Prolonged suppression of HBV in mice by a novel antibody that targets a unique epitope on hepatitis B surface antigen.
This study aimed to investigate the therapeutic potential of monoclonal antibody (mAb) against HBV as a novel treatment approach to chronic hepatitis B (CHB) in mouse models. Therapeutic effects of mAbs against various epitopes on viral surface protein were evaluated in mice mimicking persistent HBV infection. The immunological mechanisms of mAb-mediated viral clearance were systematically investigated. Among 11 tested mAbs, a novel mAb E6F6 exhibited the most striking therapeutic effects in several HBV-persistent mice. Single-dose administration of E6F6 could profoundly suppress the levels of hepatitis B surface antigen (HBsAg) and HBV DNA for several weeks in HBV-transgenic mice. E6F6 regimen efficiently prevented initial HBV infection, and reduced viral dissemination from infected hepatocytes in human-liver-chimeric mice. E6F6-based immunotherapy facilitated the restoration of anti-HBV T-cell response in hydrodynamic injection (HDI)-based HBV carrier mice. Immunological analyses suggested that the Fcγ receptor-dependent phagocytosis plays a predominant role in E6F6-mediated viral suppression. Molecular analyses suggested that E6F6 recognises an evolutionarily conserved epitope (GPCK(R)TCT) and only forms a smaller antibody-viral particle immune complex with limited interparticle crosslinking when it binds to viral particles. This unique binding characteristic of E6F6 to HBV was possibly associated with its effective in vivo opsonophagocytosis for viral clearance. These results provided new insight into understanding the therapeutic role and mechanism of antibody against persistent viral infection. The E6F6-like mAbs may provide a novel immunotherapeutic agent against human chronic HBV infection.


A case study of early experience with implementation of collaborative care in the Veterans Health Administration.
Primary care remains critically important for those who suffer from mental disorders. Although collaborative care, which integrates mental health services into primary care, has been shown to be more effective than usual care, its implementation has been slow and the experience of providers and patients with collaborative care is less well known. The objective of this case study was to examine the effects of collaborative care on patient and primary care provider (PCP) experiences and communication during clinical encounters. Participating physicians completed a self-administered visit reconstruction questionnaire in which they logged details of patient visits and described their perceptions of the visits and the influence of collaborative care. Audio recordings of visits were analyzed to assess the extent of discussion about colocated mental health services and visit time devoted to mental health topics. The main outcome measures were the extent of discussion and recommendation for collaborative care during clinical visits and providers' experiences based on their responses to the visit reconstruction questionnaire. Providers surveyed expressed enthusiasm about collaborative care and cited the time constraint of office visits and lack of specialty support as the main reasons for limiting their discussion of mental health topics with patients. Despite the availability of mental health providers at the same clinic, PCPs missed many opportunities to address mental health issues with their patients. Ongoing education for PCPs regarding how to conduct a "warm handoff" to colocated providers will need to be an integral part of the implementation of collaborative care.


Alpha tubulin genes from Leishmania braziliensis: genomic organization, gene structure and insights on their expression.
Alpha tubulin is a fundamental component of the cytoskeleton which is responsible for cell shape and is involved in cell division, ciliary and flagellar motility and intracellular transport. Alpha tubulin gene expression varies according to the morphological changes suffered by Leishmania in its life cycle. However, the objective of studying the mechanisms responsible for the differential expression has resulted to be a difficult task due to the complex genome organization of tubulin genes and to the non-conventional mechanisms of gene regulation operating in Leishmania. We started this work by analyzing the genomic organization of α-tubulin genes in the Leishmania braziliensis genome database. The genomic organization of L. braziliensis α-tubulin genes differs from that existing in the L. major and L. infantum genomes. Two loci containing α-tubulin genes were found in the chromosomes 13 and 29, even though the existence of sequence gaps does not allow knowing the exact number of genes at each locus. Southern blot assays showed that α-tubulin locus at chromosome 13 contains at least 8 gene copies, which are tandemly organized with a 2.08-kb repetition unit; the locus at chromosome 29 seems to contain a sole α-tubulin gene. In addition, it was found that L. braziliensis α-tubulin locus at chromosome 13 contains two types of α-tubulin genes differing in their 3' UTR, each one presumably containing different regulatory motifs. It was also determined that the mRNA expression levels of these genes are controlled by post-transcriptional mechanisms tightly linked to the growth temperature. Moreover, the decrease in the α-tubulin mRNA abundance observed when promastigotes were cultured at 35°C was accompanied by parasite morphology alterations, similar to that occurring during the promastigote to amastigote differentiation. Information found in the genome databases indicates that α-tubulin genes have been reorganized in a drastic manner along Leishmania speciation. In the L. braziliensis genome database, two loci containing α-tubulin sequences were found, but only the locus at chromosome 13 contains the prototypic α-tubulin genes, which are repeated in a head-to-tail manner. Also, we determined that the levels of α-tubulin mRNAs are down-regulated drastically in response to heat shock by a post-transcriptional mechanism which is dependent upon active protein synthesis.


Polyubiquitin-sensor proteins reveal localization and linkage-type dependence of cellular ubiquitin signaling.
Polyubiquitin chain topology is thought to direct modified substrates to specific fates, but this function-topology relationship is poorly understood, as are the dynamics and subcellular locations of specific polyubiquitin signals. Experimental access to these questions has been limited because linkage-specific inhibitors and in vivo sensors have been unavailable. Here we present a general strategy to track linkage-specific polyubiquitin signals in yeast and mammalian cells, and to probe their functions. We designed several high-affinity Lys63 polyubiquitin-binding proteins and demonstrate their specificity in vitro and in cells. We apply these tools as competitive inhibitors to dissect the polyubiquitin-linkage dependence of NF-κB activation in several cell types, inferring the essential role of Lys63 polyubiquitin for signaling via the IL-1β and TNF-related weak inducer of apoptosis (TWEAK) but not TNF-α receptors. We anticipate live-cell imaging, proteomic and biochemical applications for these tools and extension of the design strategy to other polymeric ubiquitin-like protein modifications.


Hypoxia-inducible factors in the first trimester human lung.
Lung development takes place in a relatively low-oxygen environment, which is beneficial for lung organogenesis, including vascular development. Hypoxia-inducible factor (HIF)-1 plays an important role in mediating oxygen-regulated events. HIF-1 is stable and initiates gene transcription under hypoxia, whereas in normoxia, interaction with the von Hippel-Lindau (VHL) tumor suppressor protein leads to rapid degradation of the HIF-1alpha subunit. Interaction with VHL requires hydroxylation of HIF-1alpha proline residues by prolyl hydroxylases (PHDs). We investigated the expression of the various components regulating HIF-1alpha stability in first trimester (8-14 weeks) human lungs. Spatial expression was assessed by immunohistochemistry and temporal expression by quantitative PCR. Immunoreactivity for PHD1, PHD3, and seven in absentia homolog (SIAH)1 was noted in the pulmonary epithelium. PHD2 was not expressed in the airway epithelium, but in the lung parenchyma. HIF-1alpha and vascular endothelial growth factor (VEGF) immunoreactivity were primarily detected in the branching epithelium. HIF-2alpha and ARNT proteins localized to the developing epithelium as well as mesenchymal, most likely vascular, structures in the parenchyma. VEGF receptor 2 (VEGFR2) was found in the subepithelium as well as in vascular structures of the mesenchyme. All components of the VEC complex (VHL, NEDD8, and Cullin2) were found in the epithelium. Quantitative PCR analysis demonstrated that VEGF, VEGFR1, HIF-1alpha, HIF-2alpha, ARNT, PHD1, PHD2, PHD3, and SIAH1 gene expression was constant during early pulmonary organogenesis. Cumulatively, the data suggest that the lung develops in a low-oxygen environment that allows for proper vascular development through HIF-regulated pathways.


Mouse B-type lamins are required for proper organogenesis but not by embryonic stem cells.
B-type lamins, the major components of the nuclear lamina, are believed to be essential for cell proliferation and survival. We found that mouse embryonic stem cells (ESCs) do not need any lamins for self-renewal and pluripotency. Although genome-wide lamin-B binding profiles correlate with reduced gene expression, such binding is not directly required for gene silencing in ESCs or trophectoderm cells. However, B-type lamins are required for proper organogenesis. Defects in spindle orientation in neural progenitor cells and migration of neurons probably cause brain disorganizations found in lamin-B null mice. Thus, our studies not only disprove several prevailing views of lamin-Bs but also establish a foundation for redefining the function of the nuclear lamina in the context of tissue building and homeostasis.


Is robotic gait training effective for individuals with cerebral palsy? A systematic review and meta-analysis of randomized controlled trials.
To determine if robotic gait training for individuals with cerebral palsy is more effective than the standard of care for improving function. PubMed, Embase, Scopus, and Cochrane databases were searched from 1980-January, 2022 for articles that investigated robotic gait training versus standard of care (i.e. physical therapy or standard gait training) for individuals with cerebral palsy. Articles were included if a randomized controlled trial design was used, and excluded if robotic gait training was combined with another neuromuscular intervention, such as functional electrical stimulation. A meta-analysis of outcomes measured in at least four studies was conducted. Eight citations met all criteria for full-text review and inclusion in the meta-analysis. A total of 188 individuals with cerebral palsy, ages four to 35, and Gross Motor Function Classification System levels I-IV were studied. Level of evidence ranged from 2b-1b. All studies utilized a tethered, assistive device for robotic gait training. The overall effect was not significantly different between the robotic gait training and control interventions for six minute walk test performance (95% CI: -0.17, 0.73; P = 0.22), free walking speed (95% CI: -0.18, 0.57; P = 0.30), or Gross Motor Function Measures D (Standing) (95% CI: -0.29, 0.39; P = 0.77) and E (Walking, Running and Jumping) (95% CI: -0.11, 0.57; P = 0.19). Tethered robotic devices that provide assistive gait training for individuals with cerebral palsy do not provide a greater benefit for improving mobility than the standard of care.


Modulation of esophageal peristalsis by vagal efferent stimulation in opossum.
Experiments were performed on anesthetized opossums to study the influence of vagal efferent stimulation on peristalsis in the esophageal smooth muscle using various stimulus parameters. Current intensity, pulse duration, frequency, and train duration were varied systematically. Electrical and mechanical activities were recorded simultaneously at 5, 3, and 1 cm above the lower esophageal sphincter (LES). Vagal efferent stimulation produced a spike burst and contraction with a latency after the termination of the stimulus. This latency varied at different sites with the same stimulus parameters. For example, a stimulus of 5 mA, 0.5 ms, 10 Hz, and 1-s train produced latencies for the electrical response of 1.48 +/- 0.04, 2.2 +/- 0.12, and 3.5 +/- 0.09 s (+/- SEM) at 5, 3, and 1 cm above LES, respectively. The differences in latency were statistically significant (P less than 0.01). The latency of response at any one site also changed with different stimulus parameters; e.g. at 1 cm above LES, the latency of electrical response at 10 Hz was 3.5 +/- 0.09 s, but at 20 Hz the latency was 2.01 +/- 0.06 s when current intensity, pulse, and train duration remained at 5 mA, 0.5 ms, and 1 s. This decrease in latency with increasing frequency was statistically significant (P less than 0.01). By changing stimulus parameters, antiperistalsis or peristalsis with different speeds of propagation could be induced. Antiperistalsis or simultaneous responses occurred near threshold stimulus parameters. Suprathreshold stimuli produced peristaltic responses. Speed of peristalsis in the distal esophagus was 1.82 +/- 0.08 cm/s with swallowing, which was not different from 1.98 +/- 0.14 cm/s (P greater than 0.05) with vagal stimulation of 5 mA, 0.5 ms, 10 Hz, and 1-s train. These studies suggest that: (a) peristalsis in the smooth muscle part of the esophagus can be explained entirely on the basis of peripheral mechanisms, and (b) the central nervous system may modulate the occurrence, polarity, and speed of propagation by modifying the intensity and frequency of vagal activation.


You probably think this paper's about you: narcissists' perceptions of their personality and reputation.
Do narcissists have insight into the negative aspects of their personality and reputation? Using both clinical and subclinical measures of narcissism, the authors examined others' perceptions, self-perceptions, and meta-perceptions of narcissists across a wide range of traits for a new acquaintance and close other (Study 1), longitudinally with a group of new acquaintances (Study 2), and among coworkers (Study 3). Results bring 3 surprising conclusions about narcissists: (a) they understand that others see them less positively than they see themselves (i.e., their meta-perceptions are less biased than are their self-perceptions), (b) they have some insight into the fact that they make positive first impressions that deteriorate over time, and (c) they have insight into their narcissistic personality (e.g., they describe themselves as arrogant). These findings shed light on some of the psychological mechanisms underlying narcissism.


Treatment responses in Asians and Caucasians with chronic hepatitis C infection.
To conduct a multicentre retrospective review of virological response rates in Asians infected with genotype 1 chronic hepatitis C (CHC) treated with combination interferon and ribavirin and then to compare their responses to that among Caucasians. Asian patients infected with genotype 1 CHC treated at 4 Australian centres between 2001 to 2005 were identified through hospital databases. Baseline demographic characteristics, biochemical, virological and histological data and details of treatment were collected. Sustained virological responses (SVR) in this cohort were then compared to that in Caucasian subjects, matched by genotype, age, gender and the stage of hepatic fibrosis. A total of 108 Asians with genotype 1 CHC were identified. The end of treatment response (ETR) for the cohort was 79% while the SVR was 67%. Due to the relatively advanced age of the Asian cohort, only sixty-four subjects could be matched with Caucasians. The ETR among matched Asians and Caucasians was 81% and 56% respectively (P = 0.003), while the SVR rates were 73% and 36% (P < 0.001) respectively. This difference remained significant after adjusting for other predictive variables. Genotype 1 CHC in Asian subjects is associated with higher rates of virological response compared to that in Caucasians.


Identification and analysis of the minimal promoter activity of a novel noncoding nuclear RNA gene, AncR-1, from the honeybee (Apis mellifera L.).
Previously, we identified a gene for a noncoding nuclear RNA, termed Ks-1, that is expressed preferentially in a restricted set of neurons in the honeybee brain. In the present study, we identified another novel gene, termed AncR-1, whose transcripts were localized to nuclei in the whole cortex region of the honeybee brain, as a candidate novel noncoding nuclear RNA gene. RNA fluorescent in situ hybridization revealed that AncR-1 and Ks-1 transcripts were located in a distinct portion of a single neural nucleus, suggesting that they have distinct functions in brain neurons. cDNA cloning revealed that the AncR-1 transcripts were up to 7 kb in size, had mRNA-like structures, and were alternatively spliced. The reporter assay using Drosophila SL-2 cells demonstrated that a TATA box-like sequence located -30 bp upstream of the 5' end of AncR-1 cDNA had promoter activity. None of the alternatively spliced AncR-1 cDNA variants contained significant open reading frames, strongly suggesting that AncR-1 transcripts function as novel noncoding nuclear RNAs. Furthermore, in situ hybridization revealed that AncR-1 was expressed not only in the brain but also in the sex organs in the queen and drones and in the hypopharyngeal glands and oenocytes of the worker bees, suggesting that AncR-1 is involved in diverse organ functions. Some of the AncR-1 transcripts enriched in the nuclei of the hypopharyngeal glands were polyadenylated, indicating the presence of mRNA-like AncR-1 transcripts in the nuclei.


Behaviour of carbonyl groups in several clinical conditions: Analysis of our survey.
Protein carbonylation is a marker of oxidative protein damage, that is likely involved in the pathogenesis of several diseases. The aim of this study was to evaluate the protein carbonyl (PC) groups in different clinical conditions. It included different groups of subjects: 81 trained subjects; 23 subjects with mild essential hypertension; 31 middle-aged subjects with metabolic syndrome (MS); 106 subjects with MS not selected for age (subdivided into two subgroups, with and without diabetes mellitus); 91 obese adults subdivided in two subgroups (BMI 30-35 Kg/m2 and BMI > 35 kg/m2); 48 subjects with obstructive sleep apnea syndrome (OSAS) subdivided in accordance with the apnea/hypopnea index (AHI); 27 subjects with chronic kidney disease (CKD) on conservative therapy; 31 subjects with CKD on haemodialysis treatment; and 50 subjects with juvenile myocardial infarction. PC groups were reduced in trained subjects in comparison with sedentary controls, while no variation was observed in mild essential hypertension. PC groups were increased in MS subjects and in adult obese subjects. In MS subjects the PC groups were not influenced by the presence of diabetes mellitus and in adult obese subjects were not influenced by the obesity degree. In OSAS subjects only those with AHI > 30 showed an increase of PC groups. PC groups increased in CKD subjects undergoing conservative treatment and haemodialysis therapy. In dialyzed subjects, after a standard dialysis session, there was a marked increase in PC groups. In juvenile myocardial infarction PC groups were higher than in controls; there was no difference between STEMI and NSTEMI and their concentration was unaffected by the number of cardiovascular risk factors or stenosed coronary vessels.


Characterization of the modes of action of anti-Pbs21 malaria transmission-blocking immunity: ookinete to oocyst differentiation in vivo.
The impact of immune sera, and peripheral blood cells (PBC) from mice immunized with Plasmodium berghei ookinetes; and of purified immunoglobulin or Fab fragments from anti-Pbs21 monoclonal antibody 13.1, upon establishment of oocyst infections in the mosquito was studied. Infections were initiated either from gametocyte-infected mice, or membrane feeders which contained either gametocytes or mature ookinetes. PBC from ookinete-immunized mice presented with non-immune serum failed to show any transmission-blocking activity. Anti-ookinete serum, intact anti-Pbs21 monoclonal antibody 13.1 or its Fab fragments, all inhibited oocyst formation significantly. When gametocyte-infected mice or gametocytes in membrane feeds were used, inhibition did not directly correlate with antibody concentration. In membrane feeders that contained ookinetes and antibody, concentration-dependent inhibition usually occurred. The efficacy of purified 13.1 IgG was dependent upon the ookinete concentration. The ookinete plasmalemma and cytoplasm were significantly disturbed after 12 h in bloodmeals that contained antibody 13.1, but not in the isotype controls. These changes may have caused the observed failure of the ookinete to migrate as rapidly as the controls from the destructive environment of the bloodmeal.


Impact of a self-instruction CPR kit on 7th graders' and adults' skills and CPR performance.
Norwegian 7th graders (12 years old) trained CPR using a 30 min DVD, an inflatable manikin and teachers as facilitators. They were instructed to take the kit home and expose as many family members and friends (adults) as possible to the training. We wanted to document the number of lay rescuers trained by this distribution model and CPR skills achieved. The school principals reported the total number trained using the kit of both 7th graders (first tier) and adults (second tier). They were divided into four groups (first and second tier before and after training) of 50 subjects and CPR was performed on a Skillmeter Anne manikin in order to be electronically documented and their performances were videotaped. A total of 54,876 7th graders and 119,758 adults were trained, representing 87% of the 7th graders in Norway and approximately 4% of the Norwegian population. The skill test of 200 subjects divided into four groups respectively showed that second tier scored significantly lower than first tier in all CPR variables except compression depth. The present model of CPR kit distribution via 7th graders enabled a significant percentage of the population to focus on CPR training. We believe the model secure discipline through the training program for first tier but not for second tier and speculate that 12-year-old children are too young to have an impact of adults commitment and motivation for training.


Spectroscopic probe of distribution of the non-intercalating drug netropsin between DNA and heparin.
Circular dichroism has been used as a monitoring tool to probe the distribution of the non-intercalating drug netropsin (NTPS) between the two biomolecules DNA and heparin. The stoichiometry of the interaction of the individual biomolecules and the drug is determined from conductometric titrations; the titration in each case shows two breaks corresponding to two stoichiometries of interaction. Though netropsin is non-intercalating, DNA wins over heparin in binding the drug due to strong hydrogen bonding capability of NTPS in the minor grooves of DNA through its greater than NH donor groups. Potential hydrogen bond breakers like KF and urea reduce the induced dichroism of NTPS-DNA system, probably dislodging some drug from DNA through hydrogen bond breaking.


Transperineal bulboprostatic anastomotic repair of pelvic fracture urethral distraction defect and role of ancillary maneuver: A retrospective study in 172 patients.
We present our experience with transperineal bulboprostatic anastomosis procedure and compare the results with age of patients, length of urethral stricture, effect of previous treatment and need for ancillary procedures. We retrospectively reviewed the outcome of 172 patients who underwent perineal urethroplasty procedure for traumatic stricture in our institute. Simple perineal anastomosis was done in 92 patients. Perineal anastomosis and corporal separation were done in 52 patients. Perineal anastomosis with inferior pubectomy was done in 25 patients. Perineal anastomosis with rerouting was done in three patients. Age, prior treatment, length of stricture, and ancillary techniques required during reconstruction were compiled. The clinical outcome was considered as failure when any postoperative instrumentation was needed. Out of 172 cases that underwent transperineal urethroplasty procedure, 157 (91.28%) were successful. Simple perineal urethroplasty procedure showed a success rate of 93.4%, perineal anastomosis with separation of corporal bodies had a success rate of 90.4%, perineal anastomosis with inferior pubectomy had a success rate of 88% and perineal anastomosis with rerouting of urethra around the corpora had a success rate of 66.7%. The success rate of delayed progressive perineal urethroplasty procedure for post-traumatic stricture urethra is excellent and majority of the failures occurs in prepubescent boys and in those undergoing secondary repair.


Metal-organic framework gels and monoliths.
The synthesis of metal-organic frameworks (MOFs) has, to date, largely been in the form of crystalline powders. However, interest in different physical morphologies of this class of materials is growing. In this perspective, we provide an overview of the structure, properties and applications of MOF monoliths. In particular, we explore the complex synthetic landscapes associated with MOF crystallization and discuss the synthetic factors leading to the formation of MOF gels, i.e. the precursor to sol-gel MOF monoliths. Finally, we provide our thoughts on the future development of this field, and attempt to highlight the importance of the MOF gel state in the discovery of new functional materials.


Timing of renal replacement therapy and clinical outcomes in critically ill patients with severe acute kidney injury.
The aim of this study is to evaluate the relationship between timing of renal replacement therapy (RRT) in severe acute kidney injury and clinical outcomes. This was a prospective multicenter observational study conducted at 54 intensive care units (ICUs) in 23 countries enrolling 1238 patients. Timing of RRT was stratified into "early" and "late" by median urea and creatinine at the time RRT was started. Timing was also categorized temporally from ICU admission into early (<2 days), delayed (2-5 days), and late (>5 days). Renal replacement therapy timing by serum urea showed no significant difference in crude (63.4% for urea <or=24.2 mmol/L vs 61.4% for urea >24.2 mmol/L; odds ratio [OR], 0.92; 95% confidence interval [CI], 0.73-1.15; P = .48) or covariate-adjusted mortality (OR, 1.25; 95% CI, 0.91-1.70; P = .16). When stratified by creatinine, late RRT was associated with lower crude (53.4% for creatinine >309 micromol/L vs 71.4% for creatinine <or=309 micromol/L; OR, 0.46; 95% CI, 0.36-0.58; P < .0001) and covariate-adjusted mortality (OR, 0.51; 95% CI, 0.37-0.69; P < .001). However, for timing relative to ICU admission, late RRT was associated with greater crude (72.8% vs 62.3% vs 59%, P < .001) and covariate-adjusted mortality (OR, 1.95; 95% CI, 1.30-2.92; P = .001). Overall, late RRT was associated with a longer duration of RRT and stay in hospital and greater dialysis dependence. Timing of RRT, a potentially modifiable factor, might exert an important influence on patient survival. However, this largely depended on its definition. Late RRT (days from admission) was associated with a longer duration of RRT, longer hospital stay, and higher dialysis dependence.


Prevalence of peste des petits ruminants in the arid zone in the Republic of Niger.
The study aimed to determine the prevalence of peste des petits ruminants in the arid zone (Niamey, Tillabéry and Tahoua) of the Republic of Niger. A serological survey was conducted and 519 serum samples were collected from 253 unvaccinated sheep and 266 unvaccinated goats. The sample included 340 female animals (168 sheep and 172 goats) and 160 kids and lambs (78 lambs and 82 kids). A competitive enzyme-linked immunosorbent assay yielded an overall seroprevalence of 45.0%. The prevalence in sheep was 42.0% compared with 47.9% in goats. The seroprevalence observed amongst small ruminants in Tahoua (49.8%) and Tillabéry (46.6%) was significantly higher (p = 0.001) than that observed in animals from Niamey (25.1%). It was also higher (p = 0.04) in sheep younger than two years (51.8%) than in adults (37.6%). Conversely, the seroprevalence showed no significant difference between male animals (35.8% in sheep; 50.1% in goats) and female animals (45.1% in sheep; 46.4% in goats). The prevalence of the disease observed amongst the sheep and goat populations confirms the continued danger of this disease in the areas studied. It is therefore necessary to develop strategies such as improving livestock services, providing effective vaccines and implementing a vaccination programme for an effective control of the disease in sub-Saharan Africa.


An experimental model for the study of lymphedema and its response to therapeutic lymphangiogenesis.
Evaluation of the efficacy of molecular treatment strategies for lymphatic vascular insufficiency requires a suitable preclinical animal model. Ideally, the model should closely replicate the untreated human disease in its pathogenesis and pathological expression. We have undertaken a study of the time course of the development and resolution of acquired, experimental lymphedema and of its responses to vascular endothelial growth factor (VEGF)-C lymphangiogenesis in the mouse tail model. We provoked post-surgical lymphedema in the mouse tail model and assessed the effects of exogenously administered human recombinant VEGF-C. Quantitative assessment of immune traffic function was performed through sequential in vivo bioluminescent imaging. In untreated lymphedema, tail edema was sustained until day 21. Exogenous administration of human recombinant VEGF-C produced a significant decrease in volume. Untreated lymphedema in the mouse tail model was characterized by the presence of dilated cutaneous lymphatics, marked acute inflammatory changes, and hypercellularity; VEGF-C produced a substantial reversion to the normal pattern, with notable regression in the size and number of cutaneous lymphatic vessels that express lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1). In vivo imaging confirmed the presence of an impairment of immune traffic in lymphedema that was ameliorated after VEGF-C administration. The post-surgical murine tail model of lymphedema closely simulates attributes of human lymphedema and provides the requisite sensitivity to detect therapeutically induced functional and structural alterations. It can, therefore, be used as an investigative platform to assess mechanisms of disease and its responses to candidate therapies, such as therapeutic lymphangiogenesis.


Improve the dosimetric outcome in bilateral head and neck cancer (HNC) treatment using spot-scanning proton arc (SPArc) therapy: a feasibility study.
To explore the dosimetric improvement, delivery efficiency, and plan robustness for bilateral head and neck cancer (HNC) treatment utilizing a novel proton therapy technique - the spot-scanning proton arc (SPArc) therapy. We evaluated fourteen bilateral HNC patients retrospectively. Both SPArc and 3-field Intensity Modulated Proton Therapy (IMPT) plans were generated for each patient using the same robust optimization parameters. The prescription doses were 70Gy (relative biological effectiveness (RBE) for CTV_high and 60Gy[RBE] for CTV_low. Clinically significant dosimetric parameters were extracted and compared. Root-mean-square deviation dose (RMSDs) Volume Histogram(RVH) was used to evaluate the plan robustness. Total treatment delivery time was estimated based on the machine parameters. The SPArc plan was able to provide equivalent or better robust target coverage while showed significant dosimetric improvements over IMPT in most of the organs at risk (OARs). More specifically, it reduced the mean dose of the ipsilateral parotid, contralateral parotid, and oral cavity by 25.8%(p = 0.001), 20.8%(p = 0.001) and 20.3%(p = 0.001) respectively compared to IMPT. This technique reduced D1 (the maximum dose covering 1% volume of a structure) of cord and brain stem by 20.8% (p = 0.009) and 10.7% (p = 0.048), respectively. SPArc also reduced the average integral dose by 17.2%(p = 0.001) and external V3Gy (the volume received 3Gy[RBE]) by 8.3%(p = 0.008) as well. RVH analysis showed that the SPArc plans reduced the dose uncertainties in most OARs compared to IMPT, such as cord: 1.1 ± 0.4Gy[RBE] vs 0.7 ± 0.3Gy[RBE](p = 0.001), brain stem: 0.9 ± 0.7Gy[RBE] vs 0.7 ± 0.7Gy[RBE](p = 0.019), contralateral parotid: 2.5 ± 0.5Gy[RBE] vs 2.2 ± 0.6Gy[RBE](p = 0.022) and ipsilateral parotid: 3.1 ± 0.7Gy[RBE] vs 2.8 ± 0.6Gy[RBE](p = 0.004) respectively. The average total estimated treatment delivery time were 283.4 ± 56.2 s, 469.2 ± 62.0 s and 1294.9 ± 106.7 s based on energy-layer-switching-time (ELST) of 0.1 s, 1 s, and 5 s respectively for SPArc plans, compared to the respective values of 328.0 ± 47.6 s(p = 0.002), 434.1 ± 52.0 s(p = 0.002), and 901.7 ± 74.8 s(p = 0.001) for 3-field IMPT plans. The potential clinical benefit of utilizing SPArc will lead to a decrease in the mean probability of salivary flow dysfunction by 31.3%(p = 0.001) compared with IMPT. SPArc could significantly spare OARs while providing a similar or better robust target coverage compared with IMPT in the treatment of bilateral HNC. In the modern proton system with ELST less than 0.5 s, SPArc could potentially be implemented in the routine clinic with a practical, achievable treatment delivery efficiency.


The landscape of antibody binding in SARS-CoV-2 infection.
The search for potential antibody-based diagnostics, vaccines, and therapeutics for pandemic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has focused almost exclusively on the spike (S) and nucleocapsid (N) proteins. Coronavirus membrane (M), ORF3a, and ORF8 proteins are humoral immunogens in other coronaviruses (CoVs) but remain largely uninvestigated for SARS-CoV-2. Here, we use ultradense peptide microarray mapping to show that SARS-CoV-2 infection induces robust antibody responses to epitopes throughout the SARS-CoV-2 proteome, particularly in M, in which 1 epitope achieved excellent diagnostic accuracy. We map 79 B cell epitopes throughout the SARS-CoV-2 proteome and demonstrate that antibodies that develop in response to SARS-CoV-2 infection bind homologous peptide sequences in the 6 other known human CoVs. We also confirm reactivity against 4 of our top-ranking epitopes by enzyme-linked immunosorbent assay (ELISA). Illness severity correlated with increased reactivity to 9 SARS-CoV-2 epitopes in S, M, N, and ORF3a in our population. Our results demonstrate previously unknown, highly reactive B cell epitopes throughout the full proteome of SARS-CoV-2 and other CoV proteins.


A new approach for studying correlations between the chemical structure and the rheological properties in carboxymethyl cellulose.
Two model sodium carboxymethyl celluloses (CMC) with similar monomer composition but with significant differences in the viscoelastic properties, that could not be assigned to variations in the average molar mass or molar mass distribution, were investigated with respect to the fraction of nonsubstituted cellulose segments in the polymers. The CMCs were hydrolyzed by a purified highly selective endoglucanase. The average molar mass and molar mass distribution of the enzyme products, as measured by size-exclusion chromatography with online multi-angle light scattering and refractive index detection (SEC/MALS/RI), revealed that the enzyme-catalyzed hydrolysis was more effective on one of the CMCs. To investigate whether this was due to a higher fraction of nonsubstituted cellulose segments in the polymer, the concentrations of nonsubstituted enzyme products, e.g., cellotetraose and cellopentaose, were measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). It was concluded that the two CMCs displayed significant differences in the fraction of nonsubstituted cellulose segments. Furthermore, the CMC with the strongest attractive intermolecular interactions, according to rheometry, also contained the highest fraction of nonsubstituted cellulose segments.


Clinical dilemmas and surgical treatment of penoscrotal, scrotal and perineal hypospadias.
Hypospadia is the most common congenital malformation of the urinary tract. It is a malformation with the opening of the urethra proximally from the usual site. The meatal opening can be anywhere alone the shaft of the penis, or in more severe forms, within the scrotum, or in the perineum. Consequently the hypospadias can be distal, medial and proximal. The proximal ones can be penoscrotal (PS), scrotal (SC) and perineal (PE). The cause of hypospadias is largely unknown; however, current epidemiology and laboratory studies have shed new light into the etiology of hypospadias. With recent advancements in molecular biology, microarray technology, it appears that hypospadias is potentially related to disrupted gene expression. Currently, the only available treatment is surgery. The aim of this study was to present our results of the surgical correction of hypospadias and methods used to answer the clinical dilemmas about the gender. Authors have used two methods for a surgical resolution of the hypospadia - one-step operation suggested by Snodgrass and two-step operation, employing free graft suggested by Bracka. Clinical dilemmas regarding the gender were answered using cytogenetic assessment through lymphocyte cultivation method, suggested by Seabright. The cytogenetic assessment was carried in 23 patients with proximal hypospadia (penoscrotal, scrotal and perineal). Characteristic male cariotype (46, XY) was found in 22 patients. In one patient, with scrotal hypospadia, we found the characteristic female cariotype. This patient had testicles. The patient with female cariotype had a TDG gene that determines the differentiation of the testicles. Although surgery remains the only therapy for the treatment of the hypospadias, better understanding of the molecular and hormonal mechanisms behind the diseases may contribute to the prevention and the decrease in the incidence of the malformation. Cytogenetic testing in patients with unclear gender is important in planning further treatment.


Diminished endothelium-derived relaxing factor activity in an experimental model of chronic heart failure.
Abnormalities in vasomotor tone, including enhanced vasoconstriction at rest and diminished vasodilation in response to various stimuli, develop as a consequence of chronic heart failure. This study was undertaken to evaluate whether a specific local mechanism, namely endothelium-derived relaxing factor (EDRF) activity, might be impaired in an experimental model of chronic heart failure. Segments of thoracic aorta (TA) and pulmonary artery (PA) were isolated from a group of rats that had hemodynamic evidence of heart failure 10 weeks after ligation of the left coronary artery (n = 25) and from a group of sham-operated control rats (n = 18). Both endothelium-dependent and endothelium-independent vascular responses were assessed by exposing arterial segments to increasing concentrations of agonists. All studies were performed in the presence of 10 microM indomethacin to avoid the influence of vasoactive prostanoids. The dose-response curve for EDRF-mediated relaxation to acetylcholine was shifted rightward in rats with heart failure, and the concentrations of acetylcholine required to achieve 50% maximal relaxation (EC50) were increased compared with those of control rats in both TA and PA segments. Additionally, the dose-response curve for relaxation to ADP was shifted rightward with significantly increased EC50 in PA segments from rats with heart failure. In contrast, EDRF-mediated relaxation to the calcium ionophore A23187 was similar in the groups. Endothelium-independent relaxation to nitroglycerin was slightly increased in TA but not PA segments in the heart-failure group. Basal EDRF activity, as assessed by the increase in force after exposure to hemoglobin, was diminished in PA segments from rats with heart failure.(ABSTRACT TRUNCATED AT 250 WORDS)


Response of recurrent high-grade glioma to treatment with (90)Y-DOTATOC.
The treatment of patients with high-grade malignant glioma still represents an unsolved clinical problem. We report the treatment of 3 patients who had World Health Organization grade IV recurrent glioblastoma: a 23-y-old woman and 2 men aged 61 and 62 y. All 3 patients were treated with the somatostatin receptor radiopharmaceutical (90)Y-labeled [1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid(0)-d-Phe(1),Tyr(3)]octreotide (DOTATOC). A cumulated dose of 1.7-2.2 GBq given in 3 or 4 cycles was locally injected into a previously implanted catheter system. Treatment was successful in all 3 patients, with only minor side effects reported. After treatment, MRI and PET showed complete remission in one patient and partial remission in the other patients. These findings correlated well with clinical improvement and improved quality of life. Receptor-mediated radionuclide therapy by locally injected (90)Y-DOTATOC is feasible and well tolerated. This approach represents an attractive strategy for the treatment of locally recurring or progressing glioblastoma.


Inflammation, sleep, obesity and cardiovascular disease.
Evidence is emerging that disturbances in sleep and sleep disorders play a role in the morbidity of chronic conditions. However, the relationship between sleep processes, disease development, disease progression and disease management is often unclear or understudied. Numerous common medical conditions can have an affect on sleep. For example, diabetes or inflammatory conditions such as arthritis can lead to poor sleep quality and induce symptoms of excessive daytime sleepiness and fatigue. It has also been suggested that poor sleep may lead to the development of cardiovascular disease for which an underlying inflammatory component has been proposed. It is therefore important that the development and progression of such disease states are studied to determine whether the sleep effect merely reflects disease progression or whether it may be in some way causally related. Sleep loss can also have consequences on safety related behaviours both for the individuals and for the society, for example the increased risk of accidents when driving while drowsy. Sleep is a complex phenotype and as such it is possible that there are numerous genes which may each have a number of effects that control an individual's sleep pattern. This review examines the interaction between sleep (both quantity and quality) and parameters of cardiovascular risk. We also explore the hypothesis that inflammation plays an essential role in cardiovascular disease and that a lack of sleep may play a key role in this inflammatory process. To review current evidence regarding the endocrine, metabolic, cardiovascular and immune functions and their interactions with regard to sleep, given the current evidence that sleep disturbances may affect each of these areas.


Ki67, chemotherapy response, and prognosis in breast cancer patients receiving neoadjuvant treatment.
The pathological complete response (pCR) after neoadjuvant chemotherapy is a surrogate marker for a favorable prognosis in breast cancer patients. Factors capable of predicting a pCR, such as the proliferation marker Ki67, may therefore help improve our understanding of the drug response and its effect on the prognosis. This study investigated the predictive and prognostic value of Ki67 in patients with invasive breast cancer receiving neoadjuvant treatment for breast cancer. Ki67 was stained routinely from core biopsies in 552 patients directly after the fixation and embedding process. HER2/neu, estrogen and progesterone receptors, and grading were also assessed before treatment. These data were used to construct univariate and multivariate models for predicting pCR and prognosis. The tumors were also classified by molecular phenotype to identify subgroups in which predicting pCR and prognosis with Ki67 might be feasible. Using a cut-off value of > 13% positively stained cancer cells, Ki67 was found to be an independent predictor for pCR (OR 3.5; 95% CI, 1.4, 10.1) and for overall survival (HR 8.1; 95% CI, 3.3 to 20.4) and distant disease-free survival (HR 3.2; 95% CI, 1.8 to 5.9). The mean Ki67 value was 50.6 ± 23.4% in patients with pCR. Patients without a pCR had an average of 26.7 ± 22.9% positively stained cancer cells. Ki67 has predictive and prognostic value and is a feasible marker for clinical practice. It independently improved the prediction of treatment response and prognosis in a group of breast cancer patients receiving neoadjuvant treatment. As mean Ki67 values in patients with a pCR were very high, cut-off values in a high range above which the prognosis may be better than in patients with lower Ki67 values may be hypothesized. Larger studies will be needed in order to investigate these findings further.


Morphological maturation of tumor cells induced by ethylnitrosourea (ENU) in rat brains. II. On the tumors by administration of ENU in the mid-gestational stage.
In order to clarify the relationship between cellular characteristics of rat brain tumors and the administration time of chemical carcinogens, the incidence and morphology of microtumors in the offspring of five pregnant Sprague-Dawley rats injected with ethylnitrosourea (ENU) intraperitoneally on the 11th day of gestation (Group I) were compared with those of six pregnant rats injected with ENU on the 18th day of gestation (Group II). Twenty-four microtumors and 6 macrotumors were obtained in Group I and 58 microtumors and 5 macrotumors in Group II. Histologically, the macrotumors of Group I showed a poorly differentiated pattern which corresponded to primitive spongioblastomas in comparison with those of Group II which resembled human mature oligodendrogliomas, astrocytomas or mixed gliomas. The constituent cells of the microtumors of both groups were immature and were morphologically similar to each other. Immunohistochemically, the cells of microtumors of both groups were negative for Leu 7 and glial fibrillary acidic protein. Microtumors in Group I showed more infiltrative proliferation than those in Group II and had indistinct borders. The 3H-thymidine labeling index of the constituent cells of Group I was significantly higher than that of Group II (p less than 0.001). This study revealed that the constituent cells of microtumors of Group I, in spite of being morphologically similar with those of Group II, possess a greater growth potency than the latter. This difference in growth potency is responsible for the difference in the histological maturity of the macrotumors in each group. The difference in morphological maturation of enlarged gliomas induced by ENU appears to depend on the imperceptible difference of the target cell maturation at the initiation of oncogenesis.


Beware the perianal granuloma! A case report.
Crohn's disease (CD) can often present initially with extraintestinal manifestations and/or perianal disease. Cytology of such a lesion helps arrive at the diagnosis. A 27-year-old woman presented with recurrent, painful, discharging perianal lesions. On examination, multiple perianal abscesses, sinuses, skin tags and healed scars were noted. Fine needle aspiration cytology (FNAC) of the perianal lesions performed as a first line investigation revealed epithelioid granulomas in a suppurative background. The possibility of tuberculosis vs. extraintestinal manifestation of CD was considered. Endoscopic studies revealed involvement of distal ileum and the entire colon by deep, serpiginous ulcers, internal fistulae and a distorted ileocecal valve. Biopsy of the intestinal lesions revealed chronic inflammation, aphthous ulcers and microgranulomas, favoring a diagnosis of CD. A granulomatous lesion in any location arouses the clinical suspicion of tuberculosis, given its prevalence in the Indian subcontinent. In view of the increasing incidence of CD in our setting, the finding of a perianal, granulomatous lesion on cytology should prompt a workup for CD in addition to the more common granulomatous counterpart, tuberculosis. Perianal aspiration is an underused diagnostic procedure that can help in narrowing the differential diagnostic possibilities.


The effects of an awareness-raising program for patients and primary care physicians on the early detection of gastro-oesophageal cancer.
To test the efficacy of education by home letter, together with improved specialist/GP liaison, in promoting the detection of treatable gastro-oesophageal cancer in patients over 40. Prospective non-randomized trial with matched control group. 37,500 individuals over 40, registered with 12 general practices, were sent yearly letters for 3 consecutive years, advising them to consult a doctor with 4 specific symptoms. A matched control population of 60,500 received no intervention. Cancer cases were identified from cancer registry data, and the stage distribution and operability rates calculated from review of hospital case notes. Twenty of 59 cancer patients in the study group (34%) and 42 of 125 in the control group (34%) underwent resections; 24% (14/59) of study group and 18% (23/125) of control patients had "curable" disease (odds ratio [OR]= 1.51 [95%CI 0.71-3.23], X(2) = 1.164, P = 0.28). In year 1 of the study, 11 of 20 (55%) study group patients underwent resection compared to P 10 of 38 (26%) control group patients (P = 0.045, Fisher's exact test, OR = 3.4 [95% CI 1.09-0.7]). In year 1, 8 of 20 study patients (40%) had "curable" disease compared to 10.5% (4/38) control patients (P = 0.0128 Fisher's exact test, OR = 5.67 [95% CI 1.44 - 22.3]). There was no difference in survival between the groups overall or on comparing each year of study. This intervention caused a transient marked improvement in resection rate and curability, but the longer term impact on stage and resection rate was nonsignificant, and survival was not affected. Improving the detection of curable upper GI cancer is likely to require more complex and intensive interventions.


Microvillous inclusion disease (microvillous atrophy).
Microvillous inclusion disease (MVID) or microvillous atrophy is a congenital disorder of the intestinal epithelial cells that presents with persistent life-threatening watery diarrhea and is characterized by morphological enterocyte abnormalities. MVID manifests either in the first days of life (early-onset form) or in the first two months (late-onset form) of life. MVID is a very rare disorder of unknown origin, probably transmitted as an autosomal recessive trait. To date, no prevalence data are available. Ultrastructural analyses reveal: 1) a partial to total atrophy of microvilli on mature enterocytes with apical accumulation of numerous secretory granules in immature enterocytes; 2) the highly characteristic inclusion bodies containing rudimentary or fully differentiated microvilli in mature enterocytes. Light microscopy shows accumulation of PAS-positive granules at the apical pole of immature enterocytes, together with atrophic band indicating microvillus atrophy and, in parallel, an intracellular PAS or CD10 positive line (marking the microvillous inclusion bodies seen on electron microscopy). Intestinal failure secondary to diarrhea is definitive. To date, no curative therapy exists and children with MVID are totally dependent on parenteral nutrition. Long-term outcome is generally poor, due to metabolic decompensation, repeated states of dehydration, infectious and liver complications related to the parenteral nutrition. As MVID is a very rare disorder, which is extremely difficult to diagnose and manage, children with MVID should be transferred to specialized pediatric gastro-intestinal centers, if possible, a center equipped to perform small bowel transplantation. Early small bowel transplantation resulting in intestinal autonomy gives new hope for disease management and outcome.


Forced chromatin looping raises fetal hemoglobin in adult sickle cells to higher levels than pharmacologic inducers.
Overcoming the silencing of the fetal γ-globin gene has been a long-standing goal in the treatment of sickle cell disease (SCD). The major transcriptional enhancer of the β-globin locus, called the locus control region (LCR), dynamically interacts with the developmental stage-appropriate β-type globin genes via chromatin looping, a process requiring the protein Ldb1. In adult erythroid cells, the LCR can be redirected from the adult β- to the fetal γ-globin promoter by tethering Ldb1 to the human γ-globin promoter with custom-designed zinc finger (ZF) proteins (ZF-Ldb1), leading to reactivation of γ-globin gene expression. To compare this approach to pharmacologic reactivation of fetal hemoglobin (HbF), hematopoietic cells from patients with SCD were treated with a lentivirus expressing the ZF-Ldb1 or with chemical HbF inducers. The HbF increase in cells treated with ZF-Ldb1 was more than double that observed with decitabine and pomalidomide; butyrate had an intermediate effect whereas tranylcypromine and hydroxyurea showed relatively low HbF reactivation. ZF-Ldb1 showed comparatively little toxicity, and reduced sickle hemoglobin (HbS) synthesis as well as sickling of SCD erythroid cells under hypoxic conditions. The efficacy and low cytotoxicity of lentiviral-mediated ZF-Ldb1 gene transfer compared with the drug regimens support its therapeutic potential for the treatment of SCD.


Insertion of transposon Tn5 into a structural gene of the membrane-bound nitrate reductase of Thiosphaera pantotropha results in anaerobic overexpression of periplasmic nitrate reductase activity.
Chlorate-resistant mutants of the denitrifying bacterium Thiosphaera pantotropha were generated by transposon Tn5 mutagenesis. One class was deficient in membrane-bound nitrate reductase activity but retained a periplasmic nitrate reductase activity. Using transposon marker rescue it was shown that in one such mutant, M-6, the transposon was inserted in the membrane-bound nitrate reductase beta subunit structural gene (termed narH in order to be consistent with the nomenclature of the Escherichia coli major nitrate reductase operon). The translated sequence (total of 106 amino acids) from around the point of transposon insertion showed approximately 90% amino acid identity with the beta subunits of the E. coli nitrate reductases. Under anaerobic growth conditions M-6 overproduced the periplasmic nitrate reductase activity allowing anaerobic growth with nitrate as electron acceptor. A regulatory link was inferred between the presence of the membrane-bound nitrate reductase and expression of the periplasmic nitrate reductase. This is the first demonstration of full denitrification in an organism possessing only a periplasmic nitrate reductase.


The World Summit on Sustainable Development: reaffirming the centrality of health.
The World Summit on Sustainable Development (WSSD) was held in Johannesburg in 2002 to review progress since the Rio conference in 1992, and to agree a new global deal on sustainable development. Unlike its predecessor, it was primarily concerned with implementation rather than with new treaties and targets, although a number of new targets were agreed, for example one on sanitation. Failure to agree a target on renewable energy was regarded as a major disappointment of the conference. While relatively modest in its achievements, and with difficulties in achieving consensus in key areas such as energy, trade, finance and globalisation, WSSD nevertheless succeeded in placing sustainable development back on the political agenda, giving new impetus, in particular to the environment and development needs of Africa, with a strong focus on local issues like household energy, water and sanitation. Health was singled out as one of five priority areas, along with water, energy, agriculture and biodiversity, and was devoted a separate chapter in the resulting Plan of Implementation, which highlighted a range of environmental health issues as well as issues relating to health services, communicable and non-communicable diseases. A number of new partnerships were formed at WSSD, including the Healthy Environments for Children Alliance (HECA) launched by WHO, which will form an important platform for implementation. The Commission on Sustainable Development has been designated main responsibility for monitoring and follow up, with its programme of work reorganised to focus on thematic clusters of issues. From the perspective of health, WSSD must be seen as a reaffirmation of the central place of health on the sustainable development agenda, and in the broader context of a process which began in Rio and was given added impetus with the Monterrey Financing for Development conference and the World Trade Organisation meeting held in Doha. Translating policies into action at all levels- global to local - remains the single biggest challenge in the years that lie ahead.


Diurnal Variation in PDK4 Expression Is Associated With Plasma Free Fatty Acid Availability in People.
Many biological pathways involved in regulating substrate metabolism display rhythmic oscillation patterns. In rodents, clock genes regulate circadian rhythms of metabolic genes and substrate metabolism. However, the interrelationships among substrate metabolism, metabolic genes, and clock genes have not been fully explored in people. We tested the hypothesis that the diurnal expression pattern of pyruvate dehydrogenase kinase 4 (PDK4), a key metabolic enzyme involved in fuel switching between glucose and free fatty acids (FFAs), is associated with plasma FFA concentration and clock genes. We analyzed peripheral blood mononuclear cells (PBMCs), subcutaneous adipose tissue, and plasma samples obtained serially during 24 hours from metabolically healthy women (n = 10) and evaluated the interrelationships among PDK4, plasma FFA, and clock genes. We also determined the potential mechanisms responsible for PDK4 transcriptional regulation by using primary human PBMCs and adipocytes. We found that PDK4 diurnal expression patterns were similar in PBMCs and adipose tissue (ρ = 0.84, P < 0.001). The diurnal variation in PBMC PDK4 expression correlated more strongly with plasma FFA and insulin (ρ = 0.86 and 0.63, respectively, both P < 0.001) concentrations than clock genes. Data obtained from primary culture experiments demonstrated that FFAs directly induced PDK4 gene expression, at least in part through activation of peroxisome proliferator-activated receptor α. Our results suggest that plasma FFA availability is an important regulator of diurnal expression patterns of PDK4, and we identify a novel interaction between plasma FFA and cellular diurnal rhythms in regulating substrate metabolism.


[Genetic complexity of ciliopathies and novel genes identification].
Ciliopathies are a large group of human disorders caused by dysfunction of primary or motile cilia and unified by their overlapping clinical features (brain malformations, retinal dystrophy, cystic kidney disease, liver fibrosis and skeletal abnormalities). Ciliopathies are mendelian disorders with prominent genetic heterogeneity and marked allelism between different clinical entities, which are in part explained by the recently identified functional modules and multi-protein complexes formed by ciliopathy-associated gene products. The current review provides an updated snapshot of this complex evolving field, highlighting the key phenotypic features and causative genes for commonly-studied ciliopathies and summarizing our emerging understanding of the correlations between the functions of subgroups of genes and clinical sub-types of ciliopathies. Using the example of Joubert syndrome, a ciliopathy characterized by a distinctive hindbrain malformation and caused by mutations in more than 20 different genes, this work also reviews the principal methods used for new gene identification, including candidate gene approaches, homozygosity mapping as well as high throughput next-generation and exome sequencing.


Second primary cancers - causes, incidence and the future.
Thanks to continually improving screening programs, diagnostic, and treatment methods, the survival rate in newly diagnosed cancer patients is increasing. With this improvement in survival, attention is now being focused on potential longterm complications such as multiple primary tumors, which represent a leading cause of late nonrelapse mortality. The number of patients who survive cancer dia-gnosis is growing by 2% each year. Multiple primary neoplasms have become the third most common finding in oncology since 1890s, when they were first described. This review aims to summarize recent information regarding the multiple primary neoplasms, elucidate the definition, etiology, association with the primary cancer treatment, genetic and environmental dispositions and finally, it recapitulates new approaches to identification of the risk factors for multiple cancers.


Are trajectories of neck-shoulder pain associated with sick leave and work ability in workers? A 1-year prospective study.
The study aimed to determine the extent to which latent trajectories of neck-shoulder pain (NSP) are associated with self-reported sick leave and work ability based on frequent repeated measures over 1 year in an occupational population. This longitudinal study included 748 Danish workers (blue-collar, n=620; white collar, n=128). A questionnaire was administered to collect data on personal and occupational factors at baseline. Text messages were used for repeated measurements of NSP intensity (scale 0-10) over 1 year (14 waves in total). Simultaneously, self-reported sick leave (days/month) due to pain was assessed at 4-week intervals, while work ability (scale 0-10) was assessed using a single item (work ability index) at 12-week intervals over the year. Trajectories of NSP, distinguished by latent class growth analysis, were used as predictors of sick leave and work ability in generalised estimation equations with multiple adjustments. Sick leave increased and work ability decreased across all NSP trajectory classes (low, moderate, strong fluctuating and severe persistent pain intensity). In the adjusted model, the estimated number of days on sick leave was 1.5 days/month for severe persistent NSP compared with 0.1 days/month for low NSP (relative risk=13.8, 95% CI 6.7 to 28.5). Similarly, work ability decreased markedly for severe persistent NSP (OR=12.9, 95% CI 8.5 to 19.7; median 7.1) compared with low NSP (median 9.5). Severe persistent NSP was associated with sick leave and poor work ability over 1 year among workers. Preventive strategies aiming at reducing severe persistent NSP among working populations are needed.


Estimating lifetime and 10-year risk of lung cancer.
Lung cancer is the commonest cancer worldwide. Mortality and incidence rates are traditionally used to assess cancer burden and as public health indicators. However, these metrics are difficult to interpret at an individual level. Providing the lifetime and 10-year risks of cancer could improve risk communication. Our aim was to estimate current lifetime and 10-year risks of lung cancer by smoking status and changes in these risks between 1995 and 2013 in a Swiss population. We used all lung cancer cases recorded between 1995 and 2013 by two population-based cancer registries in the contiguous cantons of Vaud and Valais, in Western Switzerland. We estimated sex-specific lifetime risk and 10-year risk of lung cancer using the current probability method, accounting for competing risk of death. Estimates were also provided by smoking status. Between 1995 and 2013, 9623 cases of lung cancer were recorded. During this period, the lifetime risk decreased in men from 7.1% to 6.7% and increased in women from 2.5% to 4.1%. In both sexes, the 10-year risk of lung cancer increased with age until the age of 60-70 and decreased thereafter. Difference in the cumulative risk between current, former, and never smokers were very large and reported in user-friendly charts to ease risk communication. These lifetime and 10-year risk estimates could be used systematically as public health indicators. Regularly updating risk estimations are necessary for conditions like lung cancer whose incidence has changed substantially.


Why Great Britain's success in Beijing could have been anticipated and why it should continue beyond 2012.
Home advantage in the summer Olympic Games is well known. What is not so well known is that countries that host the Olympic Games perform better in the games before and after the games in which they were hosts. To model/quantify the significance associated with these "hosting" effects and to explain the likely causes of Great Britain's improved medals haul in Beijing, while examining implications for London 2012 and beyond. Using all hosting cities/countries since World War II and analysing the number of medals awarded to competitors as a binomial proportion (p) response variable within a logit model, we identified a significant increase in the probability/odds of a country obtaining a medal in the Olympic Games before, during and after hosting the Olympics. Funding appears to be an important factor when explaining these findings. Almost all countries that have been awarded the games after World War II would appear to have invested heavily in sport before being awarded the games. A second factor in Great Britain's success is the legacy of hosting the Commonwealth Games in 2002 (a post-hosting games effect) that undoubtedly provided an infrastructure that benefited, in particular, cycling. Whether the International Olympics Committee either consciously or subconsciously take these factors into account is unclear when awarding the games to a city. What is clear is that based on these findings, Great Britain's prospects of maintaining the Olympic success achieved in Beijing is likely to continue to London 2012 and beyond.


Attenuation of Slc27a5 gene expression followed by LC-MS measurement of bile acid reconjugation using metabolomics and a stable isotope tracer strategy.
The purpose of this study was to evaluate the use of high resolution LC-MS together with metabolomics and D(4)-cholic acid (D(4)-CA) as a metabolic tracer to measure the metabolism and reconjugation of bile acids (BAs) in vitro and in vivo. Metabolic tracers are very important because they allow for the direct detection (substrate-to-product) of small and significant biological perturbations that may not be apparent when monitoring "static" endogenous levels of particular metabolites. Slc27a5, also known as fatty acid transport protein 5 (FATP5), is the hepatic BA-CoA ligase involved in reconjugating BAs during enterohepatic BA recycling. Using Slc27a5-cKD mice, silencing of ∼90% gene expression was achieved followed by reduction in the reconjugation of D(4)-CA to D(4)-taurocholic acid (D(4)-TCA), as well as other conjugated BA metabolites in plasma (p = 0.0031). The method described allowed a rapid measure of many D(4) and endogenous BA. Analysis of bile resulted in the detection of 39 BA metabolites from a 13 min analytical run. Finally, the utilization of a novel high resolution mass spectrometry method in combination with metabolomics and a stable isotope metabolic tracer allowed for the detection of targeted and untargeted BAs following silencing of the Slc27a5 gene in primary hepatocytes and in mice.


Triple-phase helical computed tomography in dogs with hepatic masses.
The purpose of this study was to determine the utility of triple-phase helical computed tomography (CT) for differentiating canine hepatic masses. Seventy dogs with hepatic masses underwent triple-phase CT followed by surgical removal of the hepatic masses. Triple-phase helical CT scans for each dog included precontrast, arterial phase, portal venous phase, and delayed phase studies. The removed hepatic masses were histopathologically classified as hepatocellular carcinoma (n = 47), nodular hyperplasia (n = 14), and hepatic metastatic tumors (n = 9) in dogs. Of the 47 hepatocellular carcinomas, the most common CT findings included a heterogeneous pattern with hyper-, iso-, and hypoenhancement in both the arterial and portal venous phases (40/47, 85.1%). Of the 14 nodular hyperplasias, the most common CT findings were a homogeneous pattern with hyper- and isoenhancement in both the portal venous and delayed phases (13/14, 92.9%). Of nine hepatic metastatic tumors, the most common CT findings included a homogeneous hypoenhancement pattern in both the arterial and portal venous phases (8/9, 88.9%). In addition, 5 (55.6%) showed homogeneous hypoenhancement patterns in the delayed phase. Findings from our study indicated that triple-phase CT is a useful tool for preoperative differentiation of hepatocellular carcinoma, nodular hyperplasia, and hepatic metastatic tumors in dogs.


Physical exercise enhances cognitive flexibility as well as astrocytic and synaptic markers in the medial prefrontal cortex.
Physical exercise enhances a wide range of cognitive functions in humans. Running-induced cognitive enhancement has also been demonstrated in rodents but with a strong emphasis on tasks that require the hippocampus. Additionally, studies designed to identify mechanisms that underlie cognitive enhancement with physical exercise have focused on running-induced changes in neurons with little attention paid to such changes in astrocytes. To further our understanding of how the brain changes with physical exercise, we investigated whether running alters performance on cognitive tasks that require the prefrontal cortex and whether any such changes are associated with astrocytic, as well as neuronal, plasticity. We found that running enhances performance on cognitive tasks known to rely on the prefrontal cortex. By contrast, we found no such improvement on a cognitive task known to rely on the perirhinal cortex. Moreover, we found that running enhances synaptic, dendritic and astrocytic measures in several brain regions involved in cognition but that changes in the latter measures were more specific to brain regions associated with cognitive improvements. These findings suggest that physical exercise induces widespread plasticity in both neuronal and nonneuronal elements and that both types of changes may be involved in running-induced cognitive enhancement.


Spatial profiles of electrical mismatch determine vulnerability to conduction failure across a host-donor cell interface.
Electrophysiological mismatch between host cardiomyocytes and donor cells can directly affect the electrical safety of cardiac cell therapies; however, the ability to study host-donor interactions at the microscopic scale in situ is severely limited. We systematically explored how action potential (AP) differences between cardiomyocytes and other excitable cells modulate vulnerability to conduction failure in vitro. AP propagation was optically mapped at 75 μm resolution in micropatterned strands (n=152) in which host neonatal rat ventricular myocytes (AP duration=153.2±2.3 ms, conduction velocity=22.3±0.3 cm/s) seamlessly interfaced with genetically engineered excitable donor cells expressing inward rectifier potassium (Kir2.1) and cardiac sodium (Na(v)1.5) channels with either weak (conduction velocity=3.1±0.1 cm/s) or strong (conduction velocity=22.1±0.4 cm/s) electrical coupling. Selective prolongation of engineered donor cell AP duration (31.9-139.1 ms) by low-dose BaCl2 generated a wide range of host-donor repolarization time (RT) profiles with maximum gradients (∇RT(max)) of 5.5 to 257 ms/mm. During programmed stimulation of donor cells, the vulnerable time window for conduction block across the host-donor interface most strongly correlated with ∇RT(max). Compared with well-coupled donor cells, the interface composed of poorly coupled cells significantly shortened the RT profile width by 19.7% and increased ∇RT(max) and vulnerable time window by 22.2% and 19%, respectively. Flattening the RT profile by perfusion of 50 μmol/L BaCl2 eliminated coupling-induced differences in vulnerability to block. Our results quantify how the degree of electrical mismatch across a cardiomyocyte-donor cell interface affects vulnerability to conduction block, with important implications for the design of safe cardiac cell and gene therapies.


Reproductive compensation in broad-nosed pipefish females.
The differential allocation hypothesis assumes that animals should weigh costs and benefits of investing into reproduction with a current mate against the expected quality of future mates, and predicts that they should invest more into reproduction when pairing with a high-quality mate. In the broad-nosed pipefish (Syngnathus typhle), males care for the embryos in a brood pouch and females compete for access to male mating partners. Both sexes prefer mating with large partners. In the present study, we show that the same female provides both large and small mating partners with eggs of similar size, weight and lipid content when mated to two males in succession. Importantly, however, eggs provided to small males (less preferred) had higher egg protein content (11% more) than those provided to large males (preferred). Thus, contrary to the differential allocation hypothesis, eggs did not contain more resources when females mated with a larger male. Instead, the pattern observed in our results is consistent with a compensatory reproductive strategy.


Genome-scale MicroRNA target prediction through clustering with Dirichlet process mixture model.
MicroRNA regulation is fundamentally responsible for fine-tuning the whole gene network in human and has been implicated in most physiological and pathological conditions. Studying regulatory impact of microRNA on various cellular and disease processes has resulted in numerous computational tools that investigate microRNA-mRNA interactions through the prediction of static binding site highly dependent on sequence pairing. However, what hindered the practical use of such target prediction is the interplay between competing and cooperative microRNA binding that complicates the whole regulatory process exceptionally. We developed a new method for improved microRNA target prediction based on Dirichlet Process Gaussian Mixture Model (DPGMM) using a large collection of molecular features associated with microRNA, mRNA, and the interaction sites. Multiple validations based on microRNA-mRNA interactions reported in recent large-scale sequencing analyses and a screening test on the entire human transcriptome show that our model outperformed several state-of-the-art tools in terms of promising predictive power on binding sites specific to transcript isoforms with reduced false positive prediction. Last, we illustrated the use of predicted targets in constructing conditional microRNA-mediated gene regulation networks in human cancer. The probability-based binding site prediction provides not only a useful tool for differentiating microRNA targets according to the estimated binding potential but also a capability highly important for exploring dynamic regulation where binding competition is involved.


DNA sequence polymorphism at the human tumor necrosis factor (TNF) locus. Numerous TNF/lymphotoxin alleles tagged by two closely linked microsatellites in the upstream region of the lymphotoxin (TNF-beta) gene.
TNF-alpha and lymphotoxin (LT, TNF-beta) genes are tandemly arranged and map within the MHC centromeric to HLA-B and telomeric to the class III genes. Both cytokines encoded by these genes are potent immunomodulators. On the other hand, some MHC-linked autoimmune diseases are characterized by abnormal levels of their expression or inducibility. A search for the putative disease-associated TNF/LT alleles depends on the informative genetic markers at the TNF locus. Previously, a low degree of genetic polymorphism at the human TNF locus has been reported, mostly bi-allelic RFLP. To localize and define additional polymorphic markers, we probed the collection of genomic clones with synthetic tandemly repeated dinucleotides, corresponding to the sequences known as microsatellites. We mapped and characterized three (TC/GA) and one (AC/GT) repeats within cloned 40-kb DNA comprising the human TNF locus. Using a polymerase chain reaction-based technique, we analyzed three of these four microsatellites and observed their length of polymorphism. Using DNA samples from blood donors, two families, and three human cell lines, we detected 13 distinct alleles of the AC/GT microsatellite neighboring human TNF genes. The variability was further increased by simultaneous analysis of the second linked microsatellite. This linked TC/GA repeat showed at least five alleles, whereas the least polymorphic TC/GA repeat located in the first intron of LT (TNF-beta) gene had two alleles. TNF alleles defined by microsatellites were stably inherited and segregated in the Mendelian way. Therefore, we describe thus far the most informative level of DNA sequence polymorphism in this part of human MHC. We propose a nomenclature for microsatellite tagged LT/TNF alleles based on their size and variability, which could also be extended to include RFLP and other not yet identified polymorphic markers. Microsatellite tagged polymorphism described here can be used in systematic linkage studies of HLA-associated diseases.


Landscape genetics of high mountain frog metapopulations.
Explaining functional connectivity among occupied habitats is crucial for understanding metapopulation dynamics and species ecology. Landscape genetics has primarily focused on elucidating how ecological features between observations influence gene flow. Functional connectivity, however, may be the result of both these between-site (landscape resistance) landscape characteristics and at-site (patch quality) landscape processes that can be captured using network based models. We test hypotheses of functional connectivity that include both between-site and at-site landscape processes in metapopulations of Columbia spotted frogs (Rana luteiventris) by employing a novel justification of gravity models for landscape genetics (eight microsatellite loci, 37 sites, n = 441). Primarily used in transportation and economic geography, gravity models are a unique approach as flow (e.g. gene flow) is explained as a function of three basic components: distance between sites, production/attraction (e.g. at-site landscape process) and resistance (e.g. between-site landscape process). The study system contains a network of nutrient poor high mountain lakes where we hypothesized a short growing season and complex topography between sites limit R. luteiventris gene flow. In addition, we hypothesized production of offspring is limited by breeding site characteristics such as the introduction of predatory fish and inherent site productivity. We found that R. luteiventris connectivity was negatively correlated with distance between sites, presence of predatory fish (at-site) and topographic complexity (between-site). Conversely, site productivity (as measured by heat load index, at-site) and growing season (as measured by frost-free period between-sites) were positively correlated with gene flow. The negative effect of predation and positive effect of site productivity, in concert with bottleneck tests, support the presence of source-sink dynamics. In conclusion, gravity models provide a powerful new modelling approach for examining a wide range of both basic and applied questions in landscape genetics.


Novel Human Podocyte Cell Model Carrying G2/G2 APOL1 High-Risk Genotype.
Apolipoprotein L1 (APOL1) high-risk genotypes (HRG), G1 and G2, increase the risk of various non-diabetic kidney diseases in the African population. To date, the precise mechanisms by which APOL1 risk variants induce injury on podocytes and other kidney cells remain unclear. Trying to unravel these mechanisms, most studies have used animal or cell models created by gene editing. We developed and characterised conditionally immortalised human podocyte cell lines derived from urine of a donor carrying APOL1 HRG G2/G2. Following induction of APOL1 expression by polyinosinic-polycytidylic acid (poly(I:C)), we assessed functional features of APOL1-induced podocyte dysfunction. As control, APOL1 wild type (G0/G0) podocyte cell line previously generated from a Caucasian donor was used. Upon exposure to poly(I:C), G2/G2 and G0/G0 podocytes upregulated APOL1 expression resulting in podocytes detachment, decreased cells viability and increased apoptosis rate in a genotype-independent manner. Nevertheless, G2/G2 podocyte cell lines exhibited altered features, including upregulation of CD2AP, alteration of cytoskeleton, reduction of autophagic flux and increased permeability in an in vitro model under continuous perfusion. The human APOL1 G2/G2 podocyte cell model is a useful tool for unravelling the mechanisms of APOL1-induced podocyte injury and the cellular functions of APOL1.


Physiological responses to heat stress in two genetically distinct chicken inbred lines.
High ambient temperature is one of the most important environmental factors negatively impacting poultry production and health. Genetics is an important contributor in mitigating the stress response to heat. Two genetically distinct highly inbred lines of similar body size (Leghorn and Fayoumi) were characterized for phenotypic differences in response to heat. At 14 days of age, birds were exposed to 38°C with 50% humidity for 4 hours, then 35°C until the conclusion of the experiment. Non-treated individuals were kept at 29.4°C for the first week and then 25°C throughout the experiment. Birds in the heat-stress group were inoculated at day (d) 21 with Newcastle disease virus (NDV) La Sota strain to investigate the effects of heat stress and NDV infection. Thirteen blood parameters were measured using the iSTAT blood analyzer at three stages: 4 h, 6 d, and 9 d post heat-stress treatment, representing acute heat (AH) exposure, chronic heat (CH1) exposure, and chronic heat exposure after virus infection (CH2), respectively. Most blood parameters were significantly changed with heat stress in Leghorns at AH and in Fayoumis at CH1 and CH2. The Leghorn line had significant acute responses with disrupted acid-base balance and metabolic disorders. The heat-resilient Fayoumis maintained a relatively well-balanced acid-base balance. The current study provides the comprehensive profile of biomarker signatures in blood associated with heat tolerance and suggests that PO2, TCO2, HCO3, and base excess can be served as potential biomarkers that can be used to genetically improve heat tolerance in poultry.


Developing community indicators to promote the health and well-being of older people.
As the US population ages and the number of older people who are "aging in place" increases, communities will face new opportunities and challenges in responding to this population's desires and needs. Qualitative research was conducted to inform the development of a model of an "elder-friendly community" and a set of indicators to measure and help improve community capacity to promote the health and well-being of older residents. Focus groups were conducted in four US cities with younger and older adults and community leaders who identified attributes that make a community a good place in which to grow old. The group interactions were videotaped, transcribed, and analyzed to identify common themes. Results were remarkably similar across sites. Participants said that a community could be considered elder-friendly if it helped older residents continue active participation in the community, sustain their independence, and reduce the risk of isolation. A model of an elder-friendly community, along with corresponding indicators, was created on the basis of focus group results. These tools are being tested in 10 pilot communities to determine their usefulness for measuring older people's health and well-being, prioritizing aging issues, and stimulating and informing action strategies to improve community "elder-friendliness."


Bilateral internal thoracic artery grafting: in situ or composite?
Bilateral internal thoracic artery (BITA) grafting is considered a superior choice for coronary artery bypass grafting (CABG). While the 10-year outcomes of BITA grafting from the recent Arterial Revascularization Trial (ART) are still pending, numerous observational studies have demonstrated the advantages of BITA grafting. These include better long-term graft patency and freedom from arteriosclerosis, in addition to higher survival rate compared to CABG using only the left internal thoracic artery (ITA). The different BITA configurations are in situ and composite-the choice of optimal grafting configuration is challenging. Patient factors such as coronary anatomy, presence of a diseased ascending aorta and the potential need for a future redo sternotomy will influence the choice of the grafting strategy. In situ BITA grafting is associated with excellent clinical outcomes and has been extensively described in the literature. However, uncertainties remain regarding the ideal in situ configuration and design. Composite BITA grafting is the other option that maximizes right ITA (RITA) utilization. In this configuration, the RITA is able to reach the distal circumflex and right coronary artery branches. This approach decreases the need for a third graft conduit.


Nuclear factor-kappaB and STAT3 are constitutively active in CD138+ cells derived from multiple myeloma patients, and suppression of these transcription factors leads to apoptosis.
Chemoresistance is a major problem in the treatment of patients with multiple myeloma (MM). Because of the central role of the nuclear transcription factors nuclear factor-kappaB (NF-kappaB) and signal transducer and activator of transcription 3 (STAT3) in chemoresistance, cell survival, and proliferation, we investigated whether MM cells derived from patients express activated NF-kappaB and STAT3 and if their suppression induces apoptosis. We assayed CD138+ cells from the bone marrow of 22 MM patients and checked for the activated forms of NF-kappaB and STAT3 by immunocytochemistry. We found that MM cells from all the patients expressed the activated forms of NF-kappaB and STAT3 but to a variable degree (NF-kappaB: low, 3 of 22; moderate, 5 of 22; or high, 14 of 22; STAT3: none, 1 of 22; low, 3 of 22; moderate, 5 of 22; or high, 14 of 22). Constitutive activation of NF-kappaB was in some cases also independently confirmed by electrophoretic mobility gel shift assay. In contrast to MM patients, activated forms of NF-kappaB and STAT3 were absent in cells from healthy individuals. Suppression of NF-kappaB and STAT3 activation in MM cells by ex vivo treatment with curcumin (diferuloylmethane) resulted in a decrease in adhesion to bone marrow stromal cells, cytokine secretion, and in the viability of cells. When compared with curcumin, dexamethasone was less effective in suppression of NF-kappaB activation and induction of apoptosis in myeloma cells. Overall, our results indicate that fresh cells from MM patients express constitutively active NF-kappaB and STAT3, and suppression of these transcription factors inhibits the survival of the cells.


Dopamine, working memory, and training induced plasticity: implications for developmental research.
Cognitive deficits and particularly deficits in working memory (WM) capacity are common features in neuropsychiatric disorders. Understanding the underlying mechanisms through which WM capacity can be improved is therefore of great importance. Several lines of research indicate that dopamine plays an important role not only in WM function but also for improving WM capacity. For example, pharmacological interventions acting on the dopaminergic system, such as methylphenidate, improve WM performance. In addition, behavioral interventions for improving WM performance in the form of intensive computerized training have recently been associated with changes in dopamine receptor density. These two different means of improving WM performance--pharmacological and behavioral--are thus associated with similar biological mechanisms in the brain involving dopaminergic systems. This article reviews some of the evidence for the role of dopamine in WM functioning, in particular concerning the link to WM development and cognitive plasticity. Novel data are presented showing that variation in the dopamine transporter gene (DAT1) influences improvements in WM and fluid intelligence in preschool-age children following cognitive training. Our results emphasize the importance of the role of dopamine in determining cognitive plasticity.


Chemsex and the city: sexualised substance use in gay bisexual and other men who have sex with men attending sexual health clinics.
The objective of this study was to analyse associations between sexualised substance use (chemsex), STI diagnoses and sexual behaviour among gay bisexual and other men who have sex with men accessing sexual health clinics to better inform clinical pathways. A retrospective case notes review was undertaken following the introduction of more detailed and holistic profomas for all gay bisexual and other men who have sex with men attending two London sexual health clinics between 1 June 2014 and 31 January 2015. Chemsex status was documented for 655/818. Overall, 30% disclosed recreational drug use of whom 113 (57%) disclosed chemsex and 27 (13.5%) injecting drugs. HIV-positive gay bisexual and other men who have sex with men were more likely to disclose chemsex (AOR 6.68; 95% CI 3.91-11.42; p < 0.001). Those disclosing chemsex had a higher incidence of acute bacterial STIs (AOR 2.83 CI 1.79-4.47; p < 0.001), rectal STIs (AOR 3.10 CI 1.81-5.32; p < 0.001) or hepatitis C (AOR 15.41 CI 1.50-158.17; p = 0.021). HIV incidence in the study period was 1.8% (chemsex) vs. 0.9% (no chemsex) (p = 0.61). Chemsex was associated with having more sexual partners, transactional sex, group sex, fisting, sharing sex toys, injecting drug use, higher alcohol consumption and the use of 'bareback' sexual networking applications (p < 0.004). Chemsex participants were also more likely to have accessed post-exposure prophylaxis for HIV in the study period and report sex with a discordant HIV or hepatitis C-infected partner (p < 0.001). Chemsex disclosure is associated with higher risk-taking behaviours, acute bacterial STIs, rectal STIs and hepatitis C incidence. HIV incidence was higher but not significantly so in the study period. Chemsex disclosure in sexual health clinics should prompt an opportunity for prevention, health promotion and wellbeing interventions.


'I really tried': management of normative issues in accounts of responses to infertility.
This paper examines how people account for health-seeking behaviour regarding infertility in developing countries, where infertility is a serious problem. Traditionally, studies of health related behaviours focus on how individuals' decisions depend upon a combination of demographic and cognitive characteristics and practical inhibitors. However, there are calls for more social approaches to health behaviour which address the social context and an individual's own rationale for their actions. Therefore, we use discursive psychology to examine how Malawian interviewees describe and explain actions (not) taken to solve their fertility problem, and the interpersonal functions (e.g., blaming and justifying) thereby served. We focus in particular on respondents' descriptions of inaction since this is neglected in the literature, which emphasises instead intense health-seeking behaviour in response to infertility. We show how respondents attend to inaction as accountable, that is, as deviation from normative expectations and thus requiring explanation, and how they emphasise the reasonable and blameless character of their decisions and (in)action. The analysis highlights how explanations of health-seeking perform social functions rather than being neutral reflections of decision-making processes. Insofar as orientations to normative expectations inform people's relentless search for a cure, this may be problematic for health care seekers and providers alike.


Delineating Repetitive Behavior Profiles across the Lifespan in Fragile X Syndrome.
Restricted repetitive behaviors (RRBs) are a core area of impairment in autism spectrum disorder (ASD), but also affect several other neurodevelopmental disorders including fragile X syndrome (FXS). Current literature has begun to describe the RRB profile in FXS up through adolescence; however, little is known about the subtypes of RRBs in adolescents and adults. Further, literature on the RRB profile of females with FXS is limited. The present study examines the RRB profile across subtypes and specific items in both males and females with FXS while assessing for differences based on age, ASD diagnosis and the impact of IQ. Participants included 154 individuals with FXS (ages 2 to 50 years old). Results revealed a peak in RRB severity in FXS between 7-12 years for the majority of RRB subscales with the exception of Sensory-Motor behaviors peaking between 2 and 12 years before declining. Distinct RRB profiles in males and females with FXS emerged in addition to significant overlap among the item and subscale levels of RRBs across gender. Further, an added diagnosis of ASD significantly increased rates of RRBs across all subscale levels, but not necessarily across all items. Lastly, IQ did not solely account for the presence of RRBs in FXS, with Sensory-Motor behaviors being driven by comorbid ASD in males with FXS, and Restricted Interest behaviors being driven by comorbid ASD regardless of gender. These findings build on the current understanding of RRBs in FXS based on gender and comorbid ASD and lay important groundwork for the development of targeted behavioral and pharmacological treatments.


Mathematically optimized cryoprotectant equilibration procedures for cryopreservation of human oocytes.
Simple and effective cryopreservation of human oocytes would have an enormous impact on the financial and ethical constraints of human assisted reproduction. Recently, studies have demonstrated the potential for cryopreservation in an ice-free glassy state by equilibrating oocytes with high concentrations of cryoprotectants (CPAs) and rapidly cooling to liquid nitrogen temperatures. A major difficulty with this approach is that the high concentrations required for the avoidance of crystal formation (vitrification) also increase the risk of osmotic and toxic damage. We recently described a mathematical optimization approach for designing CPA equilibration procedures that avoid osmotic damage and minimize toxicity, and we presented optimized procedures for human oocytes involving continuous changes in solution composition. Here we adapt and refine our previous algorithm to predict piecewise-constant changes in extracellular solution concentrations in order to make the predicted procedures easier to implement. Importantly, we investigate the effects of using alternate equilibration endpoints on predicted protocol toxicity. Finally, we compare the resulting procedures to previously described experimental methods, as well as mathematically optimized procedures involving continuous changes in solution composition. For equilibration with CPA, our algorithm predicts an optimal first step consisting of exposure to a solution containing only water and CPA. This is predicted to cause the cells to initially shrink and then swell to the maximum cell volume limit. To reach the target intracellular CPA concentration, the cells are then induced to shrink to the minimum cell volume limit by exposure to a high CPA concentration. For post-thaw equilibration to remove CPA, the optimal procedures involve exposure to CPA-free solutions that are predicted to cause swelling to the maximum volume limit. The toxicity associated with these procedures is predicted to be much less than that of conventional procedures and comparable to that of the corresponding procedures with continuous changes in solution composition. The piecewise-constant procedures described in this study are experimentally facile and are predicted to be less toxic than conventional procedures for human oocyte cryopreservation. Moreover, the mathematical optimization approach described here will facilitate the design of cryopreservation procedures for other cell types.