Polyamines inhibit NADPH oxidase-mediated superoxide generation and putrescine prevents programmed cell death induced by polyamine oxidase-generated hydrogen peroxide.
Our previous results indicate that during protoplast isolation an oxidative burst occurs [A.K. Papadakis and KA Roubelakis-Angelakis (1999) Plant Physiol 127:197-205] and that suppression of totipotency is correlated with reduced antioxidant activity and low redox state [A.K. Papadakis et al. (2001b) Plant Physiol 126:434-444]. Polyamines are known to affect cell development and to act as antioxidants. Polyamines applied during isolation of tobacco (Nicotiana tabacum L.) protoplasts reduced the accumulation of O2*- but not that of H2O2. This antioxidant effect is probably due to the inhibition of microsomal membrane NADPH oxidase, which occurred in a concentration-dependent manner, with spermine exerting the highest inhibitory effect. However, during protoplast culture, polyamine oxidase activity increased severalfold in spermidine- and spermine-treated protoplasts, concomitant with H2O2 titers. A cell death program was executed in untreated protoplasts, as documented by membrane malfunction, induced DNase activity, DNA fragmentation and a positive TUNEL reaction. Protoplast cell death was prevented in protoplasts treated with putrescine, but not by treatment with spermidine or spermine, which rather had the opposite effect. The data presented suggest that PAs may be implicated in the expression of plant protoplast totipotency.


[Clinical trial of SM-108 in myeloproliferative disorders].
SM-108 (4-carbamoylimidazolium-5-olate) is a new purine antagonist which blocks IMP dehydrogenase, the key enzyme of de novo GMP biosynthesis. Eleven patients were entered into a clinical study of SM-108 for the treatment of myeloproliferative disorders. These included 10 cases of chronic myelogenous leukemia (CML) and one case of essential thrombocythemia (ET). Two cases of CML had received prior chemotherapy. SM-108 was given orally at a dose of 400-600mg/m2 daily. Of nine evaluable cases, four cases of CML achieved complete responses and two cases of CML achieved partial responses (response rate = 66.7%). The duration of drug action was short, and the daily dosage required adjustment at frequent intervals. Long-term response without therapy has not been seen with SM-108 in CML. There was no response in the case of ET. The side effects included constipation (1/11), diarrhea (1/11) and mild hepatic toxicity (1/11). No cardiac or renal toxicity was observed.


Novel residues lining the CFTR chloride channel pore identified by functional modification of introduced cysteines.
Substituted cysteine accessibility mutagenesis (SCAM) has been used widely to identify pore-lining amino acid side chains in ion channel proteins. However, functional effects on permeation and gating can be difficult to separate, leading to uncertainty concerning the location of reactive cysteine side chains. We have combined SCAM with investigation of the charge-dependent effects of methanethiosulfonate (MTS) reagents on the functional permeation properties of cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channels. We find that cysteines substituted for seven out of 21 continuous amino acids in the eleventh and twelfth transmembrane (TM) regions can be modified by external application of positively charged [2-(trimethylammonium)ethyl] MTS bromide (MTSET) and negatively charged sodium [2-sulfonatoethyl] MTS (MTSES). Modification of these cysteines leads to changes in the open channel current-voltage relationship at both the macroscopic and single-channel current levels that reflect specific, charge-dependent effects on the rate of Cl(-) permeation through the channel from the external solution. This approach therefore identifies amino acid side chains that lie within the permeation pathway. Cysteine mutagenesis of pore-lining residues also affects intrapore anion binding and anion selectivity, giving more information regarding the roles of these residues. Our results demonstrate a straightforward method of screening for pore-lining amino acids in ion channels. We suggest that TM11 contributes to the CFTR pore and that the extracellular loop between TMs 11 and 12 lies close to the outer mouth of the pore.


Dosimetric study of Cs-131, I-125, and Pd-103 seeds for permanent prostate brachytherapy.
As a well-established single-modality approach for early-stage prostate cancer, transperineal interstitial permanent prostate brachytherapy (TIPPB) has gained increasing popularity due to its favorable clinical results. Currently, three isotopes, namely Cs-131, I-125, and Pd-103, are commercially available for TIPPB. This is the first study to systematically explore the dosimetric difference of these three isotopes for TIPPB. In total, 25 patients with T1-T2c prostate cancer previously implanted with I-125 seeds were randomly selected and replanned with Cs-131, I-125, and Pd-103 seeds to the prescription doses of 115, 145, and 125 Gy, respectively. The planning goals attempted were prostate V(p)100 approximately 95%, D(p)90 >or= 100%, and prostatic urethra D(u)10 <or= 150%. The dosimetric parameters, as well as the number of seeds and needles required, were analyzed and compared. The mean homogeneity index (HI) was 0.59, 0.56, and 0.46 for Cs-131, I-125, and Pd-103 plans, respectively. The average D(u)10 was 124.6%, 125.7%, and 129.7%, respectively. The average rectum V(r)100 was 0.19, 0.22, and 0.31 cc, respectively. In addition, the average number of seeds was 57.9, 63.0, and 63.7, and the average number of needles required was 31.6, 32.9, and 33.6 for Cs-131, I-125, and Pd-103 seeds, respectively. This study demonstrates that TIPPB, utilizing Cs-131 seeds, allows for better dose homogeneity, while providing comparable prostate coverage and sparing of the urethra and rectum, with a comparable number of, or fewer, seeds and needles required, compared to I-125 or Pd-103 seeds. Further biological and clinical studies associated with Cs-131 are warranted.


Electrical feedback in the cone pedicle: a computational analysis.
One of the fundamental principles of neuroscience is that direct electrical interactions between neurons are not possible without specialized electrical contacts, gap junctions, because the transmembrane resistance of neurons is typically much higher than the resistance of the adjacent extracellular space. However it has been proposed that in the retina direct electrical interactions between cones and second-order neurons occur due to the specific morphology of the cone synaptic terminal. This electrical mechanism could potentially explain the phenomenon of "negative feedback" from horizontal cells to cones and the recent finding that the tips of horizontal cell dendrites contain hemichannels has rekindled interest in the idea. We quantitatively evaluated the possibility that hemichannels and/or glutamate channels mediate electrical feedback from horizontal cells to cones. The calculations show that it is unlikely that an electrical mechanism plays a significant functional role because 1) the necessity of preserving adequate cone-to-horizontal-cell synaptic transmission limits the extracellular space resistance and the horizontal-cell dendritic transmembrane resistances to values at which the effectiveness of electrical feedback is very low and its electrical effect on the cone presynaptic membrane is negligible, 2) electrical feedback is most effective in the dark and weaker during light adaptation, which contradicts the experimental data, and 3) electrical negative feedback is associated with much stronger electrical positive feedback from horizontal cells to cones, a phenomenon that has never been reported. Therefore it is likely that negative feedback from horizontal cells to cones is chemical in nature.


Detection of SARS-CoV-2 RNA in wastewater and importance of population size assessment in smaller cities: An exploratory case study from two municipalities in Latvia.
Wastewater-based epidemiology (WBE) has regained global importance during the COVID-19 pandemic. The mobility of people and other factors, such as precipitation and irregular inflow of industrial wastewater, are complicating the estimation of the disease prevalence through WBE, which is crucial for proper crisis management. These estimations are particularly challenging in urban areas with moderate or low numbers of inhabitants in situations where movement restrictions are not adopted (as in the case of Latvia) because residents of smaller municipalities tend to be more mobile and less strict in following the rules and measures of disease containment. Thus, population movement can influence the outcome of WBE measurements significantly and may not reflect the actual epidemiological situation in the respective area. Here, we demonstrate that by combining the data of detected SARS-CoV-2 RNA copy number, 5-hydroxyindoleacetic acid (5-HIAA) analyses in wastewater and mobile call detail records it was possible to provide an accurate assessment of the COVID-19 epidemiological situation in towns that are small (COVID-19 28-day cumulative incidence r = 0.609 and 35-day cumulative incidence r = 0.89, p < 0.05) and medium-sized towns (COVID-19 21-day cumulative incidence r = 0.997, 28-day cumulative incidence r = 0.98 and 35-day cumulative incidence r = 0.997, p < 0.05). This is the first study demonstrating WBE for monitoring COVID-19 outbreaks in Latvia. We demonstrate that the application of population size estimation measurements such as total 5-HIAA and call detail record data improve the accuracy of the WBE approach.


Caveolin-1 loss of function accelerates glucose transporter 4 and insulin receptor degradation in 3T3-L1 adipocytes.
Caveolae are a specialized type of lipid rafts that are stabilized by oligomers of caveolin protein. Caveolae are particularly enriched in adipocytes. Here we analyzed the effects of caveolin-1 knockdown and caveolae ablation on adipocyte function. To this end, we obtained several multiclonal mouse 3T3-L1 cell lines with a reduced expression of caveolin-1 (95% reduction) by a small interfering RNA approach using lentiviral vectors. Control cell lines were obtained by lentiviral infection with lentiviral vectors encoding appropriate scrambled RNAs. Caveolin-1 knockdown adipocytes showed a drastic reduction in the number of caveolae (95% decrease) and cholera toxin labeling was reorganized in dynamic plasma membrane microdomains. Caveolin-1 depletion caused a specific decrease in glucose transporter 4 (GLUT4) and insulin receptor protein levels. This reduction was not the result of a generalized defect in adipocyte differentiation or altered gene expression but was explained by faster degradation of these proteins. Caveolin-1 knockdown adipocytes showed reductions in insulin-stimulated glucose transport, insulin-triggered GLUT4 recruitment to the cell surface, and insulin receptor activation. In all, our data indicate that caveolin-1 loss of function reduces maximal insulin response through lowered stability and diminished expression of insulin receptors and GLUT4. We propose that caveolin-1/caveolae control insulin action in adipose cells.


Healthcare Supply Chain Network Coordination Through Medical Insurance Strategies with Reference Price Effect.
China has established the universal medical insurance system and individual out of pocket costs have decreased, however, the average healthcare expenditure of the Chinese population and the expenses of the whole society have increased substantially. One major challenge which impedes the progress of attaining sustainable development of the social healthcare system in China is that the number of hospital admissions is disproportionate. Superior hospitals are overcrowded, whereas subordinate hospitals are experiencing low admissions. In this paper, we apply the game theory model to coordinate the healthcare supply chain network, which is composed of the government, medical insurance fund, superior hospitals, subordinate hospitals and patients. Especially by taking the reference price effect into account, this paper analyzes different medical insurance reimbursement strategies and their influence on patient choice and the healthcare supply chain network. The result shows that the reference price effect increases the leverage of medical insurance, guides patients' choice, optimizes the allocation of medical resources and reduces the medical expends. In comparison to a decentralized decision- making strategy, a centralized decision- making strategy can stimulate both superior hospital and subordinate hospital's cooperative intentions which benefits the social healthcare system.


Plasma and Vitreous Metabolomics Profiling of Proliferative Diabetic Retinopathy.
To determine the differences of metabolites and metabolic pathways between patients with proliferative diabetic retinopathy (PDR) and without diabetes (nondiabetic controls) in plasma and vitreous, respectively, and to characterize the relationship between plasma and vitreous metabolic profiles. Liquid chromatography/tandem mass spectrometry technology was performed to distinct metabolite profiles of plasma and vitreous. A total of 139 plasma samples from 88 patients with PDR and 51 nondiabetic controls, as well as 74 vitreous samples from 51 patients with PDR and 23 nondiabetic controls, were screened. Pathway analysis was performed using MetaboAnalyst 5.0. Pearson correlation analysis was used to investigate the correlation of metabolites in vitreous and plasma. After adjusting for age, fasting blood glucose, and urea, in vitreous metabolomes, a total of 76 features distinguished patients with PDR from controls. Fifteen differential metabolites were found in plasma metabolites. Pantothenate and CoA biosynthesis was the common metabolic pathway altered in both plasma and vitreous. Aromatic amino acid metabolism pathways were dysregulated in vitreous of PDR. For four metabolic features, there were positive correlations between vitreous and plasma. Despite great differences between the metabolic profiles of plasma and vitreous in PDR cases, there are also similarities in the change of metabolites and metabolic pathways. Exploring the relationship of metabolomics between vitreous and plasma may help provide new understanding of the mechanism of PDR.


Contextually relevant resources in speech-language therapy and audiology in South Africa--are there any?
In this editorial introduction we aim to explore the notion of contextually relevant resources. We argue that it is the responsibility of speech-language therapists (SLTs) and audiologists (As) working in South Africa to develop contextually relevant resources, and not to rely on the countries or cultures where the professions originated to do so. Language is often cited as the main barrier to contextually relevant resources: most SLTs and As are aware of the need for more resources in the local languages. However, the issue is not as straightforward as translating resources from English into other languages. The challenges related to culture, e.g. formal education and familiarity with the test situation, have to be considered, as well as the population on which norms were obtained and the nature of vocabulary or picture items. This paper introduces four original research papers that follow in this edition of the journal, and showcases them as examples of innovative development in our field. At the same time we call for the further development of assessment materials, intervention resources, and contributions to the evidence base in our context. We emphasise the importance of local knowledge to drive the development of these resources in innovative and perhaps unexpected ways, and suggest that all clinicians have an important role to play in this process.


Modeling the combined effects of enterocins A and B, lactate, and EDTA on the growth of Salmonella at different temperatures.
The effects of enterocins A and B (produced by Enterococcus faecium CTC492), lactate, and ethylenediaminetetraacetic acid (EDTA) on the growth of Salmonella were modeled together with temperature using the response surface methodology. Six serovars of Salmonella enterica were inoculated (ca. 10(3) cells/ml) in brain-heart infusion broth with different levels of the studied factors and then incubated at different temperatures. The results showed that while Salmonella growth was affected by all the factors, temperature was the most important factor influencing the time to detection of the pathogen. All factors, including temperature, showed significant two-way interactions. The presence of enterocins A and B, lactate, and EDTA had an inhibitory effect that was enhanced at suboptimal temperatures for growth, thus delaying the time to detection. Moderate-low concentrations of lactate and EDTA increased the inhibitory effect of enterocins A and B. The effectiveness of these bacteriocins was not further enhanced by high concentrations of lactate (>3.6%) or EDTA (>200 mg/l). The mathematical model obtained from these analyses provides a useful tool to assess the effects of natural antimicrobials and their interactions with other growth-related factors on the growth response of Salmonella. The results can be applied to determine the most effective combination of hurdles to be used in the preservation of food products.


Noninvasive estimation of left ventricular filling pressures in patients with heart failure after surgical ventricular restoration and restrictive mitral annuloplasty.
Doppler echocardiography, including tissue Doppler imaging, is widely applied to assess diastolic left ventricular function using early transmitral flow velocity combined with mitral annular velocity as a noninvasive estimate of left ventricular filling pressures. However, the accuracy of early transmitral flow velocity/mitral annular velocity in patients with heart failure, particularly after extensive cardiac surgery, is debated. Global diastolic strain rate during isovolumic relaxation obtained with 2-dimensional speckle-tracking analysis was recently proposed as an alternative approach to estimate left ventricular filling pressures. We analyzed diastolic function in patients with heart failure after surgical ventricular restoration and/or restrictive mitral annuloplasty. Echocardiography, including tissue Doppler imaging and speckle-tracking analysis, was performed to determine early transmitral flow velocity/atrial transmitral flow velocity, isovolumetric relaxation time, deceleration time, early transmitral flow velocity/mean mitral annular velocity, strain rate during isovolumic relaxation, and early transmitral flow velocity/strain rate during isovolumic relaxation. These noninvasive indices were correlated with relaxation time constant Tau, peak rate of pressure decline, and left ventricular end-diastolic pressure obtained in the catheterization room using high-fidelity pressure catheters. Twenty-three patients were analyzed 6 months after restrictive mitral annuloplasty (n = 8), surgical ventricular restoration (n = 4), or a combined procedure (n = 11). The strongest correlation with invasive indices, in particular left ventricular end-diastolic pressure, was found for strain rate during isovolumic relaxation (r = -0.76, P < .001). Early transmitral flow velocity/mean mitral annular velocity did not correlate significantly with any of the invasive indices. Strain rate during isovolumic relaxation (cutoff value < 0.38 s(-1)) accurately predicted left ventricular end-diastolic pressure of 16 mm Hg or more with 100% sensitivity and 93% specificity. In a group of patients with heart failure who were investigated 6 months after cardiac surgery, early transmitral flow velocity/mean mitral annular velocity correlated poorly with invasively obtained diastolic indexes. Global strain rate during isovolumic relaxation, however, correlated well with left ventricular end-diastolic pressure and peak rate of pressure decline. Our data suggest that global strain rate during isovolumic relaxation is a promising noninvasive index to assess left ventricular filling pressures in patients with heart failure after extensive cardiac surgery, including restrictive mitral annuloplasty and surgical ventricular restoration.


[Good for your health? An analysis of the requirements for scientific substantiation in European health claims regulation].
To identify the various types of evidence, as well as their relative importance in European health claims regulation, in order to analyze the consequences for consumer protection of the requirements for scientific substantiation in this regulation. Qualitative analysis of various documents relevant to the regulatory process, particularly as to the implications of the standards of proof for the functional food market, as well as consumer behavior. European regulation defines a hierarchy of evidence that turns randomized controlled trials into a necessary and sufficient condition for health claim authorizations. Consumer protection can be interpreted in different manners. High standards of proof protect consumers from false information about the health outcomes of functional foods, while lower standards lead to more, albeit less accurate information about such outcomes being available to consumers.


Molecular targeting of inhibitor of apoptosis proteins based on small molecule mimics of natural binding partners.
An assay based on a solvent-sensitive fluorogenic dye molecule, badan, is used to test the binding affinity of a library of tetrapeptide molecules for the BIR3 (baculovirus IAP repeat) domain of XIAP (X-linked inhibitor of apoptosis protein). The fluorophore is attached to a tetrapeptide, Ala-Val-Pro-Cys-NH(2), through a thiol linkage and, upon binding to XIAP, undergoes a solvatochromic shift in fluorescence emission. When a molecule (e.g., a natural protein known to bind to XIAP or a tetrapeptide mimic) displaces the dye, the emission shifts back to the spectrum observed in water. As emission intensity is related to the binding of the tetrapeptide, the intensity can be used to determine the equilibrium constant, K, for the displacement of the dye by the tetrapeptide. The results permit residue-specific analysis of the interaction. Furthermore, we show that hydrophobic effects in the fourth position are general and can effectively increase overall affinity.


Go ahead, grow a head! A planarian's guide to anterior regeneration.
The unique ability of some planarian species to regenerate a head de novo, including a functional brain, provides an experimentally accessible system in which to study the mechanisms underlying regeneration. Here, we summarize the current knowledge on the key steps of planarian head regeneration (head-versus-tail decision, anterior pole formation and head patterning) and their molecular and cellular basis. Moreover, instructive properties of the anterior pole as a putative organizer and in coordinating anterior midline formation are discussed. Finally, we highlight that regeneration initiation occurs in a two-step manner and hypothesize that wound-induced and existing positional cues interact to detect tissue loss and together determine the appropriate regenerative outcomes.


Demographics, Changes in Treatment Patterns, and Outcomes of Bone and Soft Tissue Sarcomas in Korea-A Sarcoma-Specific, Institutional Registry-Based Analysis.
Because of the heterogeneity of sarcomas, establishing a well-collected, sarcoma-specific database is important for sarcoma research. We analyzed the first histology-based, sarcoma-specific institutional registry in Korea, which collected 28 years of patient data according to a predefined data format. Adult bone and soft tissue sarcoma patients who were treated from June 1989 to January 2017 were identified and analyzed, based on the ICD-O-3 codes. Among the 3420 patients included, soft tissue and bone sarcomas comprised 77.8% (n = 2661) and 22.2% (n = 759), respectively. Median age at diagnosis was 50 (range, 16-98) in soft tissue sarcomas and 37 (range, 16-85) in bone sarcomas. Males and females comprised 45.5% and 54.5% of soft tissue sarcomas and 52.7% and 47.3% of bone sarcomas, respectively. Among the 3407 patients with treatment data available, 90.5% of the patients with soft tissue sarcomas and 80.8% of the patients with bone sarcomas received surgery first, of which 57.8% and 71.7% did not receive any subsequent treatment, respectively. Overall, the proportion of patients who received surgery alone decreased from 85.7% to 60.5% from the pre-2000 period to the 2010-2017 period. However, the use of adjuvant chemotherapy increased in patients with soft tissue sarcomas (from 8.0% to 17.2% in the same period), and the use of perioperative radiotherapy also increased in both groups (from 1.4% to 22.7% in soft tissue sarcomas, and 0% to 14.5% in bone sarcomas in the same period). In both soft tissue and bone sarcomas, old age (≥65 years) and diagnosis in the early study period were associated with poorer survival. We presented a comprehensive summary of our sarcoma registry, including the demographics, changes in treatment patterns, and survival outcomes. This study will provide a framework for future studies.


People who report religious experiences may also display enhanced temporal-lobe signs.
Religious and god-related experiences have been hypothesized to be a portion of the continuum of phenomena that are generated by endogenous, transient electrical stimulation within deep structures of the temporal lobe. According to this hypothesis, normal people, without psychiatric history, who report intense religious experiences should also demonstrate a wide range of temporal lobe-related private behaviors. To test this prediction, a self-report inventory that contained 140 temporal-lobe-relevant information, opinion-belief, and sampled MMPI statements was administered to two separate groups (n = 108; n = 41) of male and female first-year university students. In Study I, subjects who had reported religious experiences, particularly those who did not attend church regularly, scored significantly higher on a variety of statement clusters (n = 7 to 14 items) that contained temporal-lobe symptomology relative to groups who did not report religious experiences and did not attend church regularly. In Study II subjects, regardless of church attendance, who reported religious experiences scored significantly higher on the temporal-lobe clusters. People who reported religious experiences were more likely to have kept a dairy and to enjoy poetry reading or writing. However, religious experiments and churchgoers did not score higher (in either experiment) on clusters that contained mundane psychological or proprioceptive statements, descriptions of odd sensations, or modified portions of the Lie scale from the MMPI.


The distribution of plaque and gingivitis and the influence of toothbrushing hand in a group of South Wales 11-12 year-old children.
Numerous factors may affect the distribution of plaque and gingivitis in any individuals mouth. Of considerable importance must be the oral hygiene habits of each person, which will be influenced by compliance and dexterity with tooth cleaning methods. The pattern of gingivitis seen at a young age may, with time, reflect the eventual distribution of attachment loss. This in part, could explain the considerable variation in chronic periodontal disease seen between individuals and at different sites within the same mouth. This study reports the baseline data for the distribution of plaque and gingivitis in 1105, 11-12-year-old children in South Wales. The children were selected by disproportionate stratified random sampling and examined by a multidisciplinary group with the long-term aim of evaluating the importance of malocclusion to dental health and psychosocial variables. Toothbrushing frequency had a very low but significant correlation with the distribution of plaque and gingivitis, accounting therefore for only a small % of the variance in the group. For the total group and right-handed toothbrushers, buccal plaque and gingivitis was significantly increased on right contralateral teeth. No specific pattern for plaque and gingivitis distribution by side was seen for 100 left-handed toothbrushers. Plaque and gingivitis also showed significant differences dependent upon arch, tooth number, and surface. The population is being followed at 4-year intervals to monitor the pattern of periodontal disease with time and correlate changes with these baseline findings.


Biphasic Activation of Ribulose Bisphosphate Carboxylase in Spinach Leaves as Determined from Nonsteady-State CO(2) Exchange.
The activation kinetics of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) following an increase in photon flux density (PFD) were studied by analyzing CO(2) assimilation time courses in spinach leaves (Spinacia oleracea). When leaves were exposed to 45 minutes of darkness before illumination at 690 micromoles per square meter per second, Rubisco activation followed apparent first-order kinetics with a relaxation time of about 3.8 minutes. But when leaves were illuminated for 45 minutes at 160 micromoles per square meter per second prior to illumination at 690 micromoles per square meter per second the relaxation time for Rubisco activation was only 2.1 minutes. The kinetics of this change in relaxation times were investigated by exposing dark-adapted leaves to 160 micromoles per square meter per second for different periods before increasing the PFD to 690 micromoles per square meter per second. It was found that the apparent relaxation time for Rubisco activation changed from 3.8 to 2.1 minutes slowly, requiring at least 8 minutes for completion. This result indicates that at least two sequential, slow processes are involved in light-mediated activation of Rubisco in spinach leaves and that the relaxation times characterizing these two processes are about 4 and 2 minutes, respectively. The kinetics of the first process in the reverse direction and the dependence of the relaxation time for the second process on the magnitude of the increase in PFD were also determined. Evidence that the first slow process is activation of the enzyme Rubisco activase and that the second slow process is the catalytic activation of Rubisco by activase is discussed.


Telomerase inhibition enhances the response to anticancer drug treatment in human breast cancer cells.
Breast cancer is the most common malignancy among women. Current therapies for breast tumors are based on the use of chemotherapeutic drugs that are quite toxic for the patients and often result in resistance. Telomerase is up-regulated in 95% of breast carcinomas but not in adjacent normal tissues. Therefore, it represents a very promising target for anticancer therapies. Unfortunately, the antiproliferative effects of telomerase inhibition require extensive telomere shortening before they are fully present. Combining telomerase inhibition with common chemotherapeutic drugs can be used to reduce this lag phase and induce tumor cell death more effectively. Few studies have analyzed the effects of telomerase inhibition in combination with anticancer drugs in breast cancer cells. In this study, we inhibited telomerase activity in two breast cancer cell lines using a dominant-negative human telomerase reverse transcriptase and analyzed cell viability after treatment with different anticancer compounds. We found that dominant-negative human telomerase reverse transcriptase efficiently inhibits telomerase activity and causes telomere shortening over time. Moreover, cells in which telomerase was suppressed were more sensitive to anticancer agents independently of their mechanism of action and this sensitization was dependent on the presence of shorter telomeres. Altogether, our data show that blocking telomere length maintenance in combination with anticancer drugs can be used as an effective way to induce death of breast cancer cells.


Antibody responses to Borrelia burgdorferi detected by western blot vary geographically in Canada.
Lyme disease is emerging in eastern and central Canada, and most cases are diagnosed using the two-tier serological test (Enzyme Immuno Assay [EIA] followed by Western blot [WB]). Simplification of this algorithm would be advantageous unless it impacts test performance. In this study, accuracy of individual proteins of the IgG WB algorithm in predicting the overall test result in samples from Canadians was assessed. Because Borrelia burgdorferi strains vary geographically in Canada, geographic variations in serological responses were also explored. Metrics of relative sensitivity, specificity and the kappa statistic measure of concordance were used to assess the capacity of responses to individual proteins to predict the overall IgG WB result of 2524 EIA (C6)-positive samples from across Canada. Geographic and interannual variations in proportions of samples testing positive were explored by logistic regression. No one protein was highly concordant with the IgG WB result. Significant variations were found amongst years and geographic regions in the prevalence of samples testing positive using the overall IgG WB algorithm, and for individual proteins of the algorithm. In most cases the prevalence of samples testing positive were highest in Nova Scotia, and lower in samples from Manitoba westwards. These findings suggest that the current two tier test may not be simplified and continued use of the current two-tier test method and interpretation is recommended. Geographic and interannual variations in the prevalence of samples testing positive may be consistent with B. burgdorferi strain variation in Canada, and further studies are needed to explore this.


Endoscopic injection sclerotherapy for esophagogastric variceal bleeding in children with biliary atresia.
To determine the safety and effectiveness of endoscopic injection sclerotherapy (EIS) for children with biliary atresia. Subjects were 7 patients with biliary atresia with esophagogastric varices and variceal bleeding. Intravariceal injection using 5% ethanolamine oleate was performed under fluoroscopy until varices were eradicated. Endoscopic examination revealed that bleeding occurred in the junctional gastric varices in most of the cases. The mean number of EIS sessions required for obliteration of the varices was 2.3. In the observation period (mean: 21 months), recurrent esophagogastric varices occurred in 2 patients. One had variceal bleeding that was treated successfully by additional EIS. There were no severe complications associated with EIS. EIS under fluoroscopy was safe and effective for variceal bleeding in children with biliary atresia.


Design of (Gd-DO3A)n-polydiamidopropanoyl-peptide nucleic acid-D(Cys-Ser-Lys-Cys) magnetic resonance contrast agents.
We hypothesized that chelating Gd(III) to 1,4,7-tris(carboxymethylaza)cyclododecane-10-azaacetylamide (DO3A) on peptide nucleic acid (PNA) hybridization probes would provide a magnetic resonance genetic imaging agent capable of hybridization to a specific mRNA. Because of the low sensitivity of Gd(III) as an magnetic resonance imaging (MRI) contrast agent, a single Gd-DO3A complex per PNA hybridization agent could not provide enough contrast for detection of cancer gene mRNAs, even at thousands of mRNA copies per cell. To increase the Gd(III) shift intensity of MRI genetic imaging agents, we extended a novel DO3An-polydiamidopropanoyl (PDAPm) dendrimer, up to n = 16, from the N-terminus of KRAS PNA hybridization agents by solid phase synthesis. A C-terminal D(Cys-Ser-Lys-Cys) cyclized peptide analog of insulin-like growth factor 1 (IGF1) was included to enable receptor-mediated cellular uptake. Molecular dynamic simulation of the (Gd-DO3A-AEEA)16-PDAP4-AEEA2-KRAS PNA-AEEA-D(Cys-Ser-Lys-Cys) genetic imaging nanoparticles in explicit water yielded a pair correlation function similar to that of PAMAM dendrimers, and a predicted structure in which the PDAP dendron did not sequester the PNA. Thermal melting measurements indicated that the size of the PDAP dendron included in the (DO3A-AEEA)n-PDAPm-AEEA2-KRAS PNA-AEEA-D(Cys-Ser-Lys-Cys) probes (up to 16 Gd(III) cations per PNA) did not depress the melting temperatures (Tm) of the complementary PNA/RNA hybrid duplexes. The Gd(III) dendrimer PNA genetic imaging agents in phantom solutions displayed significantly greater T1 relaxivity per probe (r1 = 30.64 +/- 2.68 mM(-1) s(-1) for n = 2, r1 = 153.84 +/- 11.28 mM(-1) s(-1) for n = 8) than Gd-DTPA (r1 = 10.35 +/- 0.37 mM(-1) s(-1)), but less than that of (Gd-DO3A)32-PAMAM dendrimer (r1 = 771.84 +/- 20.48 mM(-1) s(-1)) (P < 0.05). Higher generations of PDAP dendrimers with 32 or more Gd-DO3A residues attached to PNA-D(Cys-Ser-Lys-Cys) genetic imaging agents might provide greater contrast for more sensitive detection.


[Chronic prostatitis symptom index of Chinese].
To establish the chronic prostatitis symptom index which more suitable for Chinese to refine and standardize evaluation of current symptoms in men with "chronic prostatitis". The literature of previous work was reviewed to develop a symptom index instrument. There were 18 questions representing three broad categories: pain symptoms, urinary symptoms and other symptoms. To evalute the clinical utility and applicability for Chinese people of this tool, we used it in 100 patients with chronic prostatitis and 100 control patients (40 with benign prostatic hyperplasia, BPH; 30 with infertility; 30 with erectile dysfunction). Of the three aspects symptoms, the primary component was pain. But pain in lower back and lower abdomen had less specificity. Pain in the rectal area was not very common but it could well distinguish patients with prostatitis from those BPH, infertility and ED. Urinary symptoms were also common. We used 5 symptom questions in urinary aspect in the last version of this index. In other symptoms, we dropped the question of uncomfortable of waist and back at last. It could not distinguish patients with prostatitis from those infertility and erectile dysfunction. Finally we analyzed the result and modified the new version of chronic prostatitis symptom index. It included 5 pain symptom questions, 5 urinary symptom questions and 2 other symptom questions. There are 12 questions in total. The chronic prostatitis symptom index we developed was validated and useful in clinical practice as well as research protocols. Moreover, it was more suitable for Chinese people.


Study on the Knowledge, Attitude, and Practice (KAP) of Nursing Staff and Influencing Factors on COVID-19.
The aim of this study is to investigate the knowledge, attitude, and practice (KAP) on Coronavirus Disease 2019 (COVID-19) care among nursing staff and analyze its influencing factors. The survey was conducted on February 18, 2020, among 7,716 voluntary participants from 143 medical institutions in Zhejiang, China. The findings indicated that KAP of nursing staff scored well. However, the accuracy of psychological nursing knowledge was much lower, 14.3% only. Nursing staff working in isolation wards have higher knowledge (OR = 1.776, 95% CI: 1.491-2.116), attitude (OR = 1.542, 95% CI: 1.298-1.832), and practice (OR = 1.902, 95% CI: 1.590-2.274) scores than those in general wards. In terms of KAP, nursing staff with working experience ≤ 10 years scored lower than those with working experience ≥ 20 years, with OR values of 0.490 (95% CI: 0.412-0.583), 0.654 (95% CI: 0.551-0.775), and 0.747 (95% CI: 0.629-0.886), respectively. It is necessary to take measures to enhance the training on COVID-19, especially for KAP of junior nurses in general wards.


An Exploratory Study of the Relationships Between Diesel Engine Exhaust Particle Inhalation, Pulmonary Inflammation and Anxious Behavior.
Recent technical developments brought negative side effects such as air pollution and large-scale fires, increasingly exposing people to diesel engine exhaust particles (DEP). Testing how DEP inhalation triggers pathophysiology in animal models could be useful in determining how it affects humans. To this end, the aim of this study was to investigate the effects of pulmonary exposure to DEP for seven consecutive days in experimental male C5BL6/N mice. Twenty-four C5BL6/N mice were treated with one of the three test materials: distilled water for control, a low DEP exposure (5 mg/kg), or a high DEP exposure (15 mg/kg). Exposure to DEP induced decreased body weight; however, it gradually increased pulmonary weight in a DEP-dose-dependent manner. DEP exposure significantly elevated soot accumulation in the lungs, with the alteration of pulmonary homeostasis. It also elevated infiltrated immune cells, thus significantly increasing inflammatory cytokine mRNA and protein production in the lungs and broncho-alveolar lavage fluid, respectively. Pulmonary DEP exposure also altered behavioral responses in the open field test (OFT). Low exposure elevated moving distance and speed, while significantly decreasing the number of trials to enter the central zone. Different concentrations of DEP resulted in different behavioral changes; however, while anxiety levels increased, their degree was independent of DEP concentrations. Results suggest that DEP exposure may possess pro-inflammatory responses in the lungs and trigger anxiety.


Genetic variation of porcine circovirus type 2 (PCV2) and its relevance to vaccination, pathogenesis and diagnosis.
Porcine circovirus-associated disease (PCVAD) encompasses a group of complex, multi-factorial syndromes, which are dependent on infection with porcine circovirus type 2 (PCV2). Current strains of PCV2 circulating in the field are classified into two groups, termed PCV2a and PCV2b. Outbreaks of severe PCVAD in North America and other countries are often linked to a shift from PCV2a to PCV2b as the predominant genotype. Therefore, genotype-specific differences in pathogenesis and antigenicity have been suggested. Overall, evidence suggests that virulence is a function of the specific PCV2 isolate, regardless of genotype. In addition, only minor antigenic differences have been reported. In terms of immunopathogenesis, a conserved decoy epitope, located in the C-terminal region of the capsid protein, provides an explanation for the inability to identify pathogenic differences between genotypes. Finally, genetic variation in PCV2 and the resulting consequences with respect to vaccination and diagnostics are discussed.


Modification of isothermal test for detecting DNA of pathogens of tick-borne borreliosis.
Early diagnosis of tick-borne borreliosis determines the indications for etiotropic therapy, and the detection of borrelia in a tick that has bitten you serves as the basis for antibiotic prophylaxis. To determine the causative agent of borreliosis, PCR methods are most widely used, which requires special conditions for organizing the work of laboratories and the use of expensive equipment. In addition, the procedure for isolating bacterial DNA and subsequent amplification takes several hours of working time. At the same time, methods for detecting borrelia in the isothermal LAMP-reaction are described, which makes it possible to significantly speed up the diagnosis, does not require complex equipment and highly qualified personnel. It is also known that LAMP in some cases allows analysis without prior extraction of nucleic acids. The purpose was a development of a modified test for isothermal detection of DNA of borreliosis pathogens for an accelerated result and the possibility of excluding the stage of nucleic acid extraction. We used 40 samples of Borrelia DNA and 11 Ixodes persulcatus ticks. To shorten the detection time for Borrelia, the previously described LAMP method was modified by the introduction of additional loop primers. The copy number of the positive DNA sample of the borrelia plasmid was estimated using digital PCR. The results of the LAMP reaction were compared with those of the commercial PRC-RT test. The additional use of loop primers approximately halved the detection time for Borrelia DNA without affecting the comparative diagnostic efficiency. The analytical sensitivity limit of the modified LAMP method was 4 copies/μl or 21 molecules of the plasmid standard added to the reaction. In comparative testing with RT-PCR, the sensitivity of the LAMP method is 90%, and the specificity is 100%. The possibility of detecting borrelia in ticks without the stage of DNA extraction has been demonstrated for the first time. A modified isothermal method for the detection of pathogens of tick-borne borreliosis has been developed, which allows analysis within 20-30 minutes, including in ticks without preliminary DNA extraction.


Organomercurial Lyase (MerB)-Mediated Demethylation Decreases Bacterial Methylmercury Resistance in the Absence of Mercuric Reductase (MerA).
The mer operon encodes enzymes that transform and detoxify methylmercury (MeHg) and/or inorganic mercury [Hg(II)]. Organomercurial lyase (MerB) and mercuric reductase (MerA) can act sequentially to demethylate MeHg to Hg(II) and reduce Hg(II) to volatile elemental mercury (Hg0) that can escape from the cell, conferring resistance to MeHg and Hg(II). Most identified mer operons encode either MerA and MerB in tandem or MerA alone; however, microbial genomes were recently identified that encode only MerB. However, the effects of potentially producing intracellular Hg(II) via demethylation of MeHg by MerB, independent of a mechanism to further detoxify or sequester the metal, are not well understood. Here, we investigated MeHg biotransformation in Escherichia coli strains engineered to express MerA and MerB, together or separately, and characterized cell viability and Hg detoxification kinetics when these strains were grown in the presence of MeHg. Strains expressing only MerB are capable of demethylating MeHg to Hg(II). Compared to strains that express both MerA and MerB, strains expressing only MerB exhibit a lower MIC with MeHg exposure, which parallels a redistribution of Hg from the cell-associated fraction to the culture medium, consistent with cell lysis occurring. The data support a model whereby intracellular production of Hg(II), in the absence of reduction or other forms of demobilization, results in a greater cytotoxicity than the parent MeHg compound. Collectively, these results suggest that in the context of MeHg detoxification, MerB must be accompanied by an additional mechanism(s) to reduce, sequester, or redistribute generated Hg(II). IMPORTANCE Mercury is a globally distributed pollutant that poses a risk to wildlife and human health. The toxicity of mercury is influenced largely by microbially mediated biotransformation between its organic (methylmercury) and inorganic [Hg(II) and Hg0] forms. Here, we show in a relevant cellular context that the organomercurial lyase (MerB) enzyme is capable of MeHg demethylation without subsequent mercuric reductase (MerA)-mediated reduction of Hg(II). Demethylation of MeHg without subsequent Hg(II) reduction results in a greater cytotoxicity and increased cell lysis. Microbes carrying MerB alone have recently been identified but have yet to be characterized. Our results demonstrate that mer operons encoding MerB but not MerA put the cell at a disadvantage in the context of MeHg exposure, unless subsequent mechanisms of reduction or Hg(II) sequestration exist. These findings may help uncover the existence of alternative mechanisms of Hg(II) detoxification in addition to revealing the drivers of mer operon evolution.


Synthesis, Characterization, DNA/HSA Interactions, and Anticancer Activity of Two Novel Copper(II) Complexes with 4-Chloro-3-Nitrobenzoic Acid Ligand.
The purpose of this study was to identify new metal-based anticancer drugs; to this end, we synthesized two new copper(II) complexes, namely [Cu(ncba)4(phen)] (1) and [Cu(ncba)4(bpy)] (2), comprised 4-chloro-3-nitrobenzoic acid as the main ligand. The single-crystal XRD approach was employed to determine the copper(II) complex structures. Binding between these complexes and calf thymus DNA (CT-DNA) and human serum albumin (HSA) was explored by electronic absorption, fluorescence spectroscopy, and viscometry. Both complexes intercalatively bound CT-DNA and statically and spontaneously quenched DNA/HSA fluorescence. A CCK-8 assay revealed that complex 1 and complex 2 had substantial antiproliferative influences against human cancer cell lines. Moreover, complex 1 had greater antitumor efficacy than the positive control cisplatin. Flow cytometry assessment of the cell cycle demonstrated that these complexes arrested the HepG2 cell cycle and caused the accumulation of G0/G1-phase cells. The mechanism of cell death was elucidated by flow cytometry-based apoptosis assays. Western blotting revealed that both copper(II) complexes induced apoptosis by regulating the expression of the Bcl-2(Bcl-2, B cell lymphoma 2) protein family.


A Hybrid Knowledge-Based and Empirical Scoring Function for Protein-Ligand Interaction: SMoG2016.
We present the third generation of our scoring function for the prediction of protein-ligand binding free energy. This function is now a hybrid between a knowledge-based potential and an empirical function. We constructed a diversified set of ∼1000 complexes from the PDBBinding-CN database for the training of the function, and we show that this number of complexes generates enough data to build the potential. The occurrence of 420 different types of atomic pairwise interactions is computed in up to five different ranges of distances to derive the knowledge-based part. All of the parameters were optimized, and we were able to considerably improve the accuracy of the scoring function with a Pearson correlation coefficient against experimental binding free energies of up to 0.57, which ranks our new scoring function as one of the best currently available and the second-best in terms of standard deviation (SD = 1.68 kcal/mol). The function was then further improved by inclusion of different terms taking into account repulsion and loss of entropy upon binding, and we show that it is capable of recovering native binding poses up to 80% of the time. All of the programs, tools, and protein sets are released in the Supporting Information or as open-source programs.


Induction of Fas-mediated apoptosis on circulating lymphocytes by surgical stress.
The authors determined whether the decrease in lymphocytes after surgery is related to apoptosis. Surgery induces a profound but transient depletion of circulating lymphocytes, However, the mechanism underlying this phenomenon is unclear. Peripheral blood mononuclear cells were obtained from 18 patients before and after elective surgery and studied for morphologic and biochemical markers of apoptosis, DNA fragmentation, and Fas expression. The DNA staining of peripheral blood mononuclear cells obtained after surgery, which had been cultured for 24 hours in vitro, showed chromatin condensation and fragmentation of cells into collapsed spheres. Moreover, DNA isolated from these peripheral blood mononuclear cells formed a ladder of oligonucleosomal fragments. However, peripheral blood mononuclear cells obtained before surgery showed neither of these changes. The observation that none of these apoptotic cells ingested latex suggested that they were of lymphocytic origin. Fas-positive lymphocytes increased significantly 2 hours after the start of surgery and returned to preoperative levels by postoperative day 7. Anti-Fas antibody augmented apoptosis, whereas ZB4, a Fas antagonist, inhibited apoptosis in lymphocytes after surgery. These results indicate that circulating lymphocytes in the early perioperative period are susceptible to Fas-mediated apoptosis, which may cause depletion of circulating lymphocytes after surgery.


Interplay of Phonon and Exciton-Mediated Superconductivity in Hybrid Semiconductor-Superconductor Structures.
We predict a strong enhancement of the critical temperature in a conventional Bardeen-Cooper-Schrieffer (BCS) superconductor in the presence of a bosonic condensate of exciton polaritons. The effect depends strongly on the ratio of the cutoff frequencies for phonon and exciton-polariton mediated BCS superconductivity, respectively. We also discuss a possible design of hybrid semiconductor-superconductor structures suitable for the experimental observation of such an effect.


BSP-SLIM: a blind low-resolution ligand-protein docking approach using predicted protein structures.
We developed BSP-SLIM, a new method for ligand-protein blind docking using low-resolution protein structures. For a given sequence, protein structures are first predicted by I-TASSER; putative ligand binding sites are transferred from holo-template structures which are analogous to the I-TASSER models; ligand-protein docking conformations are then constructed by shape and chemical match of ligand with the negative image of binding pockets. BSP-SLIM was tested on 71 ligand-protein complexes from the Astex diverse set where the protein structures were predicted by I-TASSER with an average RMSD 2.92 Å on the binding residues. Using I-TASSER models, the median ligand RMSD of BSP-SLIM docking is 3.99 Å which is 5.94 Å lower than that by AutoDock; the median binding-site error by BSP-SLIM is 1.77 Å which is 6.23 Å lower than that by AutoDock and 3.43 Å lower than that by LIGSITE(CSC) . Compared to the models using crystal protein structures, the median ligand RMSD by BSP-SLIM using I-TASSER models increases by 0.87 Å, while that by AutoDock increases by 8.41 Å; the median binding-site error by BSP-SLIM increase by 0.69Å while that by AutoDock and LIGSITE(CSC) increases by 7.31 Å and 1.41 Å, respectively. As case studies, BSP-SLIM was used in virtual screening for six target proteins, which prioritized actives of 25% and 50% in the top 9.2% and 17% of the library on average, respectively. These results demonstrate the usefulness of the template-based coarse-grained algorithms in the low-resolution ligand-protein docking and drug-screening. An on-line BSP-SLIM server is freely available at http://zhanglab.ccmb.med.umich.edu/BSP-SLIM.


The effect of NaOCl dentin treatment on nanoleakage formation.
The term "nanoleakage" has been introduced to explain a penetration pathway within hybrid layers of the dentin-composite junction in the absence of gap formation. This phenomenon is argued in the literature to be a risk factor for the quality of the dentin bonding. NaOCl is a well-known agent used to remove collagen layers that are exposed after acid etching. The purpose of this study was to determine the influence of an NaOCl treatment of dentin to the formation of nanoleakage. Class V cavities were prepared in 40 human molars with the cervical margins located in dentin. After etching with phosphoric acid, 20 samples were treated with 10% NaOCl for 60 s, and 20 samples were used as control. Composite restorations were placed using two different bonding systems. The specimen were exposed in silver nitrate solution and then in developer. After embedding, the teeth were sectioned, dried, coated with gold, and analyzed by SEM. For all the specimen in the control group, backscatter images indicated a silver penetration within the hybrid layer. There was no penetration of silver detectable in the teeth that were treated with NaOCl. We conclude that the removal of the collagen layer by using NaOCl avoids the formation of nanoleakages with the materials tested.


Enzymatic and electron paramagnetic resonance studies of anabolic pyruvate synthesis by pyruvate: ferredoxin oxidoreductase from Hydrogenobacter thermophilus.
Pyruvate: ferredoxin oxidoreductase (POR; EC 18.104.22.168) catalyzes the thiamine pyrophosphate-dependent oxidative decarboxylation of pyruvate to form acetyl-CoA and CO(2). The thermophilic, obligate chemolithoautotrophic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus TK-6, assimilates CO(2) via the reductive tricarboxylic acid cycle. In this cycle, POR acts as pyruvate synthase catalyzing the reverse reaction (i.e. reductive carboxylation of acetyl-CoA) to form pyruvate. The pyruvate synthesis reaction catalyzed by POR is an energetically unfavorable reaction and requires a strong reductant. Moreover, the reducing equivalents must be supplied via its physiological electron mediator, a small iron-sulfur protein ferredoxin. Therefore, the reaction is difficult to demonstrate in vitro and the reaction mechanism has been poorly understood. In the present study, we coupled the decarboxylation of 2-oxoglutarate catalyzed by 2-oxoglutarate: ferredoxin oxidoreductase (EC 184.108.40.206), which generates sufficiently low-potential electrons to reduce ferredoxin, to drive the energy-demanding pyruvate synthesis by POR. We demonstrate that H. thermophilus POR catalyzes pyruvate synthesis from acetyl-CoA and CO(2), confirming the operation of the reductive tricarboxylic acid cycle in this bacterium. We also measured the electron paramagnetic resonance spectra of the POR intermediates in both the forward and reverse reactions, and demonstrate the intermediacy of a 2-(1-hydroxyethyl)- or 2-(1-hydroxyethylidene)-thiamine pyrophosphate radical in both reactions. The reaction mechanism of the reductive carboxylation of acetyl-CoA is also discussed.


Venous Leg Ulcers: Advanced Therapies and New Technologies.
The prevalence of venous leg ulcers (VLUs) differs between 1.5% and 3% in the general population. The challenge in treating VLUs is common recurrence. Moreover, VLUs can be resistant to healing, despite appropriate treatment. In these cases, advanced wound therapies should be considered. The number of new technologies, applied in VLUs treatment, has increased in the last years. These therapies include biophysical interventions such as ultrasound therapy, electrical stimulations, electromagnetic therapy, or phototherapy. Furthermore, stem cell therapies, biologic skin equivalents, platelet-rich plasma therapy, oxygen therapies, anti-TNF therapy, or negative pressure wound therapy are advanced venous ulcer therapeutic methods that may support the standard of care. Medical devices, such as a muscle pump activator, or intermittent pneumatic compression device, may be especially useful for specific subgroups of patients suffering from VLUs. Some of the above-mentioned technologies require broader evidence of clinical efficacy and are still considered experimental therapies in dermatology.


Maintaining volunteer commitment to local watershed initiatives.
Australia's Landcare program is advanced as a successful international example of local watershed groups and governments working together to improve natural resource management. One of the aspects considered critical in the success of watershed groups is engaging widespread participation. This paper draws on two regional surveys that explored burnout, or loss of engagement, among Landcare participants in the state of Victoria using the Maslach Burnout Inventory. Survey findings indicated that a large proportion of respondents were experiencing high burnout in terms of low personal accomplishment and suggested that there was potential for burnout to increase. The authors suggest that the expectations of watershed groups must be based around a realistic assessment of the capacity for volunteer groups to deliver improved environmental and social outcomes.


The California Violence Prevention Initiative: advancing policy to ban Saturday night specials.
The California Violence Prevention Initiative (VPI) was conceived in 1993 as a 5-year, $35 million comprehensive community, media, research, and policy advocacy effort to reduce violence among youth. The VPI included an emphasis on three broad policy areas: shifting society's definition of violence to include a public health perspective, reducing access to alcohol and other drugs, and limiting availability of handguns. For the first 3 years of the VPI, the policy focus was on reducing the availability of handguns to youth through efforts to ban the manufacture and sale of Saturday night specials (SNSs). Prior to the VPI, there were no local SNS bans. Now, there are bans in 41 California jurisdictions, including major population centers. After two vetoes of a statewide legislative ban by the former governor, an SNS ban was signed by a newly elected governor.


RIMB-1/RIM-Binding Protein and UNC-10/RIM Redundantly Regulate Presynaptic Localization of the Voltage-Gated Calcium Channel in Caenorhabditis elegans.
Presynaptic active zones (AZs) contain many molecules essential for neurotransmitter release and are assembled in a highly organized manner. A network of adaptor proteins known as cytomatrix at the AZ (CAZ) is important for shaping the structural characteristics of AZ. Rab3-interacting molecule (RIM)-binding protein (RBP) family are binding partners of the CAZ protein RIM and also bind the voltage-gated calcium channels (VGCCs) in mice and flies. Here, we investigated the physiological roles of RIMB-1, the homolog of RBPs in the nematode Caenorhabditis elegans RIMB-1 is expressed broadly in neurons and predominantly localized at presynaptic sites. Loss-of-function animals of rimb-1 displayed slight defects in motility and response to pharmacological inhibition of synaptic transmission, suggesting a modest involvement of rimb-1 in synapse function. We analyzed genetic interactions of rimb-1 by testing candidate genes and by an unbiased forward genetic screen for rimb-1 enhancer. Both analyses identified the RIM homolog UNC-10 that acts together with RIMB-1 to regulate presynaptic localization of the P/Q-type VGCC UNC-2/Cav2. We also find that the precise localization of RIMB-1 to presynaptic sites requires presynaptic UNC-2/Cav2. RIMB-1 has multiple FN3 and SH3 domains. Our transgenic rescue analysis with RIMB-1 deletion constructs revealed a functional requirement of a C-terminal SH3 in regulating UNC-2/Cav2 localization. Together, these findings suggest a redundant role of RIMB-1/RBP and UNC-10/RIM to regulate the abundance of UNC-2/Cav2 at the presynaptic AZ in C. elegans, depending on the bidirectional interplay between CAZ adaptor and channel proteins.SIGNIFICANCE STATEMENT Presynaptic active zones (AZs) are highly organized structures for synaptic transmission with characteristic networks of adaptor proteins called cytomatrix at the AZ (CAZ). In this study, we characterized a CAZ protein RIMB-1, named for RIM-binding protein (RBP), in the nematode Caenorhabditis elegans Through systematic analyses of genetic interactions and an unbiased genetic enhancer screen of rimb-1, we revealed a redundant role of two CAZ proteins RIMB-1/RBP and UNC-10/RIM in regulating presynaptic localization of UNC-2/Cav2, a voltage-gated calcium channel (VGCC) critical for proper neurotransmitter release. Additionally, the precise localization of RIMB-1/RBP requires presynaptic UNC-2/Cav2. These findings provide new mechanistic insight about how the interplay among multiple CAZ adaptor proteins and VGCC contributes to the organization of presynaptic AZ.


Green Extraction of Antioxidant Polyphenols from Green Tea (Camellia sinensis).
In this study, the feasibility of improving the extraction yield of green tea antioxidant polyphenols by the combination of ultrasound-assisted extraction (UAE) and deep eutectic solvents (DESs) was investigated. Choline chloride (ChCl)-glycerol was selected as the best DES among 12 ChCl-based DESs to extract tea antioxidant polyphenols. Subsequently, the influences of extraction parameters on total phenolic content (TPC) values were investigated, and liquid/solid ratio, ultrasonic power, and ultrasonic time were optimized based on the response surface methodology. The optimal extraction conditions were a liquid to solid ratio of 36:1 (mL/g), ultrasonic power of 461.5 W, and ultrasonic time of 21 min, with the highest TPC value of 243 ± 7 mg gallic acid equivalent (mg GAE)/g dry weight (DW), which was 13% higher than that before optimization. In addition, under the optimal extraction conditions, tea polyphenolic extract exhibited higher antioxidant activity compared with conventional extraction methods. Four major catechins in the green tea extracts, including (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG) and (-)-epigallocatechin gallate (EGCG) were identified and quantified by high-performance liquid chromatography. In addition, scanning electron microscopy (SEM) analysis revealed that UAE-DES effectively disrupted the green tea leaf cells, thereby improving tea polyphenol yield. In summary, UAE-DES is an ideal green extraction method for the extraction of tea antioxidant polyphenols.


Inlays and onlays.
Dentists must meet the increased demands of patients for esthetic dentistry. Because the substitution of amalgam with direct composite restorations is somewhat problematic and restricted to special indications only, the most suitable method is the esthetic inlay. All available systems are reviewed and described. These restorations must be inserted with adhesive techniques. The performance and the problems associated with these techniques are described in detail. Improvements in luting techniques, eg, the use of ultrasonic energy, may further simplify the application technique for esthetic inlays. For clinical success, the margin quality of adhesively luted inlays is essential. In vitro and in vivo studies with esthetic inlays report good clinical behavior of such restorations. However, until the results of long-term clinical studies are available, these systems should not be used on a routine basis as a substitute for amalgam.


Lung dendritic cells in respiratory viral infection and asthma: from protection to immunopathology.
Lung dendritic cells (DCs) bridge innate and adaptive immunity, and depending on context, they also induce a Th1, Th2, or Th17 response to optimally clear infectious threats. Conversely, lung DCs can also mount maladaptive Th2 immune responses to harmless allergens and, in this way, contribute to immunopathology. It is now clear that the various aspects of DC biology can be understood only if we take into account the functional specializations of different DC subsets that are present in the lung in homeostasis or are attracted to the lung as part of the inflammatory response to inhaled noxious stimuli. Lung DCs are heavily influenced by the nearby epithelial cells, and a model is emerging whereby direct communication between DCs and epithelial cells determines the outcome of the pulmonary immune response. Here, we have approached DC biology from the perspective of viral infection and allergy to illustrate these emerging concepts.


[The use of an underlay polypropylene mesh in complicated incisional hernias: sucessful French surgical technique].
Incisional hernia repair with conventional techniques is associated with high recurrence rates of 30-50%. Surgical repair using different prosthetic biomaterials is becoming increasingly popular. On the basis of the favourable results by French surgeons, the results of underlay prosthetic mesh repair using polypropylene mesh in complicated and recurrent incisional hernias have been studied. After preparation and excision of the entire hernia sac, the peritoneum and posterior rectus sheath are closed with a continuous looped polyglyconate suture. The prosthesis used for the midline hernias is positioned on the posterior rectus sheath and extends far beyond the borders of the myoaponeurotic defect. The prosthesis for lumbar and subcostal hernias is placed in a prepared space between the transverse and oblique muscles. Intraperitoneal placement of the mesh must be avoided. Between January 1997 and September 1998 a total of 57 incisional hernia repair (25 primary hernias, 32 recurrent hernias) have been performed using this technique (28 women, 29 men, mean age 56+/-13 years). Local complications occurred in 6 patients (11%). One patient suddenly died on the 3rd postoperative day from severe pulmonary embolism (mortality 1,7%). Thirthy-seven patients with a minimum follow-up of 6 months were reexamined clinically (follow up time 6-33 months). Till now one recurrent hernia has been observed. There were only minor complaints like a feeling of tension in the abdominal wall (n = 3) and slight pain under physical stress (n = 9). The aforementioned technique of underlay prosthetic repair allows an anatomical and consolidated reconstruction of the damaged abdominal wall with excellent results and low complication rates especially in high risk patients and complicated hernias.


Downregulation of PHYTOCHROME-INTERACTING FACTOR 4 Influences Plant Development and Fruit Production.
Plant development is highly dependent on the ability to perceive and cope with environmental changes. In this context, PIF proteins are key players in the cellular hub controlling responses to fluctuating light and temperature conditions. Reports in various plant species show that manipulation of the PIF4 level affects important agronomical traits. In tomato (Solanum lycopersicum), SlPIF1a and SlPIF3 regulate fruit nutraceutical composition. However, the wider role of this protein family, and the potential of their manipulation for the improvement of other traits, has not been explored. Here we report the effects of constitutive silencing of tomato SlPIF4 on whole-plant physiology and development. Ripening anticipation and higher carotenoid levels observed in SlPIF4-silenced fruits revealed a redundant role of SlPIF4 in the accumulation of nutraceutical compounds. Furthermore, silencing triggered a significant reduction in plant size, flowering, fruit yield, and fruit size. This phenotype was most likely caused by reduced auxin levels and altered carbon partitioning. Impaired thermomorphogenesis and delayed leaf senescence were also observed in silenced plants, highlighting the functional conservation of PIF4 homologs in angiosperms. Overall, this work improves our understanding of the role of PIF proteins-and light signaling-in metabolic and developmental processes that affect yield and composition of fleshy fruits.


Intestinal stem cells and epithelial-mesenchymal interactions in the crypt and stem cell niche.
The intestinal epithelium contains a rapidly proliferating and perpetually differentiating epithelium. The principal functional unit of the small intestine is the crypt-villus axis. Stem cells located in the crypts of Lieberkühn give rise to proliferating progenitor or transit amplifying cells that differentiate into the 4 major epithelial cell types. The study of adult gastrointestinal tract stem cells has progressed rapidly with the recent discovery of several putative stem cell markers. Substantial evidence suggests 2 populations of stem cells: long-term quiescent (reserved) and actively cycling (primed) stem cells. These cells are in adjoining locations and are presumably maintained by the secretion of specific proteins generated in a unique microenvironment or stem cell niche surrounding each population. The relationship between these 2 populations, as well as the cellular sources and composition of the surrounding environment, remains to be defined, and is an active area of research. In this review, we will outline progress in identifying stem cells and in defining epithelial-mesenchymal interactions in the crypt. We will summarize early advances using stem cells for therapy of gastrointestinal disorders.


[Conservative treatment of ectopic pregnancies: comparison of velocity of decrease of beta-HCG between spontaneous and methotrexate induced resorptions].
The evolution of the process can take two different directions. If an Intact pregnancy continues to grow, there is a tubal abortion or a rupture of the Fallopian tube. In the oppsote case, there is spontaneous resorption of the tubal pregnancy. In certain cases with the application of methotrexate, the ectopic pregnancy growth can be interrupted and spontaneous resorption can be induced. Nowadays, the international standards of treating patients with early ectopic gravidities and no peritoneal effusions, with beta-HCG values below 6000 UI/l, gestational sac diameter below 3 cm, and without any visible heart action, imply methotrexate treatment. The objective of this study is to prove the efficiency of methotrexate use in patients who meet the above criteria. The first group did not manifest satisfying decrease of [beta-HCG after two measurements, so methotrexate was used, while the second group showed satisfying decrease of beta-HCG, measured on the third day. Comparing the velocity of decrease of beta-HCG level among both groups of patients, it was proved that iatrogenically induced decrease, that is to say the achieved resorption, was equal to the spontaneous resorption, which justified the use of methotrexate in ectopic pregnancy treatment.


Recombinant forms of tetanus toxin engineered for examining and exploiting neuronal trafficking pathways.
Tetanus toxin is a fascinating, multifunctional protein that binds to peripheral neurons, undergoes retrograde transport and trans-synaptic transfer to central inhibitory neurons where it blocks transmitter release, thereby, causing spastic paralysis. As a pre-requisite for exploiting its unique trafficking properties, a novel recombinant single chain was expressed at a high level in Escherichia coli as a soluble, easily purifiable protein. It could be activated with enterokinase to produce a dichain that matched native toxin in terms of proteolytic and neuroinhibitory activities, as well as induction of spastic paralysis in mice. Importantly, nicking was not essential for protease activity. Substitution of Glu(234) by Ala created a protease-deficient atoxic form, which blocked the neuroparalytic action of tetanus toxin in vitro, with equal potency to its heavy chain; but, the mutant proved >30-fold more potent in preventing tetanus in mice. This observation unveils differences between the intoxication processes resulting from retrograde transport of toxin in vivo and its local uptake into peripheral or central nerves in vitro, dispelling a popularly held belief that the heavy chain is the sole determinant for efficient trafficking. Thus, this innocuous mutant may be a useful vehicle, superior to the heavy chain, for drug delivery to central neurons.


Case study of anti-1-amino-3-F-18 fluorocyclobutane-1-carboxylic acid (anti-[F-18] FACBC) to guide prostate cancer radiotherapy target design.
Anti-1-amino-3-F-18 fluorocyclobutane-1-carboxylic acid (FACBC) is a novel radiotracer, which has shown some promise for use with positron emission tomography (PET)/computed tomography (CT) for visualizing prostate cancer. Here we describe a case of a prostate cancer patient who underwent radiation treatment and had an FACBC scan obtained as part of a pilot study. We explored the potential impact of FACBC on treatment planning. We registered the FACBC acquisition with the PET/CT, which required a simple translation. Then, we did a deformable image registration of the PET/CT with the planning CT-this process allowed the FACBC-defined gross tumor volume (GTVFACBC) to be projected into the planning CT. An intensity-modulated radiotherapy (IMRT) plan (plan A) not including GTVFACBC (with final dose to 81.0 Gy) was generated, as was an IMRT plan including the GTVFACBC to a final dose of 86.4 Gy (plan B). Target coverage and normal tissue dose volume histogram (DVH) endpoints were tabulated. In this particular patient, bladder constraints could not be met on any plan due to anatomic limitations. However, the impact on the rectal DVH could be assessed, and inclusion of the GTVFACBC did permit rectal DVH constraints to be met in plan B while maintaining target coverage and inhomogeneity constraints. In our test case, it was feasible to use FACBC to guide IMRT, and highlights the role of deformable image registration of the PET/CT with the planning CT. These findings can guide future studies incorporating FACBC into treatment planning.


Cooperativity and Steep Voltage Dependence in a Bacterial Channel.
This paper reports on the discovery of a novel three-membrane channel unit exhibiting very steep voltage dependence and strong cooperative behavior. It was reconstituted into planar phospholipid membranes formed by the monolayer method and studied under voltage-clamp conditions. The behavior of the novel channel-former, isolated from Escherichia coli, is consistent with a linearly organized three-channel unit displaying steep voltage-gating (a minimum of 14 charges in the voltage sensor) that rivals that of channels in mammalian excitable membranes. The channels also display strong cooperativity in that closure of the first channel permits the second to close and closure of the second channel permits closure of the third. All three have virtually the same conductance and selectivity, and yet the first and third close at positive potentials whereas the second closes at negative potentials. Thus, is it likely that the second channel-former is oriented in the membrane in a direction opposite to that of the other two. This novel structure is named "triplin." The extraordinary behavior of triplin indicates that it must have important and as yet undefined physiological roles.


On the general mechanism of selective induction of cytochrome P450 enzymes by chemicals: some theoretical considerations.
The CYP isoforms that are selectively induced following exposure to structurally-diverse chemicals often are the ones capable of metabolizing these chemicals. However, the molecular mechanism underlying this apparent functional coupling is not understood at present. Three hypotheses are developed to explain the complex process of selective chemical induction of CYPs: i) each inducible CYP may have a corresponding intracellular receptor that interacts with the inducer chemical and mediates the selective induction of this CYP; ii) each inducible CYP and its corresponding receptor may share highly similar steric structures for their substrate/inducer-binding sites and iii) each chemically-inducible CYP gene may have distinct genomic response element(s) that interact selectively with the corresponding receptor. The readers are introduced to a novel theoretical framework that offers a plausible mechanistic explanation at the molecular level concerning the complex process of how an organism selectively activates the biosynthesis of certain CYP isoform(s) that can effectively metabolize a chemical to which the organism is exposed. The theoretical framework developed herein seeks to ignite additional critical thinking on this important research subject as well as to promote experimental testing of the proposed theories in the future. Undoubtedly, these studies will enhance the understanding of the molecular mechanisms for the selective induction of CYP enzymes by chemicals.


Thrombocytosis: too much of a good thing?
Thrombocytosis is due to (a) a variety of disorders that cause reactive stimulation of platelet production, (b) familial mutations, or (c) essential thrombocythemia (ET) and other myeloproliferative disorders (MPDs). The MPDs are clonal abnormalities of the pluripotent hematopoietic stem cell. Dysregulation of megakaryocytopoiesis in ET involves defective binding of thrombopoietin by platelets and megakaryocytes resulting in increased levels of plasma free thrombopoietin, and increased sensitivity of megakaryocytes to thrombopoietin leading to their increased proliferation. Bleeding and thrombosis are the major causes of morbidity and mortality in ET and the other MPDs. The elevated platelet count and qualitative platelet defects have been implicated in the pathophysiology of these hemostatic problems. However, these platelet abnormalities do not correlate well with clinical complications. It is proposed that bleeding and thrombosis could be due to vascular abnormalities that result from dysfunctional hematopoietic stem cell-derived endothelial cells.


Glomerular hyperfiltration in hypertensive African Americans.
The incidence of end-stage renal disease attributable to hypertension is 5-fold greater in African Americans than in whites. To determine whether glomerular hyperfiltration is an antecedent to renal failure, we compared responses of renal blood flow and glomerular filtration rate to graded infusions of norepinephrine (0. 01, 0.025, and 0.05 microg. kg(-1). min(-1) for 30 minutes each) in 29 African Americans and 33 age-matched French Canadian whites with essential hypertension. Renal blood flow and glomerular filtration rate were measured by using a constant-infusion technique of PAH and inulin, respectively. Studies were conducted on an inpatient clinical research center, and antihypertensive medications had been discontinued for at least 1 week. Based on 24-hour blood pressure monitoring, nighttime blood pressures decreased (P<0.01) in the French Canadians but not in the African Americans. Baseline renal blood flow was higher (P<0.05) in the African Americans (1310+/-127 mL. min(-1) per 1.73 m(2)) than in the French Canadians (1024+/-42 mL. min(-1) per 1.73 m(2)); baseline glomerular filtration rate was also higher (P<0.01) in the African Americans (140+/-4 versus 121+/-4 mL. min(-1) per 1.73 m(2)). In response to norepinephrine-induced blood pressure increases, renal blood flow was autoregulated and did not change in either patient group. In the African Americans, glomerular filtration rate increased (P<0.01) to 167 mL. min(-1) per 1.73 m(2) during the first norepinephrine infusion, without subsequent change. In contrast, glomerular filtration rate did not change with norepinephrine-induced increases of blood pressure in the French Canadians. In the African Americans, the elevation of baseline glomerular filtration rate, with a further increase in response to norepinephrine, may be indicative of glomerular hyperfiltration. Glomerular hyperfiltration and lack of nocturnal blood pressure decline may contribute to the higher incidence of end-stage renal disease in hypertensive African Americans.


CT angiography for brain death diagnosis.
Lack of cerebral circulation is an important confirmatory test for brain death (BD). Conventional angiography remains the standard imaging method, but CT angiography (CTA) is emerging as an alternative. France accepts BD diagnoses relying on a score based on lack of opacification of 7 intracerebral vessels in CTA images. The purpose of this study was to validate the efficiency of this score and to evaluate the sensitivity of a novel 4-point CTA score in confirming BD. A prospective multicentric study was conducted during 12 months with 105 patients referred for CTA to confirm a clinical diagnosis of BD. Clinical data were recorded. CTA images were interpreted first by local radiologists at the referent center, resulting in a 7-point score based on lack of opacification of the pericallosal and cortical segments of the middle cerebral arteries (MCAs), internal cerebral veins (ICVs), and 1 great cerebral vein per patient and, second, by a consensus panel of 3 expert radiologists, blinded to the initial scores, resulting in novel 4-point scores based on the lack of opacification of the cortical segments of the MCAs and ICVs. Injection of contrast medium did not alter renal function. With the initial 7-point score, sensitivity was 62.8%. With the simplified 4-point score, sensitivity was 85.7% and specificity was 100%. Opacification of ICVs was absent in 98.1% of patients. Lack of opacification in the cortical segments of the MCAs and internal veins in CTA is efficient and reliable for confirming BD.


Towards the phylogenetic placement of the enigmatic African genus Prolabeops Schultz, 1941.
The small cyprinid genus Prolabeops Schultz, 1941 is restricted to the Nyong and Sanaga River systems in Cameroon. In the past, the genus had been suggested to be either a member of the Labeoninae, Torinae or the Smiliogastrinae mainly on the basis of morphological similarities, and it is nowadays considered as incertae sedis within the Cypriniformes. This study provides the first attempt to reveal the phylogenetic position of Prolabeops using molecular data. For this purpose, the authors sequenced a large fraction of the mitochondrial genome (c. 13,600 bp), including all mitochondrial protein coding genes, of two Prolabeops melanhypopterus specimens and an additional four Enteromius specimens. The large-scale phylogenetic analysis was based on an alignment including all mitochondrial protein coding genes of 902 specimens representing c. 899 cypriniform species. Prolabeops was clearly recovered within the African Smiliogastrinae, forming a weakly supported clade together with Enteromius jae, Enteromius hulstaerti and Barboides gracilis. The study data underline the urgent need of a thorough taxonomic revision of the small African barbs collectively placed in the genus Enteromius.


Phage Peptide Libraries As a Source of Targeted Ligands.
One of the dominant trends in modern pharmacology is the creation of drugs that act directly on the lesion focus and have minimal toxicity on healthy tissues and organs. This problem is particularly acute in relation to oncologic diseases. Short tissue- and organ-specific peptides capable of delivering drugs to the affected organ or tissue are considered promising targeted agents that can be used in the diagnosis and therapy of diseases, including cancer. The review discusses in detail the technology of phage display as a method for obtaining specific targeted peptide agents and offers examples of their use in diagnostic and clinical practice.


Melatonin Lowers HIF-1α Content in Human Proximal Tubular Cells (HK-2) Due to Preventing Its Deacetylation by Sirtuin 1.
Although melatonin is widely known for its nephroprotective properties, there are no reports clearly pointing at its impact on the activity of hypoxia-inducible factor-1 (HIF-1), the main mediator of metabolic responses to hypoxia, in kidneys. The aim of the present study was to elucidate how melatonin affects the expression of the regulatory subunit HIF-1α in renal proximal tubules. HK-2 cells, immortalized human proximal tubular cells, were cultured under hypoxic conditions (1% O2). Melatonin was applied at 100 μM concentration. Protein and mRNA contents were determined by Western blot and RT-qPCR, respectively. HIF-1α acetylation level was established by means of immunoprecipitation followed by Western blot. Melatonin receptors MT1 and MT2 localization in HK-2 cells was visualized using immunofluorescence confocal analysis. It was found that melatonin in HK-2 cells (1) lowered HIF-1α protein, but not mRNA, content; (2) attenuated expression of HIF-1 target genes; (3) increased HIF-1α acetylation level; and (4) diminished sirtuin 1 expression (both protein and mRNA). Sirtuin 1 involvement in the regulation of HIF-1α level was confirmed applying cells with silenced Sirt1 gene. Moreover, the presence of membrane MT1 and MT2 receptors was identified in HK-2 cells and their ligand, ramelteon, turned out to mimic melatonin action on both HIF-1α and sirtuin 1 levels. Thus, it is concluded that the mechanism of melatonin-evoked decline in HIF-1α content in renal proximal tubular cells involves increased acetylation of this subunit which results from the attenuated expression of sirtuin 1, an enzyme reported to deacetylate HIF-1α. This observation provides a new insight to the understanding of melatonin action in kidneys.


Risk to household contacts of tuberculous patients based on Mantoux test and antibody titre.
Tuberculosis, being an infectious disease, carries a risk of infection to contacts attending tuberculous patients. This study was conducted to evaluate the risk for household contacts of tuberculous patients as compared to non-contacts. The study was conducted at PGMI, Gulab Devi Hospital and Defence Housing Authority Lahore. The study included 120 household contacts and 80 non-contacts. A Cross sectional study for evaluation of antituberculous antibodies levels by ELISA method in two groups; Mantoux positive household contacts 49, Mantoux negative household contacts 71 and normal healthy persons 'non contacts' 80. Routine Haematological investigations like HB, TLC and ESR were done by conventional methods and all the sera of 200 subjects included in the study were tested for IgM, IgG and IgA anti tuberculous antibodies by enzyme linked immunosorbant assay (ELISA). Purified protein derivative 0.1 ml containing 5 TU was injected intradermally. The test was read after 72 hours by measuring the induration around injection site of forearm. There was no difference in the average age of the household contacts and non-contacts. The complaints of pyrexia, night sweats and weight loss were more in house hold contacts as compared to non-contacts. The awareness about BCG vaccination was equal in both. There were 49 contacts with positive Mantoux test while negative Mantoux test was found in 71 contacts. There were only three Mantoux positive among eighty non-contacts. There was no significant difference in the presence of IgM among household contacts as compared to non-contacts. However both IgG and IgA were present in significantly higher number of household contacts compared to non-contacts. Household contacts of patients suffering from active pulmonary tuberculosis have more chances of being infected with Mycobacterium tuberculosis as compared to the healthy non-contact, as shown by the higher levels of antituberculous antibodies & positivity of Mantoux test.


Medicaid Tape-to-Tape findings: California, New York, and Michigan, 1981.
Presented in this report is an overview of Medicaid enrollment, utilization, and expenditures in California during 1981. The California Medicaid program, called Medi-Cal, is the largest in the Nation in terms of program beneficiaries. During 1981, California had one of the most generous Medicaid programs in the country in terms of eligibility and covered services. At the same time, there were benefit limitations and reimbursement restrictions in place that were designed to restrict program expenditures. The data in this report were provided by the State to the Health Care Financing Administration as part of the Medicaid Tape-to-Tape Project. Data from Michigan and New York are also included for comparison purposes.


Simultaneous infection with Legionella pneumophila and Legionella micdadei in an immunologically intact host. A case report.
The Authors reported a case of dual infection with Legionella Pneumophila and Legionella micdadei in an immunologically intact host. The bacteriological and serological data are compatible with a simultaneous recent infection.


[Nature of the epidemic process in Sonne and Flexner dysenteries and the causes of its induction].
A comparative study of the epidemic process in Sh. sonnei and Sh. flexneri dysentery in different regions of the USSR revealed that the morbidity level of Sh. sonnei dysentery changed simultaneously in the regions under study at intervals of 2-3 years. Sh. flexneri dysentery showed morbidity rises occurring at intervals 6-8 years, and their occurrence did not coincide with the periods of elevated morbidity in Sh. sonnei dysentery. The data obtained in the cohort analysis and in the study of recurrent morbidity suggest that Sh. flexneri dysentery produces more pronounced postinfection immunity than Sh. sonnei dysentery, and the immunological factor probably affects the dynamics of the epidemic of these Shigella infections.


Segmental analysis of thallium 201 myocardial perfusion scintigraphy: its value in a community hospital.
In a community hospital, we correlated results of thallium 201 myocardial scintigraphy with coronary arteriographic data in 79 patients. Scintigraphy was 92% sensitive and 85% specific in detecting coronary artery disease. There were no false-negative scintigrams in patients with double or triple vessel disease. The most important factors determining sensitivity of the method in detecting individual coronary stenoses were (1) location of the stenosis in the coronary tree, (2) number of vessels involved, and (3) degree of obstruction. Higher prevalence of perfusion defects in areas of 90% to 99% stenosis as compared with 50% to 89% lesions was of borderline statistical significance (86% vs 59%; P = .06). Myocardial perfusion scintigraphy was unable to predict the number of significantly narrowed coronary vessels. Predictive value of a perfusion defect for a significant coronary stenosis was 87% for anterior, 88% for septal, 90% for lateral, 89% for posterior, and 78% for inferior segment. We conclude that segmental analysis of myocardial scintigrams may be of value in a community hospital.


Highly conserved NIKS tetrapeptide is functionally essential in eukaryotic translation termination factor eRF1.
Class-1 polypeptide chain release factors (RFs) play a key role in translation termination. Eukaryotic (eRF1) and archaeal class-1 RFs possess a highly conserved Asn-Ile-Lys-Ser (NIKS) tetrapeptide located at the N-terminal domain of human eRF1. In the three-dimensional structure, NIKS forms a loop between helices. The universal occurrence and exposed nature of this motif provoke the appearance of hypotheses postulating an essential role of this tetrapeptide in stop codon recognition and ribosome binding. To approach this problem experimentally, site-directed mutagenesis of the NIKS (positions 61-64) in human eRF1 and adjacent amino acids has been applied followed by determination of release activity and ribosome-binding capacity of mutants. Substitutions of Asn61 and Ile62 residues of the NIKS cause a decrease in the ability of eRF1 mutants to promote termination reaction in vitro, but to a different extent depending on the stop codon specificity, position, and nature of the substituting residues. This observation points to a possibility that Asn-Ile dipeptide modulates the specific recognition of the stop codons by eRF1. Some replacements at positions 60, 63, and 64 cause a negligible (if any) effect in contrast to what has been deduced from some current hypotheses predicting the structure of the termination codon recognition site in eRF1. Reduction in ribosome binding revealed for Ile62, Ser64, Arg65, and Arg68 mutants argues in favor of the essential role played by the right part of the NIKS loop in interaction with the ribosome, most probably with ribosomal RNA.


Echocardiographic parameters in athletes of different sports.
Competitive athletics is often associated with moderate left ventricular (LV) hypertrophy, and it has been hypothesized that training mode and type of exercise modulates long-term cardiac adaptation. The purpose of the study was to compare cardiac structure and function among athletes of various sports and sedentary controls. Standard transthoracic two-dimensional M-mode and Doppler echocardiography was performed at rest in Caucasian male canoe/kayak paddlers (n = 9), long distance runners (LDR, n = 18), middle distance runners (MDR, n = 17), basketball players (BP, n = 31), road cyclists (n = 8), swimmers (n = 10), strength/power athletes (n = 9) of similar age (range, 15 to 31 yrs), training experience (4 to 9 years), and age-matched healthy male sedentary controls (n = 15). Absolute interventricular septum (IVS) thickness and LV wall thickness, but not LV diameter, were greater in athletes than sedentary controls. Left ventricular mass of all athletes but relative wall thickness of only BP, swimmers, cyclists, and strength/power athletes were higher as compared with controls (p < 0.05). Among athletes, smaller IVS thickness was observed in MDR than BP, cyclists, swimmers or strength/power athletes, while LDR had higher body size-adjusted LV diameter as compared to BP, cyclists and strength/power athletes. In conclusion, relative LV diameter was increased in long distance runners as compared with basketball players, cyclists, and strength/power athletes. Basketball, road cycling, strength/power, and swimming training were associated with increased LV concentricity as compared with paddling or distance running. Key pointsThe type of cardiac hypertrophy seems to be only moderately exercise-specific.Long-distance runners develop larger left ventricular dilation as compared with basketball players, cyclists, and strength/power athletes.Myocardial wall thickening is triggered by different sporting activities that involve large muscle groups.


Modulation of ionizing radiation induced oxidative imbalance by semi-fractionated extract of Piper betle: an in vitro and in vivo assessment.
The study was planned to evaluate modulatory effect of aqueous extract of Piper betle leaf (PBL) on ionizing radiation mediated oxidative stress leading to normal tissues damage during radiotherapy and other radiation exposures. The total polyphenols and flavonoids known as free radical scavenger (chelators) were measured in the extract. To ascertain antioxidant potential of PBL extract we studied free radical scavenging, metal chelation, reducing power, lipid peroxidation inhibition and ferric reducing antioxidant properties (FRAP) using in vitro assays. Mice were exposed to varied radiation doses administered with the same extract prior to irradiation to confirm its oxidative stress minimizing efficacy by evaluating ferric reducing ability of plasma, reduced glutathione, lipid peroxidation and micro-nuclei frequency. PBL extract was effective in scavenging DPPH (up to 92% at 100 microg/ml) and superoxide radicals (up to 95% at 80 microg/ml), chelated metal ions (up to 83% at 50 microg/ml) and inhibited lipid peroxidation (up to 55.65% at 500 microg/ml) in a dose dependant manner using in vitro model. Oral administration of PBL extract (225 mg/kg body weight) 1 hr before irradiation in mice significantly enhanced (p < 0.01) radiation abated antioxidant potential of plasma and GSH level in all the observed organs. The treatment with extract effectively lowered the radiation induced lipid peroxidation at 24 hrs in all the selected organs with maximum inhibition in thymus (p < 0.01). After 48 hrs, lipid peroxidation was maximally inhibited in the group treated with the extract. Frequency of radiation induced micronucleated cells declined significantly (34.78%, p < 0.01) at 24 hrs post-irradiation interval by PBL extract administration. The results suggest that PBL extract has high antioxidant potential and relatively non-toxic and thus could be assertively used to mitigate radiotherapy inflicted normal tissues damage and also injuries caused by moderate doses of radiation during unplanned exposures.