Pif1 Activity is Modulated by DNA Sequence and Structure.
The gene encoding the Pif1 helicase was first discovered in a Saccharomyces cerevisiae genetic screen as a mutant that reduces recombination between mitochondrial respiratory mutants and was subsequently rediscovered in a screen for genes affecting the telomere length in the nucleus. It is now known that Pif1 is involved in numerous aspects of DNA metabolism. All known functions of Pif1 rely on binding to DNA substrates followed by ATP hydrolysis, coupling the energy released to translocation along DNA to unwind duplex DNA or alternative DNA secondary structures. The interaction of Pif1 with higher-order DNA structures, like G-quadruplex DNA, as well as the length of single-stranded (ss)DNA necessary for Pif1 loading have been widely studied. Here, to test the effects of ssDNA length, sequence, and structure on Pif1's biochemical activities in vitro, we used a suite of oligonucleotide-based substrates to perform a basic characterization of Pif1 ssDNA binding, ATPase activity, and helicase activity. Using recombinant, untagged S. cerevisiae Pif1, we found that Pif1 preferentially binds to structured G-rich ssDNA, but the preferred binding substrates failed to maximally stimulate ATPase activity. In helicase assays, significant DNA unwinding activity was detected at Pif1 concentrations as low as 250 pM. Helicase assays also demonstrated that Pif1 most efficiently unwinds DNA fork substrates with unstructured ssDNA tails. As the chemical step size of Pif1 has been determined to be 1 ATP per translocation or unwinding event, this implies that the highly structured DNA inhibits conformational changes in Pif1 that couple ATP hydrolysis to DNA translocation and unwinding.


A transiently expressed SK current sustains and modulates action potential activity in immature mouse inner hair cells.
From just after birth, mouse inner hair cells (IHCs) expressed a Ca(2+)-activated K(+) current that was reduced by intracellular BAPTA at concentrations >or= 1 mM. The block of this current by nifedipine suggests the direct involvement of Ca(v)1.3 Ca(2+) channels in its activation. On the basis of its high sensitivity to apamin (K(D) 360 pM) it was identified as a small-conductance Ca(2+)-activated K(+) current (SK), probably SK2. A similar current was also found in outer hair cells (OHCs) from the beginning of the second postnatal week. In both cell types the appearance of the SK current coincided with their becoming responsive to acetylcholine (ACh), the main efferent neurotransmitter in the cochlea. The effect of ACh on IHCs was abolished when they were simultaneously superfused with strychnine, consistent with the presence of nicotinic ACh receptors (nAChRs). Extracellular Ca(2+) either potentiated or blocked the nAChR current depending on its concentration, as previously reported for the recombinant alpha9alpha10 nAChR. Outward currents activated by ACh were reduced by blocking the SK current with apamin or by preventing SK current activation with intracellular BAPTA (>or= 10 mM). The endogenous mobile Ca(2+) buffer concentration was estimated to be equivalent to about 1 mM BAPTA, suggesting that in physiological conditions the SK channel is significantly activated by Ca(2+) influx through both Ca(v)1.3 Ca(2+) channels and alpha9alpha10 nAChRs. Current clamp experiments showed that in IHCs the SK current is required for sustaining a train of action potentials and also modulates their frequency when activated by ACh.


Late Luteal Subphase Food Craving Is Enhanced in Women with Obesity and Premenstrual Dysphoric Disorder (PMDD).
Dysregulated food craving is a complex weight-related behavior. To identify novel targets for enhancing the efficacy of weight loss interventions, we examined whether food craving varies across the menstrual cycle according to the abdominal obesity type and premenstrual dysphoric disorder (PMDD) diagnosis, and, if so, whether it is related to affective symptomatology. Reproductive-age women were classified into one of the four study groups according to whether they have abdominal obesity (AO) or are abdominally lean (AL), and the presence of PMDD: (1) AO:PMDD+ (n = 13), (2) AL:PMDD+ (n = 14), (3) AO:PMDD- (n = 15), and (4) AL:PMDD- (n = 16). Self-report measures as well as urinary luteinizing hormone (LH) tests were provided by the participants across 2-3 menstrual cycles. The ratings of food cravings were similar across the menstrual cycle, except the last, late luteal subphase as the AO:PMDD+ participants had the highest food craving rating. Irritability and depression were correlated with food cravings, but not in a distinctive manner across the menstrual cycle by group. Our study found that women with abdominal obesity and PMDD display a temporal vulnerability to a food-related behavior. The possibility of shared neurobiology between the two conditions is discussed and should be examined in future studies.


Effectiveness of trained community volunteers in improving knowledge and management of childhood malaria in a rural area of Rivers State, Nigeria.
Malaria accounts for 70% of illnesses and 30% of deaths among children under 5 years in Nigeria. This study was aimed at determining the effectiveness of trained community volunteers in delivering multiple anti-malaria interventions to achieve rapid reduction in morbidity and mortality among under 5 children. A quasi-experimental study was carried out in two rural communities in Rivers State, Nigeria among 368 mothers/caregivers. A set of 184 of the mothers/caregivers (experimental group) were trained on malaria and provided with bed nets and drugs (artemisinin-lumefantrine) to treat children under 5 years who developed fever during the period of the experiment. Another set of 184 mothers/caregivers (controls) did not receive similar training and drugs. Both groups were compared at baseline and after 6 months of the experiment on their knowledge of malaria prevention and treatment. Level of significance was set at P = 0.05. In the experimental group: Adequate knowledge about malaria increased from 115 (62.5%) to 175 (95.1%) (P < 0.0001), early commencement of treatment of fever increased from 68 (37.0%) to 131 (75.7%) (P < 0.0001), and children cured of malaria increased from 87 (47.3%) to 146 (84.4%) (P < 0.0001). Insecticide-treated bed nets use also increased from 86 (46.7%) to 161 (87.5%) (P < 0.0001). There were no significant changes in the control group. The study demonstrated the inherent potentials in using community-based volunteers in malaria prevention and control for those in rural areas with poor health service delivery. We advocate its adaptation for far-reaching reduction in childhood morbidity and mortality and rapid attainment of millennium development goals 4.


Tuberculosis surveillance and control in Germany - An application of the Berlin Declaration Monitoring and Evaluation Framework.
In 2007, the 'Berlin Declaration on Tuberculosis' (BD) was signed by the Ministers of the WHO European Region Member States (MS) to address the re-emerging threat of tuberculosis (TB) by fully implementing the Stop TB strategy. WHO Euro developed a Monitoring and Evaluation (M&E) Framework using programmatic and epidemiological indicators to follow-up the BD from 2013 onwards. Ahead of this, we explored the Framework performance for Germany, a low TB incidence country. We selected from a surveillance perspective on five country-specific indicators asked for all MS. Information sources included guidelines, notification data and laboratory quality assurance results. As to national TB guidelines (indicator 1.1.1), up-to-date guidance documents are available. Regarding external quality assurance (3.2) proficiency testing confirms high quality of culture and drug-susceptibility testing. M/XDR-TB prevention and control is reflected by a 1.7% MDR-TB rate in 2010 with decreasing tendency (4.1.2). As to treatment success (5), the WHO target of 85% is reached for all age groups except the elderly ≥60 years. Germany disposes of an electronic case-based surveillance providing evidence for TB policy and practices (5.2.2). The selected M&E Framework indicators proved well applicable to a low-incidence country. The results show no specific gaps in TB surveillance and control in Germany.


Laparoscopic approach reduces likelihood of perioperative complications in patients undergoing adrenalectomy.
Laparoscopy has been reported advantageous over the conventional open technique for adrenalectomy. However, most comparative series include the relatively more challenging cases in the open group. The aim of this study is to assess the actual role of laparoscopy in reducing perioperative complications compared to open surgery in patients undergoing adrenalectomy. Between January 1992 and December 2002, we performed 148 adrenalectomies in 138 patients. Depending on the approach, patients were divided into laparoscopic (LA) or open adrenalectomy (OA) groups. Demographics, tumor characteristics, operative data, and outcomes were analyzed. Linear and logistic regressions identified factors influencing perioperative outcomes. Multivariate-adjusted logistic regression assessed independent relationship between factors and perioperative outcomes. A total of 78 cases were performed laparoscopically and 70 open. Patients were matched for age and sex. Tumor size was smaller (3 +/- 2 vs 5 +/- 3 cm), operative time was shorter (133 +/- 65 vs 165 +/- 100 min), estimated blood loss was less (114 +/- 152 vs 350 +/- 417 cc), length of stay was shorter (3 +/- 2 vs 7 +/- 3 days), and overall complication rate was lower (7% vs 20%) in the LA compared to the OA group. The incidence of cancer in tumors > or = 6 cm (31%) was higher than in those < 6 cm (4%). All patients with cancer underwent OA. LA was the only factor independently associated with a decreased likelihood of intraoperative bleeding and postoperative pulmonary complications. Large and malignant adrenal tumors are more frequently removed through an open approach. However, this fact has no influence on the advantages of the LA over the OA. Laparoscopy reduces perioperative adrenalectomy perioperative complication rates. It has a positive impact on intraoperative bleeding and postoperative pulmonary complications.


Drug prophylaxis in atherosclerosis.
Heparin and heparinoids constitute the one drug group shown to arrest and reverse atherosclerotic changes in rabbits on a high cholesterol/fat diet. Little use has been made of this finding because heparin has been administered by routes which produce anticoagulation and thus bleeding. Inhalation once a fortnight results in a high concentration of heparin in endothelium with low plasma concentrations. No toxicity has been demonstrated with long term heparin but this is not true for heparinoids.


Desalination of brackish groundwater by direct contact membrane distillation.
The direct contact membrane distillation (DCMD) applied for desalination of brackish groundwater with self-made polyvinylidene fluoride (PVDF) membranes was presented in the paper. The PVDF membrane exhibited high rejection of non-volatile inorganic salt solutes and a maximum permeate flux 24.5 kg m(-2) h(-1) was obtained with feed temperature at 70 degrees C. The DCMD experimental results indicated that the feed concentration had no significant influence on the permeate flux and the rejection of solute. When natural groundwater was used directly as the feed, the precipitation of CaCO(3) would be formed and clog the hollow fibre inlets with gradual concentration of the feed, which resulted in a rapid decline of the module efficiency. The negative influence of scaling could be eliminated by acidification of the feed. Finally, a 250 h DCMD continuous desalination experiment of acidified groundwater with the concentration factor at constant 4.0 was carried out. The permeate flux kept stable and the permeate conductivity was less than 7.0 microS cm(-1) during this process. Furthermore, there was no deposit observed on the membrane surface. All of these demonstrated that DCMD could be efficiently used for production of high-quality potable water from brackish groundwater with water recovery as high as 75%.


Scintigraphic manifestations of thyrotoxicosis.
The term thyrotoxicosis refers to the clinical syndrome of increased systemic metabolism that results when the serum concentrations of free thyroxine, free triiodothyronine, or both are elevated. The term hyperthyroidism refers to overactivity of the thyroid gland with a resultant increase in thyroid hormone synthesis and release into the systemic circulation. These terms are not interchangeable, since thyrotoxicosis can develop in thyroid conditions that are not associated with increased thyroid function, such as thyroiditis, or in so-called factitious hyperthyroidism. The clinical signs and symptoms of thyrotoxicosis are virtually identical regardless of the cause. However, in a given patient, every attempt should be made to determine the exact cause of the thyrotoxicosis, as this in turn determines the prognosis and treatment. Since thyroid scintigraphy demonstrates the functional state of the thyroid gland, it should be used, in conjunction with determination of radioactive iodine uptake, as the imaging modality of choice for diagnosis of thyrotoxicosis. Although the scintigraphic features of several of the thyroid disorders that cause thyrotoxicosis may overlap, their recognition helps narrow the differential diagnosis, thereby guiding the referring physician in the work-up and management of this disorder.


Fructose catabolism in Xanthomonas campestris pv. campestris. Sequence of the PTS operon, characterization of the fructose-specific enzymes.
In Xanthomonas campestris pv. campestris, fructose is transported and phosphorylated into fructose 1-phosphate through a phosphoenolpyruvate-dependent phosphotransferase system. The nucleotide sequence of the fruA gene encoding the phosphotransferase system permease specific of fructose (EIIFru) was determined. The fructose 1-phosphate produced by the phosphotransferase system is phosphorylated into fructose 1,6-bisphosphate by a 1-phosphofructokinase. This enzyme was characterized and the corresponding gene (fruK) was sequenced. Sequence comparisons revealed that FruK is a member of a new family of ATP-binding proteins composed of sugar (or sugar-phosphate) kinases. In phosphotransferase system-deficient strains, fructose can still be transported by an unidentified permease. The intracellular fructose is then phosphorylated by a multimeric fructokinase of 135 kDa specific for fructose and inhibited by fructose, fructose 1,6-bisphosphate, and mannose. Several other enzymes of fructose metabolism were assayed and a potential pathway for fructose catabolism is presented.


[Spatial reorganization of the organelles of catecholamine-synthesizing group A1 neurons during emotional stress].
Dynamics of spatial reorganization of organellas in catecholamine-synthesizing neurons of A1 group, that participate in reactions of the cardiovascular control, have been studied under normal conditions and under stimulation of the ventromedial nucleus in the hypothalamus. During transition of the hypertensive reaction into the hypotensive one, concentration of various organellas in the Golgi complex area and in the perinuclear zone is intensively increasing. Deep mutual nucleocytoplasmic invaginations appear. The spatial reorganization of the organellas in the neuron demonstrates that some new integrations take place in its metabolic system and reflects its another qualitative state. This serves as an impulse for structural adaptive modifications of the synaptic apparatus. There is a reason to suppose that one of the ways for organization of the alive system stability to an emotional stress is the way of reorganization of the connections, creation of another physiological architecture of the functional system, that can ensure the adaptive reactions of the organism to emotional overstrains.


The test of Lasègue: systematic review of the accuracy in diagnosing herniated discs.
A systematic review of the literature including statistical meta-analysis. To evaluate published methods of the test of Lasègue or straight leg raising test and the cross straight leg raising test by using a recently developed criteria list and to summarize and explore reasons for variation in diagnostic accuracy. Little evidence exists on the diagnostic accuracy of the widely used straight leg raising test and the cross straight leg raising test in diagnosing herniated discs in patients with low back pain. MEDLINE and EMBASE searches up to 1997 showed 17 diagnostic publications evaluating the straight leg raising test with surgery as reference standard. Quality of methods was assessed with a specific checklist. Eleven studies were selected for statistical pooling. Sources of variation and heterogeneity were studied by meta-regression of the diagnostic odds ratio. All studies were surgical case-series at nonprimary care level. Verification-bias was obvious in one study. Pooled sensitivity for straight leg raising test was 0. 91 (95% CI 0.82-0.94), pooled specificity 0.26 (95% CI 0.16-0.38). Pooled diagnostic odds ratio was 3.74 (95% CI 1.2-11.4). Discriminative power was lower in recent studies, in studies with only inclusion of primary hernias, and with blind assessment of both the index-test (straight leg raising test) and the reference (surgery). For the cross straight leg raising test pooled sensitivity was 0.29 (95% CI 0.24-0.34), pooled specificity was 0.88 (95% CI 0.86-0.90), and the pooled diagnostic odds ratio 4.39 (95% CI 0.74-25.9). The diagnostic accuracy of the straight leg raising test is limited by its low specificity. Discriminative power decreased with a more valid design, a more homogenous case-mix, and year of publication. Although the studies may reflect everyday clinical practice, they do not enable a valid evaluation of the diagnostic accuracy of both tests. Diagnostic research should evaluate the validity of the complete diagnostic process and study the evidence of the added value of the different tests used. [Key words: sensitivity, specificity, diagnosis, meta-analysis, test of Lasègue, straight leg raising test]


Two distinct strategies of cotton and soybean differing in leaf movement to perform photosynthesis under drought in the field.
This paper reports an experimental test of the hypothesis that cotton and soybean differing in leaf movement have distinct strategies to perform photosynthesis under drought. Cotton and soybean were exposed to two water regimes: drought stressed and well watered. Drought-stressed cotton and soybean had lower maximum CO2 assimilation rates than well-watered (control) plants. Drought reduced the light saturation point and photorespiration of both species - especially in soybean. Area-based leaf nitrogen decreased in drought-stressed soybean but increased in drought-stressed cotton. Drought decreased PSII quantum yield (ΦPSII) in soybean leaves, but increased ΦPSII in cotton leaves. Drought induced an increase in light absorbed by the PSII antennae that is dissipated thermally via ΔpH- and xanthophylls-regulated processes in soybean leaves, but a decrease in cotton leaves. Soybean leaves appeared to have greater cyclic electron flow (CEF) around PSI than cotton leaves and drought further increased CEF in soybean leaves. In contrast, CEF slightly decreased in cotton under drought. These results suggest that the difference in leaf movement between cotton and soybean leaves gives rise to different strategies to perform photosynthesis and to contrasting photoprotective mechanisms for utilisation or dissipation of excess light energy. We suggest that soybean preferentially uses light-regulated non-photochemical energy dissipation, which may have been enhanced by the higher CEF in drought-stressed leaves. In contrast, cotton appears to rely on enhanced electron transport flux for light energy utilisation under drought, for example, in enhanced nitrogen assimilation.


Vibrio cholerae soluble hemagglutinin/protease is a metalloenzyme.
A soluble hemagglutinin/protease produced by Vibrio cholerae, which has previously been shown to hydrolyze fibronectin and ovomucin and to cleave lactoferrin and the A subunit of the heat-labile enterotoxin of Escherichia coli, appears to be a zinc metalloendopeptidase. Both its hemagglutinative and protease functions are inhibited by chelating agents, including Zincov, a hydroxamic acid derivative specifically designed to inhibit zinc metalloproteases. Thermolysin, a known zinc-containing protease, also causes hemagglutination of responder chicken erythrocytes. This activity is inhibited by Zincov, which does not affect the hemagglutination activity of trypsin and pronase. The hemagglutinin/protease is active on furylacryloyl-Gly-Leu-NH2, a synthetic substrate for thermolysin and other similar proteases. The hemagglutination activity of V. cholerae-infected or cholera toxin-treated infant rabbit intestinal fluid is not inhibited by Zincov, which suggests that this activity is not due to the hemagglutinin/protease, as formerly proposed.


Recent advances in Chinese patent medicines entering the international market.
As an indispensable part of Traditional Chinese medicine (TCM), Chinese patent medicines have played an important role in preventing and treating diseases in China. Since they are easy to use, easy to store, and cost-effective, Chinese patent medicines have been generally accepted and widely used in Chinese clinical practice as a vital medical resource. In recent years, as TCM has developed and it has been accepted around the world, many Chinese patent medicine companies have gained international market access and successfully registered several Chinese patent medicines as over-the-counter (OTC) or prescription drugs in regions and countries that primarily use Western medicine such as the EU, Russia, Canada, Singapore, and Vietnam. Moreover, several Chinese patent medicines have been obtained the US Food and Drug Administration (FDA) approval conducting phase II or III clinical trials in the US. The current work has focused on several Chinese patent medicines that have been successfully registered or that have been submitted for registration abroad. Summarized here are recent advances in the efficacy and molecular mechanisms of these Chinese patent medicines to treat respiratory infectious diseases (Lianhua Qingwen capsules, Jinhua Qinggan granules, and Shufeng Jiedu Capsules), cardiovascular and cerebrovascular diseases (Compound Danshen Dripping Pills, Huatuo Zaizao pills, and Tongxinluo Capsules), cancers (a Kanglaite injection and a Shenqi Fuzheng Injection), and gynecological diseases (Guizhi Fuling Capsules). The hope is that this review will contribute to a better understanding of Chinese patent medicines by people around the world.


Treatment of gingivitis and periodontitis. Research, Science and Therapy Committee of the American Academy of Periodontology.
This paper is one of three in a series prepared by the Research, Science and Therapy Committee of The American Academy of Periodontology and is intended for the information of the dental profession. It represents the position of the Academy regarding the current state of knowledge about treatment of gingivitis and periodontitis. The other papers are entitled The Etiology and Pathogenesis of Periodontal Diseases and Diagnosis of Periodontal Diseases.


Adolescent tobacco use and its determinants: evidence from Global Youth Tobacco Survey, Bangladesh 2007.
Adolescent tobacco use (ATU) is on the rise worldwide and the problem is particularly severe in developing countries. Based on nationally representative data, this study aims to investigate the association between ATU and its possible correlates for Bangladesh, where the prevalence rate of ATU is high. The data set is extracted from the Global Youth Tobacco Survey for Bangladesh conducted in 2007. The survey collected information from a total of 3113 students from 52 schools, with a response rate of 100% at the school level, while a response rate of 88.9% was achieved from the students. Students covered in the survey were in grades 7, 8, 9, and 10, with age ranging from 11 to 17 years. The prevalence rate of ATU at the time of the survey was 8.4%, while 35.6% of the students had used at least a type of tobacco products before. Logistic regressions were used to obtain the odds ratios (ORs) in favor of ATU for each of the possible determinants and the confidence intervals (CIs) of these ratios. Use of tobacco among friends (OR = 3.46; CI = 2.37-5.05), the experience of seeing others smoking at home (OR = 2.10; CI = 1.36-3.22) or other places (OR = 1.6; CI = 1.02-2.57), receiving pocket money (OR = 7.6; CI = 4.59-13.28), receiving free tobacco from vendors (OR = 2.3; CI = 1.44-3.78), and exposure to advertisements and promotions of tobacco products (OR = 1.83; CI = 1.23-2.79) were associated with a higher likelihood of ATU. Increased awareness of health hazards of tobacco use through education in schools helped mitigate the problem of ATU. The findings of this study have ramifications for tobacco control prevention strategies in Bangladesh.


[Factors associated with involuntary hospital admissions in technology-dependent children].
To identify the factors associated with involuntary hospital admissions of technology-dependent children, in the municipality of Ribeirão Preto, São Paulo State, Brazil. A cross-sectional study, with a quantitative approach. After an active search, 124 children who qualified under the inclusion criteria, that is to say, children from birth to age 12, were identified. Data was collected in home visits to mothers or the people responsible for the children, through the application of a questionnaire. Analysis of the data followed the assumptions of the Generalized Linear Models technique. 102 technology-dependent children aged between 6 months and 12 years participated in the study, of whom 57% were male. The average number of involuntary hospital admissions in the previous year among the children studied was 0.71 (±1.29). In the final model the following variables were significantly associated with the outcome: age (OR=0.991; CI95%=0.985-0.997), and the number of devices (OR=0.387; CI95%=0.219-0.684), which were characterized as factors of protection and quantity of medications (OR=1.532; CI95%=1.297-1.810), representing a risk factor for involuntary hospital admissions in technology-dependent children. The results constitute input data for consideration of the process of care for technology-dependent children by supplying an explanatory model for involuntary hospital admissions for this client group.


The comparison of two recovery room warming methods for hypothermia patients who had undergone spinal surgery.
The purpose of this study was to compare the time needed to reach a specified temperature and the efficiency of two warming methods-warm cotton blankets and a radiant warmer-for hypothermia patients in a postanesthetic care unit (PACU) after spinal surgery. This study was conducted according to a quasi-experimental design. Data were collected from a medical referral center in northern Taiwan. A total of 130 post-spinal surgery patients with hypothermia were recruited in the study. Of the 130 patients in the PACU, 65 were warmed by the radiant warmer (group R); whereas the other 65 patients were warmed by warm cotton blankets (group B). Tympanic temperature was measured for each patient every 10 min until it reached 36 °C in the PACU. Analysis of covariance and generalized estimating equation regression analysis were performed to compare the time needed to reach a specified temperature and the efficiency of the two warming methods, respectively. Both groups were similar in their baseline characteristics. After adjusting for temperature upon arrival at the PACU, group R needed a significantly shorter time for rewarming to 36 °C than group B (F [1, 125]= 58.17, p < .001). The results of the generalized estimating equation also showed that the radiant warmer was more efficient than warm cotton blankets in increasing patients' body temperatures to 36 °C (χ2 = 37.44, p < .001). None of the patients appeared to have wound infections, and there were no differences in the length of hospital stay or medical costs for current hospitalization in both groups. Using the radiant warmer may be a more efficient method than providing warm cotton blankets for warming post-spinal surgery hypothermia patients in the PACU. For hospitals that are unable to use forced-air warming to warm postsurgical hypothermia patients in the PACU, the radiant warmer is a more efficient device to rewarm patients.


Association between hospice care and psychological outcomes in Alzheimer's spousal caregivers.
Dementia care giving can lead to increased stress, physical and psychosocial morbidity, and mortality. Anecdotal evidence suggests that hospice care provided to people with dementia and their caregivers may buffer caregivers from some of the adverse outcomes associated with family caregiving in Alzheimer's Disease (AD). This pilot study examined psychological and physical outcomes among 32 spousal caregivers of patients with AD. It was hypothesized that caregivers who utilized hospice services would demonstrate better outcomes after the death of their spouse than caregivers who did not utilize hospice. The charts of all spousal caregivers enrolled in a larger longitudinal study from 2001 to 2006 (N=120) were reviewed, and participants whose spouse had died were identified. Of these, those who received hospice care (n=10) were compared to those who did not (n=22) for various physiological and psychological measures of stress, both before and after the death of the care recipient. An Analysis of Covariance (ANCOVA), with postdeath scores as the dependent variable and pre-death scores as covariates, was used for all variables. Significant group differences were found in postdeath depressive symptoms (HAM-D; F(1,29)=6.10, p<0.05) and anxiety symptoms (HAM-A; F(1,29)=5.71, p<0.05). Most psychological outcome variables demonstrated moderate effect sizes with a Cohen's d of>0.5 between groups. These data suggest that hospice enrollment may ameliorate the detrimental psychological effects in caregivers who have lost a spouse with Alzheimer's Disease. Based on these pilot data, further prospective investigation is warranted.


Stakeholder participation in management of invasive vertebrates.
Stakeholders are increasingly involved in species conservation. We sought to understand what features of a participatory conservation program are associated with its ecological and social outcomes. We conducted a case study of the management of invasive vertebrates in Australia. Invasive vertebrates are a substantial threat to Australia's native species, and stakeholder participation in their management is often necessary for their control. First, we identified potential influences on the ecological and social outcomes of species conservation programs from the literature. We used this information to devise an interview questionnaire, which we administered to managers of 34 participatory invasive-vertebrate programs. Effects of invasive species were related to program initiator (agency or citizen), reasons for use of a participatory approach, and stakeholder composition. Program initiator was also related to the participation methods used, level of governance (i.e., governed by an agency or citizens), changes in stakeholder interactions, and changes in abundance of invasive species. Ecological and social outcomes were related to changes in abundance of invasive species and stakeholder satisfaction. We identified relations between changes in the number of participants, stakeholder satisfaction, and occurrence of conflict. Potential ways to achieve ecological and social goals include provision of governmental support (e.g., funding) to stakeholders and minimization of gaps in representation of stakeholder groups or individuals to, for example, increase conflict mitigation. Our findings provide guidance for increasing the probability of achieving ecological and social objectives in management of invasive vertebrates and may be applicable to other participatory conservation programs.


Prenatal expression patterns of genes associated with neuropsychiatric disorders.
Neurodevelopmental disorders presumably involve events that occur during brain development. The authors hypothesized that neuropsychiatric disorders considered to be developmental in etiology are associated with susceptibility genes that are relatively upregulated during fetal life (i.e., differentially expressed). The authors investigated the presence of prenatal expression enrichment of susceptibility genes systematically, as composite gene sets associated with six neuropsychiatric disorders in the microarray-based "BrainCloud" dorsolateral prefrontal cortex transcriptome. Using a fetal/postnatal log2-fold change threshold of 0.5, genes associated with syndromic neurodevelopmental disorders (N=31 genes, p=3.37×10-3), intellectual disability (N=88 genes, p=5.53×10-3), and autism spectrum disorder (N=242 genes, p=3.45×10-4) were relatively enriched in prenatal transcript abundance, compared with the overall transcriptome. Genes associated with schizophrenia by genome-wide association studies were not preferentially fetally expressed (N=106 genes, p=0.46), nor were genes associated with schizophrenia by exome sequencing (N=212 genes, p=0.21), but specific genes within copy-number variant regions associated with schizophrenia were relatively enriched in prenatal transcript abundance, and genes associated with schizophrenia by meta-analysis were functionally enriched for some neurodevelopmental processes. In contrast, genes associated with neurodegenerative disorders were significantly underexpressed during fetal life (N=46 genes, p=1.67×10-3). The authors found evidence for relative prenatal enrichment of putative susceptibility genes for syndromic neurodevelopmental disorders, intellectual disability, and autism spectrum disorder. Future transcriptome-level association studies should evaluate regions other than the dorsolateral prefrontal cortex, at other time points, and incorporate further RNA sequencing analyses.


The Eye Color Experiment: From Berlin to Auschwitz and Back.
In an effort to alter eye color during World War II, devout Nazi researcher Karin Magnussen had adrenaline eye drops administered to inmates at the concentration camp Auschwitz-Birkenau. A Sinti family, with a high prevalence of heterochromia iridis, was forced to participate in this study. Members of this family, as well as other victims, were later killed and had their eyes enucleated and sent to Magnussen for examination. Magnussen articulated the findings of these events in a manuscript that has never been published. The author is the first ophthalmologist to review this manuscript. The generation who experienced the atrocities of World War II will soon be gone and awareness of what happened during this tragic chapter of world history is fading. To describe these events to raise awareness among future generations. A literature review and archival search was conducted. Magnussen's research was based on an animal study published in 1937. For Magnussen's study, adrenaline drops were administered to inmates, including a 12-year-old girl from the Sinti family. As there was a reported case of deaf-mutism within the family, Waardenburg syndrome seems to be the most plausible explanation for this family's heritable heterochromia. The effort to change eye color was doomed to fail from the beginning because there was a probable diagnosis of Waardenburg syndrome. Extinction of humans for ophthalmological research is an insane act beyond imagination. For the sake of these victims, and for the generations who still feel their pain, it is imperative to tell their stories.


Pediatric Project ECHO®: A Virtual Community of Practice to Improve Palliative Care Knowledge and Self-Efficacy among Interprofessional Health Care Providers.
Background: Health care providers (HCPs) require ongoing training and mentorship to fully appreciate the palliative care needs of children. Project ECHO® (Extension for Community Healthcare Outcomes) is a model for delivering technology-enabled interprofessional education and cultivating a community of practice among HCPs who care for children with life-limiting illness. Objectives: To develop, implement, and evaluate the Project ECHO model within the pediatric palliative care (PPC) context. Specific objectives were to evaluate (1) participation levels, (2) program acceptability, (3) HCP knowledge changes, (4) HCP self-efficacy changes, and (5) perceived practice changes after six months. Intervention: An interprofessional PPC curriculum was informed by a needs assessment. The curriculum was delivered through monthly virtual 90-minute TeleECHO sessions (didactic presentation and case-based learning) from January 2018 to December 2019. The program was freely available to all HCPs wishing to participate. Design: A mixed-methods design with repeat measures was used. Surveys were distributed at baseline and six months to assess outcomes using 7-point Likert scales. Descriptive and inferential statistical analyses were conducted. The study was approved by the Research Ethics Board at the Hospital for Sick Children. Results: Twenty-four TeleECHO sessions were completed with a mean of 32 ± 12.5 attendees. Acceptability scores (n = 43) ranged from 5.1 ± 1.1 to 6.5 ± 0.6. HCPs reported improvements in knowledge and self-efficacy across most topics (11 out of 12) and skills (8 out of 10) with demonstrated statistical significance (p < 0.05). Most participants reported positive practice impacts, including enhanced ability to provide PPC in their practice. Conclusion: Project ECHO is a feasible and impactful model for fostering a virtual PPC-focused community of practice among interprofessional HCPs.


[Orbital manifestation of microscopic polyangiitis (mPA)--presentation of clinical cases].
Microscopic polyangiitis (mPA) is one of the ANCA-associated primary systemic vasculitides. It is defined as necrotic vasculitis without granuloma development and without minimal immuno-depots in situ. The clinical picture is characterised by destruction of the small vessels with the main manifestations in the upper and lower respiratory tract, in the lungs and kidneys. The eye manifestations were noted yet not so often. There were 2 clinical cases, aged 5-12 years, with primary eye manifestations of the mPA. The initial oedema and the reddening of the lid area were common to all the patients. The diagnostic excisions of the inflammatory tumour of the anterior orbital area have shown histologically vasculitis of the small vessels. mPA was diagnosed by clinical and immunological parameters, including the examination of the respiratory tract and kidneys. In inflammatory orbital pseudotumour, diagnostic excision and evaluation of the immunological parameters are useful for the exclusion of a systemic vasculitis. Proper examinations must be performed to determine other possible organ manifestations. Prognosis is characterised by the severity of kidney involvement. Early immunosuppressive therapy is crucially important for survival prognosis.


The association between clinical, sociodemographic, familial, and environmental factors and treatment resistance in schizophrenia: A machine-learning-based approach.
Prediction of treatment resistance in schizophrenia (TRS) would be helpful to reduce the duration of ineffective treatment and avoid delays in clozapine initiation. We applied machine learning to identify clinical, sociodemographic, familial, and environmental variables that are associated with TRS and could potentially predict TRS in the future. Baseline and follow-up data on trait(-like) variables from the Genetic Risk and Outcome of Psychosis (GROUP) study were used. For the main analysis, we selected patients with non-affective psychotic disorders who met TRS (n = 200) or antipsychotic-responsive criteria (n = 423) throughout the study. For a sensitivity analysis, we only selected patients who met TRS (n = 76) or antipsychotic-responsive criteria (n = 123) at follow-up but not at baseline. Random forest models were trained to predict TRS in both datasets. SHapley Additive exPlanation values were used to examine the variables' contributions to the prediction. Premorbid functioning, age at onset, and educational degree were most consistently associated with TRS across both analyses. Marital status, current household, intelligence quotient, number of moves, and family loading score for substance abuse also consistently contributed to the prediction of TRS in the main or sensitivity analysis. The diagnostic performance of our models was modest (area under the curve: 0.66-0.69). We demonstrate that various clinical, sociodemographic, familial, and environmental variables are associated with TRS. Our models only showed modest performance in predicting TRS. Prospective large multi-centre studies are needed to validate our findings and investigate whether the model's performance can be improved by adding data from different modalities.


Improved response to nab-paclitaxel compared with cremophor-solubilized paclitaxel is independent of secreted protein acidic and rich in cysteine expression in non-small cell lung cancer.
The secreted protein acidic and rich in cysteine (SPARC) is a matricellular glycoprotein that is produced by tumor and/or neighboring stroma. SPARC expression is thought to facilitate the intracellular accumulation of nanoparticle albumin-bound paclitaxel (nab-paclitaxel, abraxane [ABX]). Gene hypermethylation is a common mechanism for loss of SPARC expression in non-small cell lung cancer (NSCLC). We aim to demonstrate the role of SPARC expression as biomarker for treatment selection using ABX in NSCLC and to evaluate the presence of synergistic antitumor effect when a demethylating agent is combined with ABX. We analyzed the SPARC messenger RNA expression and SPARC gene methylation status in 13 NSCLC cell lines and 22 minimally passaged patient-derived (PD) NSCLC tumors using real-time (RT) polymerase chain reaction. The effect of ABX on tumor growth was compared with cremophor-solubilized paclitaxel (taxol) in severe combined immunodeficiency mice bearing SPARC-positive PD xenografts. The effect of pretreatment with a demethylating agent, 5-Aza-2'-deoxycytidine (DEC) in SPARC-negative tumors was assessed. SPARC expression was weak to absent in 62% of established NSCLC cell lines and 68% of PD NSCLC tumor xenografts. SPARC expression could be up-regulated/restored by DEC treatment in both SPARC-negative cell lines and PD xenografts in vitro and in vivo. ABX demonstrated better antitumor efficacy than equitoxic dose of taxol in SPARC-expressing xenografts and some SPARC-negative xenografts. At equimolar doses in vitro, there was similar increased cytotoxicity on DEC pretreatment with either ABX or taxol in SPARC-negative cell lines. At equitoxic doses, there was similar additive antitumor activity of DEC with either ABX or taxol in SPARC-negative PD xenografts. Endogenous SPARC status is somewhat uncorrelated with response to ABX in NSCLC. The greater antitumor effect of ABX compared with equitoxic dose of taxol observed in SPARC-expressing NSCLC tumors can also be seen in some SPARC-negative tumors. DEC pretreatment similarly enhanced antitumor activity with either ABX or taxol in SPARC-negative tumors.


The impact of utilizing different optical coherence tomography devices for clinical purposes and in multiple sclerosis trials.
Optical coherence tomography (OCT) derived retinal measures, particularly peri-papillary retinal nerve fiber layer (RNFL) thickness, have been proposed as outcome measures in remyelinating and neuroprotective trials in multiple sclerosis (MS). With increasing utilization of multiple centers to improve power, elucidation of the impact of different OCT technologies is crucial to the design and interpretation of such studies. In this study, we assessed relation and agreement between RNFL thickness and total macular volume (in MS and healthy controls) derived from three commonly used OCT devices: Stratus time-domain OCT, and Cirrus HD-OCT and Spectralis, two spectral-domain (SD) OCT devices. OCT was performed on both Cirrus HD-OCT and Stratus in 229 participants and on both Cirrus HD-OCT and Spectralis in a separate cohort of 102 participants. Pearson correlation and Bland-Altman analyses were used to assess correlation and agreement between devices. All OCT retinal measures correlated highly between devices. The mean RNFL thickness was 7.4 µm lower on Cirrus HD-OCT than Stratus, indicating overall poor agreement for this measurement between these machines. Further, the limits of agreement (LOA) between Cirrus HD-OCT and Stratus were wide (-4.1 to 18.9 µm), indicating poor agreement at an individual subject level. The mean RNFL thickness was 1.94 µm (LOA: -5.74 to 9.62 µm) higher on Spectralis compared to Cirrus HD-OCT, indicating excellent agreement for this measurement across this cohort. Although these data indicate that these three devices agree poorly at an individual subject level (evidenced by wide LOA in both study cohorts) precluding their co-utilization in everyday practice, the small difference for mean measurements between Cirrus HD-OCT and Spectralis indicate pooled results from these two SD-devices could be used as outcome measures in clinical trials, provided patients are scanned on the same machine throughout the trial, similar to the utilization of multiple different MRI platforms in MS clinical trials.


Allegheny college hosts neuroscience and humanities summer institute.
The Neuroscience and Humanities Summer Institute, hosted by Allegheny College, opened doors of opportunity, perception, and creativity for faculty and students across the nation. Offered first in 2002, and a second time in June of 2004, this weeklong event was designed to provide a medium for fostering development of interdisciplinary courses linking neuroscience and the humanities (e.g., the fine arts, philosophy and language). During the Institute, participants attended presentations by Allegheny faculty introducing the six courses of this type that they have developed starting in 2000, lectures by guest speakers, workshops, and discussion modules. Participants were encouraged to gather ideas about Allegheny's neuroscience and humanities courses and formulate specific plans to take back to their schools. These opportunities and experiences resulted in the formation of valuable connections and the development of ideas around the links between neuroscience and humanities.


Investigating the antioxidant and acetylcholinesterase inhibition activities of Gossypium herbaceam.
Our previous research showed that standardized extract from the flowers of the Gossypium herbaceam labeled GHE had been used in clinical trials for its beneficial effects on brain functions, particularly in connection with age-related dementia and Alzheimer's disease (AD). The aim of this work was to determine the components of this herb and the individual constituents of GHE. In order to better understand this herb for AD treatment, we investigated the acetylcholinesterase (AChE) inhibition and antioxidant activity of GHE as well as the protective effects to PC12 cells against cytotoxicity induced by tertiary butyl hydroperoxide (tBHP) using in vitro assays. The antioxidant activities were assessed by measuring their capabilities for scavenging 1,1-diphenyl-2-picylhydrazyl (DPPH) and 2-2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical as well as in inhibiting lipid peroxidation. Our data showed that GHE exhibited certain activities against AChE and also is an efficient free radical scavenger, which may be helpful in preventing or alleviating patients suffering from AD.


The evolution of human rights in World Health Organization policy and the future of human rights through global health governance.
The World Health Organization (WHO) was intended to serve at the forefront of efforts to realize human rights to advance global health, and yet this promise of a rights-based approach to health has long been threatened by political constraints in international relations, organizational resistance to legal discourses, and medical ambivalence toward human rights. Through legal research on international treaty obligations, historical research in the WHO organizational archives, and interview research with global health stakeholders, this research examines WHO's contributions to (and, in many cases, negligence of) the rights-based approach to health. Based upon such research, this article analyzes the evolving role of WHO in the development and implementation of human rights for global health, reviews the current state of human rights leadership in the WHO Secretariat, and looks to future institutions to reclaim the mantle of human rights as a normative framework for global health governance.


Soluble CD40 ligand is elevated in type 1 diabetic nephropathy but not predictive of mortality, cardiovascular events or kidney function.
Soluble CD40 ligand (sCD40L) derived from platelets mediates atherothrombosis, leading to proinflammatory and proatherosclerotic responses. We investigated the predictive value of plasma sCD40L for all-cause mortality, cardiovascular mortality and morbidity, progression towards end-stage renal disease (ESRD) and rate of decline in glomerular filtration rate (GFR) in patients with type 1 diabetes (T1DM) and nephropathy. The study was a prospective, observational follow-up study of 443 T1DM patients with diabetic nephropathy (274 men; age 42.1 ± 10.5 years [mean ± SD], duration of diabetes 28.3 ± 8.9 years, GFR 76 ± 33 ml/min/1.73 m2) and a control group of 421 patients with longstanding type 1 diabetes and persistent normoalbuminuria (232 men; age 45.4 ± 11.5 years, duration of diabetes 27.7 ± 10.1 years) at baseline. sCD40L was measured by ELISA. Plasma sCD40L levels were higher in patients with diabetic nephropathy compared to normoalbuminuric patients (median (range) 1.54 (0.02-13.38) vs. 1.30 (0.04-20.65) µg/L, respectively p = 0.004). The patients were followed for 8.1 (0.0-12.9) years (median (range)). Among normoalbuminuric patients, sCD40L levels did not predict all-cause mortality (p = 0.33) or combined fatal and non-fatal cardiovascular disease (CVD) (p = 0.27). Similarly, among patients with diabetic nephropathy, the covariate adjusted sCD40L levels did not predict all-cause mortality (p = 0.86) or risk of fatal and non-fatal CVD (p = 0.08). Furthermore, high levels of sCD40L did not predict development of ESRD (p = 0.85) nor rate of decline in GFR (p = 0.69). Plasma sCD40L is elevated in T1DM nephropathy but is not a predictor of all-cause mortality, cardiovascular mortality and morbidity or deterioration of kidney function


Brain mechanisms impacted by psychological therapies for pain: identifying targets for optimization of treatment effects.
Psychological therapies, such as cognitive behavioral therapy, are widely used multifaceted approaches that have been shown to improve pain-related functioning. A small but growing number of studies have used brain imaging to support the use of psychological therapies for pain. Although these studies have led to an increased understanding of how therapies may engage neural systems, there are multiple technical and conceptual challenges to consider. Based on the current literature, several components of effective psychological therapies for pain may be supported by changes in neural circuitry, which are most consistently represented by diminished activation and/or reduced hyperconnectivity in brain regions related to pain processing, emotion, and cognitive control. Findings may vary based on methodological approaches used and may also differ depending on targets of treatment. To provide a nuanced understanding of the current literature, specific targets and components of effective treatments for which a neural basis has been investigated are reviewed. These treatment components include catastrophic thinking about pain, increasing self-efficacy, mindfulness, anxiety symptom reduction, and exposure-based approaches. In general, such strategies have the potential to normalize regional hyperactivations and reduce hyperconnectivity in brain regions associated with nociceptive processing, cognition, and emotion, although additional research is needed. By determining if there are indeed distinct brain mechanisms engaged by different components of psychological therapy and evidence for specific changes in neural function after these interventions, future therapies may be more optimally tailored for individuals afflicted with chronic pain.


Purine (N)-Methanocarba Nucleoside Derivatives Lacking an Exocyclic Amine as Selective A3 Adenosine Receptor Agonists.
Purine (N)-methanocarba-5'-N-alkyluronamidoriboside A3 adenosine receptor (A3AR) agonists lacking an exocyclic amine resulted from an unexpected reaction during a Sonogashira coupling and subsequent aminolysis. Because the initial C6-Me and C6-styryl derivatives had unexpectedly high A3AR affinity, other rigid nucleoside analogues lacking an exocyclic amine were prepared. Of these, the C6-Me-(2-phenylethynyl) and C2-(5-chlorothienylethynyl) analogues were particularly potent, with human A3AR Ki values of 6 and 42 nM, respectively. Additionally, the C2-(5-chlorothienyl)-6-H analogue was potent and selective at A3AR (MRS7220, Ki 60 nM) and also completely reversed mouse sciatic nerve mechanoallodynia (in vivo, 3 μmol/kg, po). The lack of a C6 H-bond donor while maintaining A3AR affinity and efficacy could be rationalized by homology modeling and docking of these hypermodified nucleosides. The modeling suggests that a suitable combination of stabilizing features can partially compensate for the lack of an exocyclic amine, an otherwise important contributor to recognition in the A3AR binding site.


Transcriptional regulation of the HIV-1 inhibitory factor human mannose receptor 1 by the myeloid-specific transcription factor PU.1.
HIV-1 infection of cells results in the modulation of cellular gene expression by virus-encoded proteins in a manner that benefits the virus. We reported that HIV-1 transactivator of transcription (Tat) dysregulates the expression of the human mannose receptor 1 (hMRC1). hMRC1 is involved in the innate immune response of macrophages to foreign pathogens. Tat does not act directly on the hMRC1 promoter but instead inhibits PU.1, a cellular transcription factor regulating hMRC1 gene expression. Here, we characterize the PU.1-dependent regulation of hMRC1 expression. We identified four potential PU.1 binding sites in the hMRC1 promoter region but found that only one, PU.1(b), functioned as a true binding site for PU.1. Transfer of the PU.1(b) box to a heterologous promoter did not activate this promoter per se but rendered it responsive to PU.1. Our results support the view that PU.1 acts as a transcriptional co-factor whose activity can be regulated by HIV-1 Tat.


Indices of cholesterol metabolism and relative responsiveness to ezetimibe and simvastatin.
The level and duration of exposure to circulating low-density lipoprotein-cholesterol (LDL-C) are major contributors to coronary atherosclerosis. Therefore, optimal prevention will require long-term LDL-C reduction, making it important to select the most effective agent for each individual. We tested the hypothesis that individuals with high fractional absorption of cholesterol respond better to the cholesterol absorption inhibitor ezetimibe than to simvastatin, whereas low absorbers, who have elevated rates of cholesterol synthesis, respond better to simvastatin. A randomized, double-blind, placebo-controlled, crossover trial was performed in 215 African- and European-American men. Participants were randomized to placebo, ezetimibe (10 mg/d), simvastatin (10 mg/d), and both drugs for 6 wk each. Plasma levels of LDL-C, surrogate markers for cholesterol absorption (campesterol) and synthesis (lathosterol), and proprotein convertase subtilisin-like kexin type 9 were measured at baseline and after treatment. LDL-C levels were reduced by 19% (ezetimibe), 25% (simvastatin), and 41% (ezetimibe+simvastatin) from a baseline of 146 +/- 20 mg/dl; results were similar between ethnic groups. Reduction in LDL-C correlated poorly with baseline levels of noncholesterol sterols and proprotein convertase subtilisin-like kexin type 9. Although individual responses varied widely, change in LDL-C on ezetimibe correlated with response to simvastatin (r = 0.46, P < 0.001). Combination therapy lowered LDL-C by 15% or greater in more than 95% of participants. Baseline cholesterol absorption and synthesis did not predict responsiveness to LDL-lowering drugs. Responsiveness to simvastatin and ezetimibe were highly correlated, suggesting that factors downstream of the primary sites of action of these drugs are a major determinant of response.


Mangiferin protects against alcoholic liver injury via suppression of inflammation-induced adipose hyperlipolysis.
Adipose dysfunction is closely associated with alcoholic liver disease. The impact of mangiferin on ethanol-induced liver injury and the probable underlying molecular mechanism has not been sufficiently addressed. In the present study, mice were subjected to a chronic plus a single binge ethanol feeding to induce liver injury. In addition, the differentiated adipocytes from primary mouse adipocytes were isolated and used for the mechanism studies. Our study demonstrated that mangiferin protects against ethanol induced adipose hyperlipolysis by restoring PDE3B stability, which is associated with activating the AMPK/TBK1 signaling and suppressing the noncanonical NF-κB activation, leading to the reduction of free fatty acid release and the amelioration of ethanol-induced liver injury. Our findings identify that mangiferin ameliorates alcoholic liver injury via suppression of inflammation-induced adipose hyperlipolysis, suggesting that mangiferin might be a potential effective agent for the management of alcoholic liver injury.


Sensitizing antigen-specific CD8+ T cells for accelerated suicide causes immune incompetence.
Death receptor-mediated activation-induced apoptosis of antigen-specific T cells is a major mechanism of peripheral tolerance induction and immune homeostasis. Failure to undergo activation-induced cell death (AICD) is an important underlying cause of many autoimmune diseases. Thus, enhancing the T cell's own suicide mechanism may provide an efficient therapy for the treatment of autoimmune diseases. Bisindolylmaleimide VIII (Bis VIII), a PKC inhibitor, can sensitize T cells for death receptor-induced apoptosis and thus can inhibit the development of T cell-mediated autoimmune disease in vivo. In this study, we have analyzed the functional consequences of accelerated suicide for a protective CD8+ T cell-mediated immune response. Our data indicate that CD8+ T cells are sensitized by Bis VIII to AICD, both in vitro and in vivo. The sensitizing effect of Bis VIII appears to be mediated by specific downmodulation of the antiapoptotic molecule cellular FLICE-like inhibitory protein (cFLIP(L)). Importantly, Bis VIII administration during an acute lymphocytic choriomeningitis virus (LCMV) infection causes the depletion of virus-specific CD8+ T cells and subsequently impaired cytotoxicity and virus clearance. We conclude that resistance to death receptor-induced apoptosis is crucial for the efficient induction of a protective immune response, and that Bis VIII-based immunotherapies have to be applied under well-controlled conditions to avoid the induction of immune incompetence and the inability to respond to pathogen infection.


Thalidomide and a Dipeptidyl Peptidase 4 Inhibitor in a Rat Model of Experimental Autoimmune Myocarditis.
Myocarditis is a potentially fatal disease, but curative treatments have not yet been established. Myocardial inflammation is an important pathogenesis of this disease, and immunosuppressants such as methylprednisolone and immunoglobulin have been used for treatment; however, the effectiveness needs to be improved. Thalidomide and dipeptidyl peptidase (DPP) 4 inhibitors were recently investigated regarding their immunomodulatory properties. This study aimed to test whether thalidomide or a DPP4 inhibitor (evogliptin) can improve the effectiveness of myocarditis treatment using a rat model of experimental autoimmune myocarditis (EAM). Rats with or without myocarditis were administered thalidomide at 100 mg/kg/day and DPP4 inhibitor at 10 mg/kg/day orally. Measurement of echocardiography, serum inflammatory cytokines, myocardial histopathological examination, and immunohistochemical staining for leukocytes, macrophages, CD4+ T cells, and cytoskeleton were performed after 3 weeks, and the fibrosis area was measured after 3 and 6 weeks. Thalidomide and DPP4 inhibitor did not reduce the severity of myocarditis compared with the EAM without treatment rats by comparing the echocardiographic data, myocardial CD4+, macrophages, neutrophil infiltrations, and the heart weight/body weight ratio in 3 weeks. The levels of inflammatory cytokines were not lower in the thalidomide and DPP4 inhibitor-treated group than in the untreated group in 3 weeks. In 6 weeks, thalidomide and DPP4 inhibitors did not reduce the fibrosis area compared to untreated groups. Although thalidomide and the DPP4 inhibitor had an immunomodulatory effect and are used against inflammatory diseases, they did not ameliorate myocardial inflammation and fibrosis in this rat model of EAM.


An atypical form of alphaB-crystallin is present in high concentration in some human cataractous lenses. Identification and characterization of aberrant N- and C-terminal processing.
Two unique polypeptides, 22.4 and 16.4 kDa, were prominent in some human cataracts. Both proteins were identified as modified forms of the small heat shock protein, alphaB-crystallin. The concentration of total alphaB-crystallin in most of these cataracts was significantly increased. The 22.4-kDa protein was subsequently designated as alphaB(g). Mass spectrometric analyses of tryptic and Asp-N digests showed alphaB(g) is alphaB-crystallin minus the C-terminal lysine. alphaB(g) constituted 10-90% of the total alphaB-crystallin in these cataracts and was preferentially phosphorylated over the typical form of alphaB-crystallin. Human alphaB(g) and alphaB-crystallin were cloned and expressed in Escherichia coli. The differences in electrophoretic mobility and the large difference in native pI values suggest some structural differences exist. The chaperone-like activity of recombinant human alphaB(g) was comparable to that of recombinant human alphaB-crystallin in preventing the aggregation of lactalbumin induced by dithiothreitol. The mechanism involved in generating alphaB(g) is not known, but a premature termination of the alphaB-crystallin gene was ruled out by sequencing the polymerase chain reaction products of the last exon for the alphaB-crystallin gene from lenses containing alphaB(g). The 16.4-kDa protein was an N-terminally truncated fragment of alphaB(g). The high concentration of alphaB-crystallin in these cataracts is the first observation of this kind in human lenses.


The impact of successful learning of self-regulation on reward processing in children with ADHD using fMRI.
Neurofeedback (NF) is a non-pharmacological treatment for attention-deficit/hyperactivity disorder (ADHD) that is targeting self-regulation, is efficacious when standard protocols are used and induces partly specific neurophysiological changes in the inhibitory network. However, its effects on reward processing, which is also considered an important aspect of ADHD and has been linked to neurophysiological deficits, remain unknown. Children with ADHD (N = 15, mean age 11.8, SD 1.52) were randomly assigned to either slow cortical potential NF (n = 8) or EMG biofeedback control training (n = 7) and received 20 sessions of training under comparable conditions. Learning was defined as the slope of successful training runs across all transfer sessions. Whole brain analysis, region-of-interest analysis of anticipatory ventral striatal (VS) activation, and analysis of behavioral data were performed. Clinically, the NF group improved more than the EMG group. Whole brain analysis indicated increased activation in the left superior frontal gyrus in the control group only, and in medial prefrontal cortex and dorsolateral prefrontal gyrus (DLPFC) after treatment across all groups. Only successful learners of self-regulation (n = 8) showed increased left inferior frontal gyrus and DLPFC activation after treatment. Left VS activation was increased after treatment and showed a significant time*medication-status interaction. Specific treatment effects were found in left frontal regions for the control treatment and successful learners. Also, unmedicated participants, irrespective of treatment type or successful learning, showed treatment-induced improvement in reward processing. The results suggest no prominent specific effect of NF on reward processing. However, cautious interpretation is warranted due to the small sample.


Rationale for the NCCLS standard H20-T for differential leukocyte counting.
The National Committee for Clinical Laboratory Standards (NCCLS) tentative standard for Leukocyte Differential Counting (H20-T) was developed in the United States as a reference method for differential leukocyte counting as well as a method for the evaluation of the performance of differential leukocyte counters. The key features of this document are outlined and the reasons for their inclusion in the standard are given.


Pancreatic cysts: Diagnostic accuracy and risk of inappropriate resections.
Pancreatic cystic neoplasms (PCN) frequently undergo surgery, given malignant potential. Pancreatic cyst surgery is associated with significant rates of morbidity and mortality. It is crucial to accurately characterize these lesions pre-operatively to avoid unnecessary surgery in patients with benign pancreatic cysts. We aimed to assess the correlation between pre-operative (pre-op) diagnosis based on imaging and clinical presentation, and post-operative (post-op) diagnosis based on histopathology in patients undergone pancreatic cyst surgery. From January 2000 to January 2012, we randomly selected 2000 patients with ICD-9 code 211.6 and 577.2. Amongst these we identified 281 patients undergone pancreas surgery. Patients with no pre-op imaging or non-cyst indication for surgery were excluded (n = 107). Imaging details, demographics, pre-operative physician diagnosis and histopathologic details of pancreatic cysts were recorded in 174 patients. There was a discrepancy between the pre- and post-operative pancreatic cyst diagnosis in 54 (31%) patients. There was no difference in the proportion of various imaging studies (CT, EUS or MRI) between patients with a correct and patients with an incorrect pre-op diagnosis. The pre-op diagnosis was confirmed at pathology in 87.5% of the presumed SCNs, in 80% of the presumed pseudocysts, in 73.3% of the presumed BD-IPMNs, in 66.7% of the presumed MD/mixed-IPMNs and in 53.6% of the presumed MCNs. The accuracy of the pre-operative diagnosis of presumed MCN was significantly lower compared to the non-MCN cysts (53.6% vs. 75%; p = 0.037). Fourteen percent of resections were performed for asymptomatic benign cysts, preoperatively suspected to be potentially pre-malignant cysts. In nearly 1 out of 3 patients undergone pancreas cyst surgery, there is a discrepancy between pre- and post-op diagnosis. Pre-op diagnosis of presumed MCN is more likely to be incorrect, compared to the other cysts.


[Ineffectiveness of hyperbaric oxygen therapy in multiple sclerosis. A randomized placebo-controlled double-blind study].
Seventeen patients with definite and progressive multiple sclerosis entered a double-blind randomized placebo-controlled therapeutic trial of hyperbaric oxygen (HBO). Exposure in monoplace chamber was 90 minutes long each time, 5 days a week, for 4 weeks. The treatment and placebo groups received 100% oxygen at 1.5 bar constant pressure or normal air at 0.1-0.2 bar, respectively. The clinical status of the patients in both groups were compared until one year after treatment. There was no benefit of HBO versus placebo according to the Kurtzke disability status scale. A lesser proportion of patients with deterioration of bowel/bladder function 12 months after therapy was the only benefit of HBO versus placebo according to Kurtzke functional systems scales. On the whole however, HBO is useless in the management of progressive forms of multiple sclerosis.


Epstein-Barr virus latent infection membrane protein alters the human B-lymphocyte phenotype: deletion of the amino terminus abolishes activity.
A latent infection membrane protein (LMP) encoded by the Epstein-Barr virus (EBV) genome in latently infected, growth-transformed lymphocytes alters the phenotype of a human EBV-negative B-lymphoma cell line (Louckes) when introduced by gene transfer. These LMP-expressing cells exhibit increased homotypic adhesion due to increased expression of the adhesion molecules LFA-1 and ICAM-1. Increased homotypic adhesion could foster B-cell growth by facilitating autocrine growth factor effects. LFA-3 expression is also induced. The induction of LFA-3 and ICAM-1 results in increased heterotypic adhesion to T lymphocytes. This could result in more effective T-cell immune surveillance. Since LMP is expressed in EBV-transformed lymphocytes and has been demonstrated to transform rodent fibroblasts in vitro, a wide range of possible effects on B-lymphoma cell growth were assayed. In the Louckes B-lymphoma cell line, EBV LMP causes increased cell size, acid production, plasma membrane ruffling, and villous projections. Although cell proliferation rate was not greatly affected, the steady-state intracellular free calcium level, transforming growth factor beta responsiveness, and expression of the lymphocyte activation markers (CD23 and transferrin receptor) were increased. Thus, LMP appears to be a mediator of EBV effects on B-cell transformation. In transfected lymphoma cells, LMP localizes to patches at the cell periphery and associates with the cytoskeleton as it does in EBV-transformed B lymphocytes or in rodent fibroblasts. A partially deleted form of LMP (D1LMP) does not aggregate in patches or associate with the cytoskeleton and had little effect on B-cell growth. Thus, cytoskeletal association may be integral to LMP activity.


Arsenic poisoning.
This discussion was selected from the weekly Grand Rounds in the Department of Medicine, University of New Mexico School of Medicine, Albuquerque. Taken from a transcription, it has been edited by Ralph C. Williams, Jr, MD, Professor and Chair of the Department of Medicine.


Angiotensin-converting enzyme inhibitors in essential hypertension.
At the Cambridge Conference, the pros and cons of various classes of antihypertensive drugs were extensively discussed, including the place of angiotensin-converting enzyme inhibitors in today's spectrum. A consensus was easily reached that the most important advantage of having the present broad armamentarium of antihypertensive drugs is that a wide choice is now available from which to find the most suitable agent for the individual patient. A revised stepped-care design was proposed, in which the doctor has the choice of starting antihypertensive therapy with a thiazide diuretic, a beta-blocker, an angiotensin-converting enzyme inhibitor, or a calcium antagonist. Small doses of any agent should be used to start with, and doses should not subsequently be increased beyond that at which side effects appear. Should such symptoms occur, the doctor has the choice of either switching to another first-step compound or reducing the dose of the first agent and combining it with one of other available drugs.


30-Day outcomes analysis of NSQIP-pediatric for surgical management of head and neck lymphatic malformations in children.
Identify adverse events associated with surgical management of head and neck lymphatic malformations in children by investigating a US national database. Pediatric American College of Surgeons (ACS) National Surgical Quality Improvement Program® (ACS NSQIP® - pediatric), years 2012-2016. Subjects included children under 18 years with a postoperative diagnosis of International Classification of Diseases (ICD), 9 th revision code: 228.1 - lymphangioma, any site or ICD-10 code D18.1. 163 patients were identified. 67 (41.1%) were female and 96 (58.9%) were male. The median age was 4.2 years (interquartile range [IQR] 1.6-11.3 years). Sixty-five (39.9%) underwent head and neck procedures. 19 (11.7%) patients with 30-day adverse events were identified, with unplanned reoperation being the most common; in the head and neck surgery group, there were 5 (7.7%). There was no significant difference in the occurrence of adverse events between head and neck surgery and the non-head and neck surgery group (p = 0.2238). No individual patient clinical factors were identified to be associated with increased rates of 30-day adverse event rates for surgical management of lymphatic malformations in children. Our investigation highlights the need for quality improvement to help decrease 30-day adverse events for surgery for this group, as the rate was quite high overall (11.7%). The wound complication rate was very low, but we observed a non-negligible number of reoperations and readmissions. Improving quality of care for surgical management of lymphatic malformations in children should focus future investigations on limiting reoperations and readmissions within the first 30 days postoperatively.


Inducible and reversible Clock gene expression in brain using the tTA system for the study of circadian behavior.
The mechanism of circadian oscillations in mammals is cell autonomous and is generated by a set of genes that form a transcriptional autoregulatory feedback loop. While these "clock genes" are well conserved among animals, their specific functions remain to be fully understood and their roles in central versus peripheral circadian oscillators remain to be defined. We utilized the in vivo inducible tetracycline-controlled transactivator (tTA) system to regulate Clock gene expression conditionally in a tissue-specific and temporally controlled manner. Through the use of Secretogranin II to drive tTA expression, suprachiasmatic nucleus- and brain-directed expression of a tetO::Clock(Delta19) dominant-negative transgene lengthened the period of circadian locomotor rhythms in mice, whereas overexpression of a tetO::Clock(wt) wild-type transgene shortened the period. Low doses (10 mug/ml) of doxycycline (Dox) in the drinking water efficiently inactivated the tTA protein to silence the tetO transgenes and caused the circadian periodicity to return to a wild-type state. Importantly, low, but not high, doses of Dox were completely reversible and led to a rapid reactivation of the tetO transgenes. The rapid time course of tTA-regulated transgene expression demonstrates that the CLOCK protein is an excellent indicator for the kinetics of Dox-dependent induction/repression in the brain. Interestingly, the daily readout of circadian period in this system provides a real-time readout of the tTA transactivation state in vivo. In summary, the tTA system can manipulate circadian clock gene expression in a tissue-specific, conditional, and reversible manner in the central nervous system. The specific methods developed here should have general applicability for the study of brain and behavior in the mouse.


Interferon-beta-1 b decreased matrix metalloproteinase-9 serum levels in primary progressive multiple sclerosis.
Recent reports have shown that matrix-metalloproteinases (MMPs) facilitate T-cell migration into the CNS and play a role in disruption of the blood-brain-barrier and myelin break-down. An increase of MMP-9 serum levels predicts disease activity in relapsing remitting multiple sclerosis (RRMS). Interferon-beta (IFN-beta), which is an established treatment for RRMS, inhibits T-cell migration in vitro in parallel with the downregulation of MMP expression. Only limited data are available for primary progressive multiple sclerosis (PPMS) which differs in demographic and immunological aspects as well as in MRI criteria from RRMS. In this study, 19 patients with laboratory-supported definite PPMS were treated with 8 x 10(6) IU IFN-beta1b (Betaferon) subcutaneously every other day. Serum was collected before treatment and on months 1, 2, 3, 6 and 9 during treatment. Levels of MMP-9 and of its natural inhibitor known as tissue-inhibitor of matrix-metalloproteinase-1 (TIMP-1) were quantified by ELISA. In addition MMP-2 serum levels were determined by zymography. 19 healthy volunteers served as controls. Before treatment serum levels of MMP-9 were elevated in patients with PPMS compared with controls, whereas there was no difference in TIMP-1 serum levels. During treatment with IFN- beta1b the concentration of MMP-9 in the serum of 18 out of 19 PPMS patients decreased,whereas serum levels of MMP-2 and TIMP-1 remained nearly unaffected. Our results demonstrate that the MMP-9 to TIMP-1 ratio in patients with PPMS is elevated in comparison with healthy controls. The suppression of MMP-9 by IFN-beta1b indicates that this drug is immunomodulatory active in PPMS patients. Further studies are necessary to test if IFN-beta exerts a beneficial effect in PPMS.


Tailored Parameterization of the LIE Method for Calculating the Binding Free Energy of Vps34-Inhibitor Complexes.
Vps34 is the only isoform of the PI3K family in fungi, making this protein an attractive target to develop new treatments against pathogenic fungi. The high structural similarity between the active sites of the human and fungal Vps34 makes repurposing of human Vps34 inhibitors an appealing strategy. Nonetheless, while some of the cross-reactive inhibitors might have the potential to treat fungal infections, a safer approach to prevent undesired side effects would be to identify molecules that specifically inhibit the fungal Vps34. This study presents the parameterization of four LIE models for estimating the binding free energy of Vps34-inhibitor complexes. Two models are parameterized using a multiparametric linear regression leaving one or more free parameters, while the other two are based on the LIE-D model. All of the models show good predictive capacity (R 2 > 0.7, r > 0.85) and a low mean absolute error (MAE < 0.71 kcal/mol). The current study highlights the advantages of LIE-D-derived models when predicting the weight of the different contributions to the binding free energy. It is expected that this study will provide researchers with a valuable tool to identify new Vps34 inhibitors for relevant applications such as cancer treatment and the development of new antimicrobial agents.


Utility of smart watches for identifying arrhythmias in children.
Arrhythmia symptoms are frequent complaints in children and often require a pediatric cardiology evaluation. Data regarding the clinical utility of wearable technologies are limited in children. We hypothesize that an Apple Watch can capture arrhythmias in children. We present an analysis of patients ≤18 years-of-age who had signs of an arrhythmia documented by an Apple Watch. We include patients evaluated at our center over a 4-year-period and highlight those receiving a formal arrhythmia diagnosis. We evaluate the role of the Apple Watch in arrhythmia diagnosis, the results of other ambulatory cardiac monitoring studies, and findings of any EP studies. We identify 145 electronic-medical-record identifications of Apple Watch, and find arrhythmias confirmed in 41 patients (28%) [mean age 13.8 ± 3.2 years]. The arrythmias include: 36 SVT (88%), 3 VT (7%), 1 heart block (2.5%) and wide 1 complex tachycardia (2.5%). We show that invasive EP study confirmed diagnosis in 34 of the 36 patients (94%) with SVT (2 non-inducible). We find that the Apple Watch helped prompt a workup resulting in a new arrhythmia diagnosis for 29 patients (71%). We note traditional ambulatory cardiac monitors were worn by 35 patients (85%), which did not detect arrhythmias in 10 patients (29%). In 73 patients who used an Apple Watch for recreational or self-directed heart rate monitoring, 18 (25%) sought care due to device findings without any arrhythmias identified. We demonstrate that the Apple Watch can record arrhythmia events in children, including events not identified on traditionally used ambulatory monitors.


Enhancement of antitumor immunity by expression of CD70 (CD27 ligand) or CD154 (CD40 ligand) costimulatory molecules in tumor cells.
CD70 (CD27 ligand (CD27L)), CD153 (CD30L), and CD154 (CD40L) are members of the tumor necrosis factor family of costimulatory molecules and expressed on the surface of T cells that are important for both T- and B-cell help. We examined the capacity for expression of these tumor necrosis factor family members on tumor cells to induce an antitumor response either in the presence or absence of interleukin 12. Retroviral vectors were constructed that expressed high levels of membrane bound CD70, CD153, or CD154 following infection and selection of the murine tumor lines MCA 207 or TS/A. The genetically modified tumor cells expressing these molecules were able to stimulate splenic cell proliferation, demonstrating that the expressed costimulatory molecules were biologically active. When tested for tumor establishment, the expression of either CD70 or CD154 was able to slow tumor growth. Similarly, CD70 or CD154 were able to induce antitumor immunity at a higher frequency when tested in vaccination and therapy models. CD70 was able to induce antitumor immunity at a level similar to CD80 when tested either in the presence or absence of interleukin 12. Moreover, coexpression of CD70 and CD80 was able to synergize the induction of a higher frequency of antitumor immunity in a vaccination model. Taken together, our results suggest that CD154 and in particular CD70 are able to contribute to the induction of the immune response to tumor in murine models and thus may be of use for human clinical trials.


Effect of opioids on Ca2+/cAMP responsive element binding protein.
Ca2+/cAMP response element binding protein (CREB) is an important factor linking the opioid-regulated secondary messenger systems to alterations in gene expression. Opioids regulate CREB level, its phosphorylation and binding to its corresponding response element in the promoters of several genes implicated in drug addiction. CREB mediates the action of opioids on the expression of several genes in brain regions responsible for drug-seeking behavior and manifestation of signs of dependence. Moreover, alterations in CREB level can effect the rewarding properties of morphine and regulate the self-administration of cocaine. At the cellular level CREB acts as convergence point for different cellular pathways. Opioids affect two different intracellular mediator systems: inhibitory--connected with cAMP, and stimulatory--involving calcium and the PKC pathway. Both can affect CREB but in different phases of opiate action. The presence of this biphasic mechanism can explain the phenomenon of the induction of some CRE-controlled genes after both acute and chronic morphine administration. Cellular studies also highlight the relevance of other ATF/CREB family members which can affect Ca2+/cAMP response element (CRE) controlled transcription as well as other transcription factors which make the opioid induction longer lasting.


NK Cell Responses to Human Tick-Borne Encephalitis Virus Infection.
Tick-borne encephalitis virus (TBEV) is a flavivirus that is transferred to humans by infected ticks. The virus causes tick-borne encephalitis, a severe infection of the CNS with a high risk for long-lasting sequelae. Currently, no treatment exists for the disease. Understanding the cellular immune response to this infection is important to gain further understanding into the pathogenesis, treatment, and prevention of the disease. NK cells are known to participate in the control of viral infections. We performed a longitudinal analysis of the human NK cell response to TBEV infection in a cohort of infected individuals from the onset of severe clinical symptoms to the convalescence phase. NK cell activation, as measured by expression of Ki67, was apparent at the time of hospitalization. By 3 wk after hospitalization, it decreased to levels seen in healthy controls. Concomitant with the increase in NK cell activation, augmented levels of IL-12, IL-15, IL-18, IFN-γ, and TNF were detected in patient plasma. This TBEV-induced NK cell activation was restricted predominantly to differentiated CD57(+)CD56(dim) NK cells. Functionally, CD56(dim) NK cells responded poorly to target cells at the time of hospitalization, but they recovered functional capacity to control levels during the convalescent phase. In contrast, the responsiveness of NK cells to cytokine stimulation remained intact throughout the disease. These findings demonstrate that NK cells respond to TBEV infection with characteristics that are distinct from those of other human viral infections and provide insights into the NK cell response to clinical TBEV infection.


Modeling genetic inheritance of copy number variations.
Copy number variations (CNVs) are being used as genetic markers or functional candidates in gene-mapping studies. However, unlike single nucleotide polymorphism or microsatellite genotyping techniques, most CNV detection methods are limited to detecting total copy numbers, rather than copy number in each of the two homologous chromosomes. To address this issue, we developed a statistical framework for intensity-based CNV detection platforms using family data. Our algorithm identifies CNVs for a family simultaneously, thus avoiding the generation of calls with Mendelian inconsistency while maintaining the ability to detect de novo CNVs. Applications to simulated data and real data indicate that our method significantly improves both call rates and accuracy of boundary inference, compared to existing approaches. We further illustrate the use of Mendelian inheritance to infer SNP allele compositions in each of the two homologous chromosomes in CNV regions using real data. Finally, we applied our method to a set of families genotyped using both the Illumina HumanHap550 and Affymetrix genome-wide 5.0 arrays to demonstrate its performance on both inherited and de novo CNVs. In conclusion, our method produces accurate CNV calls, gives probabilistic estimates of CNV transmission and builds a solid foundation for the development of linkage and association tests utilizing CNVs.


The modulation of lymphocyte functions by molecules secreted by macrophages. II. Conditions leading to increased secretion.
Cultures of peritoneal exudate cells rich in macrophages were studied for the secretion of lymphostimulatory molecules. Two conditions produced increased secretion: (a) addition to the cultures of various agents that readily interacted with macrophages, such as latex particles, antibody-coated red cells, endotoxin, Listeria organisms, or Be salt; or (b) addition of activated lymphocytes. In the first case the increased activity was found during the first 24 or 48 h after uptake of the stimuli. Increased activity was found in normal or peptone-stimulated macrophages but not in macrophages after injection of endotoxin or thioglycollate. The addition of T lymphocytes from Listeria-infected mice to macrophage cultures increased greatly the activities. This increase was also produced by addition to antigen-primed T cells together with antigen. The lymphocytes by themselves did not secrete active factors. The lymphostimulatory activities were tested on thymocyte DNA synthesis and on antibody formation in vitro. The latter assay was done on spleen cells from immunized mice where one striking effect was the stimulation of differentiation to antibody-secreting cells. Some dissociation of both activities (thymocyte DNA synthesis and B-cell differentiation) was observed with selected culture fluids.


Rationalising germplasm collections: a case study for wheat.
In total 70 genebank accessions comprising 50 hexaploid, 12 tetraploid and 8 diploid wheats of the Gatersleben collection were selected based on the screening of the passport data for identical cultivar names or accession numbers of the donor genebanks. Twelve potential duplicate groups consisting of three to nine accessions with identical names/numbers were selected and analysed with DNA markers (microsatellites). A bootstrap approach based on re-sampling of both microsatellite markers and alleles within marker loci was used to test for homogeneity. Although several homogeneous groups were identified it became clear that cultivar name identity alone did not allow the determination of duplicates. A combination of SSR-analysis followed by the bootstrap method and database survey considering the botanical classification and other data (origin, growth habit and donor) available is recommended in order to determine duplicates. A procedure for the identification of duplicates and their further handling in ex situ genebanks is discussed.


Quantitative community fingerprinting methods for estimating the abundance of operational taxonomic units in natural microbial communities.
Molecular fingerprinting techniques offer great promise for analyzing changes in microbial community structure, especially when dealing with large number of samples. However, a serious limitation has been the lack of quantification offered by such techniques since the relative abundances of the identified operational taxonomic units (OTUs) in the original samples are not measured. A quantitative fingerprinting approach designated "qfingerprinting" is proposed here. This method involves serial dilutions of the sample of interest and further systematic fingerprinting of all dilution series. Using the ultimate dilutions for which OTU are still PCR amplifiable and taking into account peak size inaccuracy and peak reproducibility, the relative abundance of each OTU is then simultaneously determined over a scale spanning several orders of magnitude. The approach was illustrated by using a quantitative version of automated ribosomal intergenic spacer analysis (ARISA), here called qARISA. After validating the concept with a synthetic mixture of known DNA targets, qfingerprinting was applied to well-studied marine sediment samples to examine specific changes in OTU abundance associated with sediment depth. The new strategy represents a major advance for the detailed quantitative description of specific OTUs within complex communities. Further ecological applications of the new strategy are also proposed.


Cellular control of protein levels: A systems biology perspective.
How cells regulate protein levels is a central question of biology. Over the past decades, molecular biology research has provided profound insights into the mechanisms and the molecular machinery governing each step of the gene expression process, from transcription to protein degradation. Recent advances in transcriptomics and proteomics have complemented our understanding of these fundamental cellular processes with a quantitative, systems-level perspective. Multi-omic studies revealed significant quantitative, kinetic and functional differences between the genome, transcriptome and proteome. While protein levels often correlate with mRNA levels, quantitative investigations have demonstrated a substantial impact of translation and protein degradation on protein expression control. In addition, protein-level regulation appears to play a crucial role in buffering protein abundances against undesirable mRNA expression variation. These findings have practical implications for many fields, including gene function prediction and precision medicine.


Dynamic Response of the Lumbar Spine to Whole-body Vibration Under a Compressive Follower Preload.
A finite element study of dynamic response of the lumbar spine to whole-body vibration. The aim of this study was to develop and validate a finite element model for exploring the impact of whole-body vibration on the entire lumbar spine with a compressive follower preload applied. Several finite element studies have investigated the biodynamic characteristics of the human lumbar spine when exposed to whole-body vibration. However, very limited studies have been performed to quantitatively describe dynamic response in time domain of the entire lumbar spine to vibration loading under a compressive follower preload. A three-dimensional nonlinear finite element model of the human lumbar spine (L1-sacrum) subjected to the compressive follower preload was created. Transient dynamic analysis was conducted on the model to compute the spinal response to a sinusoidal vertical vibration load of ±40 N under a 400 N preload. The obtained dynamic response results at all spinal levels were collected and plotted as a function of time. As a comparison, the corresponding results for vertical static loads (-40 and 40 N) under the preload (400 N) were also computed. Plots of the dynamic response at all levels showed a cyclic response with time, and their vibration amplitudes (peak-to-bottom variations) were markedly higher than the corresponding changing amplitudes of static load cases. The increasing effect of the vibration load reached 314.5%, 263.2%, 242.4%, and 232.7%, respectively, in axial displacement of vertebral center, disc bulge, intradiscal pressure, and annulus stress (von-Mises stress). In addition, increasing the compressive follower preload led to an increase in the dynamic response and a decrease in their vibration amplitudes. This study may be useful to help quantify the effect of cyclic loading on the entire lumbar spine under physiologic compressive loading, and better understand vibration characteristics of the spine. 5.


Helping midlife women predict the onset of the final menses: SWAN, the Study of Women's Health Across the Nation.
Women approaching menopause often ask their doctors, "When are my periods going to end?" The objective of this study was to predict time to the final menstrual period (FMP). This multiethnic, observational cohort study, the Study of Women's Health Across the Nation, has been ongoing since 1996. Data collected from seven annual study visits were used. The community-based cohort from seven national sites included 3,302 white, African American, Hispanic, Chinese, and Japanese women aged 42 to 52 years at baseline with a uterus and at least one ovary, who were not pregnant or taking reproductive hormones, and had at least one menstrual period within the past 3 months at baseline. The time to the FMP was defined retrospectively after 12 months of amenorrhea. Uni- and multivariable Cox proportional hazard models, hazard ratios (HRs), and 95% CIs were computed for variables of interest. A total of 2,662 women, of whom 706 had an observed FMP, were included. Age, menstrual cycles that had become farther apart (HR = 2.56, 95% CI = 1.94-3.39) or more variable (HR = 1.79, 95% CI = 1.45-2.21), and current smoking (HR = 1.68, 95% CI = 1.35-2.08) were all associated with shorter time to the FMP. Higher (log) follicle-stimulating hormone (HR = 2.32, 95% CI = 2.02-2.67) was related to a shorter time to the FMP, but the highest estradiol category (>or=100 pg/mL [367 pmol/L]) was associated with an earlier onset of the FMP (HR = 2.16, 95% CI = 1.63-2.89). The number of vasomotor symptoms was related to an earlier FMP, whereas higher physical activity and educational levels were associated with a later FMP. Age, menstrual cycle recall, smoking status, and hormone measurements can be used to estimate when the FMP will occur, allowing for more precise estimates for older midlife women: in the most extreme cases, ie, age 54, high estradiol level, current smoking, and high follicle-stimulating hormone level, the FMP can be estimated to within 1 year.


Anaerosolibacter carboniphilus gen. nov., sp. nov., a strictly anaerobic iron-reducing bacterium isolated from coal-contaminated soil.
A strictly anaerobic, mesophilic, iron-reducing bacterial strain, IRF19(T), was isolated from coal-contaminated soil in the Republic of Korea. IRF19(T) cells were straight, rod-shaped, Gram-staining-negative and motile by means of flagella. The optimum pH and temperature for their growth were determined to be pH 7.5-8.0 and 40 °C, while the optimum range was pH 6.5-10.0 and 20-45 °C, respectively. Strain IRF19(T) did not require NaCl for growth but it tolerated up to 2% (w/v). Growth was observed with yeast extract, D-glucose, D-fructose, D-ribose, D-mannitol, D-mannose, L-serine, L-alanine and L-isoleucine. Fe(III), elemental sulfur, thiosulfate and sulfate were used as electron acceptors. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain IRF19(T) is affiliated to the family Clostridiaceae and is most closely related to Salimesophilobacter vulgaris Zn2(T) (93.5% similarity), Geosporobacter subterraneus VNs68(T) (93.2%) and Thermotalea metallivorans B2-1(T) (92.3%). The major cellular fatty acids of strain IRF19(T) were C14 : 0, iso-C15 : 0 and C16 : 0, and the profile was distinct from those of the closely related species. The major respiratory quinone of strain IRF19(T) was menaquinone MK-5 (V-H2). The main polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, an unknown phospholipid and two unknown polar lipids. The G+C content of the genomic DNA of strain IRF19(T) was determined to be 37.4 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic results, strain IRF19(T) is considered to represent a novel species of a novel genus of the family Clostridiaceae , for which we propose the name Anaerosolibacter carboniphilus gen. nov., sp. nov., with the type strain IRF19(T) ( =KCTC 15396(T) =JCM 19988(T)).