) in dogs diagnosed with mammary carcinoma. With this purpose, thirty-three (33) serum samples from female dogs with histopathological diagnosis of mammary carcinoma, without evidence of metastasis, were used (group B). The material was classified based on the degree of malignancy, as follows: subgroup B1 (low-grade malignancy; n=26) and subgroup B2 (high grade of malignancy; n=7). Serum samples from healthy females (group A; n=10) were used as negative control. Our results showed that levels of cytokines (TNF-α, INF-γ, IL-1, and IL-6), NOx (nitrite/nitrate), AOPP (protein oxidation), and FRAP (antioxidant power) were significantly (P<0.05) increased in dogs with mammary carcinoma (group B), when compared with group A. On the other hand, ADA activity was significantly decreased (P<0.05) in both subgroups B1 and B2, when compared with group A. BChE activity, however, was reduced (P<0.05) only in subgroup B2 when compared with group A and subgroup B1. Unlike other variables, NO, AOPP, and IFN-γ were influenced by the degree of tumor malignancy, i.e., their levels were even higher in subgroup B2. Therefore, based on these results, we can conclude that all variables investigated are related to the pathogenesis of this disease, since they were altered in dogs with mammary tumor. Additionally, we suggest that ADA activity had an anti-inflammatory effect on these tumor samples, probably in order to modulate the inflammatory response.


Adenosine 5'-triphosphate is the predominant source of peripheral adenosine in human B lymphoblasts.
Adenosine 5'-triphosphate (ATP) and adenosine are the crucial endogenous signaling molecules in immunity and inflammation. In this study we identified the source of extracellular adenosine in human B lymphoblasts, and evaluate the ATP release and metabolism. We observed that the B cells continuously released substantial quantities of ATP (35 pmol/10(6) cells) when subjected to slow motion in the incubation medium. The adenosine level in the B cell incubation medium was very low, and increased (5-fold) upon inhibition of adenosine deaminase activity with 10 muM of 2-deoxycoformycin (DCF). Inclusion of an inhibitor of equilibrate nucleoside transport (nitrobenzylthioinosine) in the incubation medium in the presence of DCF resulted in the elevation of adenosine level by 9-fold. Inhibition of ecto-ATPase activity with 100 muM of ARL67156 was associated with a 2-fold increase of the extracellular ATP level and a 3-fold decrease of adenosine concentration in the cell culture media. Inclusion of alpha,alpha-methyleneadenosine 5'-diphosphate, a selective inhibitor of ecto-5'-nucleotidase in the incubation medium resulted in a significant decrease (7-fold) the adenosine concentration. In conclusion, our results indicate that ATP released from the B cell is the primary source of peripheral adenosine, and that the activities of ecto enzymes and the efficiency of Ado uptake through the nucleoside transporters determine the Ado level on the B cell surface.


Myocarditis complicated by massive right ventricular thrombus and extensive pulmonary embolism: A case report.
An intracardiac thrombus may develop as a consequence of myocarditis, and in rare cases, a dominantly right ventricular thrombus develops, which may impair cardiac function and even cause life-threatening cardiovascular events. We report a 24-year-old man presented with recurrent episodes of palpitation and precordial discomfort after catching a cold 2 months ago. Transthoracic echocardiography (TTE) and computed tomography pulmonary angiogram (CTPA) revealed a mass attached to the apex of the right ventricle and extensive bilateral pulmonary artery emboli. There was no indication where the thrombi originated from in this young patient without any underlying disease except myocarditis. Pulmonary endarterectomy and embolectomy of pulmonary arteries and right ventricle were performed. Postoperative pathological results confirmed the presence of fibrinous necrosis and hemosiderin deposition. The formation of an intraventricular thrombus is closely related to myocarditis, which can affect individuals of all ages, but especially young people. Thus, patients with myocarditis should be closely monitored and followed up because of the increased risk of extensive thrombosis.


Effects of baby-friendly hospital initiative on breast-feeding practices in sindh.
To determine changes in the breastfeeding practices of mothers after receiving counseling on 'Ten Steps to Successful Breastfeeding' as defined by the Baby Friendly Hospital Initiative comparing baby friendly hospitals (BFHs) and non-baby-friendly hospitals in Sindh, Pakistan. The observational study was conducted from June 2007 to June 2009 in randomly selected baby-friendly and non-baby-friendly hospitals of Sindh, Pakistan. Non-probability purposive sampling was employed.The maternity staff was trained on 'Ten Steps to Successful Breastfeeding.'The changes in breastfeeding practices were analysed by SPSS version 15. A total of 236 women were included in the study. Of them, 196 (83.05%) were from baby-friendly hospitals and 40 (16.94%) from non-baby-friendly hospitals. Besides, 174 (88.7%) mothers in baby-friendly hospitals and 5 (12.5%) in non-baby-friendly hospitals during antenatal care received counseling by healthcare providers.There was an increase in breastfeeding practice up to 194 (98.97%) in the first category compared to 12 (30%) in the other category. Counseling under the Baby Friendly Hospital Initiative improved breastfeeding practices up to 98.97% in baby-friendly compared to non-baby-friendly hospitals.


Neonatal alloimmune thrombocytopenia due to anti-HPA-2b (anti-Koa).
Most severe cases of neonatal alloimmune thrombocytopenia (NAIT) are due to anti-HPA-1a (anti-PlA1) antibodies. We report a case of NAIT due to anti-HPA-2b that resulted in in utero intracranial hemorrhage.A 33-year-old G2P1A0 Caucasian woman had a routine ultrasound at 34 weeks. The fetus appeared to have a left hemispheric hematoma. IVIG, 1g/kg, was started immediately and administered weekly until delivery. One day after receiving the first dose of IVIG, fetal platelet count was 18 x 10(9)/L, and Hb was 116 g/L. Eleven mL of matched platelets compatible by monoclonal antibody immobilization of platelet antigens (MAIPA) assay were transfused in utero, raising the platelet count to 62 x 10(9)/L. Repeat transfusions were done later that week and 1 week later, with pretransfusion counts of 19 x 10(9)/L and 16 x 10(9)/L, respectively. Delivery by C section was done at 35.5 weeks, after the third platelet transfusion. Platelet count at birth was 77 x 10(9)/L. Drainage of the hematoma was performed after transfusion. Testing with a solid phase ELISA revealed reactivity against GP1b/IX. MAIPA testing after platelet treatment with the protease inhibitor leupeptin demonstrated the presence of anti-HPA-2b. On PCR-SSP the mother was HPA-2a homozygous, the father was HPA-2a/2b. Antibodies against the HPA-2b antigen located on the GP1b/IX complex have been reported in rare cases of NAIT. Testing is complicated by proteolytic degradation of the antigen-bearing fragment. Compatible platelets are easily found since approximately 85 percent of donors are HPA-2a/2a.


Endocrine and spermatological characteristics of 135 patients with bilateral megalotestis.
Little is known about the biological and clinical significance of bilaterally enlarged testes (bilateral megalotestes or macroorchidism). We have investigated clinical, endocrine and spermatological characteristics of 135 andrological patients with a testicular volume of more than 25 ml bilaterally. 135 patients with normal testicular volumes served as controls. Subjects with bilateral megalotestis had higher sperm counts and better sperm morphology and motility than the controls. Levels of LH, FSH, prolactin, testosterone and estradiol were lower in the megalotestis group. This was statistically significant for LH and FSH. No testicular tumours were diagnosed in either group. We conclude that in most andrological patients with bilateral megalotestes fertility parameters are excellent despite the unusually low hormone levels found in a significant minority. No specific pathology underlying the large gonadal volume could be identified.


Increased thrombin generation in women with polycystic ovary syndrome: A pilot study on the effect of metformin and oral contraceptives.
Polycystic ovary syndrome (PCOS) is associated with risk factors for cardiovascular disease (CVD) which may be modified by the use of metformin and oral contraceptives (OC). Thrombin generation (TG) measures are risk markers of CVD and address the composite of multiple factors that influence blood coagulation. This prospective, randomized, intervention study evaluated the potential influence of PCOS on TG measures and the effect of OC and/or metformin on TG measures in women with PCOS. Ninety patients with PCOS and 35 controls were included. Patients were randomized to 12 months of treatment with metformin, metformin+OC or OC alone. C-reactive protein (CRP), fibrinogen, total cholesterol, trunk fat mass, body mass index, estradiol, testosterone, sex hormone binding globulin (SHBG) as well as TG measures, i.e. the lag time for formation of thrombin, the endogenous thrombin potential (ETP), peak thrombin concentration (peak) and time to peak were determined at baseline and after 12 months of treatment. CRP and total testosterone were significantly higher and SHBG significantly lower in PCOS women than in controls (P=0.012, P<0.001 and P=0.008, respectively). The TG measures ETP, peak and lag time were increased in women with PCOS compared to controls (P<0.01). Significant correlations were observed between TG measures and fibrinogen, CRP, SHBG and fat trunk mass (P>0.01). ETP (P=0.006), peak (P=0.003) and lag time (P=0.023) remained increased after adjustment for these potential confounders. Treatment with OC and metformin+OC further increased ETP (P<0.001) and peak (P<0.005) and reduced time to peak (P<0.04). The increase in ETP was significantly lower in the metformin+OC group than in the OC group (P<0.05). Metformin alone did not affect TG significantly. PCOS is associated with increase in TG measures independent of other risk factors of CVD. OC increase TG measures further and may thus add to the increased risk of CVD already present in women with PCOS.


[Seasonal fluctuations in fluoride content in waters of the Warta oxbow lake in Luboń].
Studies were carried out in 2002-2003 at the Department of Inland Fishing and Aquaculture, Agricultural University of Poznań, on seasonal fluctuations in fluoride content in waters of the Warta oxbow lake in Luboń (52 degrees 19' N, 16 degrees 53' E). This reservoir has been loaded during several decades with fluorine compounds, originating from the deposits of post-crystallization lye of aluminum fluoride and fluorosilicic acid. The greatest amounts of fluorine compounds were deposited in the bottom sediments in the deepest part of the reservoir. On the basis of these two-year observations it was concluded that the concentration of fluoride in water is subject to significant variations (between 1.0 and 6.2 mg F(-)/L). The lowest fluoride concentration in water was recorded during vernal circulation. On the other hand, the highest values were found in the bottom water layer during the summer stagnation. Seasonal fluctuations in fluoride content in waters of the Warta oxbow lake were closely connected with mineralization in the bottom sediment as revealed by oxygen deficiencies in the bottom water layer.


[Secondary angle-block glaucoma in posterior scleritis].
Posterior scleritis is an often misdiagnosed disease of the eye. Mainly it appears in elderly women and tends to be recurrent. Symptoms leading to diagnosis are swelling of the eye lids, a red eye, disturbances of the motility and protrusio bulbi. In rare cases you find exudative choroidal or retinal detachment, edema of the macula, or the optic nerve head, and secondary angle closure glaucoma. Often posterior scleritis is associated with general illness as herpes zoster, mixed connective tissue diseases, or Boeck's disease. Differential diagnosis are choroidal tumors as for example, melanoma, hemangioma, and metastases. The typical uveal effusion can also be caused by an arterio-venous fistula, panretinal photocoagulation, buckling procedure for retinal detachment, and by intraocular surgery in general. Especially cyclitis anularis pseudotumorosa has to be considered and shut out. Most important diagnostical means are ultrasound, and CT-scan. The underlying case describes an 81 years old woman that presented with acute angle closure glaucoma, and exudative choroidal detachment of the right eye. The ultrasound and CT-scan investigations confirmed the diagnosis of scleritis posterior. The acute angle closure glaucoma, and the choroidal detachment regressed immediately under the treatment with steroids given locally and systemically. There was no impact of miotics and peripheral iridectomy which both could not avoid recurrence of angle closure glaucoma.


Type 1 Diabetes Mellitus-Related circRNAs Regulate CD4+ T Cell Functions.
Circular RNAs (circRNAs) participate in development of malignancies through its active role as a "miRNA sponge." Their roles in type 1 diabetes mellitus (T1DM) pathogenesis are elusive. Here, the important role of circRNAs in T1DM was explored. circRNA profiling was performed for isolated CD4+ T cells from blood of T1DM and healthy volunteers. There were 257 differentially expressed circRNAs. Only three upregulated DEcircRNAs (hsa_circ_0000324, hsa_circ_0001853, and hsa_circ_0068797) were consistent with the GEO database. Through KEGG analyses, it was found that the three DEcircRNAs were associated with 11 miRNAs and 8 immune-related target genes (mRNA). Further analysis found that four miRNAs, two circRNAs, and four mRNAs were associated with nine circRNA-miRNA-mRNA networks. This confirmed the requirements of sponge mechanisms. The qRT-PCR analysis revealed that circRNA000324/miRNA675-5p/MAPK14 and circRNA000324/miRNA-675-5p/SYK may be potential mechanisms in regulation of differentiation and proliferation of CD4+ T cell in patients with T1DM. Therefore, targeting circRNA to regulate cellular immune responses by regulating CD4+ T cell differentiation may be a new strategy for the treatment of T1DM.


Asymmetric conductivity of engineered porins.
Positively charged peptide segments of 16 and 18 residues were inserted at a periplasmic turn of the porin from Rhodobacter blasticus in order to form an electric field-dependent plug. The X-ray diffraction analysis of a mutant confirmed that the structure of the porin had remained intact and that the insert was mobile. Incorporation experiments of single molecules into lipid bilayers showed that the distribution of electric conduction increments depended on the field polarity. The observed distributions are explained if the porin molecules enter the bilayer preferentially with their periplasmic surface first. Furthermore, the conduction of membrane-incorporated porin mutants changed reproducibly on field reversal showing asymmetries of reverse similar 15%, while the wild-type remained constant. This asymmetry is most likely caused by the electric field pressing the charged insert onto the pore eyelet in one field direction and removing it from the eyelet in the other. The results encourage attempts to improve the inserts in order to eventually reach diode characteristics.


Aiding nature's organelles: artificial peroxisomes play their role.
A major goal in medical research is to develop artificial organelles that can implant in cells to treat pathological conditions or to support the design of artificial cells. Several attempts have been made to encapsulate or entrap enzymes, proteins, or mimics in polymer compartments, but only few of these nanoreactors were active in cells, and none was proven to mimic a specific natural organelle. Here, we show the necessary steps for the development of an artificial organelle mimicking a natural organelle, the peroxisome. The system, based on two enzymes that work in tandem in polymer vesicles, with a membrane rendered permeable by inserted channel proteins was optimized in terms of natural peroxisome properties and function. The uptake, absence of toxicity, and in situ activity in cells exposed to oxidative stress demonstrated that the artificial peroxisomes detoxify superoxide radicals and H2O2 after endosomal escape. Our artificial peroxisome combats oxidative stress in cells, a factor in various pathologies (e.g., arthritis, Parkinson's, cancer, AIDS), and offers a versatile strategy to develop other "cell implants" for cell dysfunction.


Simplicilones A and B Isolated from the Endophytic Fungus Simplicillium subtropicum SPC3.
Two new tetracyclic polyketides with a spirocenter, simplicilones A (1) and B (2) were isolated from the broth-culture of the endophytic fungus Simplicilliumsubtropicum (SPC3) in the course of our screening for new bioactive secondary metabolites. This endophytoic fungus is naturally harboured in the fresh bark of the Cameroonian medicinal plant Duguetia staudtii (Engl. and Diels) Chatrou. The planar structures of the simplicilones were elucidated by MS and 1D as well as 2D NMR spectroscopic techniques. The relative configuration was assigned by NOESY experiments in conjunction with coupling constants; subsequently, the absolute configurations were assigned by the modified Mosher's method. The compounds showed weak cytotoxic effects against the cell line KB3.1 (in vitro cytotoxicity (IC50) = 25 µg/mL for 1, 29 µg/mL for 2), but were inactive against the tested Gram-positive and Gram-negative bacteria as well as fungi.


The unique hemoglobin system of Pleuragramma antarcticum, an antarctic migratory teleost. Structure and function of the three components.
Pleuragramma antarcticum (suborder Notothenioidei, family Nototheniidae) is the most abundant fish in the antarctic shelf. This pelagic species has a circum-antarctic distribution and is characterized by spawning migration. This species displays the highest multiplicity of major hemoglobins (three); the other notothenioids have a single one (except one species, having two) with relatively low oxygen affinity regulated by pH and organophosphates. The hemoglobins of P. antarcticum display strong Bohr and Root effects; however, they reveal important functional differences in subunit cooperativity and organophosphate regulation and, above all, in the response of oxygenation to temperature. Despite the substitution ValbetaE11 --> Ile found in Hb 2, which decreases the affinity in human mutants, the hemoglobins have similar oxygen affinity, higher than that of the other notothenioids. Hb 1 has the alpha chain in common with Hb 2 and the beta in common with Hb 3. The amino acid sequence of all four chains has been established. Thus the hematological features of P. antarcticum differ remarkably from those of antarctic notothenioids. This unique and sophisticated oxygen transport system may adequately meet the requirements of the unusual mode of life of this fish.


Laser scanning confocal microscopy for in situ monitoring of alkali-silica reaction.
Alkali-silica reaction (ASR) occurs in concrete between reactive siliceous components in the aggregate and the strongly alkaline pore solution, resulting in the formation of a potentially expansive gel product. Lithium additives have been shown to reduce expansion associated with ASR, but the mechanism(s) by which lithium reduces expansion have not been understood. Therefore, development of an in situ method to observe reactions associated with ASR is highly desirable, as it will allow for non-destructive observation of the reaction product formation and damage evolution over time, as the reaction progresses. A technique to image into mortar through glass aggregate by laser scanning confocal microscopy (LSCM), producing three-dimensional representations of the sample was developed. This LSCM technique was utilized to monitor the progress of alkali-silica reaction in mortar samples prepared with alkali-reactive glass aggregate both in the presence and in the absence of lithium additives: LiNO3, LiCl or LiOH. The method proved to be effective in qualitatively monitoring crack formation and growth and product formation, within cracks and at the paste/aggregate interface. In particular, dendritic products were observed at the paste/aggregate interface only in those samples containing lithium, suggesting that these products may play a role in ASR mitigation.


[Spontaneous rupture of the esophagus in a patient with chronic obstructive pulmonary disease].
A case report of successful surgical treatment of spontaneous rupture of the esophagus in a patient with chronic obstructive pulmonary disease, bronchial asthma and obesity is presented. Rupture of the esophagus occurred during a coughing without vomiting.


Primary extragastrointestinal stromal tumor arising in the vaginal wall: Significant clinicopathological characteristics of a rare aggressive soft tissue neoplasm.
Gastrointestinal (GI) stromal tumor is the most common mesenchymal neoplasm of the GI tract but also occurs with a lower frequency in extragastrointestinal regions and is called extragastrointestinal stromal tumor (EGIST). We report an unusual case of EGIST presenting as a vaginal mass. A 41-year-old woman presented with a gradually enlarging vaginal mass for the last 2 years. Physical examination revealed an elliptical, non-tender mass about 7.5 cm × 7 cm in size in the posterior vaginal wall and was resected completely. Under histological examination, the tumor showed a spindle cell type with coagulation necrosis, hemorrhage and high mitotic count. Immunohistochemical analysis revealed tumor cells were positive for DOG1, CD117, CD34 and p53 protein. Ki-67 labeling was 8%. Genetic analysis showed a deletion of exon 11 of the c-kit gene at codons 557-558. EGISTs should be kept in mind in the differential diagnosis in patients presenting with solid mass of the vaginal wall.


Kinetics of Unimolecular Decay of Methyl Vinyl Ketone Oxide, an Isoprene-Derived Criegee Intermediate, under Atmospherically Relevant Conditions.
Isoprene is the most abundant unsaturated hydrocarbon in the atmosphere. Ozonolysis of isoprene produces methyl vinyl ketone oxide (MVKO), which may react with atmospheric SO2, formic acid, and other important species at substantial levels. In this study, we utilized ultraviolet absorption to monitor the unimolecular decay kinetics of syn-MVKO in real time at 278-319 K and 100-503 Torr. After removing the contributions of radical reactions and wall loss, the unimolecular decay rate coefficient of syn-MVKO was measured to be kuni = 70 ± 15 s-1 (1σ uncertainty) at 298 K with negligible pressure dependence. In addition, kuni increases from ca. 30 s-1 at 278 K to ca. 175 s-1 at 319 K with an effective Arrhenius activation energy of 8.3 ± 2.5 kcal mol-1, kuni(T) = (9.3 × 107)exp(-4200/T) s-1. Our results indicate that unimolecular decay is the major sink of MVKO in the troposphere. The data would improve the estimation for the steady-state concentrations of MVKO and thus its oxidizing ability.


Simultaneous screening of multiple bacterial tRNA synthetases using an Escherichia coli S30-based transcription and translation assay.
The search for novel antibiotics to combat the growing threat of resistance has led researchers to screen libraries with coupled transcription and translation systems. In these systems, a bacterial cell lysate supplies the proteins necessary for transcription and translation, a plasmid encoding a reporter protein is added as a template, and a complex mixture of amino acids and cofactors is added to supply building blocks and energy to the assay. Firefly luciferase is typically used as the reporter protein in high-throughput screens because the luminescent signal is strong and, since bacterial lysates contain no luciferase, the background is negligible. The typical coupled transcription and translation assay is sensitive to inhibitors of RNA polymerase and to compounds that bind tightly to the ribosome. We have found a way to increase the information content of the screen by making the assay more sensitive to inhibitors of tRNA synthetases. Restricting the concentration of amino acids added to the reaction mixture allows the simultaneous screening of multiple tRNA synthetase enzymes along with the classic transcription and translation targets. In addition, this assay can be used as a convenient way to determine if an antibacterial compound of unknown mechanism inhibits translation through inhibition of a tRNA synthetase, and to identify which synthetase is the target.


Regional and hemispheric determinants of language laterality: implications for preoperative fMRI.
Language is typically a function of the left hemisphere but the right hemisphere is also essential in some healthy individuals and patients. This inter-subject variability necessitates the localization of language function, at the individual level, prior to neurosurgical intervention. Such assessments are typically made by comparing left and right hemisphere language function to determine "language lateralization" using clinical tests or fMRI. Here, we show that language function needs to be assessed at the region and hemisphere specific level, because laterality measures can be misleading. Using fMRI data from 82 healthy participants, we investigated the degree to which activation for a semantic word matching task was lateralized in 50 different brain regions and across the entire cortex. This revealed two novel findings. First, the degree to which language is lateralized across brain regions and between subjects was primarily driven by differences in right hemisphere activation rather than differences in left hemisphere activation. Second, we found that healthy subjects who have relatively high left lateralization in the angular gyrus also have relatively low left lateralization in the ventral precentral gyrus. These findings illustrate spatial heterogeneity in language lateralization that is lost when global laterality measures are considered. It is likely that the complex spatial variability we observed in healthy controls is more exaggerated in patients with brain damage. We therefore highlight the importance of investigating within hemisphere regional variations in fMRI activation, prior to neuro-surgical intervention, to determine how each hemisphere and each region contributes to language processing.


[Effects of azithromycin on fecal flora of healthy adult volunteers].
New macrolide antibiotic, azithromycin (AZM), was administered to six healthy male volunteers and its effects on their intestinal microflora were investigated. Each volunteer was given 500 mg of AZM orally, once a day for 3 consecutive days. Stool samples were obtained from them prior to the medication and 1, 7, 14 and 28 days after the third day the medication. A slight decrease in the total aerobic bacterial count was observed. Also, several species of anaerobic bacteria showed slight decreases through the 14th day post medication. Individual variances were observed, however. A marked decrease in the number of Bifidobacterium was found for each of the volunteers. Clostridium difficile was detected from one of the volunteers on the 28th day post medication without diarrhea related symptoms.


The effects of neostigmine and glycopyrrolate on skin conductance as a measure of pain.
The number of fluctuations in skin conductance per second has been shown to correlate with postoperative pain. In this context, the effects of cholinesterase inhibitors and anticholinergic drugs used for the reversal of muscle relaxants were investigated. Muscle relaxant effects were reversed in 30 patients with neostigmine and glycopyrrolate. No reversal agents were given to 30 controls. Patients' level of pain was quantified using a numeric rating scale (0-10) at several time points in the recovery room. The number of fluctuations in skin conductance per second was measured simultaneously. The number of fluctuations in skin conductance per second was significantly higher in patients with no and severe pain in the control compared with the reversal group (no pain 0.19 vs. 0.12; severe pain 0.40 vs. 0.19). The number of fluctuations in skin conductance per second was less sensitive to identify time points with moderate/severe pain in the reversal group. Skin conductance-based assessment of pain is affected by reversal agents.


Human immunodeficiency virus infection of human lymph nodes in the SCID-hu mouse.
The SCID-hu mouse is a small animal in which human hematolymphoid organs can be engrafted and maintained in vivo. In this study, parameters are described for reproducible infection of SCID-hu mice after i.v. inoculation. Infection was found to be dependent upon the time after inoculation, the virus isolate, the titer of virus, and the human target organ implanted into the mouse. Ten to 14 days after the i.v. administration of HIV isolates derived freshly from patients (e.g., JR-CSF, JR-FL, SM), 100% of engrafted human lymph nodes in SCID-hu mice were infected; greater than 95% of these animals were also viremic. Implants of human thymus or connective tissue, as well as the endogenous murine hematolymphoid organs, were not infected. As demonstrated by a combination of in situ hybridization and immunohistochemistry, both T-lymphoid and myelomonocytic lineage cells were infected in this system. HIV isolates that have been adapted to growth in vitro (e.g., HTLV-IIIb) were not infectious. When either 3'-azido-3'-deoxythymidine (AZT) or 2',3'-dideoxyinosine (ddIno) was administered to SCID-hu mice before HIV infection, the animals were protected in dose ranges similar to those used in man. This animal model may now be used as an efficient intermediate step between the lab and the clinic to study the infectious process in vivo and to best select efficacious antiviral compounds against HIV.


Probing the effect of side chains on the conformation and stability of helical peptides via isotope-edited infrared spectroscopy.
The mechanism of helix stabilization or destabilization by different amino acids has been the subject of several experimental and theoretical studies; these studies suggest that large or bulky side chains may modulate helix stability by altering the hydration of the helix backbone. In this paper, we report a spectroscopic study to determine the effect of alanine to leucine substitutions on the conformation and solvation of specific segments of a model helical peptide. A 25-residue, alanine-rich, helical peptide [Ac-(AAAAK)(4)-AAAAY-NH(2) (AKA)] and its two leucine variants [Ac-LLLLK-(AAAAK)(3)-AAAAY-NH(2) (LKA) and Ac-(AAAAK)(4)-LLLLY-NH(2) (AKL)] were characterized by infrared (IR) and electronic circular dichroism (ECD) spectroscopies. Introduction of (13)C isotopes into specific, consecutive, backbone carbonyls for certain blocks of each of the peptides mentioned above allows the IR spectra to be interpreted in terms of the conformation and solvation of specific residues within the helix. These isotope-edited IR spectra of the leucine peptides do not show evidence of a decrease in the degree of backbone solvation compared to the alanines, but suggest that the peptide may adopt a distorted conformation to accommodate the larger leucine side chains at the N-terminus. These experiments demonstrate the power of isotope-edited IR in dissecting subtle changes in helix conformation at the residue level.


[Isolation, idetification and anti-HIV-1 integrase activity of culturable endophytic fungi from Tibetan medicinal plant Phlomis younghusbandii Mukerjee].
A total of 52 endophytic fungi were isolated from roots and stems of Tibetan medicinal plant Phlomis younghusbandii Mukerjee. These fungal isolates were molecularly identified based on ITS sequnces and 28S sequences distributed to 12 genera, including Phoma, Chaetosphaeronema, Fusarium and Leptosphaeria, etc. Among them, the dominant genus was Phoma. Extracts of all strains were evaluated for anti-HIV-1 integrase activity by using soluable integrase expressed in E. coli BL21 (DE3). The results showed that seven samples from five fungal endophytes PHY-24, PHY-38, PHY-40, PHY-51, PHY-53, which belonged to genus Chaetosphaeronema, inhibited strand transfer reaction catalyzed by HIV-1 integrase with IC50 values, of 6.60, 5.20, 2.86, 7.86, 4.47, 4.56 and 3.23 microg x mL(-1) respectively. In conclusion, the endophytic fungi of Phlomis younghusbandii Mukerjee are valuable for further screening anti-HIV-1 integrase agents.


Comparison of early and late treatment with a recombinant endotoxin neutralizing protein in a rat model of Escherichia coli sepsis.
To test the efficacy of a recombinant endotoxin neutralizing protein as compared with saline in rats with Escherichia coli sepsis. Prospective, controlled animal trial. Hospital animal research laboratory. Male Wistar rats challenged with intraperitoneal E. coli, O18ac K1, and treated 1 hr later with ceftriaxone and gentamicin. Recombinant endotoxin neutralizing protein, 50 mg/kg, was administered to rats 1, 2, or 3 hrs after E. coli challenge; saline was administered to control animals. Quantitative bacteremia, 1 hr after challenge and before antibiotic administration, was not significantly different between treatment groups (range geometric mean 451 to 621 colony-forming units [cfu]/mL). The endotoxin concentration, measured immediately before recombinant endotoxin neutralizing protein administration, was significantly higher in animals sampled and treated at 2 hrs (geometric mean 260 EU/mL; 95% confidence interval 140 to 480 EU/mL), or 3 hrs (geometric mean 697 EU/mL; 95% confidence interval 307 to 1585 EU/mL) after E. coli challenge, compared with animals sampled and treated at 1 hr (geometric mean 17 EU/mL; 95% confidence interval 7 to 69 EU/ mL). Survival rate was significantly greater in rats treated with recombinant endotoxin neutralizing protein at 1 hr (23/27; p < .001) or 2 hrs (8/30; p < .01) after E. coli challenge than in controls (1/32). Administration of recombinant endotoxin neutralizing protein delayed up to 2 hrs after challenge with E. coli improves survival in antibiotic-treated rats with Gram-negative sepsis.


Medically unexplained symptoms and the problem of power in the primary care consultation: a qualitative study.
Patients presenting in primary care frequently exhibit physical symptoms that may be unrelated to organic pathology. Such symptoms are commonly regarded as products of psychological or emotional problems, and their legitimacy as 'medical' matters is often called into question. Our aim was to explore GPs' attitudes to the management of patients that present with medically unexplained symptoms in primary care. Semi-structured interviews were conducted with 15 GPs in North-West England. Interviews were audio-taped and subsequently transcribed and analysed using a constant comparison technique. Subjects conceptualized patients presenting with medically unexplained symptoms as the presentation of psychological distress. They presented problems of control and authority in the consultation, and difficulties in managing this had a negative impact on the doctor-patient relationship. Such consultations were frustrating for the GP and potentially harmful to the patient. Patients with medically unexplained symptoms were seen to be presenting with inappropriate symptoms that were a manifestation of emotional or social distress. GPs felt ill-equipped to deal with the presentations and the frustrations they felt and may need help in actively and productively managing these patients.


Seeking relationships between Internet addiction and depressiveness, daytime sleepiness, as well as perceived social support in young adults.
An assessment of the scale of Internet addiction among young adults and seeking relationships between Internet addiction and chosen sociodemographic variables, perceived social support, prevalence of depressiveness, and excessive daytime sleepiness. The study was carried out using a diagnostic poll method with author's own questionnaire as well as standardized diagnostic tools: The Internet Addiction Test (IAT), the Beck Depression Inventory (BDI), The Epworth Sleepiness Scale (ESS), as well as The Multidimensional Scale of Perceived Social Support (MSPSS). The study was conducted on 567 individuals. Pathological Internet use was identified in 1.41% of the surveyed. Perceived social support was significantly lower in the group of respondents who pathologically used Internet in comparison with ones who used Internet in the proper way. Severity of depressiveness symptoms and daytime sleepiness was significantly higher in respondents who were addicted to Internet in comparison with not addicted individuals. The scale of pathological Internet use among surveyed was low. Problematic Internet use refers to women and men to the same degree, as well as to single people and individuals in relationships. Persons who work and do not study as well as residents of large and medium cities more frequently use Internet in a more pathological way than individuals who study and live-in small cities and in the countryside. Internet addiction creates a significant relationship with the prevalence of more severe symptoms of depressiveness and excessive daytime sleepiness, whereas pathological pattern of Internet use more often concerns individuals whose perception of social support is low.


A C-H triplebond O hydrogen bond stabilized polypeptide chain reversal motif at the C terminus of helices in proteins.
The serendipitous observation of a C-H cdots, three dots, centered O hydrogen bond mediated polypeptide chain reversal in synthetic peptide helices has led to a search for the occurrence of a similar motif in protein structures. From a dataset of 634 proteins, 1304 helices terminating in a Schellman motif have been examined. The C-H triplebond O interaction between the T-4 C(alpha)H and T+1 Cz doublebond O group (C triplebond O< or =3.5A) becomes possible only when the T+1 residue adopts an extended beta conformation (T is defined as the helix terminating residue adopting an alpha(L) conformation). In all, 111 examples of this chain reversal motif have been identified and the compositional and conformational preferences at positions T-4, T, and T+1 determined. A marked preference for residues like Ser, Glu and Gln is observed at T-4 position with the motif being further stabilized by the formation of a side-chain-backbone O triplebond H-N hydrogen bond involving the side-chain of residue T-4 and the N-H group of residue T+3. In as many as 57 examples, the segment following the helix was extended with three to four successive residues in beta conformation. In a majority of these cases, the succeeding beta strand lies approximately antiparallel with the helix, suggesting that the backbone C-H triplebond O interactions may provide a means of registering helices and strands in an antiparallel orientation. Two examples were identified in which extended registry was detected with two sets of C-H cdots, three dots, centered O hydrogen bonds between (T-4) C(alpha)H triplebond O (T+1) and (T-8) C(alpha)H triplebondC doublebond O (T+3).


Biocompatibility of polycations: in vitro agglutination and lysis of red blood cells and in vivo toxicity.
The effects of six polycations were studied in vitro on red blood cells (RBC) and in vivo after intravenous administration. Hemagglutination and hemolysis depended not only on the molar mass and the concentration of these polycations, but also on their chemical nature. The hemagglutination and hemolysis induced by poly(L-lysine), diethylaminoethyldextran, poly(dimethyldiallylammonium) chloride and poly[2-(dimethylamino)ethyl methacrylate] was low to moderate, whereas a severe hemolysis was induced by a partially quaternized poly[thio-1-(N,N-diethyl-aminoethyl)ethylene]. In the case of poly(epsilon-lysine), no significant hemagglutination nor hemolysis was observed. The presence of plasma proteins reduced both agglutination and hemolysis. This protective effect was enhanced when the polycations interacted with plasma proteins before contact with RBC. In the presence of albumin, the behavior depended on the polycation and on the order of addition of the three components of the suspension, namely albumin, polycation and RBC. Depending on the polycation, albumin-polycation complexes were either less active or more active on RBC than the same polycation in protein-free medium. In vivo the studied polycations induced an immediate mortality except poly(epsilon-lysine), which induced a delayed mortality. The minimal dose of polycations inducing immediate mortality paralleled their effect on RBC.


Autonomous spheroid formation by culture plate compartmentation.
Scaffold-free 3D cell cultures (e.g. pellet cultures) are widely used in medical science, including cartilage regeneration. Their drawbacks are high time/reagent consumption and lack of early readout parameters. While optimisation was achieved by automation or simplified spheroid generation, most culture systems remain expensive or require tedious procedures. The aim of this study was to establish a system for resource efficient spheroid generation with additional early readout parameters. This was achieved by a new approach for spheroid generation via self-assembly from monolayer via compartmentation of cell culture surfaces utilising laser engraving (grid plates). The compartmentation triggered contraction and rolling up of the cell monolayer, finishing in condensation into a spheroid in human adipose-derived stem cell (ASC/TERT1) and human articular chondrocytes (hACs)-ASC/TERT1 co-cultures, when cultivated on grid plates under chondrogenic conditions. Plates with 1 and 3 mm grid size yielded stable diameters (about 140µm and 300µm, respectively). ASC/TERT1 spheroids fully formed within 3 weeks while co-cultures took 1-2 weeks, forming significantly faster with increasing hAC ratio (p< 0.05 and 0.01 for 1:1 and 1:4 ASC/TERT1:hAC ratio, respectively). Co-cultures showed slightly lower spheroid diameters, due to earlier spheroid formation and incomplete monolayer formation. However, this was associated with a more homogeneous matrix distribution in the co-culture. Both showed differentiation capacity comparable to standard pellet culture in (immune-)histochemistry and RT-qPCR. To assess usability for cartilage repair, spheroids were embedded into a hydrogel (fibrin), yielding cellular outgrowth and matrix deposition, which was especially pronounced in co-cultures. The herein presented novel cell culture system is not only a promising tool for autonomous spheroid generation with the potential of experimental and clinical application in tissue engineering, but also for the generation of 'building blocks' for subsequential biofabrication strategies such as bioprinting.


A motor planning stage represents the shape of upcoming movement trajectories.
Interactions with our environment require curved movements that depend not only on the final position of the hand but also on the path used to achieve it. Current studies in motor control, however, largely focus on point-to-point movements and do not consider how movements with specific desired trajectories might arise. In this study, we examined intentionally curved reaching movements that navigate paths around obstacles. We found that the preparation of these movements incurred a large reaction-time cost. This cost could not be attributed to nonmotor task requirements (e.g., stimulus perception) and was independent of the execution difficulty (i.e., extent of curvature) of the movement. Additionally, this trajectory representation cost was not observed for point-to-point reaches but could be optionally included if the task encouraged consideration of straight trajectories. Therefore, when the path of a movement is task relevant, the shape of the desired trajectory is overtly represented as a stage of motor planning. This trajectory representation ability may help explain the vast repertoire of human motor behaviors.


Evolution of a national approach to evaluating the benefits of the electronic health record.
Demonstrating the value of eHealth investments to users and other stakeholders, and driving ongoing optimization of benefits are increasingly important components of eHealth implementations in Canada. For these reasons, Canada Health Infoway is working with provincial partners to make research and evaluation an important component in Electronic Health Record (EHR) investments. Leading provinces working with Infoway, including British Columbia, the Province of Quebec, and Newfoundland and Labrador, are making evaluation a central part of their eHealth strategies and using evaluation results to optimize the benefits of investments. We provide an overview of benefits evaluation strategies and lessons learned. Examples of early evaluations completed and those currently underway provide insights into the challenges and benefits of investments in EHR.


Concerted electron and proton transfer in ionic crystals mapped by femtosecond x-ray powder diffraction.
X-ray powder diffraction, a fundamental technique of structure research in physics, chemistry, and biology, is extended into the femtosecond time domain of atomic motions. This allows for mapping (macro)molecular structure generated by basic chemical and biological processes and for deriving transient electronic charge density maps. In the experiments, the transient intensity and angular positions of up to 20 Debye Scherrer reflections from a polycrystalline powder are measured and atomic positions and charge density maps are determined with a combined spatial and temporal resolutions of 30 pm and 100 fs. We present evidence for the so far unknown concerted transfer of electrons and protons in a prototype material, the hydrogen-bonded ionic ammonium sulfate [(NH(4))(2)SO(4)]. Photoexcitation of ammonium sulfate induces a sub-100 fs electron transfer from the sulfate groups into a highly confined electron channel along the c-axis of the unit cell. The latter geometry is stabilized by transferring protons from the adjacent ammonium groups into the channel. Time-dependent charge density maps derived from the diffraction data display a periodic modulation of the channel's charge density by low-frequency lattice motions with a concerted electron and proton motion between the channel and the initial proton binding site. Our results set the stage for femtosecond structure studies in a wide class of (bio)molecular materials.


Improving Hepatitis C Virus Screening Rates in Primary Care: A Targeted Intervention Using the Electronic Health Record.
With the advent of effective treatments for hepatitis C virus (HCV), it has become a public health priority to increase the identification of HCV carriers and link them to systems of care. As a result, in 2012, the Centers for Disease Control and Prevention recommended that all adults born between 1945 and 1965 should receive one-time testing for HCV. In response to this mandate, we sought an effective nonintrusive means to increase HCV screening rates in our busy primary care practices. We designed an HCV testing decision support module that was integrated into the electronic health record (EHR) and triggered an automatic test order for eligible patients at the time of visit. Rates of HCV screening for eligible patients were measured before and after implementation. Hepatitis C virus screening rates increased by 254% after implementation of this tool. Incorporating a clinical reminder into the EHR effectively and appropriately increased the hepatitis C testing rates among primary care patients with no previous testing. Such tools can be an effective means to operationalize health system-wide testing efforts.


Highly multiplexed oligonucleotide probe-ligation testing enables efficient extraction-free SARS-CoV-2 detection and viral genotyping.
There is an urgent and unprecedented need for sensitive and high-throughput molecular diagnostic tests to combat the SARS-CoV-2 pandemic. Here we present a generalized version of the RNA-mediated oligonucleotide Annealing Selection and Ligation with next generation DNA sequencing (RASL-seq) assay, called "capture RASL-seq" (cRASL-seq), which enables highly sensitive (down to ~1-100 pfu/ml or cfu/ml) and highly multiplexed (up to ~10,000 target sequences) detection of pathogens. Importantly, cRASL-seq analysis of COVID-19 patient nasopharyngeal (NP) swab specimens does not involve nucleic acid purification or reverse transcription, steps that have introduced supply bottlenecks into standard assay workflows. Our simplified protocol additionally enables the direct and efficient genotyping of selected, informative SARS-CoV-2 polymorphisms across the entire genome, which can be used for enhanced characterization of transmission chains at population scale and detection of viral clades with higher or lower virulence. Given its extremely low per-sample cost, simple and automatable protocol and analytics, probe panel modularity, and massive scalability, we propose that cRASL-seq testing is a powerful new technology with the potential to help mitigate the current pandemic and prevent similar public health crises.


Reduction of Raf kinase inhibitor protein expression by Bcr-Abl contributes to chronic myelogenous leukemia proliferation.
Chronic myelogenous leukemia (CML) is characterized by a reciprocal chromosomal translocation (9;22) that generates the Bcr-Abl fusion gene. The Ras/Raf-1/MEK/ERK pathway is constitutively activated in Bcr-Abl-transformed cells, and Ras activity enhances the oncogenic ability of Bcr-Abl. However, the mechanism by which Bcr-Abl activates the Ras pathway is not completely understood. Raf kinase inhibitor protein (RKIP) inhibits activation of MEK by Raf-1 and its downstream signal transduction, resulting in blocking the MAP kinase pathway. In the present study, we found that RKIP was depleted in CML cells. We investigated the interaction between RKIP and Bcr-Abl in CML cell lines and Bcr-Abl(+) progenitor cells from CML patients. The Abl kinase inhibitors and depletion of Bcr-Abl induced the expression of RKIP and reduced the pERK1/2 status, resulting in inhibited proliferation of CML cells. Moreover, RKIP up-regulated cell cycle regulator FoxM1 expression, resulting in G(1) arrest via p27(Kip1) and p21(Cip1) accumulation. In colony-forming unit granulocyte, erythroid, macrophage, megakaryocyte, colony-forming unit-granulocyte macrophage, and burst-forming unit erythroid, treatment with the Abl kinase inhibitors and depletion of Bcr-Abl induced RKIP and reduced FoxM1 expressions, and inhibited colony formation of Bcr-Abl(+) progenitor cells, whereas depletion of RKIP weakened the inhibition of colony formation activity by the Abl kinase inhibitors in Bcr-Abl(+) progenitor cells. Thus, Bcr-Abl represses the expression of RKIP, continuously activates pERK1/2, and suppresses FoxM1 expression, resulting in proliferation of CML cells.


Supplementation with Eurycoma longifolia Extract Modulates Diurnal Body Temperature Fluctuation and Sleep Rhythm in Mice.
Eurycoma longifolia (Tongkat Ali; TA) is a traditional medicinal herb, commonly known as Malaysian ginseng. The root tea has been traditionally applied to treat fevers, aches, sexual dysfunction and other ailments. We evaluated the effects of TA extract supplementation on diurnal core body temperature (BT) and sleep architecture in model mice. Dietary supplementation with TA extract for 4 wk resulted in significantly and moderately reduced BT during the rest and active phases, respectively. A high dose delayed the onset of BT elevation at the start of the active phase, indicating that the effect was dose-dependent. Electroencephalography findings revealed that dietary supplementation with TA extract changed sleep rhythms and delta power during the inactive phase of NREM sleep, indicating improved sleep quality. Our findings suggested that dietary TA extract could be a promising natural aid that alleviates sleep problems via thermoregulation.


Increasing the cisplatin cytotoxicity and cisplatin-induced DNA damage by conferone in 5637 cells.
Despite widespread application of cisplatin in treatment of transitional cell carcinomas, its efficiency is far from satisfactory due to acquired drug resistance. The present study was carried out to estimate the effects of conferone, a sesquiterpene-coumarin isolated from Ferula badrakema, on increasing cisplatin cytotoxicity in 5637 cells. In order to determine conferone effects, 5637 cells were cultured in the presence of different concentrations of conferone and cisplatin in combination. The cytotoxicity and DNA damaging effects were then studied using MTT and comet assays, respectively. The results revealed that 24 h after the combination of 1 µg mL⁻¹ cisplatin with 32 µg mL⁻¹ conferone, the cytotoxicity of cisplatin was increased by 36.76%, and comet assay analyses showed that conferone could enhance the DNA damaging effects of cisplatin by 41%.


Macroporous germanium by electrochemical deposition.
Germanium is electrodeposited in a template formed from a dried suspension of silica spheres. The germanium completely fills the pores of the silica matrix. The semiconductor, as deposited, is amorphous but can be crystallized by annealing. Selective dissolution of the silica template gives a macroporous germanium-air sphere matrix, which offers interesting possibilities for photonic applications.


Development of PARP and Immune-Checkpoint Inhibitor Combinations.
PARP inhibitors drive increased DNA damage, particularly in tumors with existing defects in DNA repair. This damage not only promotes immune priming through a range of molecular mechanisms, but also leads to adaptive upregulation of programmed death ligand 1 (PD-L1) expression. In this context, PARP inhibition and programmed cell death 1(PD-1)/PD-L1-targeting antibodies represent a rationale combination. In this review, we detail the basic and translational science underpinning this promising new combination, summarize available clinical data, and discuss the key questions that remain to be addressed during future development.


Contextual constraint and lexical competition: Revisiting biased misperception during reading.
In identifying and accessing lexical items while comprehending text, readers must rapidly select a word from visually similar words before integrating it into a sentence. It has been proposed that readers are likely to misperceive a low frequency word as a highly frequent orthographically similar alternative, particularly when the alternative is supported by previous context (Gregg & Inhoff, 2016; Perea & Pollatsek, 1998; Pollatsek, Perea, & Binder, 1999; Slattery, 2009). In such cases, the misperception may not be corrected until the reader encounters incongruent information. However, many of these studies place incongruent text directly after the critical word, confounding whether readers regress backward in text to resolve their misperception or to halt forward text progression in order resolve a lexical level conflict between the word form and its competitor. In 3 eye tracking while reading experiments, we adapted materials from previous studies to include a postcritical spillover region to address this possibility. Two of these experiments were designed to permit an ex-Gaussian analysis of the distribution of first pass reading prior to disambiguating information. The evidence suggests that postlexical competition-inhibition between orthographically similar forms can delay forward movement of the eyes as a competitor is inhibited. The possibility that misperception and postlexical competition-inhibition arise from the same set of mechanisms is discussed. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Association between the severity of chronic lung disease and first-year outcomes of very low birth weight infants.
The objective of this study was to determine if there is an association between the severity of chronic lung disease in very low birth weight infants (as assessed by the duration of supplemental oxygen requirements in the neonatal period) and first-year neurodevelopmental, sensory, and growth outcomes as well as duration of neonatal hospitalization and first-year hospital readmissions. Retrospective chart review with matched subject groups. The subjects of this study were very low birth weight infants born between 1987 and 1991 in a 17-county perinatal region of North Carolina. Infants were categorized into one of three groups on the basis of duration of supplemental oxygen requirements. Infants who were breathing room air by 28 days were classified as having no chronic lung disease; infants who required supplemental oxygen at 28 days but not at 36 weeks postmenstrual age were classified as having mild chronic lung disease; and infants who required supplemental oxygen at 36 weeks postmenstrual age were classified as having severe chronic lung disease. Infants were matched for birth weight, sex, and race. The matched groups (n = 174) were compared with respect to the incidence of first year adverse neurodevelopmental and sensory outcomes, growth patterns, and hospital readmissions during the first year. Results were analyzed with general linear models and logistic regression analyses. The incidence of any adverse neurodevelopmental or sensory outcome increased as severity of chronic lung disease increased from none (3.6%) to mild (21.4%) to severe (31.6%, p < 0.001). Growth patterns were similar in infants with no and mild chronic lung disease, but infants with severe chronic lung disease were significantly lighter (p < 0.01) and shorter (p < 0.005) at 40 weeks postmenstrual age and significantly lighter at 1 year adjusted age (p < 0.05). The duration of the initial hospitalization increased with chronic lung disease severity (p < 0.001). Infants with severe chronic lung disease were readmitted to the hospital significantly more often (p < 0.005) than infants with no chronic lung disease or mild chronic lung disease. Very low birth weight infants who required supplemental oxygen at or beyond 28 days were at increased risk for adverse neurodevelopmental and sensory outcomes, but only those infants who continued to require supplemental oxygen at 36 weeks were at increased risk for poor growth and readmission to the hospital.


Determining the causes of impairments in the Little Scioto River, Ohio, USA: part 2. Characterization of causes.
Two stream reaches in the Little Scioto River (OH, USA) were characterized for the causes of impairments measured at two locations. By inductive inference, six candidate causes were winnowed down to three and five candidate causes for each of the two stream reaches. Using a formal strength-of-evidence process, a single cause was determined. At the most upstream location, habitat alterations, including fine-textured substrates and low DO, were characterized as the probable causes for an increased percentage of anomalies of fish, a decreased percentage of mayflies, and an increased percentage of tolerant macroinvertebrates. An increase in the relative weight of fish was attributed to an artificially narrow, deepened channel. Approximately 2 km downstream, polycyclic aromatic hydrocarbon (PAH)-contaminated sediments were identified as the cause for both fish and macroinvertebrate impairments. Causal characterization using first elimination and then a strength-of-evidence approach narrowed and defined the causes of ecological impairment even in this situation, where many complex and interacting candidate causes existed. Applying a formal method highlighted types of data and associations that can strengthen and present a more convincing determination of the causes of impairment.


EphB4-targeted imaging with antibody h131, h131-F(ab')2 and h131-Fab.
Accumulating evidence suggests that overexpression of the tyrosine kinase receptor EphB4, a mediator of vascular development, is a novel target for tumor diagnosis, prognosis and therapy. Noninvasive imaging of EphB4 expression could therefore be valuable for evaluating disease course and therapeutic efficacy at the earliest stages of anti-EphB4 treatment. In this study, we systematically investigated the use of anti-EphB4 antibody h131 (150 kDa) and its fragments (h131-F(ab')2, 110 kDa; h131-Fab, 50 kDa) for near-infrared fluorescence (NIRF) imaging of EphB4 expression in vivo. h131-F(ab')2 and h131-Fab were produced through pepsin and papain digestion of h131 respectively, whose purity was confirmed by FPLC and SDS-PAGE. After conjugation with Cy5.5, in vivo characteristics of h131, h131-F(ab')2 and h131-Fab were evaluated in EphB4-positive HT29 tumor model. Although h131-Cy5.5 demonstrated highest tumor uptake among these probes, its optimal tumor uptake level was obtained at 2 days post injection (p.i.). For h131-Fab-Cy5.5, maximum tumor uptake was achieved at 4 h p.i. However, no significant difference was observed between h131-Fab-Cy5.5 and hIgG-Fab-Cy5.5, indicating the tumor accumulation was mainly caused by passive targeting. In contrast, h131-F(ab')2-Cy5.5 demonstrated prominent tumor uptake at 6 h p.i. The target specificity was confirmed by hIgG-F(ab')2-Cy5.5 control and immunofluorescent staining. Collectively, h131-F(ab')2 exhibited prominent and specific tumor uptake at early time points, which suggests it is a promising agent for EphB4-targeted imaging.


Green Tea Extract Concurrent with an Oral Nutritional Supplement Acutely Enhances Muscle Microvascular Blood Flow without Altering Leg Glucose Uptake in Healthy Older Adults.
Postprandial macro- and microvascular blood flow and metabolic dysfunction manifest with advancing age, so vascular transmuting interventions are desirable. In this randomised, single-blind, placebo-controlled, crossover trial, we investigated the impact of the acute administration of green tea extract (GTE; containing ~500 mg epigallocatechin-3-gallate) versus placebo (CON), alongside an oral nutritional supplement (ONS), on muscle macro- and microvascular, cerebral macrovascular (via ultrasound) and leg glucose/insulin metabolic responses (via arterialised/venous blood samples) in twelve healthy older adults (42% male, 74 ± 1 y). GTE increased m. vastus lateralis microvascular blood volume (MBV) at 180 and 240 min after ONS (baseline: 1.0 vs. 180 min: 1.11 ± 0.02 vs. 240 min: 1.08 ± 0.04, both p < 0.005), with MBV significantly higher than CON at 180 min (p < 0.05). Neither the ONS nor the GTE impacted m. tibialis anterior perfusion (p > 0.05). Leg blood flow and vascular conductance increased, and vascular resistance decreased similarly in both conditions (p < 0.05). Small non-significant increases in brachial artery flow-mediated dilation were observed in the GTE only and middle cerebral artery blood flow did not change in response to GTE or CON (p > 0.05). Glucose uptake increased with the GTE only (0 min: 0.03 ± 0.01 vs. 35 min: 0.11 ± 0.02 mmol/min/leg, p = 0.007); however, glucose area under the curve and insulin kinetics were similar between conditions (p > 0.05). Acute GTE supplementation enhances MBV beyond the effects of an oral mixed meal, but this improved perfusion does not translate to increased leg muscle glucose uptake in healthy older adults.


Effect of Intercalated Water on Potassium Ion Transport through Kv1.2 Channels Studied via On-the-Fly Free-Energy Parametrization.
We introduce a two-dimensional version of the method called on-the-fly free energy parametrization (OTFP) to reconstruct free-energy surfaces using Molecular Dynamics simulations, which we name OTFP-2D. We first test the new method by reconstructing the well-known dihedral angles free energy surface of solvated alanine dipeptide. Then, we use it to investigate the process of K+ ions translocation inside the Kv1.2 channel. By comparing a series of two-dimensional free energy surfaces for ion movement calculated with different conditions on the intercalated water molecules, we first recapitulate the widely accepted knock-on mechanism for ion translocation and then confirm that permeation occurs with water molecules alternated among the ions, in accordance with the latest experimental findings. From a methodological standpoint, our new OTFP-2D algorithm demonstrates the excellent sampling acceleration of temperature-accelerated molecular dynamics and the ability to efficiently compute 2D free-energy surfaces. It will therefore be useful in large variety complex biomacromolecular simulations.


Language skills in a child with Leber hereditary optic neuropathy following intrathecal chemotherapy for acute lymphoblastic leukemia.
The language skills of a male child with Leber hereditary optic neuropathy (LHON) and coincidentally treated for acute lymphoblastic leukemia (ALL) with intrathecal chemotherapy at the age of 3 years 8 months were comprehensively evaluated twice over a 6-month period approximately 5½ years after diagnosis of ALL. Despite marked chemotherapy-related leukoencephalopathic changes documented on magnetic resonance imaging, the child presented with stable language skills, which were generally average to above-average based on the normative data from a comprehensive language test battery. In light of the coincidental presentation in the child of a diagnosis of LHON, which may lead to serious vision impairment and increased vulnerability to drug neurotoxicity, coupled with a history of central nervous system (CNS)-directed treatment for ALL resulting in progressive white matter pathology, the study highlights the importance of ongoing monitoring of the child's language development throughout his adolescent years.


Separation and identification of underivatized plasma acylcarnitine isomers using liquid chromatography-tandem mass spectrometry for the differential diagnosis of organic acidemias and fatty acid oxidation defects.
A simple HPLC-MS/MS method has been established to separate and identify underivatized acylcarnitine isomers. Human plasma samples were deproteinized and concentrated. Acylcarnitines were separated on a reverse phase column and detected with triple quadrupole linear ion trap mass spectrometry. Deuterium-labeled internal standards were used for quantitation. To identify acylcarnitines without pure standards, information-dependent acquisition linking to enhanced product ion scan mode was used. 112 acylcarnitines, including stereoisomers, were found in samples of patients. Dicarboxylic acylcarnitines, such as methylmalonylcarnitine and glutarylcarnitine, were detected with high sensitivity. Three stereoisomers of (R,S)2-methyl-3-hydroxy butyrylcarnitine were detected in samples of patients with β-ketothiolase deficiency. Validation results revealed excellent precision and accuracy of the method. In general the within- and between-run coefficients of variation (CV%) were less than 15%, and recoveries were in the range of 92.7-117.5%. In addition, the reference intervals of acylcarnitines for children aged 3-day to13-year old were established. Using the new method and reference intervals, we have correctly diagnosed 49 patients with fatty acid oxidation defects or organic acidemias in 176 high-risk patients.


Therapeutic alliance in psychological therapy for people with recent onset psychosis who use cannabis.
This paper examines the role of therapeutic alliance in predicting outcomes in a Randomized Controlled Trial of Motivational Interviewing and Cognitive Behavioral Therapy (MICBT) for problematic cannabis use in recent onset psychosis. All clients were participating in a three arm pragmatic rater-blind randomized controlled trial of brief MICBT plus standard care compared with longer term MICBT plus standard care and standard care alone. Participants completed measures to assess clinical symptoms, global functioning and substance misuse at baseline, 4.5months, 9months and 18months. Clients and therapists completed the Working Alliance Inventory approximately one month into therapy. Client alliance data was available for 35 participants randomized to therapy and therapist alliance data was available for 52 participants randomized to therapy. At baseline, poorer client-rated alliance was associated with more negative symptoms, poorer insight and greater cannabis use, whereas poorer therapist-rated alliance was only associated with amount of cannabis used per cannabis using day. Alliance ratings were also positively associated with amount of therapy: client-rated alliance was higher in the longer compared to the briefer therapy; therapist-rated alliance was associated with greater number of sessions attended (controlling for type of therapy) and therapy completion. In predicting outcome, client-rated alliance predicted total symptom scores and global functioning scores at follow-up. Neither client nor therapist alliance predicted changes in substance misuse at any time point. Findings demonstrate that individuals with psychosis and substance misuse who form better alliances with their therapists gain greater benefits from therapy, at least in terms of improvements in global functioning.


Preparation of chlamydomonas chloroplasts for the in vitro import of polypeptide precursors.
To study the import of polypeptide precursors we have adapted and compared two procedures for the isolation of competent chloroplasts from the green unicellular alga, Chlamydomonas reinhardtii: silicasol gradient centrifugation and elutriation. The chloroplasts actively import the precursor of the small subunit of ribulose bisphosphate carboxylase-oxygenase in vitro.


Six decades of vitiligo genetics: genome-wide studies provide insights into autoimmune pathogenesis.
Generalized vitiligo (GV) is a complex disease in which patchy depigmentation results from autoimmune loss of melanocytes from affected regions. Genetic analyses of GV span six decades, with the goal of understanding biological mechanisms and elucidating pathways that underlie the disease. The earliest studies attempted to describe the mode of inheritance and genetic epidemiology. Early genetic association studies of biological candidate genes resulted in some successes, principally HLA and PTPN22, but in hindsight many such reports now seem to be false-positives. Later, genome-wide linkage studies of multiplex GV families identified NLRP1 and XBP1, which appear to be valid GV susceptibility genes that control key aspects of immune regulation. Recently, the application of genome-wide association studies to analysis of GV has produced a rich yield of validated GV susceptibility genes that encode components of biological pathways reaching from immune cells to the melanocyte. These genes and pathways provide insights into underlying pathogenetic mechanisms and possible triggers of GV, establish relationships to other autoimmune diseases, and may provide clues to potential new approaches to GV treatment and perhaps even prevention. These results thus validate the hopes and efforts of the early investigators who first attempted to comprehend the genetic basis of vitiligo.


Development, Implementation, and Evaluation of Introductory Pharmacy Practice Experiences at an Academic Medical Center.
Introductory pharmacy practice experiences (IPPEs) for pharmacy students are required by the Accreditation Council for Pharmacy Education, but guidance for rotational structure is limited. To describe the design, implementation, and evaluation of IPPE rotations at a large, multisite, academic medical center. A large IPPE program was successfully implemented and sustained. Rapid cycle changes were made based on post rotational surveys completed by both preceptors and IPPE students to assess and modify the rotations until the average experience was rated at least a 4 on a 5 point scale, illustrating a mutually beneficial collaboration between the Medical Center and the School of Pharmacy. The IPPE program structure and capacity has continued to grow at the Medical Center in a collaborative manner.


Modification of sleep respiratory patterns by auditory stimulation: indications of a technique for preventing sudden infant death syndrome?
Sudden infant death syndrome (SIDS) has been associated with premorbid respiratory irregularities that may contribute to the subsequent occurrence of SIDS (e.g. by chronic hypoxia). Conventional apnea alarms will detect periods of extended apnea but not other chronic respiratory irregularities that could contribute to SIDS. The effect of a brief, quiet, repetitive auditory stimulus on the sleep respiratory behavior of infant dogs and pigs was assessed. Stimulus presentation was associated with an increase in the sleep respiration rate irrespective of the stimulus presentation rate being faster or slower than the baseline respiratory rate, and irrespective of the pattern of presentation being regular or random. These results suggest that apnea monitoring systems may be improved by an additional function in which less extreme respiratory irregularities initiate the time-limited presentation of a quiet, repetitive, auditory stimulus, aimed at normalizing the respiratory pattern. This function could enhance the preventative role of the alarm systems by reducing the development of pathology that may promote some instances of SIDS.


Particle-based phasor-FLIM-FRET resolves protein-protein interactions inside single viral particles.
Fluorescence lifetime imaging microscopy (FLIM) is a popular modality to create additional contrast in fluorescence images. By carefully analyzing pixel-based nanosecond lifetime patterns, FLIM allows studying complex molecular populations. At the single-molecule or single-particle level, however, image series often suffer from low signal intensities per pixel, rendering it difficult to quantitatively disentangle different lifetime species, such as during Förster resonance energy transfer (FRET) analysis in the presence of a significant donor-only fraction. In this article we investigate whether an object localization strategy and the phasor approach to FLIM have beneficial effects when carrying out FRET analyses of single particles. Using simulations, we first showed that an average of ∼300 photons, spread over the different pixels encompassing single fluorescing particles and without background, is enough to determine a correct phasor signature (SD < 5% for a 4-ns lifetime). For immobilized single- or double-labeled dsDNA molecules, we next validated that particle-based phasor-FLIM-FRET readily allows estimating fluorescence lifetimes and FRET from single molecules. Thirdly, we applied particle-based phasor-FLIM-FRET to investigate protein-protein interactions in subdiffraction HIV-1 viral particles. To do this, we first quantitatively compared the fluorescence brightness, lifetime, and photostability of different popular fluorescent protein-based FRET probes when genetically fused to the HIV-1 integrase enzyme in viral particles, and conclude that eGFP, mTurquoise2, and mScarlet perform best. Finally, for viral particles coexpressing FRET-donor/acceptor-labeled IN, we determined the absolute FRET efficiency of IN oligomers. Available in a convenient open-source graphical user interface, we believe that particle-based phasor-FLIM-FRET is a promising tool to provide detailed insights in samples suffering from low overall signal intensities.