Body composition and circulating high-molecular-weight adiponectin and IGF-I in infants born small for gestational age: breast- versus formula-feeding.
Prenatal growth restraint, if followed by postnatal overweight, confers risk for adult disease including diabetes. The mechanisms whereby neonatal nutrition may modulate such risk are poorly understood. We studied the effects of nutrition (breast-feeding [BRF] vs. formula-feeding [FOF]) on weight partitioning and endocrine state (as judged by high-molecular-weight [HMW] adiponectin and IGF-I) of infants born small for gestational age (SGA). Body composition (by absorptiometry), HMW adiponectin, and IGF-I were assessed at birth and 4 months in BRF infants born appropriate for gestational age (AGA; n = 72) and SGA infants receiving BRF (n = 46) or FOF (n = 56), the latter being randomized to receive a standard (FOF1) or protein-rich formula (FOF2). Compared with AGA-BRF infants, the catchup growth of SGA infants was confined to lean mass, independently of nutrition. Compared with AGA-BRF infants, SGA-BRF infants had normal HMW adiponectin and IGF-I levels at 4 months, whereas SGA-FOF infants had elevated levels of HMW adiponectin (particularly SGA-FOF1) and IGF-I (particularly SGA-FOF2). In conclusion, neonatal nutrition seems to influence endocrinology more readily than body composition of SGA infants. Follow-up will disclose whether the endocrine abnormalities in SGA-FOF infants can serve as early markers of an unfavorable metabolic course and whether they may contribute to design early interventions that prevent subsequent disease, including diabetes.


Red cell mass responses to daily erythropoietin and iron injections in the ovine fetus.
Anemic ovine fetuses supplemented with intra-amniotic iron undergo rapid expansion of red cell mass. The present study tested the hypothesis that nonanemic fetuses that were supplemented with daily intra-amniotic iron plus intravascular injections of erythropoietin would experience accelerated erythrocyte production. Nine late gestation ovine fetuses received 100 to 120 units of erythropoietin intravascularly plus 10 mg of iron intra-amniotically daily for 7 days (low erythropoietin dose group). Four additional fetuses received 1000 units of erythropoietin plus 10 mg iron daily for the same period (high erythropoietin dose group). Responses were compared with 9 nonsupplemented time-control fetuses. Statistical testing was by 3-factor repeated measures analysis of variance. Immediately after erythropoietin injection, plasma erythropoietin concentration was elevated approximately 25- and 250-fold in the low and high erythropoietin dose groups, respectively. Erythropoietin returned to basal levels by 24 hours after the injection. Plasma iron concentration increased in the low erythropoietin dose group but not in the control or high erythropoietin dose groups. Reticulocyte index increased in both erythropoietin supplemented groups but not in control fetuses. Hematocrit level increased above control by day 5 in the low erythropoietin dose group and by day 2 in the high erythropoietin dose group. Red cell mass increased significantly on supplement day 7 in the low erythropoietin dose group and on day 5 in the high erythropoietin dose group. Fetal blood gases and pH were unchanged with time in all 3 groups. Although daily combined erythropoietin and iron supplements in nonanemic ovine fetuses significantly increased circulating red cell mass in a dose-dependent manner, this increase was small relative to the rapid expansion of red cell mass previously observed after iron supplementation in fetuses with hemorrhage-induced anemia. We speculate that this difference in response may be due to a combination of rapid fetal clearance of erythropoietin plus a relative insensitivity to erythropoietin caused by the absence of other cytokines, which are elevated during fetal anemia.


Model-free causality analysis of cardiovascular variability detects the amelioration of autonomic control in Parkinson's disease patients undergoing mechanical stimulation.
We tested the hypothesis that causality analysis, applied to the spontaneous beat-to-beat variability of heart period (HP) and systolic arterial pressure (SAP), can identify the improvement of autonomic control linked to plantar mechanical stimulation in patients with Parkinson's disease (PD). A causality index, measuring the strength of the association from SAP to HP variability, and derived according to the Granger paradigm (i.e. SAP causes HP if the inclusion of SAP into the set of signals utilized to describe cardiovascular interactions improves the prediction of HP series), was calculated using both linear model-based (MB) and nonlinear model-free (MF) approaches. Univariate HP and SAP variability indices in time and frequency domains, and bivariate descriptors of the HP-SAP variability interactions were computed as well. We studied ten PD patients (age range: 57-78 years; Hoehn-Yahr scale: 2-3; six males, four females) without orthostatic hypotension or symptoms of orthostatic intolerance and 'on-time' according to their habitual pharmacological treatment. PD patients underwent recordings at rest in a supine position and during a head-up tilt before, and 24 h after, mechanical stimulation was applied to the plantar surface of both feet. The MF causality analysis indicated a greater involvement of baroreflex in regulating HP-SAP variability interactions after mechanical stimulation. Remarkably, MB causality and more traditional univariate or bivariate techniques could not detect changes in cardiovascular regulation after mechanical stimulation, thus stressing the importance of accounting for nonlinear dynamics in PD patients. Due to the higher statistical power of MF causality we suggest its exploitation to monitor the baroreflex control improvement in PD patients, and we encourage the clinical application of the Granger causality approach to evaluate the modification of the autonomic control in relation to the application of a pharmacological treatment, a rehabilitation procedure or external intervention.


Equiresolution catadioptric sensors.
A prominent characteristic of most catadioptric sensors is their lack of uniformity of resolution. We describe catadioptric sensors whose associated projections from the viewing sphere to the image plane have constant Jacobian determinants and so are equiresolution in the sense that any two equal solid angles are allocated the same number of pixels in the image plane. We show that in the orthographic case the catoptric component must be a surface of revolution of constant Gaussian curvature. We compare these equiresolution sensors in both the perspective and orthographic cases with other sensors that were proposed earlier for treating the uniformity-of-resolution problem.


Knowledge and attitude on sexually transmitted infections and contraceptive use among university students in Bhutan.
The unmet needs of contraception can lead to unintended pregnancy and transmission of sexually transmitted infections (STI). Therefore, this study aimed to evaluate the contraception use, knowledge, and attitude on STI among students under Royan University of Bhutan (RUB). This was a cross-sectional study using an online questionnaire. The questionnaire was developed in Google forms and the link was shared through the college WeChat groups. The questionnaire consisted of four parts on socio-demographic, sexual behaviour and contraceptive use, knowledge, and attitude on STIs. All the students under RUB were invited to participate voluntarily in this study. The socio-demography was presented in frequency and proportion. A total of 1,283 students participated in this survey and 55.0% (701) were females. Of this, 29.4% (377) were sexually active and 94.4% reported using modern contraception. Commonly used contraceptives were: condoms (83.8%, 316) and emergency contraceptives (20.6%, 78), respectively. The mean knowledge and attitude scores on STI were 9.94 (range 2-20) and 12 (range 2-14), respectively. Good knowledge and attitude on STI were reported in 53.2% (683) and 70.1% (899) of participants. Students reported average knowledge and a good attitude towards STI. Contraceptive use among university students was low. There is a need to strengthen health education on STIs in schools and universities. All forms of contraceptives especially condoms should be made easily available to sexually active people.


Triangle-programmed coulometric nanotitrations completed by continuous flow with potentiometric detection
Coulometric nanotitrations were realized in a microchannel system using a continuous-flow titration technique with a triangle current-time profile. Redox and acid-base titrations were carried out on Fe(II) and nitric acid samples, respectively, with the same nanotitrator device. A linear relation between the concentration and the coulometric current transferred to the solution was found. The advantages of this universally applicable nanotitrator are fast response, low sample volume, high sensitivity, and high reproducibility as well as the convenience of handling an automated analyzer of the flow-through type.


Snails in the desert: Species diversification of Theba (Gastropoda: Helicidae) along the Atlantic coast of NW Africa.
The spatial subdivision of species often plays a pivotal role in speciation. Across their entire range, species are rarely panmictic and crucial consequences of spatial subdivision are (1) random genetic drift including historical factors, (2) uniform selection, and (3) divergent selection. Each of these consequences may result in geographic variation and eventually reproductive isolation, but their relative importance in speciation is still unclear. In this study, we used a combination of genetic, morphological, and climatic data to obtain a comprehensive picture of differentiation among three closely related, parapatrically distributed taxa of the land snail genus Theba occurring along the Atlantic coasts of South Morocco and Western Sahara. We conducted Mantel and partial Mantel tests to relate phenotypic and genotypic variation of these species to geography and/or climate. As null hypothesis for an evolutionary scenario, we assumed nonadaptive speciation and expected a pattern of isolation by distance among taxa. Rejection of the null hypothesis would indicate isolation by environment due to adaptation. Generally, genetic drift plays an important role but is rarely considered as sole driver of speciation. It is the combination of drift and selection that predominantly drives speciation. This study, however, provides a potential example, in which nonadaptive speciation, that is, genetic drift, is apparently the main driver of shaping the diversity of Theba in NW Africa. Restriction of gene flow between populations caused by geographic isolation probably has played an important role. Climate oscillations during the Plio- and Pleistocene may have led to repeated ecological changes in NW Africa and disruptions of habitats promoting differentiation by geographic isolation. The inferred evolutionary scenario, however, did not fully explain the incongruence between the AFLP- and mtDNA-tree topologies. This incongruence might indicate past hybridization among the studied Theba forms.


Relationship between symptom over-reporting and pre- and post-combat trauma history in veterans evaluated for PTSD.
We examined the prevalence of pre- and post-combat traumatic events in the histories of 129 combat veterans referred to be evaluated for PTSD and examined the impact of these non-combat traumatic events on self-reported psychiatric symptoms. Participants were consecutive referrals to a Veterans Affairs outpatient post-traumatic stress disorder (PTSD) clinic who completed structured interviews, self-report measures (e.g., Minnesota Multiphasic Personality Inventory-2; MMPI-2), and a trauma history questionnaire as part of their routine clinical evaluations. Findings show that non-combat trauma was prevalent in this sample, with 65% (21% pre-combat) reporting physical assaults and 12% (11% pre-combat) reporting sexual assaults. Overall, history of sexual or physical assaults did not appear to be systematically related to reported symptom level. However, chi square analyses revealed a consistent over-representation of veterans who reported sexual trauma in the category suggestive of response exaggeration (i.e., MMPI-2 F-K validity index > or = 13). Finally, a hierarchical regression equation predicting F-K scores was computed, but accounted for only 15.9% of the variance in F-K. Presence of sexual assault history was the only predictor associated with a more pronounced response set suggestive of exaggeration or deception. These findings tentatively indicate that if history of sexual or physical assault has an impact on symptom reporting in combat veterans evaluated for PTSD, it is of modest magnitude.


A virtual infection model quantifies innate effector mechanisms and Candida albicans immune escape in human blood.
Candida albicans bloodstream infection is increasingly frequent and can result in disseminated candidiasis associated with high mortality rates. To analyze the innate immune response against C. albicans, fungal cells were added to human whole-blood samples. After inoculation, C. albicans started to filament and predominantly associate with neutrophils, whereas only a minority of fungal cells became attached to monocytes. While many parameters of host-pathogen interaction were accessible to direct experimental quantification in the whole-blood infection assay, others were not. To overcome these limitations, we generated a virtual infection model that allowed detailed and quantitative predictions on the dynamics of host-pathogen interaction. Experimental time-resolved data were simulated using a state-based modeling approach combined with the Monte Carlo method of simulated annealing to obtain quantitative predictions on a priori unknown transition rates and to identify the main axis of antifungal immunity. Results clearly demonstrated a predominant role of neutrophils, mediated by phagocytosis and intracellular killing as well as the release of antifungal effector molecules upon activation, resulting in extracellular fungicidal activity. Both mechanisms together account for almost [Formula: see text] of C. albicans killing, clearly proving that beside being present in larger numbers than other leukocytes, neutrophils functionally dominate the immune response against C. albicans in human blood. A fraction of C. albicans cells escaped phagocytosis and remained extracellular and viable for up to four hours. This immune escape was independent of filamentation and fungal activity and not linked to exhaustion or inactivation of innate immune cells. The occurrence of C. albicans cells being resistant against phagocytosis may account for the high proportion of dissemination in C. albicans bloodstream infection. Taken together, iterative experiment-model-experiment cycles allowed quantitative analyses of the interplay between host and pathogen in a complex environment like human blood.


Functional consequences of haem orientational disorder in sperm-whale and yellow-fin-tuna myoglobins.
Ligand-binding kinetics of native and reconstituted sperm-whale myoglobin were studied in relation to haem orientational disorder by rapid kinetic methods. In addition, native yellow-fin-tuna myoglobin with significant amount of haem disorder was also used. The O2 dissociation and association rates were found for the proteins with different degrees of haem disorder, and these results suggest that the isomers are characterized by almost identical kinetic parameters. Rates of CO recombination after photolysis were also identical for the two orientational isomers. The results clearly indicate that the rotation of the haem about the alpha-gamma meso axis has little or no effect on the ligand-binding properties of these myoglobins.


Relationship between the Korean Version of the Sniffin' Stick Test and the T&T Olfactometer in the Korean Population.
The Korean version of the Sniffin' stick (KVSS) test is widely used in Korea to evaluate olfactory function. However, its validity and reliability have not been studied well. In this study, the authors administered the KVSS and the T%T olfactometer test to evaluate olfactory function and to establish relationships between these two test measures. Two hundred and eleven patients participated in this prospective randomized study. One hundred and nine patients with no olfactory symptoms and 102 patients with decreased olfaction participated. All participants were underwent KVSS II and T&T olfactometer testing. The mean recognition threshold of the T&T olfactometer was -1.8±0.9 for patients with normal olfaction and 4.0±2.6 for patients with decreased olfaction. The mean Threshold-Discrimination-Identification score of the KVSS II was 30.0±3.8 for patients with normal olfaction and 15.9±7.1 for patients with decreased olfaction. Correlation coefficient between the two tests was significantly high (r(s)=-0.725, P<0.01). The KVSS and T&T olfactometry test are both reliable tests of olfactory function and their results are well correlated with each other.


Quantitative MRI measures of cartilage predict knee replacement: a case-control study from the Osteoarthritis Initiative.
Knee osteoarthritis commonly requires joint replacement, substantially reduces quality of life and increases healthcare utilisation and costs. This study aimed to identify whether quantitative measures of articular cartilage structure predict knee replacement, and to establish their utility as outcomes in clinical trials of disease-modifying therapy. A nested case-control study was performed in Osteoarthritis Initiative participants, a multicentre observational cohort of 4796 participants with or at risk of knee osteoarthritis. 127 knees were replaced between baseline and 4 years follow-up, and one control knee per case matched for baseline radiographic disease stage (Kellgren-Lawrence grade; KLG), gender and age. Quantitative cartilage measures were obtained from 3 T magnetic resonance images at the exam before knee replacement, and longitudinal change during the previous 12 months when available (n=110). Cartilage thickness loss in the central and total medial femorotibial compartment (primary and secondary predictor variables) was significantly greater in case than control knees (AUC=0.59/0.58). Differences in cartilage loss were greater at earlier than later radiographic disease stages (p<0.01 for interaction with KLG). Cartilage thickness loss in the central tibia was the most predictive longitudinal measure (AUC=0.64). Denuded bone areas in the medial femur were the most predictive and discriminatory cross-sectional measure between case and control knees (AUC=0.66). This study demonstrates the predictive value of quantitative, MRI-based measures of cartilage for the clinically relevant endpoint of knee replacement, providing support for their utility in clinical trials to evaluate the effectiveness of structure-modifying intervention.


Perquisites: the intrinsic form of pay.
Compensation packages, particularly for executives, are made up of five elements--salary, employee benefits, short-term incentives, long-term incentives, and perquisites. Of these, perquisites are probably the most misunderstood, according to author Bruce Ellig, vice-president of Corporate Compensation and Benefits for Pfizer Inc. The value of perquisites, or "perks," is their degree of exclusivity--that is, their worth as status symbols or "membership benefits" for those employees at or above a certain level in the company's hierarchy. And their value is enhanced when they (as some do) qualify for favorable tax treatment. True perks fall into six categories--time off with pay, executive services, nonperformance awards, health-care, survivor protection, and retirement coverage. Within these categories, perks take many forms. Time off with pay, for example, might include employment contracts, liberalized vacations, the privilege of working at home, disability benefits, sabbaticals, severance pay, or outplacement assistance. The use of perks varies among companies, but is more popular among small companies.


Optical fiber modulator derivates from hollow optical fiber with suspended core.
A fiber optic integrated modulation-depth-tunable modulator based on a type of hollow optical fiber with suspended core is proposed and investigated. We synthesized magnetic fluid containing superparamagnetic Fe(3)O(4) nanoparticles and encapsulated it in the hollow optical fiber as the cladding layer of the suspended core by fusing the hollow optical fiber with the multimode optical fibers. The light with a wavelength of 632.8 nm is coupled in and out of the modulating element by a tapering technique. Experimental results show that the light attenuation in the system can be greatly influenced by only 2.0×10(-2) μL of the magnetic fluid under different magnetic field strengths. The saturated modulation depth is 43% when the magnetic field strength is 489 Oe. The response time of the system is <120 ms. Significantly, this work presents information for the development of all-fiber modulators, including other integrated electro-optic devices, such as optical switch, optical fiber filter, and magnetic sensors utilizing the special structure of this hollow optical fiber with suspended core and superparamagnetic magnetic fluid.


Relationship of lower uterine segment cancer with Lynch syndrome: a novel case with an hMLH1 germline mutation.
Lynch syndrome is a genetic disease that often develops in patients with endometrial cancer and is caused by abnormal DNA mismatch repair (MMR) genes. In the United States, it was recently reported that the prevalence of Lynch syndrome with an hMSH2 mutation in patients with endometrial cancer in the lower uterine segment (LUS) is much greater than that in patients with endometrial cancer, although no such reports have been published in Asia. In this study, we examined the correlation between endometrial cancer in LUS and abnormalities in MMR genes. We examined 625 patients, who were diagnosed with endometrial cancer and underwent a hysterectomy. Nine patients (1.4%) had cancer based on pathological confirmation of a tumor in the lower part of the uterus and no cancer in the upper part. These cases were compared with 27 cases of sporadic endometrial (non-LUS) cancer. The age and BMI of the patients with LUS cancer were significantly lower than those of the patients with non-LUS cancer. No differences were observed in the pathological characteristics. The microsatellite instability (MSI)-positive rates were similar. Immunohistochemistry showed a decreased expression of hMLH1 and hMSH6 in patients with LUS cancer. In contrast with earlier reports from the United States, hMSH2 was expressed in all the cases. Of the 2 patients with LUS cancer who exhibited high MSI, 1 patient showed abnormal methylation of hMLH1, while the other patient was diagnosed with Lynch syndrome with a mutation in the hMLH1 gene. This is the second report on the relationship of LUS cancer and Lynch syndrome, and the first to describe an Asian patient with LUS cancer with Lynch syndrome induced by an hMLH1 mutation.


Electronically resonant third-order sum frequency generation spectroscopy using a nanosecond white-light supercontinuum.
Third-order sum frequency generation (TSFG) is one of the third-order nonlinear optical processes, and has the generation mechanism analogous to third harmonic generation (THG). By using a white-light supercontinuum, we can obtain broadband multiplex TSFG spectra. In the present study, we developed an electronically resonant TSFG spectrometer, and applied it to obtain TSFG spectra of hemoproteins. Analyzed TSFG ratio spectra clearly showed the resonant enhancement attributable to the electronic state of hemoproteins. This is a promising method for the imaging of electronic states of molecules inside living cells or tissues.


Radioactive waste management and environmental contamination issues at the Chernobyl site.
The destruction of the Unit 4 reactor at the Chernobyl Nuclear Power Plant resulted in the generation of radioactive contamination and radioactive waste at the site and in the surrounding area (referred to as the Exclusion Zone). In the course of remediation activities, large volumes of radioactive waste were generated and placed in temporary near-surface waste storage and disposal facilities. Trench and landfill type facilities were created from 1986-1987 in the Chernobyl Exclusion Zone at distances 0.5-15 km from the nuclear power plant site. This large number of facilities was established without proper design documentation, engineered barriers, or hydrogeological investigations and they do not meet contemporary waste-safety requirements. Immediately following the accident, a Shelter was constructed over the destroyed reactor; in addition to uncertainties in stability at the time of its construction, structural elements of the Shelter have degraded as a result of corrosion. The main potential hazard of the Shelter is a possible collapse of its top structures and release of radioactive dust into the environment. A New Safe Confinement (NSC) with a 100 y service life is planned to be built as a cover over the existing Shelter as a longer-term solution. The construction of the NSC will enable the dismantlement of the current Shelter, removal of highly radioactive, fuel-containing materials from Unit 4, and eventual decommissioning of the damaged reactor. More radioactive waste will be generated during NSC construction, possible Shelter dismantling, removal of fuel-containing materials, and decommissioning of Unit 4. The future development of the Exclusion Zone depends on the future strategy for converting Unit 4 into an ecologically safe system, i.e., the development of the NSC, the dismantlement of the current Shelter, removal of fuel-containing material, and eventual decommissioning of the accident site. To date, a broadly accepted strategy for radioactive waste management at the reactor site and in the Exclusion Zone, and especially for high level and long-lived waste, has not been developed.


In-vitro evaluation and comparison of the anti-microbial potency of commercially available oral hygiene products against Streptococcus mutans.
Dental caries is the destruction of enamel, dentin, or cementum of teeth due to bacterial activities, which if left untreated can cause considerable pain, discomfort, and treatment costs are very high. Of the oral bacteria, Streptococcus mutans is considered to be causative agent of dental caries in humans. This study aims at screening the antibacterial potential of available oral hygiene products against S. mutans, the primary etiological agent. A selective number of toothpaste and mouth rinse available in the nearby local market was subjected to the study. The experiments were designed in a way to determine which one of these products had the greatest anti-S.mutans activity. Antibiotic sensitivity tests against the pathogenic strains were also conducted. Pathogenic strains of S.mutans were isolated from clinical dental specimen and identified using MTCC standard strain No. 890. Of the 86.66% samples which showed positive for S.mutans growth, almost 96% conferred sensitivity to 0.08 mg/ml of penicillin. Among the toothpastes used, A showed the maximum inhibitory activity against S.mutans inhibiting its growth even at a very low concentration of 0.0156 g/ml. Similar results were seen in the case of the mouth rinses used for the study, which showed that N had the most effective activity against S.mutans even at 1:8 dilution. The oral hygiene products containing triclosan proved to be the most effective followed by those containing fluoride. Herbal products showed comparatively lesser activity in inhibiting the growth of S. mutans. Even though there are so many products available to curb the progression of dental caries in the population targeted, there is still an undoubted prevalence and incidence of caries among the general public. The need for a better alternative to help control dental caries is on the rise even today.


Artificial Intelligence: Innovation to Assist in the Identification of Sono-anatomy for Ultrasound-Guided Regional Anaesthesia.
Ultrasound-guided regional anaesthesia (UGRA) involves the targeted deposition of local anaesthesia to inhibit the function of peripheral nerves. Ultrasound allows the visualisation of nerves and the surrounding structures, to guide needle insertion to a perineural or fascial plane end point for injection. However, it is challenging to develop the necessary skills to acquire and interpret optimal ultrasound images. Sound anatomical knowledge is required and human image analysis is fallible, limited by heuristic behaviours and fatigue, while its subjectivity leads to varied interpretation even amongst experts. Therefore, to maximise the potential benefit of ultrasound guidance, innovation in sono-anatomical identification is required.Artificial intelligence (AI) is rapidly infiltrating many aspects of everyday life. Advances related to medicine have been slower, in part because of the regulatory approval process needing to thoroughly evaluate the risk-benefit ratio of new devices. One area of AI to show significant promise is computer vision (a branch of AI dealing with how computers interpret the visual world), which is particularly relevant to medical image interpretation. AI includes the subfields of machine learning and deep learning, techniques used to interpret or label images. Deep learning systems may hold potential to support ultrasound image interpretation in UGRA but must be trained and validated on data prior to clinical use.Review of the current UGRA literature compares the success and generalisability of deep learning and non-deep learning approaches to image segmentation and explains how computers are able to track structures such as nerves through image frames. We conclude this review with a case study from industry (ScanNav Anatomy Peripheral Nerve Block; Intelligent Ultrasound Limited). This includes a more detailed discussion of the AI approach involved in this system and reviews current evidence of the system performance.The authors discuss how this technology may be best used to assist anaesthetists and what effects this may have on the future of learning and practice of UGRA. Finally, we discuss possible avenues for AI within UGRA and the associated implications.


Rate of false conviction of criminal defendants who are sentenced to death.
The rate of erroneous conviction of innocent criminal defendants is often described as not merely unknown but unknowable. There is no systematic method to determine the accuracy of a criminal conviction; if there were, these errors would not occur in the first place. As a result, very few false convictions are ever discovered, and those that are discovered are not representative of the group as a whole. In the United States, however, a high proportion of false convictions that do come to light and produce exonerations are concentrated among the tiny minority of cases in which defendants are sentenced to death. This makes it possible to use data on death row exonerations to estimate the overall rate of false conviction among death sentences. The high rate of exoneration among death-sentenced defendants appears to be driven by the threat of execution, but most death-sentenced defendants are removed from death row and resentenced to life imprisonment, after which the likelihood of exoneration drops sharply. We use survival analysis to model this effect, and estimate that if all death-sentenced defendants remained under sentence of death indefinitely, at least 4.1% would be exonerated. We conclude that this is a conservative estimate of the proportion of false conviction among death sentences in the United States.


Deciphering Signaling Pathway Networks to Understand the Molecular Mechanisms of Metformin Action.
A drug exerts its effects typically through a signal transduction cascade, which is non-linear and involves intertwined networks of multiple signaling pathways. Construction of such a signaling pathway network (SPNetwork) can enable identification of novel drug targets and deep understanding of drug action. However, it is challenging to synopsize critical components of these interwoven pathways into one network. To tackle this issue, we developed a novel computational framework, the Drug-specific Signaling Pathway Network (DSPathNet). The DSPathNet amalgamates the prior drug knowledge and drug-induced gene expression via random walk algorithms. Using the drug metformin, we illustrated this framework and obtained one metformin-specific SPNetwork containing 477 nodes and 1,366 edges. To evaluate this network, we performed the gene set enrichment analysis using the disease genes of type 2 diabetes (T2D) and cancer, one T2D genome-wide association study (GWAS) dataset, three cancer GWAS datasets, and one GWAS dataset of cancer patients with T2D on metformin. The results showed that the metformin network was significantly enriched with disease genes for both T2D and cancer, and that the network also included genes that may be associated with metformin-associated cancer survival. Furthermore, from the metformin SPNetwork and common genes to T2D and cancer, we generated a subnetwork to highlight the molecule crosstalk between T2D and cancer. The follow-up network analyses and literature mining revealed that seven genes (CDKN1A, ESR1, MAX, MYC, PPARGC1A, SP1, and STK11) and one novel MYC-centered pathway with CDKN1A, SP1, and STK11 might play important roles in metformin's antidiabetic and anticancer effects. Some results are supported by previous studies. In summary, our study 1) develops a novel framework to construct drug-specific signal transduction networks; 2) provides insights into the molecular mode of metformin; 3) serves a model for exploring signaling pathways to facilitate understanding of drug action, disease pathogenesis, and identification of drug targets.


Clinical Outcome Event Adjudication in a 10-Year Prospective Study of Nucleos(t)ide Analogue Therapy for Chronic Hepatitis B.
Background and Aims: In the REALM (Randomized, Observational Study of Entecavir to Assess Long-Term Outcomes Associated with Nucleoside/Nucleotide Monotherapy for Patients with Chronic HBV Infection) study, 12,378 patients with chronic hepatitis B virus (HBV) infection received up to 10 years of randomized therapy with entecavir or another HBV nucleos(t)ide analogue. Monitored clinical outcome events (COEs) included malignant neoplasms, HBV disease progression events, and deaths. An external event adjudication committee (EAC) was convened to provide real-time review of reported COEs to optimize data quality, and minimize potential adverse effects of the large cohort, interdisciplinary outcome assessments, geographic scope, and long duration. Methods: The EAC comprised an international group of hepatologists and oncologists with expertise in diagnosis of targeted COEs. The EAC reviewed and adjudicated COEs according to prospectively defined diagnostic criteria captured in the EAC charter. Operational processes, including data collection and query procedures, were implemented to optimize efficiency of data recovery to maximize capture of adjudicated COEs, the primary study outcome measure. Results: A total of 1724 COEs were reported and 1465 of these events were adjudicated by the EAC as reported by the investigators (85.0% overall concordance). Concordance by COE type varied: deaths, 99.6%; hepatocellular carcinoma (HCC), 83.3%; non-HCC malignancies, 88.0%; non-HCC HBV disease progression, 68.2%. Reasons for lack of concordance were most commonly lack of adequate supporting data to support an adjudicated diagnosis or evidence that the event pre-dated the study. Conclusions: The REALM EAC performed a critical role in ensuring data quality and consistency; EAC performance was facilitated by well-defined diagnostic criteria, effective data capture, and efficient operational processes. Trial registration: ClinicalTrials.gov NCT00388674.


Is Pupil Diameter Influenced by Refractive Error?
To investigate the relationship between pupil diameter and refractive error and how refractive correction, target luminance, and accommodation modulate this relationship. Sixty emmetropic, myopic, and hyperopic subjects (age range, 18 to 35 years) viewed an illuminated target (luminance: 10, 100, 200, 400, 1000, 2000, and 4100 cd/m) within a Badal optical system, at 0 diopters (D) and -3 D vergence, with and without refractive correction. Refractive error was corrected using daily disposable contact lenses. Pupil diameter and accommodation were recorded continuously using a commercially available photorefractor. No significant difference in pupil diameter was found between the refractive groups at 0 D or -3 D target vergence, in the corrected or uncorrected conditions. As expected, pupil diameter decreased with increasing luminance. Target vergence had no significant influence on pupil diameter. In the corrected condition, at 0 D target vergence, the accommodation response was similar in all refractive groups. At -3 D target vergence, the emmetropic and myopic groups accommodated significantly more than the hyperopic group at all luminance levels. There was no correlation between accommodation response and pupil diameter or refractive error in any refractive group. In the uncorrected condition, the accommodation response was significantly greater in the hyperopic group than in the myopic group at all luminance levels, particularly for near viewing. In the hyperopic group, the accommodation response was significantly correlated with refractive error but not pupil diameter. In the myopic group, accommodation response level was not correlated with refractive error or pupil diameter. Refractive error has no influence on pupil diameter, irrespective of refractive correction or accommodative demand. This suggests that the pupil is controlled by the pupillary light reflex and is not driven by retinal blur.


Beliefs about the harms of long-term use of nicotine replacement therapy: perceptions of smokers in England.
Previous research has shown that a substantial proportion of smokers believe that nicotine causes serious diseases such as cancer, possibly deterring the use of nicotine replacement therapy (NRT) for smoking cessation or smoking reduction. This study examined beliefs about the harms specifically from long-term use of NRT and associations between these and its use for smoking cessation and smoking reduction. Data were collected from 1657 smokers and recent ex-smokers involved in the Smoking Toolkit Study, a series of monthly household surveys of English adults aged 16 and over. Participants were asked if they thought the use of NRT for a year or more was harmful, and if so, to volunteer what they believed the harms to be. They were also asked if they were using NRT for smoking reduction and/or if they had used NRT in the past year during a quit attempt. Six percent and 25% of smokers respectively, believed that the long term use of NRT was very or quit harmful to health; and a further 29% reported that they 'didn't know'. The most commonly reported harms were addiction and lung cancer. There was no association between these beliefs and use of NRT for smoking reduction or smoking cessation. A significant minority of smokers in England believe that the use of nicotine replacement therapy for a year or more is harmful. However, belief that long-term nicotine replacement therapy use can cause health harm does not appear to act as a deterrent to using it in a quit attempt or for smoking reduction.


Inequalities reinforced through online and distance education in the age of COVID-19: The case of higher education in Nepal.
Coronavirus disease (COVID-19) has triggered serious disruption in economic, social and cultural dynamics around the globe. Higher education has also suffered undeniable challenges as a result of the pandemic, with thousands of university students all over the world experiencing displacement, disconnect and disengagement from formal learning. In the Global South, online and distance education programmes tend to be concentrated in urban centres. In Nepal, students from rural areas, low socio-economic and gendered spaces, and those with low proficiencies in English and technological skills are experiencing inequalities in access to and participation in online and distance education. This article outlines how universities' shift to online teaching and learning modes due to the COVID-19 pandemic has reinforced social inequalities in Nepal. For the study presented here, the author collected data through netnographic research methods. These included online interviews with university executives, online focus group discussions (FGDs) with university teachers and students, observation of and participation in online classes and policy conferences and reviews of online documents. The article analyses three overriding mechanisms which are reinforcing social inequalities in higher education: (1) universities' policy trajectories in shifting teaching/learning from face-to-face to online mode; (2) infrastructural limitations challenging effective implementation of online teaching/learning; and (3) a lack of strong pedagogic support for students from disadvantaged and marginalised spaces, including those with low proficiencies in English and technological skills. The author presents a number of tangible strategies for universities to implement in order to mitigate social inequalities. He recommends the adoption of policies and practices that optimise the inclusive use of online and distance education programmes for best effect, both now and in the post-pandemic era.


Impact of Excessive Weight Gain on Cardiovascular Outcomes in Type 1 Diabetes: Results From the Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications (DCCT/EDIC) Study.
Intensive treatment (INT) of type 1 diabetes reduces the incidence of cardiovascular disease (CVD) events compared with conventional treatment (CONV), but it also results in more weight gain. Our objective was to examine whether excessive weight gain from INT of type 1 diabetes is independently associated with subsequent CVD events. Quartiles (Q) of weight gain in 1,213 participants aged 18 years and older at enrollment in the Diabetes Control and Complications Trial (DCCT) were determined within randomized treatment groups (INT vs. CONV) using change in BMI from baseline to the closeout DCCT visits. Effects of this weight gain on CVD risk factors and outcomes during an additional 20 years of observational follow-up were then determined. The Q4 INT group experienced greater proportional weight gain (median change in BMI, 6.08 kg/m2), increases in CVD risk factors, and need for medications for hypertension and lipids compared with the Q1-3 INT and comparable CONV groups. Over a mean of 26 years of follow-up, the numbers of major and total CVD events were not statistically different in Q4 compared with Q1-3 of either the INT or CONV group. By year 14, however, the incident CVD event curve became significantly higher in the Q4 INT group than in the Q1-3 INT groups (P = 0.024) and was similar to that for the CONV group. For the first 13 years after DCCT, INT for type 1 diabetes reduced macrovascular events compared with CONV, even when excessive weight gain occurred. After this, total CVD events significantly increased in the Q4 INT group, becoming equivalent to those in the CONV group. Longer follow-up is needed to determine whether this trend continues and results in more major CVD events.


Formation of Azaspiracids-3, -4, -6, and -9 via decarboxylation of carboxyazaspiracid metabolites from shellfish.
The azaspiracid (AZA) class of phycotoxins has been responsible for extended closures of shellfisheries in various locations around Europe, where levels of AZA1-3 are regulated in shellfish. Since their discovery in 1995, AZAs have been the focus of much research, resulting in the discovery of numerous analogues. During studies of procedures for processing of AZA-contaminated mussels ( Mytilus edulis ), an unusual phenomenon was observed involving AZA3. In uncooked tissues, AZA3 levels would increase significantly when heated for short periods of time in the absence of water loss. A similar increase in AZA3 concentrations occurred during storage of shellfish tissue reference materials for several months at temperatures as low as 4 degrees C. Concentrations of AZA1 and AZA2 did not change during these experiments. Several possible explanations were investigated, including an AZA3-specific matrix effect upon heating of tissues, release of AZA3 from the matrix, and formation of AZA3 from a precursor. Preliminary experiments indicated that toxin conversion was responsible, and more detailed studies focused on this possibility. LC-MS analysis of heating trials, deuterium labeling experiments, and kinetic studies demonstrated that a carboxylated AZA analogue, AZA17, undergoes rapid decarboxylation when heated to produce AZA3. Heat-induced decarboxylation of AZA19, AZA21, and AZA23 to form AZA6, AZA4, and AZA9, respectively, was also demonstrated. This finding is of great significance in terms of procedures used in the processing of shellfish for regulatory analysis, and it exemplifies the role that chemical analysis can play in understanding the contribution of metabolic processes to the toxin profiles observed in shellfish samples.


Depleted uranium induces sex- and tissue-specific methylation patterns in adult zebrafish.
We examined the effects of chronic exposure to different concentrations (2 and 20 μg L(-)(1)) of environmentally relevant waterborne depleted uranium (DU) on the DNA methylation patterns both at HpaII restriction sites (5'-CCGG-3') and across the whole genome in the zebrafish brain, gonads, and eyes. We first identified sex-dependent differences in the methylation level of HpaII sites after exposure. In males, these effects were present as early as 7 days after exposure to 20 μg L(-)(1) DU, and were even more pronounced in the brain, gonads, and eyes after 24 days. However, in females, hypomethylation was only observed in the gonads after exposure to 20 μg L(-)(1) DU for 24 days. Sex-specific effects of DU were also apparent at the whole-genome level, because in males, exposure to 20 μg L(-)(1) DU for 24 days resulted in cytosine hypermethylation in the brain and eyes and hypomethylation in the gonads. In contrast, in females, hypermethylation was observed in the brain after exposure to both concentrations of DU for 7 days. Based on our current knowledge of uranium toxicity, several hypotheses are proposed to explain these findings, including the involvement of oxidative stress, alteration of demethylation enzymes and the calcium signaling pathway. This study reports, for the first time, the sex- and tissue-specific epigenetic changes that occur in a nonhuman organism after exposure to environmentally relevant concentrations of uranium, which could induce transgenerational epigenetic effects.


Imaging with two spiral diffracting elements intermediated by a pinhole.
A pseudoscopic (inverted depth) image made with spiral diffracting elements intermediated by a pinhole is explained by its symmetry properties. The whole process is made under common white light illumination and allows the projection of images. The analysis of this projection demonstrates that the images of two objects pointing away longitudinally have the main features of standard pseudoscopic image points. An orthoscopic (normal depth) image has also been obtained with the breaking of the symmetry conditions.


Are women ready for the HPV vaccine?
An HPV (human papillomavirus) vaccine will be available soon. Because vaccination for both boys and girls will be critical to reduce the incidence of this disease, we evaluated the willingness of women to accept the HPV vaccine and to consent to having their children vaccinated. A 20-item questionnaire was distributed to women who had children between the ages of 8 and 14 years, the likely age range of the children who will be targeted to receive the vaccine. 200 women completed surveys. The median age for the respondents was 34.4 years (range, 23-53 years). 77% of the women surveyed would be willing to accept an HPV vaccine for themselves. 67% who had a daughter (n = 156) and 66% who had a son (n = 137) would consent to have their child vaccinated. Those not accepting the vaccine cited the risk of unknown adverse effects or their current lack of sexual activity. Women who would not consent to have their children vaccinated cited the risk of unknown adverse effects and their belief that their children are not sexually active. Male children not directly benefiting from the vaccine were also a reason for not considering the vaccine for sons. A history of an abnormal Papanicolaou smear was not associated with willingness to accept the vaccine. This pilot study demonstrated a willingness of women to accept the HPV vaccine for themselves and their children. Gynecologists and gynecologic oncologists must focus not only on the diagnosis and treatment of cervical cancer but also on its prevention.


Visual development and the acquisition of motion velocity sensitivities.
We consider the hypothesis that systems learning aspects of visual perception may benefit from the use of suitably designed developmental progressions during training. Four models were trained to estimate motion velocities in sequences of visual images. Three of the models were developmental models in the sense that the nature of their visual input changed during the course of training. These models received a relatively impoverished visual input early in training, and the quality of this input improved as training progressed. One model used a coarse-to-multiscale developmental progression (it received coarse-scale motion features early in training and finer-scale features were added to its input as training progressed), another model used a fine-to-multiscale progression, and the third model used a random progression. The final model was nondevelopmental in the sense that the nature of its input remained the same throughout the training period. The simulation results show that the coarse-to-multiscale model performed best. Hypotheses are offered to account for this model's superior performance, and simulation results evaluating these hypotheses are reported. We conclude that suitably designed developmental sequences can be useful to systems learning to estimate motion velocities. The idea that visual development can aid visual learning is a viable hypothesis in need of further study.


A cohort study on the risk of acute liver injury among users of ketoconazole and other antifungal drugs.
The aim of this cohort study was to estimate the risk of clinical acute liver injury among users of oral antifungals identified in the general population of the General Practice Research Database in UK. The cohort included 69 830 patients, 20-79 years old, free of liver and systemic disease, who had received at least one prescription for either oral fluconazole, griseofulvin, itraconazole, ketoconazole, or terbinafine between 1991 and 1996. Sixteen cases of acute liver injury were identified and validated. Ten cases occurred during nonuse of oral antifungals with a background rate of 0.6 per 100,000 person-months (95% confidence interval 0.3,1.1). Five cases occurred during current use of oral antifungals. Two were using ketoconazole, another two itraconazole, and one terbinafine. Incidence rates of acute liver injury were 134.1 per 100 000 person-months (36.8,488.0) for ketoconazole, 10.4 (2.9-38.1) for itraconazole, and 2.5 (0.4,13. 9) for terbinafine. The remaining case was associated with past use of fluconazole. Ketoconazole was the antifungal associated with the highest relative risk, 228.0 (95% confidence interval 33.9,933.0), when compared with the risk among nonusers, followed by itraconazole and terbinafine with relative risks of 17.7 (2.6,72.6) and 4.2 (0.2, 24.9), respectively. Ketoconazole and itraconazole were the two oral antifungal associated with a marked increase of clinical acute liver injury. The risk associated with ketoconazole should be taken into account when prescribing it as initial treatment for uncomplicated fungal infections.


Engineering of N. benthamiana L. plants for production of N-acetylgalactosamine-glycosylated proteins--towards development of a plant-based platform for production of protein therapeutics with mucin type O-glycosylation.
Mucin type O-glycosylation is one of the most common types of post-translational modifications that impacts stability and biological functions of many mammalian proteins. A large family of UDP-GalNAc polypeptide:N-acetyl-α-galactosaminyltransferases (GalNAc-Ts) catalyzes the first step of mucin type O-glycosylation by transferring GalNAc to serine and/or threonine residues of acceptor polypeptides. Plants do not have the enzyme machinery to perform this process, thus restricting their use as bioreactors for production of recombinant therapeutic proteins. The present study demonstrates that an isoform of the human GalNAc-Ts family, GalNAc-T2, retains its localization and functionality upon expression in N. benthamiana L. plants. The recombinant enzyme resides in the Golgi as evidenced by the fluorescence distribution pattern of the GalNAc-T2:GFP fusion and alteration of the fluorescence signature upon treatment with Brefeldin A. A GalNAc-T2-specific acceptor peptide, the 113-136 aa fragment of chorionic gonadotropin β-subunit, is glycosylated in vitro by the plant-produced enzyme at the "native" GalNAc attachment sites, Ser-121 and Ser-127. Ectopic expression of GalNAc-T2 is sufficient to "arm" tobacco cells with the ability to perform GalNAc-glycosylation, as evidenced by the attachment of GalNAc to Thr-119 of the endogenous enzyme endochitinase. However, glycosylation of highly expressed recombinant glycoproteins, like magnICON-expressed E. coli enterotoxin B subunit:H. sapiens mucin 1 tandem repeat-derived peptide fusion protein (LTBMUC1), is limited by the low endogenous UDP-GalNAc substrate pool and the insufficient translocation of UDP-GalNAc to the Golgi lumen. Further genetic engineering of the GalNAc-T2 plants by co-expressing Y. enterocolitica UDP-GlcNAc 4-epimerase gene and C. elegans UDP-GlcNAc/UDP-GalNAc transporter gene overcomes these limitations as indicated by the expression of the model LTBMUC1 protein exclusively as a glycoform. Plant bioreactors can be engineered that are capable of producing Tn antigen-containing recombinant therapeutics.


Controlled reduction for size selective synthesis of thiolate-protected gold nanoclusters Aun(n = 20, 24, 39, 40).
This work presents a controlled reduction method for the selective synthesis of different sized gold nanoclusters protected by thiolate (SR = SC2H4Ph). Starting with Au(III) salt, all the syntheses of Aun(SR)m nanoclusters with (n, m) = (20, 16), (24, 20), (39, 29), and (40, 30) necessitate experimental conditions of slow stirring and slow reduction of Au(I) intermediate species. By controlling the reaction kinetics for the reduction of Au(I) into clusters by NaBH4, different sized gold nanoclusters are selectively obtained. Two factors are identified to be important for the selective growth of Au20, Au24, and Au39/40 nanoclusters, including the stirring speed of the Au(I) solution and the NaBH4 addition speed during the step of Au(I) reduction to clusters. When comparing with the synthesis of Au25(SC2H4Ph)18 nanoclusters, we further identified that the reduction degree of Au(I) by NaBH4 also plays an important role in controlling cluster size. Overall, our results demonstrate the feasibility of attaining new sizes of gold nanoclusters via a controlled reduction route.


Analysis of the Cost-Effectiveness of Liquid Biopsy to Determine Treatment Change in Patients with Her2-Positive Advanced Breast Cancer in Colombia.
Breast cancer is highly prevalent worldwide and leads to high health-care costs. HER2-positive subtype represents 30% of all breast cancers and is associated with a poor prognosis. Patients treated with anti-HER2 therapies frequently develop resistance and require pharmacological treatment change. Liquid biopsy is a minimally invasive and an easily accessible technique, with high sensitivity and specificity, to detect molecular treatment resistance even before the onset of clinical manifestations and can thus be used to reduce unnecessary anti-HER2 treatment costs. To evaluate the cost-effectiveness of using liquid biopsy (ctDNA detection) to determine treatment change in women with HER2-positive advanced breast cancer in Colombia. We performed an economic evaluation using decision tree modeling and deterministic analyses based on literature search for first and second lines of treatment (trastuzumab, pertuzumab, docetaxel, and TDM1); resistance; outcomes; and sensitivity and specificity of tests detecting molecular resistance. The effectiveness was measured using quality-adjusted life year (QALY) score, and costs were obtained from databases with national validity, suppliers, the Colombian Drug Price Information System (SISMED), and local studies. The use of liquid biopsy (ctDNA detection) with conventional treatment was more expensive and less effective than conventional treatment without liquid biopsy (US $177,985.35 and 0.533889206 QALY, respectively). The incremental cost with liquid biopsy was US $7,333.17 and the incremental effectiveness was 0.00042256 QALY relative to the conventional method. Including liquid biopsy in the treatment of HER2-positive advanced breast cancer was considered currently inapplicable in Colombia because it was not cost effective. Our results open a window of opportunity to improve the development and implementation of ctDNA testing in Colombia, potentially reducing current costs. More evidence is required on the utility of this test, depending on the financial capacity of Colombia and other countries.


Pharmacokinetics of Rifampin, Isoniazid, Pyrazinamide, and Ethambutol in Infants Dosed According to Revised WHO-Recommended Treatment Guidelines.
There are limited pharmacokinetic data for use of the first-line antituberculosis drugs during infancy (<12 months of age), when drug disposition may differ. Intensive pharmacokinetic sampling was performed in infants routinely receiving antituberculosis treatment, including rifampin, isoniazid, pyrazinamide, and ethambutol, using World Health Organization-recommended doses. Regulatory-approved single-drug formulations, including two rifampin suspensions, were used on the sampling day. Assays were conducted using liquid chromatography-mass spectrometry; pharmacokinetic parameters were generated using noncompartmental analysis. Thirty-nine infants were studied; 14 (36%) had culture-confirmed tuberculosis. Fifteen (38%) were premature (<37 weeks gestation); 5 (13%) were HIV infected. The mean corrected age and weight were 6.6 months and 6.45 kg, respectively. The mean maximum plasma concentrations (Cmax) for rifampin, isoniazid, pyrazinamide, and ethambutol were 2.9, 7.9, 41.9, and 1.3 μg/ml, respectively (current recommended adult target concentrations: 8 to 24, 3 to 6, 20 to 50, and 2 to 6 μg/ml, respectively), and the mean areas under the concentration-time curves from 0 to 8 h (AUC0-8) were 12.1, 24.7, 239.4, and 5.1 μg · h/ml, respectively. After adjusting for age and weight, rifampin exposures for the two formulations used differed inCmax(geometric mean ratio [GMR],2.55; 95% confidence interval [CI], 1.47 to 4.41;P= 0.001) and AUC0-8(GMR, 2.52; 95% CI, 1.34 to 4.73;P= 0.005). HIV status was associated with lower pyrazinamideCmax(GMR, 0.85; 95% CI, 0.75 to 0.96;P= 0.013) and AUC0-8(GMR, 0.79; 95% CI, 0.69 to 0.90;P< 0.001) values. No other important differences were observed due to age, weight, prematurity, ethnicity, or gender. In summary, isoniazid and pyrazinamide concentrations in infants compared well with proposed adult target concentrations; ethambutol concentrations were lower but similar to previously reported pediatric studies. The low rifampin exposures require further investigation. (This study has been registered at ClinicalTrials.gov under registration no. NCT01637558.).


Temazepam triggers the release of vasopressin into the rat hypothalamic paraventricular nucleus: novel insight into benzodiazepine action on hypothalamic-pituitary-adrenocortical system activity during stress.
We investigated the influence of a representative classical benzodiazepine on the regulation of the hypothalamic-pituitary-adrenocortical (HPA) axis activity both under basal conditions and stress. Adult male Wistar rats were intravenously administered with temazepam (0.5, 1, and 3 mg/kg body weight) and plasma concentrations of corticotropin (ACTH) and vasopressin (AVP) were measured in blood samples collected via chronically implanted jugular venous catheters. Simultaneously, the release of AVP within the hypothalamic paraventricular nucleus (PVN) was monitored via microdialysis. Plasma AVP levels remained unaffected by the different treatment conditions. Temazepam blunted the stressor exposure-induced secretion of ACTH in a dose-dependent manner. Concurrently, and also in a dose-dependent manner temazepam enhanced the intra-PVN release of AVP, known to originate from magnocellular neurons of the hypothalamic neurohypophyseal system. Furthermore, temazepam did not affect the in vitro secretion of ACTH from the adenohypophyseal cells. Taken together, the results of this study suggest that temazepam modulates the central nervous regulation of the HPA axis by altering intra-PVN AVP release. An increasingly released AVP of magnocellular origin seems to provide a negative tonus on ACTH secretion most probably via inhibiting the release of ACTH secretagogues from the median eminence into hypophyseal portal blood.


Validity of coital diaries in a feasibility study for the Microbicides Development Programme trial among women at high risk of HIV/AIDS in Mwanza, Tanzania.
To compare coital diaries and face-to-face interviews (FFIs) in measuring sexual behaviour among women at high risk of HIV. To assess the effect of differing levels of support from researchers on reporting in coital diaries and FFIs. Three groups of 50 women were randomly selected from a cohort of food and recreational facility workers participating in a microbicide trial feasibility study and received differing levels of researcher support. Minimum support involved delivering and collecting coital diaries weekly; medium support included a weekly FFI and discussion of concerns; intensive support also included an unscheduled mid-week visit when diaries were checked and concerns addressed. All respondents participated in an exit FFI, including questions on sexual behaviour over the four-week study period and study acceptability. Sexual behaviours were generally reported more frequently in coital diaries than weekly or exit interviews. Vaginal and anal sex, male and female condom use, vaginal cleaning and lubrication, sex during menstruation and sex with irregular and regular partners were reported more frequently in coital diaries than exit interviews. In coital diaries, level of support was associated with reporting of vaginal sex and cleaning. In exit interviews, support level was associated with reporting of vaginal sex, vaginal cleaning and sex with regular, irregular and commercial partners. Women with minimum support reported least satisfaction with the research process. Women with intensive support were most likely to report that they informed someone about their study participation and that they completed diaries daily. Compared with FFIs, coital diaries resulted in higher reporting of socially stigmatised activities, and sexual behaviour reporting varied less by level of support. More researcher support enhanced study acceptability.


Primate cognition: from 'what now?' to 'what if?'.
The 'social brain' hypothesis has had a major impact on the study of comparative cognition. However, despite a strong sense, gained from both experimental and observational work, that monkeys and apes differ from each other, we are still no closer to understanding exactly how they differ. We hypothesize that the dispersed social systems characteristic of ape societies explains why monkeys and apes should differ cognitively. The increased cognitive control and analogical reasoning ability needed to cope with life in dispersed societies also suggests a possible route for human cognitive evolution. This hypothesis is supported by behavioural and neurobiological data, but we need more of both if we are to fully understand how our primate cousins see the world.


Investigation of solute-solvent interactions in phenol compounds: accurate ab initio calculations of solvent effects on 1H NMR chemical shifts.
Accurate (1)H chemical shifts of the -OH groups of polyphenol compounds can be calculated, compared to experimental values, using a combination of DFT, polarizable continuum model (PCM) and discrete solute-solvent hydrogen bond interactions. The study focuses on three molecular solutes: phenol, 4-methylcatechol and the natural product genkwanin in DMSO, acetone, acetonitrile, and chloroform. Excellent linear correlation between experimental and computed chemical shifts (with the GIAO method at the DFT/B3LYP/6-311++G(2d,p) level) was obtained with minimization of the solvation complexes at the DFT/B3LYP/6-31+G(d) and DFT/B3LYP/6-311++G(d,p) level of theory with a correlation coefficient of 0.991. The use of the DFT/B3LYP/6-31+G(d) level of theory for minimization could provide an excellent means for the accurate prediction of the experimental OH chemical shift range of over 8 ppm due to: (i) strong intramolecular and solute-solvent intermolecular hydrogen bonds, (ii) flip-flop intramolecular hydrogen bonds, and (iii) conformational effects of substituents of genkwanin. The combined use of ab initio calculations and experimental (1)H chemical shifts of phenol -OH groups provides a method of primary interest in order to obtain detailed structural, conformation and electronic description of solute-solvent interactions at a molecular level.


The hippocampus of the eastern rock sengi: cytoarchitecture, markers of neuronal function, principal cell numbers, and adult neurogenesis.
The brains of sengis (elephant shrews, order Macroscelidae) have long been known to contain a hippocampus that in terms of allometric progression indices is larger than that of most primates and equal in size to that of humans. In this report, we provide descriptions of hippocampal cytoarchitecture in the eastern rock sengi (Elephantulus myurus), of the distributions of hippocampal calretinin, calbindin, parvalbumin, and somatostatin, of principal neuron numbers, and of cell numbers related to proliferation and neuronal differentiation in adult hippocampal neurogenesis. Sengi hippocampal cytoarchitecture is an amalgamation of characters that are found in CA1 of, e.g., guinea pig and rabbits and in CA3 and dentate gyrus of primates. Correspondence analysis of total cell numbers and quantitative relations between principal cell populations relate this sengi to macaque monkeys and domestic pigs, and distinguish the sengi from distinct patterns of relations found in humans, dogs, and murine rodents. Calretinin and calbindin are present in some cell populations that also express these proteins in other species, e.g., interneurons at the stratum oriens/alveus border or temporal hilar mossy cells, but neurons expressing these markers are often scarce or absent in other layers. The distributions of parvalbumin and somatostatin resemble those in other species. Normalized numbers of PCNA+ proliferating cells and doublecortin-positive (DCX+) differentiating cells of neuronal lineage fall within the overall ranges of murid rodents, but differed from three murid species captured in the same habitat in that fewer DCX+ cells relative to PCNA+ were observed. The large and well-differentiated sengi hippocampus is not accompanied by correspondingly sized cortical and subcortical limbic areas that are the main hippocampal sources of afferents and targets of efferents. This points to intrinsic hippocampal information processing as the selective advantage of the large sengi hippocampus.


Electrophysiological and urodynamic studies to monitor surgical outcome in children with tethered spinal cords.
Children with myelodysplasia often develop tethering of their spinal cord within 2-3 years after correction of their initial birth defect, leading to either neurological or urological dysfunction. Surveillance of these children includes electrophysiologic/urodynamic (EMG/UDS) studies to monitor improvement or deterioration of their neurological function. We reviewed 39 patients who underwent surgical untethering from 1979 to 1994 to determine whether postoperative EMG/UDS studies were useful in assessing outcome after surgery. All patients had a complete neurologic examination and an EMG/UDS prior to and 3 months after surgery. Twenty patients (51%) showed improvement in their EMG/UDS study after untethering, 15 (39%) demonstrated no significant change, and 4 (10%) showed deterioration. Improvement consisted of either return of previously absent motor units, return of sacral reflexes, or improved bladder dynamics. In a further 3 patients, subsequent deterioration on serial postoperative EMG/UDS testing was the first indication of recurrent tethering. We conclude that serial EMG/UDS evaluation in patients at risk for tethering of the spinal cord may serve as a useful adjunct to close clinical observation.


Diagnosis and treatment of Cushing's disease in children.
Cushing's disease (CD) is rare in the paediatric age range, but presents a diagnostic and therapeutic challenge. Most paediatric endocrinologists have limited experience managing children or adolescents with CD and thus benefit from close consultation with adult colleagues. A diagnostic protocol for investigation is required which broadly follows the model for adult patients. Treatment strategies for CD are described and critically appraised. The management of paediatric CD patients after cure also presents challenges for optimizing growth, bone health, reproduction and body composition from childhood into and during adult life.


Studies on the mechanisms by which transforming growth factor-beta (TGF-beta) protects against allergic encephalomyelitis. Antagonism between TGF-beta and tumor necrosis factor.
Experimental allergic encephalomyelitis (EAE) is an autoimmune disease in which peripheral lymphoid cells are activated by immunization with myelin proteins and become effector cells that traverse the central nervous system (CNS) capillaries and initiate inflammatory demyelinating lesions. The administration of transforming growth factor-beta (TGF-beta) has been shown previously to decrease the incidence and severity of EAE. In our studies we have determined: 1) the effects of TGF-beta injected at different intervals after the EAE-inducing immunization; 2) the effect of TGF-beta on the development of sensitized T cells, as assayed by the proliferative responses of T cells from lymph nodes and peripheral blood; 3) the extent of lymphoid cell infiltration in CNS of TGF-beta-treated and control mice; and 4) the role of endogenous TGF-beta and TNF in determining the severity of both acute and relapsing EAE. The onset of acute-EAE in SJL mice, induced by immunization with spinal cord homogenate in CFA and pertussigen, is on days 10 to 15. Although daily i.p. injections of 0.2 to 2 micrograms TGF-beta 1 or TGF-beta 2 on days 5 to 9 after immunization are highly protective, injections on days 1 to 5 or 9 to 13 are not. Moreover, anti-TGF-beta accelerates and aggrevates EAE when given on days 5 and 9, but not on day 12. Anti-TNF, injected on days 5 and 9, provides a comparable degree of protection as does TGF-beta. Similarly, in relapsing EAE, anti-TGF-beta increases, whereas anti-TNF decreases the incidence and severity of relapses. TGF-beta treatment on days 5 to 9 does not influence the appearance of sensitized cells in peripheral blood and lymph nodes, but does prevent the accumulation of T cells in brain and spinal cord, as assayed on days 15 to 20. It is concluded that the protective effect of TGF-beta is exerted at the level of the target organ, CNS and/or its vascular endothelium, rather than through a direct effect on lymphoid cells, and that there is a small window of 4 days in which TGF-beta exerts its protective effect.


Arthropod nuclear receptors and their role in molting.
The molting process in arthropods is regulated by steroid hormones acting via nuclear receptor proteins. The most common molting hormone is the ecdysteroid, 20-hydroxyecdysone. The receptors of 20-hydroxyecdysone have also been identified in many arthropod species, and the amino acid sequences determined. The functional molting hormone receptors consist of two members of the nuclear receptor superfamily, namely the ecdysone receptor and the ultraspiracle, although the ecdysone receptor may be functional, in some instances, without the ultraspiracle. Generally, the ecdysone receptor/ultraspiracle heterodimer binds to a number of ecdysone response elements, sequence motifs that reside in the promoter of various ecdysteroid-responsive genes. In the ensuing transcriptional induction, the ecdysone receptor/ultraspiracle complex binds to 20-hydroxyecdysone or to a cognate ligand that, in turn, leads to the release of a corepressor and the recruitment of coactivators. 3D structures of the ligand-binding domains of the ecdysone receptor and the ultraspiracle have been solved for a few insect species. Ecdysone agonists bind to ecdysone receptors specifically, and ligand-ecdysone receptor binding is enhanced in the presence of the ultraspiracle in insects. The basic mode of ecdysteroid receptor action is highly conserved, but substantial functional differences exist among the receptors of individual species. Even though the transcriptional effects are apparently similar for ecdysteroids and nonsteroidal compounds such as diacylhydrazines, the binding shapes are different between them. The compounds having the strongest binding affinity to receptors ordinarily have strong molting hormone activity. The ability of the ecdysone receptor/ultraspiracle complex to manifest the effects of small lipophilic agonists has led to their use as gene switches for medical and agricultural applications.


The concurrent evolution of cooperation and the population structures that support it.
The evolution of cooperation often depends upon population structure, yet nearly all models of cooperation implicitly assume that this structure remains static. This is a simplifying assumption, because most organisms possess genetic traits that affect their population structure to some degree. These traits, such as a group size preference, affect the relatedness of interacting individuals and hence the opportunity for kin or group selection. We argue that models that do not explicitly consider their evolution cannot provide a satisfactory account of the origin of cooperation, because they cannot explain how the prerequisite population structures arise. Here, we consider the concurrent evolution of genetic traits that affect population structure, with those that affect social behavior. We show that not only does population structure drive social evolution, as in previous models, but that the opportunity for cooperation can in turn drive the creation of population structures that support it. This occurs through the generation of linkage disequilibrium between socio-behavioral and population-structuring traits, such that direct kin selection on social behavior creates indirect selection pressure on population structure. We illustrate our argument with a model of the concurrent evolution of group size preference and social behavior.


Dynamics of spore coat morphogenesis in Bacillus subtilis.
Spores of Bacillus subtilis are encased in a protective coat made up of at least 70 proteins. The structure of the spore coat has been examined using a variety of genetic, imaging and biochemical techniques; however, the majority of these studies have focused on mature spores. In this study we use a library of 41 spore coat proteins fused to the green fluorescent protein to examine spore coat morphogenesis over the time-course of sporulation. We found considerable diversity in the localization dynamics of coat proteins and were able to establish six classes based on localization kinetics. Localization dynamics correlate well with the known transcriptional regulators of coat gene expression. Previously, we described the existence of multiple layers in the mature spore coat. Here, we find that the spore coat initially assembles a scaffold that is organized into multiple layers on one pole of the spore. The coat then encases the spore in multiple co-ordinated waves. Encasement is driven, at least partially, by transcription of coat genes and deletion of sporulation transcription factors arrests encasement. We also identify the trans-compartment SpoIIIAH-SpoIIQ channel as necessary for encasement. This is the first demonstration of a forespore contribution to spore coat morphogenesis.


Electronic symptom reporting between patient and provider for improved health care service quality: a systematic review of randomized controlled trials. part 2: methodological quality and effects.
We conducted in two parts a systematic review of randomized controlled trials (RCTs) on electronic symptom reporting between patients and providers to improve health care service quality. Part 1 reviewed the typology of patient groups, health service innovations, and research targets. Four innovation categories were identified: consultation support, monitoring with clinician support, self-management with clinician support, and therapy. To assess the methodological quality of the RCTs, and summarize effects and benefits from the methodologically best studies. We searched Medline, EMBASE, PsycINFO, Cochrane Central Register of Controlled Trials, and IEEE Xplore for original studies presented in English-language articles between 1990 and November 2011. Risk of bias and feasibility were judged according to the Cochrane recommendation, and theoretical evidence and preclinical testing were evaluated according to the Framework for Design and Evaluation of Complex Interventions to Improve Health. Three authors assessed the risk of bias and two authors extracted the effect data independently. Disagreement regarding bias assessment, extraction, and interpretation of results were resolved by consensus discussions. Of 642 records identified, we included 32 articles representing 29 studies. No articles fulfilled all quality requirements. All interventions were feasible to implement in a real-life setting, and theoretical evidence was provided for almost all studies. However, preclinical testing was reported in only a third of the articles. We judged three-quarters of the articles to have low risk for random sequence allocation and approximately half of the articles to have low risk for the following biases: allocation concealment, incomplete outcome data, and selective reporting. Slightly more than one fifth of the articles were judged as low risk for blinding of outcome assessment. Only 1 article had low risk of bias for blinding of participants and personnel. We excluded 12 articles showing high risk or unclear risk for both selective reporting and blinding of outcome assessment from the effect assessment. The authors' hypothesis was confirmed for 13 (65%) of the 20 remaining articles. Articles on self-management support were of higher quality, allowing us to assess effects in a larger proportion of studies. All except one self-management interventions were equally effective to or better than the control option. The self-management articles document substantial benefits for patients, and partly also for health professionals and the health care system. Electronic symptom reporting between patients and providers is an exciting area of development for health services. However, the research generally is of low quality. The field would benefit from increased focus on methods for conducting and reporting RCTs. It appears particularly important to improve blinding of outcome assessment and to precisely define primary outcomes to avoid selective reporting. Supporting self-management seems to be especially promising, but consultation support also shows encouraging results.


Different mechanisms regulate productive herpes simplex virus 1 (HSV-1) and HSV-2 infections in adult trigeminal neurons.
Herpes simplex virus 1 (HSV-1) and HSV-2 establish latency in different neuronal subtypes (A5+ and KH10+) in murine trigeminal ganglia, results which correlate with restricted productive infection in these neurons in vitro. HSV-2 latency-associated transcript (LAT) contains a cis-acting regulatory element near the transcription start site that promotes productive infection in A5+ neurons and a second element in exon 1 that inhibits productive infection in KH10+ neurons. HSV-1 contains no such regulatory sequences, demonstrating different mechanisms for regulating productive HSV infection in neurons.


Reverse transcriptase-polymerase chain reaction amplification and partial sequence of T helper 1- and T helper 2-type lymphokine genes from the owl monkey (Aotus trivirgatus).
The reverse transcriptase-polymerase chain reaction (RT-PCR) was used to amplify selected lymphokine mRNAs from phytohemagglutinin-activated leukocytes of the owl monkey (Aotus trivirgatus). Interleukin-2 (IL-2), IL-4, IL-13, and interferon-gamma were selected as lymphokine mRNAs of interest, since expression of these cytokines helps define the type of T helper lymphocyte response (i.e., TH1 versus TH2). Because sequences for these lymphokine genes were not available for the owl monkey, multiple PCR primers for each lymphokine gene were designed based on published human sequences. Various PCR primer pairs were then used in the RT-PCR to determine the conditions for optimal amplification of each owl monkey cytokine mRNA. In addition, each PCR primer pair was compared for the ability to amplify lymphokine mRNAs from other primate species, including African green (Cercopithecus aethiops), squirrel (Saimiri sciureus), and rhesus (Macaca mulatta) monkeys. The specificity and sensitivity of optimal primer pair was also demonstrated by amplification of as little as 10 fg of each lymphokine gene in a background of 300 ng of irrelevant cDNA. Finally, partial sequences of owl monkey coding regions for IL-2, IL-13, and interferon-gamma were determined and compared for homology with their human counterparts. Together, these studies define specific and sensitive conditions for detection of lymphokine mRNA expression in the owl monkey and provide partial sequence information of the coding region for these lymphokines. This investigation should provide molecular probes to investigate the immune response against malaria and the effectiveness of malaria vaccines in the owl monkey that models this human disease.


Conductivity and current density image reconstruction using harmonic Bz algorithm in magnetic resonance electrical impedance tomography.
Magnetic resonance electrical impedance tomography (MREIT) is to provide cross-sectional images of the conductivity distribution sigma of a subject. While injecting current into the subject, we measure one component Bz of the induced magnetic flux density B = (Bx, By, Bz) using an MRI scanner. Based on the relation between (inverted delta)2 Bz and inverted delta sigma, the harmonic Bz algorithm reconstructs an image of sigma using the measured Bz data from multiple imaging slices. After we obtain sigma, we can reconstruct images of current density distributions for any given current injection method. Following the description of the harmonic Bz algorithm, this paper presents reconstructed conductivity and current density images from computer simulations and phantom experiments using four recessed electrodes injecting six different currents of 26 mA. For experimental results, we used a three-dimensional saline phantom with two polyacrylamide objects inside. We used our 0.3 T (tesla) experimental MRI scanner to measure the induced Bz. Using the harmonic Bz algorithm, we could reconstruct conductivity and current density images with 82 x 82 pixels. The pixel size was 0.6 x 0.6 mm2. The relative L2 errors of the reconstructed images were between 13.8 and 21.5% when the signal-to-noise ratio (SNR) of the corresponding MR magnitude images was about 30. The results suggest that in vitro and in vivo experimental studies with animal subjects are feasible. Further studies are requested to reduce the amount of injection current down to less than 1 mA for human subjects.


Priming of familiar and unfamiliar visual objects over delays in young and older adults.
Although priming of familiar stimuli is usually age invariant, little is known about how aging affects priming of preexperimentally unfamiliar stimuli. Therefore, this study investigated the effects of aging and encoding-to-test delays (0 min, 20 min, 90 min, and 1 week) on priming of unfamiliar objects in block-based priming paradigms. During the encoding phase, participants viewed pictures of novel objects (Experiments 1 and 2) or novel and familiar objects (Experiment 3) and judged their left-right orientation. In the test block, priming was measured using the possible-impossible object-decision test (Experiment 1), symmetric-asymmetric object-decision test (Experiment 2), and real-nonreal object-decision test (Experiment 3). In Experiments 1 and 2, young adults showed priming for unfamiliar objects at all delays, whereas older adults whose baseline task performance was similar to that of young adults did not show any priming. Experiment 3 found no effects of age or delay on priming of familiar objects; however, priming of unfamiliar objects was only observed in the young participants. This suggests that when older adults cannot rely on preexisting memory representations, age-related deficits in priming can emerge.


Commonality despite exceptional diversity in the baseline human antibody repertoire.
In principle, humans can produce an antibody response to any non-self-antigen molecule in the appropriate context. This flexibility is achieved by the presence of a large repertoire of naive antibodies, the diversity of which is expanded by somatic hypermutation following antigen exposure1. The diversity of the naive antibody repertoire in humans is estimated to be at least 1012 unique antibodies2. Because the number of peripheral blood B cells in a healthy adult human is on the order of 5 × 109, the circulating B cell population samples only a small fraction of this diversity. Full-scale analyses of human antibody repertoires have been prohibitively difficult, primarily owing to their massive size. The amount of information encoded by all of the rearranged antibody and T cell receptor genes in one person-the 'genome' of the adaptive immune system-exceeds the size of the human genome by more than four orders of magnitude. Furthermore, because much of the B lymphocyte population is localized in organs or tissues that cannot be comprehensively sampled from living subjects, human repertoire studies have focused on circulating B cells3. Here we examine the circulating B cell populations of ten human subjects and present what is, to our knowledge, the largest single collection of adaptive immune receptor sequences described to date, comprising almost 3 billion antibody heavy-chain sequences. This dataset enables genetic study of the baseline human antibody repertoire at an unprecedented depth and granularity, which reveals largely unique repertoires for each individual studied, a subpopulation of universally shared antibody clonotypes, and an exceptional overall diversity of the antibody repertoire.


UKPDS outcomes model 2: a new version of a model to simulate lifetime health outcomes of patients with type 2 diabetes mellitus using data from the 30 year United Kingdom Prospective Diabetes Study: UKPDS 82.
The aim of this project was to build a new version of the United Kingdom Prospective Diabetes Study (UKPDS) Outcomes Model (UKPDS-OM1), a patient-level simulation tool for predicting lifetime health outcomes of people with type 2 diabetes mellitus. Data from 5,102 UKPDS patients from the 20 year trial and the 4,031 survivors entering the 10 year post-trial monitoring period were used to derive parametric proportional hazards models predicting absolute risk of diabetes complications and death. We re-estimated the seven original event equations and estimated new equations for diabetic ulcer and some second events. The additional data permitted inclusion of new risk factor predictors such as estimated GFR. We also developed four new equations for all-cause mortality. Internal validation of model predictions of cumulative incidence of all events and death was carried out and a contemporary patient-level dataset was used to compare 10 year predictions from the original and the new models. Model equations were based on a median 17.6 years of follow-up and up to 89,760 patient-years of data, providing double the number of events, greater precision and a larger number of significant covariates. The new model, UKPDS-OM2, is internally valid over 25 years and predicts event rates for complications, which are lower than those from the existing model. The new UKPDS-OM2 has significant advantages over the existing model, as it captures more outcomes, is based on longer follow-up data, and more comprehensively captures the progression of diabetes. Its use will permit detailed and reliable lifetime simulations of key health outcomes in people with type 2 diabetes mellitus.


Placental site trophoblastic tumor: report of four cases and review of literature.
Placental site trophoblastic tumor (PSTT) is a rare variant of gestational trophoblastic disease that originates from the implantation site intermediate trophoblast. We report four patients with PSTT and review pertinent literature. Three patients presented with disease confined to the uterus and one patient with disease extension beyond the uterus. Antecedent pregnancy was full-term pregnancy in three patients and termination of a 21-week pregnancy in one patient. Interval from the antecedent pregnancy was <1 year in three patients and 13 years in one patient. Primary treatment was simple hysterectomy in three patients and radical hysterectomy in one patient. Overall, three patients received chemotherapy; one had EP/EMA as adjuvant chemotherapy, one had EMA/CO for rising levels of serum beta-hCG and one had BEP then VIP for recurrent disease. The three patients with disease confined to the uterus have remained after treatment alive and with no evidence of disease, whereas the one patient with disease extension beyond the uterus died of disease despite surgery and aggressive chemotherapy. It is concluded that disease extension beyond the uterus is the most important adverse prognostic factor. Other adverse prognostic factors are interval from antecedent pregnancy >2 years, age >40 years, and mitotic count >5 mitotic figures/10 high-power fields. Because of the relative insensitivity to chemotherapy, hysterectomy is the mainstay of treatment. EP/EMA seems to be the most effective first-line chemotherapy available to date for metastatic and relapsing PSTT.


Proteomic patterns and prediction of glomerulosclerosis and its mechanisms.
Protein expression profiles linked to sclerosis in the 5/6 nephrectomy (Nx) rat model of focal segmental glomerulosclerosis were investigated. Sections of control glomeruli from normal baseline Nx tissue and nonsclerotic and sclerotic glomeruli from 12 wk after 5/6 Nx were isolated by laser capture microdissection. Protein profiles were acquired directly by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Classification accuracy was 99.2% for distinguishing normal versus sclerotic glomeruli and 96.7 and 97.8% for nonsclerotic versus normal and sclerotic glomeruli, respectively. The proteomic pattern of the nonsclerotic glomeruli was more similar to sclerotic than normal glomeruli (P < 0.0001). Thymosin beta4, a protein with relevant interactions with plasminogen activator inhibitor-1, angiogenesis, and wound healing, was identified as a key differentially expressed protein. Thymosin beta4 immunostaining was increased in sclerotic glomeruli, predominantly in endothelial cells. Downregulation of thymosin beta4 by RNAi in cultured glomerular endothelial cells decreased angiotensin II-induced plasminogen activator inhibitor-1 expression. In conclusion, proteomic patterns can accurately distinguish normal versus nonsclerotic versus sclerotic glomeruli. The closely related proteomic patterns of nonsclerotic and sclerotic glomeruli suggest early activation of prosclerotic mechanisms even in seemingly intact glomeruli. Thymosin beta4 is a marker of such early events and may even contribute to sclerosis.


Interpretation of X chromosome dose at Sex-lethal requires non-E-box sites for the basic helix-loop-helix proteins SISB and daughterless.
For Drosophila melanogaster flies, sexual fate is determined by the X chromosome number. The basic helix-loop-helix protein product of the X-linked sisterlessB (sisB or scute) gene is a key indicator of the X dose and functions to activate the switch gene Sex-lethal (Sxl) in female (XX), but not in male (XY), embryos. Zygotically expressed sisB and maternal daughterless (da) proteins are known to form heterodimers that bind E-box sites and activate transcription. We examined SISB-Da binding at Sxl by using footprinting and gel mobility shift assays and found that SISB-Da binds numerous clustered sites in the establishment promoter Sxl(Pe). Surprisingly, most SISB-Da sites at Sxl(Pe) differ from the canonical CANNTG E-box motif. These noncanonical sites have 6-bp CA(G/C)CCG and 7-bp CA(G/C)CTTG cores and exhibit a range of binding affinities. We show that the noncanonical sites can mediate SISB-Da-activated transcription in cell culture. P-element transformation experiments show that these noncanonical sites are essential for Sxl(Pe) activity in embryos. Together with previous deletion analysis, the data suggest that the number, affinity, and position of SISB-Da sites may all be important for the operation of the Sxl(Pe) switch. Comparisons with other dose-sensitive promoters suggest that threshold responses to diverse biological signals have common molecular mechanisms, with important variations tailored to suit particular functional requirements.


Small intestine hexose transport in experimental diabetes. Increased transporter mRNA and protein expression in enterocytes.
The effect of insulinopenic diabetes on the expression of glucose transporters in the small intestine was investigated. Enterocytes were sequentially isolated from jejunum and ileum of normal fed rats, streptozotocin-diabetic rats, and diabetic rats treated with insulin. Facilitative glucose transporter (GLUT) 2, GLUT5, and sodium-dependent glucose transporter 1 protein content was increased from 1.5- to 6-fold in enterocytes isolated from diabetic animals in both jejunum and ileum. Insulin was able to reverse the increase in transporter protein expression seen after induction of diabetes. There was a four- to eightfold increase in the amount of enterocyte glucose transporter mRNA after diabetes with greater changes in sodium-dependent glucose transporter 1 and GLUT2 than in GLUT5 levels. In situ hybridization showed that after the induction of diabetes there was new hybridization in lower villus and crypt enterocytes that was reversed by insulin treatment. Thus, the increase in total hexose transport caused by diabetes is due to a premature expression of hexose transporters by enterocytes along the crypt-villus axis, causing a cumulative increase in enterocyte transporter protein during maturation. These changes are likely to represent an adaptive response by the organism to increase nutrient absorption in a perceived state of tissue starvation. These adaptive changes may lead to exacerbation of hyperglycemia in uncontrolled diabetes.


18F-FDG PET/CT of cervical carcinoma.
This article outlines the role of FDG PET/CT in patients with cervical carcinoma for evaluating tumor extent, assessing treatment response, and detecting disease recurrence. There is increasing evidence that FDG PET/CT has a role in the primary evaluation of cervical carcinoma-in particular, for evaluating lymph node status and distant metastatic disease. PET/CT is also helpful to determine prognosis, assess treatment response, and evaluate disease recurrence.