Relationship between ghrelin levels, alcohol craving, and nutritional status in current alcoholic patients.
Ghrelin is a peptide produced mainly by the gut and hypothalamus. Ghrelin is able to stimulate food-seeking behavior. Alcohol-craving and food-seeking behavior could share common neural circuits. Ghrelin is related to nutritional status, but few data are available in alcoholic patients on the relationship between ghrelin and nutritional disorders. Plasma ghrelin was evaluated in 15 current alcoholic male patients compared with 15 healthy male volunteers. Craving was evaluated by the Obsessive-Compulsive Drinking Scale. Body composition was assessed by dual-energy X-ray absorptiometry. Energy substrate utilization was evaluated by indirect calorimetry. Ghrelin was significantly reduced in alcohol-dependent patients with respect to healthy subjects (p=0.0278). A significant positive correlation was found between ghrelin and craving (r=0.55; p=0.034). A preferential utilization of lipids as an energy substrate with a reduction of the fat mass (p=0.01) and an increase of the free fat mass (p=0.0091) was found in alcoholic patients. Within our sample showing low ghrelin levels probably related to the impaired nutritional status; patients with higher levels of ghrelin showed higher levels of alcohol craving. These preliminary data indicate that ghrelin could be implicated in the neurobiological mechanisms of alcohol craving, other than a hormone influenced by the nutritional status.


Nanoscale zirconia as a nonmetallic catalyst for graphitization of carbon and growth of single- and multiwall carbon nanotubes.
We report that nanoparticulate zirconia (ZrO(2)) catalyzes both growth of single-wall and multiwall carbon nanotubes (CNTs) by thermal chemical vapor deposition (CVD) and graphitization of solid amorphous carbon. We observe that silica-, silicon nitride-, and alumina-supported zirconia on silicon nucleates single- and multiwall carbon nanotubes upon exposure to hydrocarbons at moderate temperatures (750 degrees C). High-pressure, time-resolved X-ray photoelectron spectroscopy (XPS) of these substrates during carbon nanotube nucleation and growth shows that the zirconia catalyst neither reduces to a metal nor forms a carbide. Point-localized energy-dispersive X-ray spectroscopy (EDAX) using scanning transmission electron microscopy (STEM) confirms catalyst nanoparticles attached to CNTs are zirconia. We also observe that carbon aerogels prepared through pyrolysis of a Zr(IV)-containing resorcinol-formaldehyde polymer aerogel precursor at 800 degrees C contain fullerenic cage structures absent in undoped carbon aerogels. Zirconia nanoparticles embedded in these carbon aerogels are further observed to act as nucleation sites for multiwall carbon nanotube growth upon exposure to hydrocarbons at CVD growth temperatures. Our study unambiguously demonstrates that a nonmetallic catalyst can catalyze CNT growth by thermal CVD while remaining in an oxidized state and provides new insight into the interactions between nanoparticulate metal oxides and carbon at elevated temperatures.


The Efficacy of Ketogenic Diet in 60 Chinese Patients With Dravet Syndrome.
Objective: To evaluate the efficacy and safety of ketogenic diet (KD) in patients with Dravet syndrome (DS). Methods: 60 DS patients receiving treatment of KD for more than 12 weeks from 2009 to 2018 were analyzed retrospectively. Modified Johns Hopkins protocol was used to initiate KD. Seizure frequency, electroencephalogram (EEG), cognition, language, and motor function of the patients were assessed. Side effects were monitored and adjusted accordingly. SPSS 23.0 software was used for all statistical analysis. Results: In total, 60 DS patients (34 boys, 26 girls) received treatment of KD for more than 12 weeks, and among them 41 (68.3%) patients remained on the diet for more than 24 weeks, 22 (36.7%) patients for more than 48 weeks. Seizures in 35 patients (58.3%) were reduced by over 50% at 12 weeks, and the KD effect was observed within 2 weeks in most of them. At 24 weeks, 61.1% (25/41) of the patients had a >50% seizure reduction. At 48 weeks, 77.3% (17/22) had an over 50% reduction in their seizure frequency. With the treatment of KD in the 60 DS patients, 10 patients had ever been seizure free for 12 months to 24 months (The median duration was 20 months). In 10 KD-effective patients, the background rhythm of their EEG showed obvious improvement, and interictal epileptic discharges decreased significantly. Cognitive function of 22 patients was improved. Language progressed in 14 patients. Motor function was improved in 13 patients. The efficacy of KD in DS patients did not correlate with the seizure onset age, the starting age of KD treatment, the SCN1A mutation and the numbers of antiepileptic drugs combined with KD treatment. The main adverse reactions of KD in the treatment process were gastrointestinal symptoms and metabolic disorders. Conclusions: KD treatment in DS patients has many advantages, including working rapidly, being effective in more than half of the DS patients and tolerable adverse reactions. Pharmoco-resistant DS patients are suggested to receive KD treatment.


Elucidation of the biosynthetic origin of the anti-inflammatory pseudopterosins.
The pseudopterosins are a family of diterpene glycosides isolated from the gorgonian coral Pseudopterogorgia elisabethae. These metabolites exhibit potent anti-inflammatory activity, and this review describes our efforts to elucidate their biosynthetic origin. A radioactivity-guided isolation was used to identify the terpene cyclase product. In addition, a detailed NMR-guided search for potential biosynthetic intermediates identified metabolites which were tested by incubating 3H-labeled analogues with a cell-free extract of the coral. All labeled metabolites were generated biosynthetically, and radiochemical purity was established by a combination of HPLC purification and derivatization. In summary, pseudopterosins are produced by a cyclization of geranylgeranyl diphosphate to elisabethatriene, aromatization to erogorgiaene, two successive oxidations to 7,8-dihydroxyerogorgiaene and a glycosylation to afford a seco-pseudopterosin as a key intermediate. A dehydrogenation leads to amphilectosins which undergo ring closures to yield the pseudopterosins.


Epidemiological trends of HIV-1 infection in blood donors from Catalonia, Spain (2005-2014).
Human immunodeficiency virus 1 (HIV-1) subtype B is predominant in Spain. However, the recent arrival of immigrant populations has increased the prevalence of non-B subtypes and circulating recombinant forms. The objective of this study was to determine the prevalence of HIV-1 subtypes and transmitted drug-resistance mutations in blood donors from the Catalonian region (northeastern Spain). HIV-1-positive blood donors identified in Catalonia from 2005 to 2014 were included. Demographic variables and risk factors for HIV-1 acquisition were recorded. HIV-1 subtyping was carried out by HIV-1 DNA polymerase region sequencing, and phylogenetic analyses were performed using the neighbor-joining method. During the study period, 2.8 million blood donations were screened, and 214 HIV-1-positive donors were identified, yielding an overall prevalence of 7.7 per 100,000 donations (89% men; mean age, 34 ± 10 years). Most HIV-1-positive donors were native to Spain (81%), and 61% were regular blood donors. When risk factors were known, 62% reportedly were men who had sex with men. HIV-1 subtyping was possible in 176 HIV-1-positive individuals: 143 (81%) had HIV-1 subtype B, and 33 (19%) had non-B subtypes. Most HIV-1 non-B subtypes were circulating recombinant forms (n = 20; 61%). Factors associated with HIV-1 subtype B were male sex (p = 0.007) and men who had sex with men (p < 0.001). The overall prevalence of transmitted drug-resistance mutations was 14%. Non-B subtypes, circulating recombinant forms, and transmitted drug-resistance mutation sequences circulate among HIV-1-positive blood donors in Catalonia. Continuous local epidemiological surveillance is required to implement optimal prevention strategies for controlling transfusion-transmitted HIV and to improve health policies regarding HIV infection.


Genomic organization and expression of the planarian homeobox genes Dth-1 and Dth-2.
We have characterized the genomic organization of Dth-1 and Dth-2, planarian homeobox-containing genes, previously described at the cDNA level (J. Garcia-Fernàndez, J. Baguñà and E. Saló (1991), Proc. Natl. Acad. Sci. USA, 88, 7338-7342). Genomic analysis shows that Dth-1 and Dth-2 genes encode proteins of 533 and 363 amino acids respectively. The open reading frame of Dth-1 is interrupted by two large introns of 8 kb and 12 kb Dth-2 also shows two introns, but these are short (42 bp and 44 bp) and the second interrupts helix III at position 44-45, as is the case with other homeobox genes from such divergent animals as Drosophila, honeybee, C. elegans, ascidians, and mouse, which suggests an ancient evolutionary relationship between these genes. The spatial distribution of transcripts in adult tissues, determined by in situ hybridization, demonstrates that Dth-1 is expressed at a high level in the gastrodermal cells, while Dth-2 is expressed in the peripheral parenchyma, at higher levels in the dorsal than the ventral regions. Their specific spatial distribution suggests a possible role for these homeobox genes in determination and/or differentiation of specific cell types. The expression pattern of both genes is more or less continuous, but in Dth-1 clustered discontinuous labelling in areas surrounding the gastrodermis may indicate a specific expression of this gene in groups of undifferentiated cells (neoblasts) already committed or determined to gastrodermal cell fates. In situ hybridization analysis during early regeneration shows expression only in the postblastema (stump) differentiated areas while no expression has been detected in the undifferentiated blastema, indicating that neither gene has a role in pattern formation mechanisms known to occur at the early stages of regeneration (0-3 days). Hence, Dth-1 and Dth-2 are planarian homeobox genes presumably involved in specific cell or tissue determination and/or differentiation.


Secondary prevention of venous thromboembolism with the oral direct thrombin inhibitor ximelagatran.
For many patients with venous thromboembolism, secondary prevention with vitamin K antagonists is not extended beyond six months, since the risk of recurrence may be outweighed by the risk of major bleeding. In a double-blind, multicenter trial, we randomly assigned 1233 patients with venous thromboembolism who had undergone six months of anticoagulant therapy to extended secondary prevention with the oral direct thrombin inhibitor ximelagatran (24 mg) or placebo, taken twice daily, for 18 months without monitoring of coagulation. At base line, bilateral ultrasonography of the legs and perfusion lung scanning were performed. Data from 612 patients in the ximelagatran group and 611 in the placebo group were analyzed. The occurrence of the primary end point, symptomatic recurrent venous thromboembolism, was confirmed in 12 patients assigned to ximelagatran and 71 patients assigned to placebo (hazard ratio, 0.16; 95 percent confidence interval, 0.09 to 0.30; P<0.001). Death from any cause occurred in 6 patients in the ximelagatran group and 7 patients in the placebo group, and bleeding occurred in 134 patients and 111 patients, respectively (hazard ratio, 1.19; 95 percent confidence interval, 0.93 to 1.53; P=0.17). The incidence of major hemorrhage was low (six events in the ximelagatran group and five in the placebo group), and none of these hemorrhages were fatal. The cumulative risk of a transient elevation of the alanine aminotransferase level to more than three times the upper limit of normal was 6.4 percent in the ximelagatran group, as compared with 1.2 percent in the placebo group (P<0.001). Oral ximelagatran was superior to placebo for the extended prevention of venous thromboembolism. There was no significant increase in the frequency of bleeding complications, but there was an increase in the number of patients with a transient elevation in the alanine aminotransferase level.


CCR5 signalling, but not DARC or D6 regulatory, chemokine receptors are targeted by herpesvirus U83A chemokine which delays receptor internalisation via diversion to a caveolin-linked pathway.
Herpesviruses have evolved chemokines and chemokine receptors, which modulate the recruitment of human leukocytes during the inflammatory response to infection. Early post-infection, human herpesvirus 6A (HHV-6A) infected cells express the chemokine receptor U51A and chemokine U83A which have complementary effects in subverting the CC-chemokine family thereby controlling anti-viral leukocyte recruitment. Here we show that, to potentiate this activity, the viral chemokine can also avoid clearance by scavenger chemokine receptors, DARC and D6, which normally regulate an inflammatory response. Conversely, U83A delays internalisation of its signalling target receptor CCR5 with diversion to caveolin rich membrane domains. This mechanism can redirect displaced human chemokines to DARC and D6 for clearance of the anti-viral inflammatory response, leaving the viral chemokine unchecked. Cell models for competitive binding assays were established using radiolabeled human chemokines and cold U83A on CCR5, DARC or D6 expressing cells. Flow cytometry was used to assess specific chemotaxis of CCR5 bearing cells to U83A, and internalisation of CCR5 specific chemokine CCL4 after stimulation with U83A. Internalisation analyses were supported by confocal microscopy of internalisation and co-localisation of CCR5 with caveosome marker caveolin-1, after virus or human chemokine stimulation. U83A displaced efficiently human chemokines from CCR5, with a high affinity of 0.01nM, but not from DARC or D6. Signalling via CCR5 resulted in specific chemoattraction of primary human leukocytes bearing CCR5. However, U83A effective binding and signalling to CCR5 resulted in delayed internalisation and recycling up to 2 hours in the absence of continual re-stimulation. This resulted in diversion to a delayed caveolin-linked pathway rather than the rapid clathrin mediated endocytosis previously shown with human chemokines CCL3 or CCL4. U83A diverts human chemokines from signalling, but not regulatory or scavenger, receptors facilitating their clearance, while occupying signalling receptors at the cell surface. This can enhance virus specific inflammation, facilitating dissemination to replication sensitive leukocytes while evading clearance; this has implications for linked neuro-inflammatory pathologies.


Plasmon-assisted Förster resonance energy transfer at the single-molecule level in the moderate quenching regime.
Metallic nanoparticles were shown to affect Förster energy transfer between fluorophore pairs. However, to date, the net plasmonic effect on FRET is still under dispute, with experiments showing efficiency enhancement and reduction. This controversy is due to the challenges involved in the precise positioning of FRET pairs in the near field of a metallic nanostructure, as well as in the accurate characterization of the plasmonic impact on the FRET mechanism. Here, we use the DNA origami technique to place a FRET pair 10 nm away from the surface of gold nanoparticles with sizes ranging from 5 to 20 nm. In this configuration, the fluorophores experience only moderate plasmonic quenching. We use the acceptor bleaching approach to extract the FRET rate constant and efficiency on immobilized single FRET pairs based solely on the donor lifetime. This technique does not require a posteriori correction factors neither a priori knowledge of the acceptor quantum yield, and importantly, it is performed in a single spectral channel. Our results allow us to conclude that, despite the plasmon-assisted Purcell enhancement experienced by donor and acceptor partners, the gold nanoparticles in our samples have a negligible effect on the FRET rate, which in turns yields a reduction of the transfer efficiency.


Injection anaesthesia with fentanyl-midazolam-medetomidine in adult female mice: importance of antagonization and perioperative care.
Injection anaesthesia is commonly used in laboratory mice; however, a disadvantage is that post-anaesthesia recovery phases are long. Here, we investigated the potential for shortening the recovery phase after injection anaesthesia with fentanyl-midazolam-medetomidine by antagonization with naloxone-flumazenil-atipamezole. In order to monitor side-effects, the depth of anaesthesia, heart rate (HR), core body temperature (BT) and concentration of blood gases, as well as reflex responses, were assessed during a 50 min anaesthesia. Mice were allowed to recover from the anaesthesia in their home cages either with or without antagonization, while HR, core BT and spontaneous home cage behaviours were recorded for 24 h. Mice lost righting reflex at 330 ± 47 s after intraperitoneal injection of fentanyl-midazolam-medetomidine. During anaesthesia, HR averaged 225 ± 23 beats/min, respiratory rate and core BT reached steady state at 131 ± 15 breaths/min and 34.3 ± 0.25℃, respectively. Positive pedal withdrawal reflex, movement triggered by tail pinch and by toe pinch, still occurred in 25%, 31.2% and 100% of animals, respectively. Arterial blood gas analysis revealed acidosis, hypoxia, hypercapnia and a marked increase in glucose concentration. After anaesthesia reversal by injection with naloxone-flumazenil-atipamezole, animals regained consciousness after 110 ± 18 s and swiftly returned to physiological baseline values, yet they displayed diminished levels of locomotion and disrupted circadian rhythm. Without antagonization, mice showed marked hypothermia (22 ± 1.9℃) and bradycardia (119 ± 69 beats/min) for several hours. Fentanyl-midazolam-medetomidine provided reliable anaesthesia in mice with reasonable intra-anaesthetic side-effects. Post-anaesthetic period and related adverse effects were both reduced substantially by antagonization with naloxone-flumazenil-atipamezole.


Comparing daily shoulder motion and frequency after anatomic and reverse shoulder arthroplasty.
Both anatomic (TSA) and reverse total shoulder arthroplasty (RTSA) are common interventions for glenohumeral arthrosis, with the goal of relieving pain and restoring mobility. Understanding shoulder arthroplasty motion and frequency is of interest in evaluating effectiveness and in predicting bearing wear for implant development and optimization. The purpose of this study was to measure and compare the total daily shoulder motion of patients after TSA and RTSA. Thirty-six human subjects who had undergone shoulder arthroplasty wore a custom instrumented garment that tracked upper extremity motion for the waking hours of 1 day. The 3-dimensional orientation of each humeral sensor was transformed with respect to the torso to calculate total joint motion and frequency, with comparison of TSA to RTSA. In addition, the yearly motion of the shoulder was extrapolated. The majority of shoulder motion occurred below 80° of elevation (P < .001), totaling on average 821 ± 45 and 783 ± 27 motions per hour for TSA and RTSA, respectively. Conversely, elevations >80° were significantly less frequent, totaling only 52 ± 44 (P < .001) and 38 ± 27 (P < .001) motions per hour for TSA and RTSA, respectively. No significant differences were detected between TSA and RTSA shoulders (P = .22) or their respective contralateral asymptomatic sides (P = .64, P = .62). When extrapolated, it was estimated that each TSA and RTSA shoulder elevated above 60° approximately 1 million and 0.75 million cycles per year, respectively. Mean shoulder motions after TSA or RTSA were not significantly different from the contralateral asymptomatic side. In addition, no significant differences were detected in shoulder motion or frequency between TSA and RTSA.


Cell-specific expression of aromatase and LH receptor mRNAs in rat ovary.
Current understanding of the endocrine and paracrine regulation of follicular oestrogen synthesis predicts that aromatase cytochrome P450 (P450arom) mRNA is inducible by FSH in granulosa cells. LH receptor mRNA is constitutively expressed in thecal/interstitial cells, and is also thought to be induced in granulosa cells in response to joint stimulation by FSH and oestrogen. This study provides direct evidence that FSH induces the ovarian P450arom gene selectively, perhaps exclusively, in the granulosa cells of Graafian follicles. FSH-induction of LH receptor mRNA occurs simultaneously but is independent of oestrogen synthesis per se.


Physical activity and the risk of breast cancer in BRCA1/2 mutation carriers.
BRCA1/2 mutation carriers have a high lifetime risk of developing breast cancer. Differences in penetrance indicate that this risk may be influenced by lifestyle factors. Because physical activity is one of the few modifiable risk factors, it may provide a target to add to breast cancer prevention in this high-risk population. We examined the association between self-reported lifetime sports activity and breast cancer risk in a nationwide retrospective cohort study, including 725 carriers, of whom 218 had been diagnosed with breast cancer within 10 years prior to questionnaire completion. We found a nonsignificantly decreased risk for ever engaging in sports activity (HR = 0.84, 95%CI = 0.57-1.24). Among women who had participated in sports, a medium versus low level of intensity and duration (i.e., between 11.0 and 22.7 mean MET hours/week averaged over a lifetime) reduced the risk of breast cancer (HR = 0.59, 95%CI = 0.36-0.95); no dose-response trend was observed. For mean hours/week of sports activity, a nonsignificant trend was observed (HR(low versus never) = 0.93, 95%CI = 0.60-1.43; HR(medium versus never) = 0.81, 95%CI = 0.51-1.29; HR(high versus never) = 0.78, 95%CI = 0.48-1.29; p (trend overall) = 0.272; p (trend active women) = 0.487). For number of years of sports activity no significant associations were found. Among women active in sports before age 30, mean MET hours/week showed the strongest inverse association of all activity measures (HR(medium versus low) = 0.60, 95%CI = 0.38-0.96; HR(high versus low) = 0.58, 95%CI = 0.35-0.94; p (trend) = 0.053). Engaging in sports activity after age 30 was also inversely associated with breast cancer risk (HR = 0.63, 95%CI = 0.44-0.91). Our results indicate that sports activity may reduce the risk of breast cancer in BRCA1/2 mutation carriers.


The pharmacokinetics and safety of ABT-751, a novel, orally bioavailable sulfonamide antimitotic agent: results of a phase 1 study.
Microtubules play a critical role in many cellular functions, including cell division and mitosis. ABT-751 is a novel sulfonamide antimitotic that binds to the colchicine site on beta-tubulin that leads to a block in the cell cycle at the G2M phase, resulting in cellular apoptosis. ABT-751 was investigated in this phase 1 trial designed to assess its maximum tolerated dose (MTD), dose-limiting toxicity (DLT), tolerability, and pharmacokinetics. ABT-751 was administered on a daily (q.d.) or twice daily (b.i.d.) oral schedule for 7 days every 3 weeks to 39 patients with refractory solid tumors. Toxicity was monitored weekly. Plasma and urine ABT-751 and metabolite pharmacokinetics were determined. The MTD for the q.d. schedule was 250 mg/d. DLTs during cycle 1 were abdominal pain, constipation, and fatigue. The MTD on the b.i.d. schedule was 150 mg. Cycle 1 of therapy with the 175 mg b.i.d. schedule was tolerated without DLT. However, six of seven patients reported grade 3 toxicity (ileus, constipation, abdominal pain, or fatigue), which occurred in cycle 2 or 3. ABT-751 was absorbed after oral administration with an overall mean T(max) of about 2 hours. The pharmacokinetics of ABT-751 were dose-proportional and time-independent. There was minimal accumulation of ABT-751 after multiple q.d. and b.i.d. doses. Efficacious concentrations, as determined from preclinical models (0.5-1.5 microg/mL), were achieved in all subjects. ABT-751 metabolism occurred primarily by glucuronidation and sulfation. No complete or partial tumor responses were noted, but one patient had a minor response, and four patients had stable disease lasting at least 6 months. The MTD and recommended phase 2 doses for ABT-751 were 250 mg q.d. and 150 mg b.i.d. on a 7-day schedule given every 3 weeks, due to subsequent cycle toxicities at 175 mg b.i.d. dosing. Toxicities were abdominal pain, constipation, and neuropathy.


COVID-19 infection and vaccines: potential triggers of Herpesviridae reactivation.
Since the onset of the COVID-19 outbreak, numerous articles have highlighted a possible link between COVID-19 vaccination or infection and Herpesviridae co-infection or reactivation. The authors conducted an exhaustive literature review on this topic, the results of which are presented individually for each member of the Herpesviridae family: Herpes Simplex Virus (HSV) types-1 (HSV-1) and 2 (HSV-2); Varicella-Zoster Virus (VZV); Epstein-Barr Virus (EBV); Cytomegalovirus (CMV); HHV-6; HHV-7; and HHV-8. These human herpesviruses can serve as prognostic markers for the COVID-19 infection and may even underlie some of the clinical manifestations initially attributed to SARS-CoV-2. In addition to SARS-CoV-2 infection, all corresponding vaccines approved to date in Europe appear capable of inducing herpesvirus reactivation. It is important to consider all viruses of the Herpesviridae family when managing patients infected with or recently vaccinated against COVID-19.


Insights into human evolution from ancient and contemporary microbiome studies.
Over the past decade, human microbiome research has energized the study of human evolution through a complete shift in our understanding of what it means to be human. The microbiome plays a pivotal role in human biology, performing key functions in digestion, mood and behavior, development and immunity, and a range of acute and chronic diseases. It is therefore critical to understand its evolution and changing ecology through time. Here we review recent findings on the microbiota of diverse human populations, non-human primates, and past human populations and discuss the implications of this research in formulating a deeper evolutionary understanding of the human holobiont.


Do Callous-Unemotional Traits and Conduct Disorder Symptoms Predict the Onset and Development of Adolescent Substance Use?
Despite strong evidence of the relationship between conduct disorder (CD) symptoms and substance use, it is unclear how callous-unemotional (CU) traits predict substance use over and above CD symptoms, and their potential interaction. This study used data from 753 participants followed from grade 7 to 2-years post-high school. Latent growth curve models showed that CU traits predicted the onset of cigarette use, alcohol misuse, and a substance use composite at grade 7 only when no CD symptoms were present. Among those without CD symptoms, boys showed greater change in the odds of using cigarettes, and were more likely to misuse alcohol or use any substance at grade 7 than girls. However, CD symptoms, CU traits, and their interaction did not predict the linear rates of growth of substance use over time. Thus, CU traits may uniquely predict adolescent substance use when CD symptoms are not present. This research has implications for predicting onset of adolescent substance use and for incorporating the assessment of CU traits into interventions targeting adolescent substance use.


Risk for tumor and other disease transmission by transplantation: a population-based study of unrecognized malignancies and other diseases in organ donors.
Organ donation may involve the risk of transmittal of unwanted host factors such as infections and malignancy. These may be concealed in the emergent donation process. It may be unavoidable if first observed in a donor postmortem. A number of reports on transferred cancers have been published, but quantification of the risk has never been reliably performed. We report here the first population-based analysis of unrecognized malignancies and other diseases in cadaveric or living-related donors and the possible consequences for the recipients. We compiled a cohort of all organ donors through 27 years (1969-1996) in one single kidney transplant center covering a population on one million people. This cohort was linked to the Danish Cancer Registry, the Danish National Hospital Register, and the Danish Register of Causes of Death by means of the unique personal identification number, and all cancers, diagnosis from hospital admissions, and causes of death were identified. Follow-up was to the end of 1996. A total of 626 donors (491 cadaveric and 135 living-related donors) was included in the study. Ten carcinoma in situ or dysplasia cervix uteri (by definition nonmalignant), and 13 malignant tumors (5 of these were detected in living-related donors after donation) were detected by linkage to the cancer registry. All together, 17 recipients received organs from donors with carcinoma in situ or dysplasia cervix uteri and 20 from donors with malignancies. Two recipients from organ donors with carcinoma in situ or dysplasia of the cervix uteri and two recipients from donors with malignancies had a cancer detected; however, these were likely unrelated. One died 1 year after transplantation from a melanoma transmitted from the donor. Two cadaveric donors had previous admissions for glomerulonephritis, five for pyelonephritis, five for nephrolithiasis or ureterolithiasis, four for cystitis, and one for hydronephrosis. Despite all efforts to secure a safe organ for transplantation, transmission of donor malignancy and other diseases nevertheless can happen, as is recorded many times in the literature. We have quantified the risk using the population-based cancer registry and found a risk of 8 in 626 (1.3%) for having a donor with undetected malignancy and a risk of 1 in 626 (0.2%) for transmitting a cancer. The risk for getting some transmitted glomerulonephritis is 2 in 626 (0.3%). None of the donors with cerebral malignancies transmitted any tumors to the recipients. Compared with the benefits of organ transplantation, these risks are small; however, if time allows, a search for additional medical information from registries could further minimize the risk of transmission of malignancies or other diseases. However, this requires updated, accurate, and accessible registries and legislation that allows access to personal data and transmission of such data across administrative borders.


Land motor vehicle-related drownings in Sweden.
Knowledge regarding drownings related to land motor vehicle events is limited although reports of these events occur globally. This study elucidates crash and injury mechanisms in motor vehicle-related drownings in a Swedish population during 1992 through 2006 in order to suggest preventive countermeasures. The cases were identified in the National Board of Forensic Medicine database and crosschecked against the official statistics. All available autopsy reports, hospital records, and police records were analyzed. In addition, corresponding in-depth analyses performed by the Swedish Road Administration were reviewed and analyzed. In total, 83 drownings occurred in 64 vehicles. The great majority of victims had no serious injuries (92% MAIS <or= 2) and would probably have survived if they had not drowned. Most events took place in waters directly adjacent to a roadway (36%) or bridge (34%). The vehicles were most often (72%) found upside down, and most drownings occurred in shallow water (65% depth <2 m). One third (32%) of the drivers tested positive for alcohol (mean BAC of 2.0 g/L; range 0.16-2.6). The majority (69%) of the fatalities could possibly have been prevented if effective guardrails had been in place. Drownings in motor vehicles are not negligible events and in many cases they are preventable. Sufficient design and placement of guardrails can minimize these events, but further experimental investigation should be conducted to better understand these events and how to optimize vehicle design, rescue operations, and self-rescue.


Modeling Molecular Interactions in Water: From Pairwise to Many-Body Potential Energy Functions.
Almost 50 years have passed from the first computer simulations of water, and a large number of molecular models have been proposed since then to elucidate the unique behavior of water across different phases. In this article, we review the recent progress in the development of analytical potential energy functions that aim at correctly representing many-body effects. Starting from the many-body expansion of the interaction energy, specific focus is on different classes of potential energy functions built upon a hierarchy of approximations and on their ability to accurately reproduce reference data obtained from state-of-the-art electronic structure calculations and experimental measurements. We show that most recent potential energy functions, which include explicit short-range representations of two-body and three-body effects along with a physically correct description of many-body effects at all distances, predict the properties of water from the gas to the condensed phase with unprecedented accuracy, thus opening the door to the long-sought "universal model" capable of describing the behavior of water under different conditions and in different environments.


Ascospores of large-spored Metschnikowia species are genuine meiotic products of these yeasts.
The asci of Metschnikowia species normally contain two ascospores (never more), raising the question of whether these spores are true meiotic products. We investigated this problem by crossing genetically-marked strains of the haploid, heterothallic taxa Metschnikowia hawaiiensis, Metschnikowia continentalis var. continentalis, and M. continentalis var. borealis. Asci were dissected and the segregation patterns for various phenotypes analyzed. In all cases (n = 47) both mating types (h+ and h-) were recovered in pairs of sister spores, casting further uncertainty as to whether normal meiosis takes place. However, the segregation patterns for cycloheximide resistance and several auxotrophic markers were random, suggesting that normal meiosis indeed occurs. To explain the lack of second-division segregation of mating types, we concluded that crossing-over does not occur between the mating-type locus and the centromere, and that meiosis I is tied to spore formation, which explains why the number of spores is limited to two. The latter assumption was also supported by fluorescence microscopy. The second meiotic division takes place inside the spores and is followed by the resorption of two nuclei, one in each spore.


[Burnout syndrome among nursing staff at a hospital in Madrid].
The term "burnout" is related to a situation arising increasingly more often among the professionals performing their duties by way of a long-term, direct, people-to-people relationship, which includes all healthcare professionals. This study is aimed at determining the prevalence of the Burnout syndrome and of the three components involved therein (emotional exhaustion, impersonalization and lack of personal fulfillment) among the nursing staff at the "Hospital Clínico Universitario San Carlos" in Madrid and the relationship thereof to certain socio-demographic, job-related and institutional factors. A cross-sectional study was conducted among the professionals assigned to the nursing staff at the above-mentioned hospital. The variables involved were gathered by means of a questionnaire prepared by those conducting this study. The Burnout syndrome was measured by means of the Maslach Burnout Inventory, in the validated Spanish version thereof. The nursing staff is more impersonalized (p=0.004) and less fulfilled (p=0.036) than the nursing assistant/technician group. When the results of the four scales by units were analyzed, burnout was found to be greater among the nursing staff assigned to oncology and emergency care units (p=0.001), the impersonalization in the emergency rooms (p=0.007), and Burnout is once again greater in the oncology and emergency units (p=0.000). Those professionals who answered that there was little recognition of their nursing care scored worst regarding Burnout and the three aspects thereof (p =0.000). The lower the degree of on-the-job satisfaction, the higher the scores on the four scales (p=0.000). The conclusion which may be drawn from this study is that the profile of a person affected by Burnout is that of a professional with on-the-job experience who nevertheless considers very little recognition to be given to their caregiving and a high degree of dissatisfaction with the way in which their workplaces are managed.


Woody plant encroachment into grasslands: spatial patterns of functional group distribution and community development.
Woody plant encroachment into grasslands has been globally widespread. The woody species invading grasslands represent a variety of contrasting plant functional groups and growth forms. Are some woody plant functional types (PFTs) better suited to invade grasslands than others? To what extent do local patterns of distribution and abundance of woody PFTs invading grasslands reflect intrinsic topoedaphic properties versus plant-induced changes in soil properties? We addressed these questions in the Southern Great Plains, United States at a subtropical grassland known to have been encroached upon by woody species over the past 50-100 years. A total of 20 woody species (9 tree-statured; 11 shrub-statured) were encountered along a transect extending from an upland into a playa basin. About half of the encroaching woody plants were potential N2-fixers (55% of species), but they contributed only 7% to 16 % of the total basal area. Most species and the PFTs they represent were ubiquitously distributed along the topoedaphic gradient, but with varying abundances. Overstory-understory comparisons suggest that while future species composition of these woody communities is likely to change, PFT composition is not. Canonical correspondence analysis (CCA) ordination and variance partitioning (Partial CCA) indicated that woody species and PFT composition in developing woody communities was primarily influenced by intrinsic landscape location variables (e.g., soil texture) and secondarily by plant-induced changes in soil organic carbon and total nitrogen content. The ubiquitous distribution of species and PFTs suggests that woody plants are generally well-suited to a broad range of grassland topoedaphic settings. However, here we only examined categorical and non-quantitative functional traits. Although intrinsic soil properties exerted more control over the floristics of grassland-to-woodland succession did plant modifications of soil carbon and nitrogen concentrations, the latter are likely to influence productivity and nutrient cycling and may, over longer time-frames, feed back to influence PFT distributions.


Chemically-defined camelid antibody bioconjugate for the magnetic resonance imaging of Alzheimer's disease.
Today, molecular imaging of neurodegenerative diseases is mainly based on small molecule probes. Alternatively, antibodies are versatile tools that may be developed as new imaging agents. Indeed, they can be readily obtained to specifically target any antigen of interest and their scaffold can be functionalized. One of the critical issues involved in translating antibody-based probes to the clinic is the design and synthesis of perfectly-defined conjugates. Camelid single-domain antibody-fragments (VHHs) are very small and stable antibodies that are able to diffuse in tissues and potentially cross the blood brain barrier (BBB). Here, we selected a VHH (R3VQ) specifically targeting one of the main lesions of Alzheimer's disease (AD), namely the amyloid-beta (Aß) deposits. It was used as a scaffold for the design of imaging probes for magnetic resonance imaging (MRI) and labeled with the contrastophore gadolinium using either a random or site-specific approach. In contrast to the random strategy, the site-specific conjugation to a single reduced cysteine in the C-terminal part of the R3VQ generates a well-defined bioconjugate in a high yield process. This new imaging probe is able to cross the BBB and label Aß deposits after intravenous injection. Also, it displays improved r1 and r2 relaxivities, up to 30 times higher than a widely used clinical contrast agent, and it allows MRI detection of amyloid deposits in post mortem brain tissue of a mouse model of AD. The ability to produce chemically-defined VHH conjugates that cross the BBB opens the way for future development of tailored imaging probes targeting intracerebral antigens.


NADPH Oxidase 2-Derived Reactive Oxygen Species Promote CD8+ T Cell Effector Function.
Oxidants participate in lymphocyte activation and function. We previously demonstrated that eliminating the activity of NADPH oxidase 2 (NOX2) significantly impaired the effectiveness of autoreactive CD8+ CTLs. However, the molecular mechanisms impacting CD8+ T cell function remain unknown. In the present study, we examined the role of NOX2 in both NOD mouse and human CD8+ T cell function. Genetic ablation or chemical inhibition of NOX2 in CD8+ T cells significantly suppressed activation-induced expression of the transcription factor T-bet, the master transcription factor of the Tc1 cell lineage, and T-bet target effector genes such as IFN-γ and granzyme B. Inhibition of NOX2 in both human and mouse CD8+ T cells prevented target cell lysis. We identified that superoxide generated by NOX2 must be converted into hydrogen peroxide to transduce the redox signal in CD8+ T cells. Furthermore, we show that NOX2-generated oxidants deactivate the tumor suppressor complex leading to activation of RheB and subsequently mTOR complex 1. These results indicate that NOX2 plays a nonredundant role in TCR-mediated CD8+ T cell effector function.


Abnormal contrast responses in the extrastriate cortex of blindsight patients.
When the human primary visual cortex (V1) is damaged, the dominant geniculo-striate pathway can no longer convey visual information to the occipital cortex. However, many patients with such damage retain some residual visual function that must rely on an alternative pathway directly to extrastriate occipital regions. This residual vision is most robust for moving stimuli, suggesting a role for motion area hMT+. However, residual vision also requires high-contrast stimuli, which is inconsistent with hMT+ sensitivity to contrast in which even low-contrast levels elicit near-maximal neural activation. We sought to investigate this discrepancy by measuring behavioral and neural responses to increasing contrast in patients with V1 damage. Eight patients underwent behavioral testing and functional magnetic resonance imaging to record contrast sensitivity in hMT+ of their damaged hemisphere, using Gabor stimuli with a spatial frequency of 1 cycle/°. The responses from hMT+ of the blind hemisphere were compared with hMT+ and V1 responses in the sighted hemisphere of patients and a group of age-matched controls. Unlike hMT+, neural responses in V1 tend to increase linearly with increasing contrast, likely reflecting a dominant parvocellular channel input. Across all patients, the responses in hMT+ of the blind hemisphere no longer showed early saturation but increased linearly with contrast. Given the spatiotemporal parameters used in this study and the known direct subcortical projections from the koniocellular layers of the lateral geniculate nucleus to hMT+, we propose that this altered contrast sensitivity in hMT+ could be consistent with input from the koniocellular pathway.


Potential role of an endothelium-specific growth factor, hepatocyte growth factor, on endothelial damage in diabetes.
Endothelial cells are known to secrete various antiproliferative and vasodilating factors. Although injury of endothelial cells has been postulated as an initial trigger of the progression of atherosclerosis in patients with diabetes, the mechanisms of endothelial injury in diabetes are not yet clarified. Therefore, it is important to know the effects of high glucose on the factors that may influence endothelial cell growth. A novel member of endothelium-specific growth factors, hepatocyte growth factor (HGF), is produced in vascular cells. To investigate the effects of high glucose on vascular cells, we examined 1) the effects of high glucose on endothelial cell and vascular smooth muscle cell (VSMC) growth and 2) the effects of high glucose on local HGF production in endothelial cell and VSMC. Treatment of human aortic endothelial cell with a high concentration of D-glucose, but not mannitol and L-glucose, resulted in a significant decrease in cell number. Interestingly, addition of recombinant HGF attenuated high D-glucose-induced endothelial cell death. Therefore, we measured local HGF secretion of endothelial cell. Importantly, local HGF production was significantly decreased by high D-glucose treatment. In contrast, high D-glucose treatment resulted in a significant increase in the number of human aortic VSMCs, whereas local HGF production was significantly decreased in accordance with increase in D-glucose concentration. No significant changes in numbers were observed in VSMC treated with high mannitol and L-glucose. We also studied the mechanisms of local HGF suppression by high D-glucose. High D-glucose treatment stimulated transforming growth factor-beta (TGF-beta) concentration in endothelial cell and VSMC. Decreased local vascular HGF production was abolished by addition of anti-TGF-beta antibody. As TGF-beta inhibited local HGF production in endothelial cell and VSMC, increased TGF-beta induced by high D-glucose may suppress local HGF production. This study demonstrated that high D-glucose induced endothelial cell death, stimulated VSMC growth, and decreased local HGF production through the stimulation of TGF-beta production both in endothelial cell and VSMC. Overall, decrease in a local endothelial stimulant, HGF, by high D-glucose may be a trigger of endothelial injury in diabetes, potentially resulting in the progression of atherosclerosis.


Chemical and morphological studies of bacterial spore formation. II. Spore and parasporal protein formation in Bacillus cereus var. alesti.
The development of both the spore and parasporal protein crystal of Bacillus cereus var. alesti was followed using chemical and cytological techniques. The changes which led to the formation of the fore-spore were similar to those already described for Bacillus cereus. However, adjacent to the developing fore-spore a small inclusion became discernible in phase contrast. This protein inclusion during its growth was differentiated from the chromatin and lipid-containing inclusions by sequential staining techniques. During spore and crystal formation no net synthesis of either nucleic acid was detected. Tracer studies with radioactive phosphorus confirmed that the spore chromatin was derived from that in the vegetative cell. These same studies also indicated that a turnover of ribonucleic acid occurred during the sporulation process. During their formation both the spore and crystal incorporated methionine-(35)S from the medium and from cellular material into a bound form. Sequential extractions with alkali and with alkaline-thioglycollate reagent revealed that the solubility characteristics of the mature crystal were possibly related to the presence of intermolecular disulphide bonds which developed after the major synthesis of the crystal was complete. The synthetic nature of sporogenesis and crystal formation is discussed with reference to the concept of "endotrophic" sporulation.


Additive manufacturing of biologically-inspired materials.
Additive manufacturing (AM) technologies offer an attractive pathway towards the fabrication of functional materials featuring complex heterogeneous architectures inspired by biological systems. In this paper, recent research on the use of AM approaches to program the local chemical composition, structure and properties of biologically-inspired materials is reviewed. A variety of structural motifs found in biological composites have been successfully emulated in synthetic systems using inkjet-based, direct-writing, stereolithography and slip casting technologies. The replication in synthetic systems of design principles underlying such structural motifs has enabled the fabrication of lightweight cellular materials, strong and tough composites, soft robots and autonomously shaping structures with unprecedented properties and functionalities. Pushing the current limits of AM technologies in future research should bring us closer to the manufacturing capabilities of living organisms, opening the way for the digital fabrication of advanced materials with superior performance, lower environmental impact and new functionalities.


Purification of a novel MHC class I element binding activity from thymus nuclear extracts reveals that thymic RBP-Jkappa/CBF1 binds to NF-kappaB-like elements.
We purified a DNA binding protein that recognizes a portion of the MHC class I regulatory element region 1/NF-kappaB binding site whose expression correlates with the expression of a MHC class I transgene in the thymus. The N-terminal amino acid sequence and the molecular size matched the RBP-Jkappa protein, also known as the EBV C-promoter binding factor, CBF1. Anti-peptide sera reactive with RBP-Jkappa/CBF1 also reacted with this protein in gel mobility shift assays. Although RBP-Jkappa/CBF1 is ubiquitously expressed, binding to the MHC class Ia NF-kappaB site was limited to the thymus. Comparison of the DNA binding specificities of RBP-Jkappa/CBF1 in thymic and splenic nuclear extracts revealed strong binding from both extracts to an IFN-beta kappaB site containing the RBP-Jkappa/CBF1 consensus sequence (CGTGGGAA). In contrast, only the thymic nuclear extract showed strong DNA binding activity with probes containing the NF-kappaB recognition sequences present in the MHC class Ia, IL-2Ralpha, and granulocyte-macrophage CSF promoters. Thus, RBP-Jkappa/CBF1 in thymic extracts demonstrates a clearly distinguishable DNA binding specificity that correlates with tissue-specific expression of a class I transgene. This, coupled with the fact that our previous study showed enhanced expression of the transgene in CD4+CD8+ thymocytes, suggests that RBP-Jkappa/CBF1 may play a role in the development of the immune system.


Experience of a network of transfusion coordinators for blood conservation (Ontario Transfusion Coordinators [ONTraC]).
The experiences of the development of a provincial program to promote blood conservation are herein reported. Transfusion coordinators were placed in 23 Ontario hospitals. Anonymized laboratory and clinical information was collected in a defined number of all consecutive patients admitted for three designated procedures: knee arthroplasty, abdominal aortic aneurysm (AAA), and coronary artery bypass graft (CABG) surgery (n approximately 1100, 300, and 300 at each time period, respectively). Considerable interinstitutional variation was observed in the proportion of patients who received transfusions. At 12 months, and over the 24-month period of the project, most hospitals demonstrated decreased use of allogeneic blood; at 12 months an approximate 24 percent reduction in patients undergoing knee surgery, 14 percent in AAA, and 23 percent in CABG was obtained. In addition, patients who received transfusions received less allogeneic blood. Patients who did not receive allogeneic transfusions had lower postoperative infection rates (p < 0.05) and length of stay (p < 0.0001); allogeneic transfusion was an independent predictor of increased length of stay. The main blood conservation measures employed during this time were education, preoperative autologous donation, erythropoietin, and cell salvage. The implementation of a provincial network of transfusion coordinators was feasible and allogeneic transfusion rates declined over the period the program has been in place.


Genetic evidence for a protective role of the peritrophic matrix against intestinal bacterial infection in Drosophila melanogaster.
The peritrophic matrix (PM) forms a layer composed of chitin and glycoproteins that lines the insect intestinal lumen. This physical barrier plays a role analogous to that of mucous secretions of the vertebrate digestive tract and is thought to protect the midgut epithelium from abrasive food particles and microbes. Almost nothing is known about PM functions in Drosophila, and its function as an immune barrier has never been addressed by a genetic approach. Here we show that the Drosocrystallin (Dcy) protein, a putative component of the eye lens of Drosophila, contributes to adult PM formation. A loss-of-function mutation in the dcy gene results in a reduction of PM width and an increase of its permeability. Upon bacterial ingestion a higher level of expression of antibacterial peptides was observed in dcy mutants, pointing to an influence of this matrix on bacteria sensing by the Imd immune pathway. Moreover, dcy-deficient flies show an increased susceptibility to oral infections with the entomopathogenic bacteria Pseudomonas entomophila and Serratia marcescens. Dcy mutant flies also succumb faster than wild type upon ingestion of a P. entomophila toxic extract. We show that this lethality is due in part to an increased deleterious action of Monalysin, a pore-forming toxin produced by P. entomophila. Collectively, our analysis of the dcy immune phenotype indicates that the PM plays an important role in Drosophila host defense against enteric pathogens, preventing the damaging action of pore-forming toxins on intestinal cells.


A ligand's-eye view of protein binding.
Docking tools created for structure-based design and virtual screening have also been used to automate ligand alignment for comparative molecular field analysis (CoMFA). Models based on such alignments have been compared with those obtained based solely on shared ligand substructures, but such comparisons have generally failed to distinguish between conformational specification (alignment in the internal coordinate space) and embedding in a shared external frame of reference (Cartesian alignment). Here, large sets of inhibitors were docked into two cyclooxygenase and two reverse transcriptase crystal structures, and the poses generated were evaluated in terms of the CoMFA models they produced. Realigning the conformers obtained by docking by rigid-body rotation and translation to overlay their common substructures improved model statistics and interpretability, provided the protein structure used for docking was reasonably appropriate to the ligands being considered.


Chaparral-associated hepatotoxicity.
Personal health care practices that may include the use of dietary supplements are common in the United States. Products marketed as dietary supplements are diverse and may include botanicals, vitamins, and/or minerals. Chaparral (Larrea tridentata) is a botanical dietary supplement made from a desert shrub and used for its antioxidant properties. Several reports of chaparral-associated hepatitis have been published since 1990, but a complete picture of the clinical presentation is still unclear. We reviewed the 18 case reports of adverse events associated with the ingestion of chaparral reported to the Food and Drug Administration between 1992 and 1994. These reports were from health care professionals, state health departments, and individual consumers. Of 18 reports of illnesses associated with the ingestion of chaparral, there was evidence of hepatotoxicity in 13 cases. Clinical presentation, characterized as jaundice with a marked increase in serum liver chemistry values, occurred 3 to 52 weeks after the ingestion of chaparral, and it resolved 1 to 17 weeks after most individuals stopped their intake of chaparral. The predominant pattern of liver injury was characterized as toxic or drug-induced cholestatic hepatitis; in 4 individuals, there was progression to cirrhosis; and in 2 individuals, there was acute fulminant liver failure that required liver transplants. These data indicate that the use of chaparral may be associated with acute to chronic irreversible liver damage with fulminant hepatic failure, and they underscore the potential for certain dietary supplement ingredients to cause toxic effects on the liver. Health professionals should be encouraged to inquire routinely about the use of dietary supplements and other products, to be alert to potential adverse effects that may be associated with these products, and, finally, to report any serious adverse events associated with these products through the MEDWatch Program of the Food and Drug Administration.


Inositol pyrophosphates as mammalian cell signals.
Inositol pyrophosphates are highly energetic inositol polyphosphate molecules present in organisms from slime molds and yeast to mammals. Distinct classes of enzymes generate different forms of inositol pyrophosphates. The biosynthesis of these substances principally involves phosphorylation of inositol hexakisphosphate (IP₆) to generate the pyrophosphate IP₇. Initial insights into functions of these substances derived primarily from yeast, which contain a single isoform of IP₆ kinase (yIP₆K), as well as from the slime mold Dictyostelium. Mammalian functions for inositol pyrophosphates have been investigated by using cell lines to establish roles in various processes, including insulin secretion and apoptosis. More recently, mice with targeted deletion of IP₆K isoforms as well as the related inositol polyphosphate multikinase (IPMK) have substantially enhanced our understanding of inositol polyphosphate physiology. Phenotypic alterations in mice lacking inositol hexakisphosphate kinase 1 (IP₆K1) reveal signaling roles for these molecules in insulin homeostasis, obesity, and immunological functions. Inositol pyrophosphates regulate these processes at least in part by inhibiting activation of the serine-threonine kinase Akt. Similar studies of IP₆K2 establish this enzyme as a cell death inducer acting by stimulating the proapoptotic protein p53. IPMK is responsible for generating the inositol phosphate IP₅ but also has phosphatidylinositol 3-kinase activity--that participates in activation of Akt. Here, we discuss recent advances in understanding the physiological functions of the inositol pyrophosphates based in substantial part on studies in mice with deletion of IP₆K isoforms. These findings highlight the interplay of IPMK and IP₆K in regulating growth factor and nutrient-mediated cell signaling.


Aggressive chemotherapy and the selection of drug resistant pathogens.
Drug resistant pathogens are one of the key public health challenges of the 21st century. There is a widespread belief that resistance is best managed by using drugs to rapidly eliminate target pathogens from patients so as to minimize the probability that pathogens acquire resistance de novo. Yet strong drug pressure imposes intense selection in favor of resistance through alleviation of competition with wild-type populations. Aggressive chemotherapy thus generates opposing evolutionary forces which together determine the rate of drug resistance emergence. Identifying treatment regimens which best retard resistance evolution while maximizing health gains and minimizing disease transmission requires empirical analysis of resistance evolution in vivo in conjunction with measures of clinical outcomes and infectiousness. Using rodent malaria in laboratory mice, we found that less aggressive chemotherapeutic regimens substantially reduced the probability of onward transmission of resistance (by >150-fold), without compromising health outcomes. Our experiments suggest that there may be cases where resistance evolution can be managed more effectively with treatment regimens other than those which reduce pathogen burdens as fast as possible.


Workers Without Paid Sick Leave Less Likely To Take Time Off For Illness Or Injury Compared To Those With Paid Sick Leave.
Paid sick leave is an important employer-provided benefit that helps people obtain health care for themselves and their dependents. But paid sick leave is not universally available to US workers. Little is known about paid sick leave and its relationship to health behaviors. Contrary to public health goals to reduce the spread of illness, our findings indicate that in 2013 both full- and part-time working adults without paid sick leave were more likely than workers with that benefit to attend work when ill. Those without paid sick leave were 3.0 times more likely to forgo medical care for themselves and 1.6 times more likely to forgo medical care for their family compared to working adults with paid sick leave benefits. Moreover, the lowest-income group of workers without paid sick leave were at the highest risk of delaying and forgoing medical care for themselves and their family members. Policy makers should consider the potential public health implications of their decisions when contemplating guaranteed sick leave benefits.


RF dosimetry: a comparison between power absorption of female and male numerical models from 0.1 to 4 ghz.
Realistic numerical models of human subjects and their surrounding environment represent the basic points of radiofrequency (RF) electromagnetic dosimetry. This also involves differentiating the human models in men and women, possibly with different body shapes and postures. In this context, the aims of this paper are, firstly, to propose a female dielectric anatomical model (fDAM) and, secondly, to compare the power absorption distributions of a male and a female model from 0.1 to 4 GHz. For realizing the fDAM, a magnetic resonance imaging tomographer to acquire images and a recent technique which avoids the discrete segmentation of body tissues into different types have been used. Simulations have been performed with the FDTD method by using a novel filtering-based subgridding algorithm. The latter is applied here for the first time to dosimetry, allowing an abrupt mesh refinement by a factor of up to 7. The results show that the whole-body-averaged specific absorption rate (WBA-SAR) of the female model is higher than that of the male counterpart, mainly because of a thicker subcutaneous fat layer. In contrast, the maximum averaged SAR over 1 g (1gA-SAR) and 10 g (10gA-SAR) does not depend on gender, because it occurs in regions where no subcutaneous fat layer is present.


Unified modeling of gene duplication, loss, and coalescence using a locus tree.
Gene phylogenies provide a rich source of information about the way evolution shapes genomes, populations, and phenotypes. In addition to substitutions, evolutionary events such as gene duplication and loss (as well as horizontal transfer) play a major role in gene evolution, and many phylogenetic models have been developed in order to reconstruct and study these events. However, these models typically make the simplifying assumption that population-related effects such as incomplete lineage sorting (ILS) are negligible. While this assumption may have been reasonable in some settings, it has become increasingly problematic as increased genome sequencing has led to denser phylogenies, where effects such as ILS are more prominent. To address this challenge, we present a new probabilistic model, DLCoal, that defines gene duplication and loss in a population setting, such that coalescence and ILS can be directly addressed. Interestingly, this model implies that in addition to the usual gene tree and species tree, there exists a third tree, the locus tree, which will likely have many applications. Using this model, we develop the first general reconciliation method that accurately infers gene duplications and losses in the presence of ILS, and we show its improved inference of orthologs, paralogs, duplications, and losses for a variety of clades, including flies, fungi, and primates. Also, our simulations show that gene duplications increase the frequency of ILS, further illustrating the importance of a joint model. Going forward, we believe that this unified model can offer insights to questions in both phylogenetics and population genetics.


Molecular epidemiology: on the path to prevention?
Cancer prevention has been the stated goal of molecular cancer epidemiology for the past 17 years. In this review, progress toward that goal is evaluated by using as examples well-studied environmental exposures-i.e., tobacco smoke, polycyclic aromatic hydrocarbons, aflatoxin B(1), benzene, and hepatitis B virus-and their roles in lung, breast, and liver cancers and leukemia. The contributions of molecular epidemiology discussed here include providing evidence that environmental agents pose carcinogenic risks, helping establish the causal roles of environmental factors in cancer, identifying environment-susceptibility interactions and populations at greatest risk, and developing new intervention strategies. Molecular epidemiologic and other data indicate that assessment of carcinogenic risks should address both the range of risk across the population and the risk to subgroups who may be at high risk because of genetic or acquired susceptibilities, including young children. However, for the most part, research results have not yet been effectively translated into risk assessments and preventive health policies. An infrastructure linking scientists, policy makers, and other constituencies is needed to facilitate this process. To extend our knowledge, the second generation of molecular epidemiologic research should include large-scale, collaborative studies incorporating validated biomarkers and automated technologies. An incentive to make the necessary investment is the recognition that prevention of only 20% of cancer in the United States would result in 200000 fewer new cases diagnosed each year and an annual savings of $21.4 billion in direct costs alone.


Diagnosis and management of pemphigus: Recommendations of an international panel of experts.
Several European countries recently developed international diagnostic and management guidelines for pemphigus, which have been instrumental in the standardization of pemphigus management. We now present results from a subsequent Delphi consensus to broaden the generalizability of the recommendations. A preliminary survey, based on the European Dermatology Forum and the European Academy of Dermatology and Venereology guidelines, was sent to a panel of international experts to determine the level of consensus. The results were discussed at the International Bullous Diseases Consensus Group in March 2016 during the annual American Academy of Dermatology conference. Following the meeting, a second survey was sent to more experts to achieve greater international consensus. The 39 experts participated in the first round of the Delphi survey, and 54 experts from 21 countries completed the second round. The number of statements in the survey was reduced from 175 topics in Delphi I to 24 topics in Delphi II on the basis of Delphi results and meeting discussion. Each recommendation represents the majority opinion and therefore may not reflect all possible treatment options available. We present here the recommendations resulting from this Delphi process. This international consensus includes intravenous CD20 inhibitors as a first-line therapy option for moderate-to-severe pemphigus.


Isolation and SAR studies of bicyclic iminosugars from Castanospermum australe as glycosidase inhibitors.
We report the isolation and structural determination of fourteen iminosugars, containing five pyrrolizidines and five indolizidines, from Castanospermum australe. The structure of a new alkaloid was elucidated by spectroscopic methods as 6,8-diepi-castanospermine (13). Our side-by-side comparison between bicyclic and corresponding monocyclic iminosugars revealed that inhibition potency and spectrum against each enzyme are clearly changed by their core structures. Castanospermine (10) and 1-deoxynojirimycin (DNJ) have a common d-gluco configuration, and they showed the expected similar inhibition potency and spectrum. In sharp contrast, 6-epi-castanospermine (12) and 1-deoxymannojirimycin (manno-DNJ) both have the d-manno configuration but the α-mannosidase inhibition of 6-epi-castanospermine (12) was much better than that of manno-DNJ. 6,8-Diepi-castanospermine (13) could be regarded as a bicyclic derivative of talo-DNJ, but it showed a complete loss of α-galactosidase A inhibition. This behavior against α-galactosidase A is similar to that observed for 1-epi-australine (6) and altro-DMDP.


Empirical investigation of the benefits of partial Lamarckianism.
Genetic algorithms (GAs) are very efficient at exploring the entire search space; however, they are relatively poor at finding the precise local optimal solution in the region in which the algorithm converges. Hybrid GAs are the combination of improvement procedures, which are good at finding local optima, and GAs. There are two basic strategies for using hybrid GAs. In the first, Lamarckian learning, the genetic representation is updated to match the solution found by the improvement procedure. In the second, Baldwinian learning, improvement procedures are used to change the fitness landscape, but the solution that is found is not encoded back into the genetic string. This paper examines the issue of using partial Lamarckianism (i.e., the updating of the genetic representation for only a percentage of the individuals), as compared to pure Lamarckian and pure Baldwinian learning in hybrid GAs. Multiple instances of five bounded nonlinear problems, the location-allocation problem, and the cell formation problem were used as test problems in an empirical investigation. Neither a pure Lamarckian nor a pure Baldwinian search strategy was found to consistently lead to quicker convergence of the GA to the best known solution for the series of test problems. Based on a minimax criterion (i.e., minimizing the worst case performance across all test problem instances), the 20% and 40% partial Lamarckianism search strategies yielded the best mixture of solution quality and computational efficiency.


African independent churches in Mozambique: healing the afflictions of inequality.
The recent explosive proliferation of African Independent Churches (AICs) in central Mozambique coincided with rapid growth of economic disparity in the 1990s produced by privatization, cuts in government services, and arrival of foreign aid promoted by Mozambique's World Bank/International Monetary Fund Structural Adjustment Program. Drawing on ethnographic research in the city of Chimoio, this article argues that growing inequality has led to declining social cohesion, heightened individual competition, fear of interpersonal violence, and intensified conflict between spouses in poor families. This perilous social environment finds expression in heightened fears of witchcraft, sorcery, and avenging spirits, which are often blamed in Shona ideology for reproductive health problems. Many women with sick children or suffering from infertility turn to AICs for treatment because traditional healers are increasingly viewed as dangerous and too expensive. The AICs invoke the "Holy Spirit" to exercise malevolent agents and then provide a community of mutual aid and ongoing protection against spirit threats.


The spectrum of gastrointestinal toxicity and effect on disease activity of selective cyclooxygenase-2 inhibitors in patients with inflammatory bowel disease.
The safety and toxicity associated with the use of selective cyclooxygenase-2 (COX-2) inhibitors in patients with inflammatory bowel disease (IBD) has not been extensively studied. Thirty-three patients with IBD who were prescribed celecoxib or rofecoxib were identified from questionnaire during their clinic visit at the Cedars-Sinai IBD Center between 1999 and 2002. Twenty-six had Crohn's disease (CD), 6 had ulcerative colitis (UC), and 1 had indeterminate colitis (IC). Twenty-one received rofecoxib, 10 celecoxib, and 2 received both medications at different time points. Overall, 13 (39%) patients experienced disease exacerbation, 7 of which had received celecoxib and six rofecoxib. IBD exacerbation associated with COX-2 treatment did not correlate with age, disease activity, or use of immunosuppressive medications. All patients experienced flare-up of their underlying IBD within 6 weeks of initiating COX-2 therapy. Five of 13 (38%) patients had resolution of their symptoms after discontinuing the COX-2 inhibitor, but the remaining patients required additional medical therapy to control their disease. Six other patients (18%) experienced GI side effects not associated with their underlying IBD. Five developed abdominal pain, and one developed a duodenal ulcer and a circumferential ileo-colonic ulceration with GI bleeding. Treatment with COX-2 inhibitors is associated with a high incidence of exacerbation of the underlying IBD and GI-related complications.


Effects of Pre-Activation with Variable Intra-Repetition Resistance on Throwing Velocity in Female Handball Players: a Methodological Proposal.
The purpose of this study was to investigate the acute effect of pre-activation with Variable Intra-Repetition Resistance and isometry on the overhead throwing velocity in handball players. Fourteen female handball players took part in the study (age: 21.2 ± 2.7 years, experience: 10.9 ± 3.5 years). For Post-Activation Potentiation, two pre-activation methods were used: (I) Variable Intra-Repetition Resistance: 1 x 5 maximum repetitions at an initial velocity of 0.6 m·s-1 and a final velocity of 0.9 m·s-1; (II) Isometry: 1 x 5 s of maximum voluntary isometric contraction. Both methods were "standing unilateral bench presses" with the dominant arm, using a functional electromechanical dynamometer. The variable analysed was the mean of the three overhead throws. Ball velocity was measured with a radar (Stalker ATS). The statistical analysis was performed using ANOVA with repeated measures. No significant differences were found for either method (variable resistance intra-repetition: p = 0.194, isometry: p = 0.596). Regarding the individual responses, the analysis showed that 86% of the sample increased throwing velocity with the variable resistance intra-repetition method, while 93% of the sample increased throwing velocity with the isometric method. Both the variable intra-repetition resistance and isometric methods show improvements in ball velocity in female handball players. However, the authors recommend checking individual responses, since the results obtained were influenced by the short rest interval between the pre-activation and the experimental sets.


Brain tumor tandem targeting using a combination of monoclonal antibodies attached to biopoly(beta-L-malic acid).
Tumor-specific targeting using achievements of nanotechnology is a mainstay of increasing efficacy of anti-tumor drugs. To improve drug targeting we covalently conjugated for the first time two different monoclonal antibodies, an anti-mouse transferrin receptor antibody and a mouse autoimmune anti-nucleosome antibody 2C5, onto the drug delivery nanoplatform, poly(beta-L-malic acid). The active anti-tumor drug components attached to the same carrier molecule were antisense oligonucleotides to vascular protein laminin-8. The resulting drug, a new Polycefin variant, was administered intravenously into glioma-bearing xenogeneic animals. The drug delivery system was targeted across mouse endothelial system by the anti-mouse transferring receptor antibody and to the tumor cell surface by the anti-nucleosome antibody 2C5. The targeting efficacies of the Polycefin variants bearing either two antibodies or each single antibody were compared in vitro and in vivo. ELISA confirmed the co-existence of two antibodies on the same nanoplatform molecule and their functional activities. Fluorescence imaging analysis after 24 h of intravenous injection demonstrated significantly higher tumor accumulation of Polycefin variants with the tandem configuration of antibodies than with single antibodies. The results suggest improved efficacy for tandem configuration of antibodies than for single configurations carried by a drug delivery vehicle.


Molecular cloning and characterization of acrA and acrE genes of Escherichia coli.
The DNA fragment containing the acrA locus of the Escherichia coli chromosome has been cloned by using a complementation test. The nucleotide sequence indicates the presence of two open reading frames (ORFs). Sequence analysis suggests that the first ORF encodes a 397-residue lipoprotein with a 24-amino-acid signal peptide at its N terminus. One inactive allele of acrA from strain N43 was shown to contain an IS2 element inserted into this ORF. Therefore, this ORF was designated acrA. The second downstream ORF is predicted to encode a transmembrane protein of 1,049 amino acids and is named acrE. Genes acrA and acrE are probably located on the same operon, and both of their products are likely to affect drug susceptibilities observed in wild-type cells. The cellular localizations of these polypeptides have been analyzed by making acrA::TnphoA and acrE::TnphoA fusion proteins. Interestingly, AcrA and AcrE share 65 and 77% amino acid identity with two other E. coli polypeptides, EnvC and EnvD, respectively. Drug susceptibilities in one acrA mutant (N43) and one envCD mutant (PM61) have been determined and compared. Finally, the possible functions of these proteins are discussed.


The effectiveness of vision screening by school nurses in secondary school.
Vision screening of 13 and 15 year old children by school nurses in Oxfordshire has declined in recent years due to the pressure of other commitments. A study was carried out to evaluate the likely consequences of failing to provide vision screening for secondary schoolchildren, in order to make a decision about the future of the service. Parents were also asked to complete a questionnaire about their children's past eye histories. Eight schools that had had no vision screening for at least three years participated in the study. Overall, 3.8% of children between 13 and 15 years old had a visual acuity of worse than 6/12 in one or both eyes, that is they failed vision screening; there was no evidence that this percentage increased significantly across this age range. Less than 1% of children were prescribed and wore spectacles as a consequence of failing vision screening and no new cases of eye pathology were detected. Questionnaire responses suggested that about 50% of children who did not wear spectacles had had an eye examination in the previous two years. The results indicate that vision screening is not the best way to meet the eye/vision needs of secondary schoolchildren. Strategies for targeting the school nursing resource more appropriately are being considered.


Evolution of repeated DNA sequences by unequal crossover.
It is often supposed that highly repetitious DNA's arise only as a result of unusual mechanisms or in response to selective pressure. My arguments and simulations suggest, by contrast, that a pattern of tandem repeats is the natural state of DNA whose sequence is not maintained by selection. The simulations show that periodicities can develop readily from nonreptitious DNA as a result of the random accumulation of random mutations and random homology-dependent unequal crossovers. The lengths of these periodicities, and the patterns of subrepeats within them, would fluctuate in evolution, with the probability of a given pattern being dependent on the unknown exact nature of the crossover mechanism. Qualitatively, then, unequal crossover provides a reasonable and uncontrived explanation for the prevalence of highly repeated sequences in DNA and for the patterns of periodicity they evince.


Emerging Chemistry Strategies for Engineering Native Chromatin.
Chromosomes present one of most challenging of all substrates for biochemical study. This is because genomic DNA is physically associated with an astonishing collection of nuclear factors, which serve to not only store the nucleic acid in a stable form, but also grant access to the information it encodes when needed. Understanding this complex molecular choreography is central to the field of epigenetics. One of the great challenges in this area is to move beyond correlative type information, which is now in abundant supply, to the point where we can truly connect the dots at the molecular level. Establishing such causal relationships requires precise manipulation of the covalent structure of chromatin. Tools for this purpose are currently in short supply, creating an opportunity that, as we will argue in this Perspective, is well suited to the sensibilities of the chemist.


Aluminium in human brain tissue from donors without neurodegenerative disease: A comparison with Alzheimer's disease, multiple sclerosis and autism.
A burgeoning number of studies are demonstrating aluminium in human brain tissue. While research has both quantified and imaged aluminium in human brain tissue in neurodegenerative and neurodevelopmental disease there are few similar data for brain tissue from non-neurologically impaired donors. We have used microwave assisted acid digestion and transversely heated graphite furnace atomic absorption spectrometry to measure aluminium in twenty brains from donors without recognisable neurodegenerative disease. The aluminium content of 191 tissue samples was invariably low with over 80% of tissues having an aluminium content below 1.0 μg/g dry weight of tissue. The data for these control tissues were compared with data (measured using identical procedures) for sporadic Alzheimer's disease, familial Alzheimer's disease, autism spectrum disorder and multiple sclerosis. Detailed statistical analyses showed that aluminium was significantly increased in each of these disease groups compared to control tissues. We have confirmed previous conclusions that the aluminium content of brain tissue in Alzheimer's disease, autism spectrum disorder and multiple sclerosis is significantly elevated. Further research is required to understand the role played by high levels of aluminium in the aetiology of human neurodegenerative and neurodevelopmental disease.


Climate change and shifts in spring phenology of three horticultural woody perennials in northeastern USA.
We evaluated spring phenology changes from 1965 to 2001 in northeastern USA utilizing a unique data set from 72 locations with genetically identical lilac plants (Syringa chinensis, clone "Red Rothomagensis"). We also utilized a previously validated lilac-honeysuckle "spring index" model to reconstruct a more complete record of first leaf date (FLD) and first flower date (FFD) for the region from historical weather data. In addition, we examined mid-bloom dates for apple (Malus domestica) and grape (Vitis vinifera) collected at several sites in the region during approximately the same time period. Almost all lilac sites with significant linear trends for FLD or FFD versus year had negative slopes (advanced development). Regression analysis of pooled data for the 72 sites indicated an advance of -0.092 day/year for FFD (P=0.003). The slope for FLD was also negative (-0.048 day/year), but not significant (P=0.234). The simulated data from the "spring index" model, which relies on local daily temperature records, indicated highly significant (P<0.001) negative slopes of -0.210 and -0.123 day/year for FLD and FFD, respectively. Data collected for apple and grape also indicated advance spring development, with slopes for mid-bloom date versus year of -0.20 day/year (P=0.01) and -0.146 (P=0.14), respectively. Collectively, these results indicate an advance in spring phenology ranging from 2 to 8 days for these woody perennials in northeastern USA for the period 1965 to 2001, qualitatively consistent with a warming trend, and consistent with phenology shifts reported for other mid- and high-latitude regions.


Depression and anxiety in late life: diagnostic insights and therapeutic options.
Depression and anxiety represent a sizeable public-health problem for older Americans. Effective treatment of depression and anxiety in late-life patients can improve outcomes directly related to those diseases as well as to comorbid medical diseases. A variety of approaches has been shown to be effective in older adults and is easily accessible to the primary care, specialty, and inpatient practitioner. Because of the etiologic complexity associated with mood disturbance in late life, it is helpful to consider depression and anxiety in late life as a geriatric syndrome similar to frailty, falls, incontinence, and impaired cognition. These syndromes have multiple causes of associated disability, yielding a multitude of avenues for intervention. However, translating the science of care for depression and anxiety into improved mental-health services for older adults and their families remains a challenge to providers and policy-makers alike. In the Tables, the authors offer measures for screening and response evaluation as well as guidance for pharmacotherapy and psychotherapeutic interventions that may be incorporated into primary care.


Delayed Giant Pseudoaneurysm With Left-to-Right Shunt Following Postinfarct Ventricular Septal Perforation Repair.
Left ventricular pseudoaneurysm with a left-to-right shunt is extremely rare, requiring surgery if symptomatic; however, surgery has a high risk. Here, the case of a 77-year-old man with heart failure symptoms is reported, in which he develops a giant left ventricular pseudoaneurysm 16 months after ventricular septal perforation repair as a result of acute myocardial infarction, with mild shunt blood flow from the pseudoaneurysm to the right ventricle. Intraoperative findings showed a free wall rupture along the area where the patch was secured during the initial surgery. The patient was discharged on postoperative day 13, and postoperative examination revealed no abnormalities.


Digital signaling network drives the assembly of the AIM2-ASC inflammasome.
The AIM2-ASC inflammasome is a filamentous signaling platform essential for mounting host defense against cytoplasmic dsDNA arising not only from invading pathogens but also from damaged organelles. Currently, the design principles of its underlying signaling network remain poorly understood at the molecular level. We show here that longer dsDNA is more effective in inducing AIM2 assembly, its self-propagation, and downstream ASC polymerization. This observation is related to the increased probability of forming the base of AIM2 filaments, and indicates that the assembly discerns small dsDNA as noise at each signaling step. Filaments assembled by receptor AIM2, downstream ASC, and their joint complex all persist regardless of dsDNA, consequently generating sustained signal amplification and hysteresis. Furthermore, multiple positive feedback loops reinforce the assembly, as AIM2 and ASC filaments accelerate the assembly of nascent AIM2 with or without dsDNA. Together with a quantitative model of the assembly, our results indicate that an ultrasensitive digital circuit drives the assembly of the AIM2-ASC inflammasome.


Comparison of Immediate With Delayed Stenting Using the Minimalist Immediate Mechanical Intervention Approach in Acute ST-Segment-Elevation Myocardial Infarction: The MIMI Study.
Delayed stent implantation after restoration of normal epicardial flow by a minimalist immediate mechanical intervention aims to decrease the rate of distal embolization and impaired myocardial reperfusion after percutaneous coronary intervention. We sought to confirm whether a delayed stenting (DS) approach (24-48 hours) improves myocardial reperfusion, versus immediate stenting, in patients with acute ST-segment-elevation myocardial infarction undergoing primary percutaneous coronary intervention. In the prospective, randomized, open-label minimalist immediate mechanical intervention (MIMI) trial, patients (n=140) with ST-segment-elevation myocardial infarction ≤12 hours were randomized to immediate stenting (n=73) or DS (n=67) after Thrombolysis In Myocardial Infarction 3 flow restoration by thrombus aspiration. Patients in the DS group underwent a second coronary arteriography for stent implantation a median of 36 hours (interquartile range 29-46) after randomization. The primary end point was microvascular obstruction (% left ventricular mass) on cardiac magnetic resonance imaging performed 5 days (interquartile range 4-6) after the first procedure. There was a nonsignificant trend toward lower microvascular obstruction in the immediate stenting group compared with DS group (1.88% versus 3.96%; P=0.051), which became significant after adjustment for the area at risk (P=0.049). Median infarct weight, left ventricular ejection fraction, and infarct size did not differ between groups. No difference in 6-month outcomes was apparent for the rate of major cardiovascular and cerebral events. The present findings do not support a strategy of DS versus immediate stenting in patients with ST-segment-elevation infarction undergoing primary percutaneous coronary intervention and even suggested a deleterious effect of DS on microvascular obstruction size. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01360242.


Quantitative autoradiographic delineation of the distribution of beta-adrenergic receptors in canine and feline left ventricular myocardium.
The distribution of adrenergic receptors in specific components of the heart such as vessels and myocytes cannot be determined easily with assays of membranes prepared from homogenates of whole tissue. Accordingly, we characterized the binding of the potent nonsubtype selective antagonist [125iodo]cyanopindolol to beta-receptors in unfixed transmural slices of feline and canine left ventricle. Specific binding ratios greater than 90% were achieved at radioligand concentrations near Kd and greater than 80% at saturating ligand concentrations. Binding of radioligand to receptors in transmural slices was rapid, saturable, stereoselective, and displaceable by antagonists and agonists with the rank order of potency expected of beta-adrenergic receptors. Analysis of binding isotherms indicated maximum binding capacities of 27.8 +/- 6.6 and 40.6 +/- 5.1 fmol/mg tissue protein and dissociation constants of 10.1 +/- 1.8 and 21.3 +/- 1.6 pM in feline and canine ventricular slices, respectively. The distribution of beta-receptors in myocytes and selected vascular components of the heart was determined with quantitative film autoradiography and high resolution computer-based analysis and display of the density of binding sites, maximum binding capacity, and binding affinity measurements. The results of autoradiographic analysis revealed a uniform transmural distribution of receptors in regions composed primarily of ventricular myocytes but an inverse relation between the density of beta-receptors and the diameter of coronary vessels. Large epicardial conductance arteries had half the receptor density of subjacent myocytes; small mural arteries had approximately 60% of the beta-receptor density of nearby myocytes, and the coronary resistance arterioles had the highest receptor density of any vascular compartment, which was equivalent to that of myocytes. The methods developed should be of particular value in characterizing the distribution and function of receptor subtypes and mechanisms of regulation of adrenergic responsiveness in intact myocardium.


ASTCT, CIBMTR, and EBMT clinical practice recommendations for transplant and cellular therapies in mantle cell lymphoma.
Autologous (auto-) or allogeneic (allo-) hematopoietic cell transplantation (HCT) are accepted treatment modalities for mantle cell lymphoma (MCL). Recently, chimeric antigen receptor (CAR) T-cell therapy received approval for MCL; however, its exact place and sequence in relation to HCT is unclear. The ASTCT, CIBMTR, and the EBMT, jointly convened an expert panel to formulate consensus recommendations for role, timing, and sequencing of auto-, allo-HCT, and CAR T-cell therapy for patients with newly diagnosed and relapsed/refractory (R/R) MCL. The RAND-modified Delphi method was used to generate consensus statements. Seventeen consensus statements were generated; in the first-line setting auto-HCT consolidation represents standard-of-care in eligible patients, whereas there is no clear role of allo-HCT or CAR T-cell therapy, outside of a clinical trial. In the R/R setting, the preferential option is CAR T-cell therapy especially in MCL failing or intolerant to at least one Bruton's tyrosine kinase inhibitor, while allo-HCT is recommended if CAR T-cell therapy has failed or is not feasible. In the absence of contemporary evidence-based data, the panel found RAND-modified Delphi methodology effective in providing a formal framework for developing consensus recommendations for the timing and sequence of cellular therapies for MCL.


Muscle strength, endurance, and aerobic capacity in rheumatoid arthritis: a comparative study with healthy subjects.
Isometric/isokinetic muscle strength and isokinetic endurance of the lower extremities as well as aerobic capacity were evaluated in 67 patients (43 female, 24 male; mean age 53 years, range 23-65) with classical/definite rheumatoid arthritis (RA) of functional class II. Results obtained were compared with those of a healthy reference group matched for age and sex. Disease characteristics of the group with RA were registered and lifestyle characteristics, such as work load, exercise, diet, smoking, and alcohol habits, were reported by both groups. Generally, results showed that the group with RA had decreased functional capacity. Isometric hip and knee muscle strength of the rheumatoid group was reduced to about 75% of normal function, isokinetic knee muscle strength at the velocities of 60 and 180 degrees/s to about 65% and 75% of normal function respectively, isokinetic endurance of the knee muscle groups to about 45%, and aerobic capacity to about 80% of the results obtained for the healthy reference group. Analyses of variance showed that the rheumatoid group, compared with the healthy group, had significantly reduced function on all isometric and isokinetic tests of the extensors and flexors of the knee. Results for isometric hip muscle strength were similar--all tests but one yielding highly significant differences. To avoid unnecessary functional deficits it seems important to include muscular training in rehabilitation programmes for patients with RA.


Influence of imide-substituents on the H-type aggregates of perylene diimides bearing cetyloxy side-chains at bay positions.
A series of perylene-3,4:9,10-tetracarboxylic acid diimides (PDIs, namely TYR-PDI, AEP-PDI, CET-PDI, ANP-PDI and KOD-PDI), comprising long linear cetyloxy side-chains functionalized at the 1,7-bay positions and the different substituents (i.e., hydrophobic/hydrophilic segments) symmetrically linked at the two imide-positions of the perylene core were synthesized to investigate the influence of imide-substituent patterns on the aggregation behaviours of PDIs. The photophysical properties of these PDIs were studied by UV-Vis absorption, fluorescence and time-resolved photoluminescence spectroscopy. The differences in the photophysical properties of the PDIs indicate (i) blue-shifted and broadening absorption properties in both solution and thin-films, (ii) red-shifted and broadening fluorescence behavior at their emission maximum in solution, however, blue-shifted fluorescence behavior in thin-films, and (iii) obviously longer fluorescence life-times corresponding to the existence of rotationally displaced H-type aggregates. The formation of short-range ordered rod-like microstructures through face-to-face alignment of columnar rectangular H-type PDI aggregates was rationalized by scanning electron microscopy. The X-ray diffraction study revealed that the formation of well-defined columnar rectangular (Colrp) H-type PDI aggregates indicated a nearly constant intracolumnar stacking distance of ∼3.9 Å for all PDIs. All of these findings were consistent with the formation of hydrophobic/hydrophilic interactions between the imide-substituents in addition to the strong hydrophobic π-π stacking interactions between the conjugated perylene cores, which were enforced in the H-type PDI aggregates that spontaneously self-organized into Colrp structures.


CD40 ligand-CD40 interactions are necessary for the initiation of insulitis and diabetes in nonobese diabetic mice.
The nonobese diabetic (NOD) mouse spontaneously develops T cell-dependent autoimmune diabetes. Here, we investigate the role of CD40 ligand (CD40L)-CD40 costimulation in the initiation and progression of this disease. Anti-CD40L mAb treatment of 3- to 4-wk-old NOD females (the age at which insulitis typically begins) completely prevented the insulitis and diabetes. In contrast, treatment of such mice with anti-CD40L at >9 wk of age did not inhibit the disease process. These results suggest that a costimulatory signal by CD40L is required early but not in the effector phase of disease development. Anti-CD40L treatment affected the priming of islet Ag-specific T cell responses in vivo. Cytokine analysis revealed a dramatic decrease in IFN-gamma and IL-2 release without a concomitant increase in IL-4 production by T cells from anti-CD40L-treated mice. Thus, anti-CD40L impaired the islet Ag-specific Th1 cell response in vivo, and the prevention of diabetes by anti-CD40L was not associated with switching of the response from a Th1 to a Th2 profile. Cotransfer of splenocytes from anti-CD40L-treated mice with splenocytes from diabetic NOD mice into NOD/scid mice did not inhibit the transfer of disease, indicating that anti-CD40L does not prevent the disease by inducing regulatory cells. Since anti-CD40L clearly prevented the insulitis by inhibiting the development and further accumulation of pathogenic Th1 cells to islets of Langerhans, we conclude that CD40L-CD40 costimulation is required for early events in the development of spontaneous autoimmune diabetes.