PKCζ-Mitogen-Activated Protein Kinase Signaling Mediates Crotalphine-Induced Antinociception.
Crotalphine (CRP) is a structural analogue to a peptide that was first identified in the crude venom from the South American rattlesnake Crotalus durissus terrificus. This peptide induces a potent and long-lasting antinociceptive effect that is mediated by the activation of peripheral opioid receptors. The opioid receptor activation regulates a variety of intracellular signaling, including the mitogen-activated protein kinase (MAPK) pathway. Using primary cultures of sensory neurons, it was demonstrated that crotalphine increases the level of activated ERK1/2 and JNK-MAPKs and this increase is dependent on the activation of protein kinase Cζ (PKCζ). However, whether PKCζ-MAPK signaling is critical for crotalphine-induced antinociception is unknown. Here, we biochemically demonstrated that the systemic crotalphine activates ERK1/2 and JNK and decreases the phosphorylation of p38 in the lumbar spinal cord. The in vivo pharmacological inhibition of spinal ERK1/2 and JNK, but not of p38, blocks the antinociceptive effect of crotalphine. Of interest, the administration of a PKCζ pseudosubstrate (PKCζ inhibitor) prevents crotalphine-induced ERK activation in the spinal cord, followed by the abolishment of crotalphine-induced analgesia. Together, our results demonstrate that the PKCζ-ERK signaling pathway is involved in crotalphine-induced analgesia. Our study opens a perspective for the PKCζ-MAPK axis as a target for pain control.


Evaluation of 3 approaches for increasing patient engagement in clinical research: Feedback from a community engagement advisory board.
The purpose of this study was to obtain feedback from a diverse group of community advisory board members about different clinic or hospital-based approaches to increasing research participation. Members of an established community engagement advisory board (n=16) provided qualitative and survey data regarding attitudes and preferences for 3 hospital and clinic system strategies to recruit patients into clinical research including universal consent for research, patient registries, and patient portals. Overall, there was moderate support for each of the 3 approaches discussed. Board members described advantages and disadvantages of each method. Based on the qualitative data, universal consent was viewed as the best strategy for consenting high volumes of patients for research. However, patient registries and portals were seen as more acceptable, less-intrusive and more likely to result in higher participation rates. Survey data were consistent with qualitative findings. Input from community stakeholders is needed to identify strategies to enhance participation and increase diversity in clinical research. Members of our CEAB identified patient registries and portals as feasible and nonintrusive approaches to increasing research participation. Additional research is needed to confirm these findings and to establish best practices for supporting patients in using registry approaches.


A streamlined failure mode and effects analysis.
Explore the feasibility and impact of a streamlined failure mode and effects analysis (FMEA) using a structured process that is designed to minimize staff effort. FMEA for the external beam process was conducted at an affiliate radiation oncology center that treats approximately 60 patients per day. A structured FMEA process was developed which included clearly defined roles and goals for each phase. A core group of seven people was identified and a facilitator was chosen to lead the effort. Failure modes were identified and scored according to the FMEA formalism. A risk priority number,RPN, was calculated and used to rank failure modes. Failure modes with RPN > 150 received safety improvement interventions. Staff effort was carefully tracked throughout the project. Fifty-two failure modes were identified, 22 collected during meetings, and 30 from take-home worksheets. The four top-ranked failure modes were: delay in film check, missing pacemaker protocol/consent, critical structures not contoured, and pregnant patient simulated without the team's knowledge of the pregnancy. These four failure modes had RPN > 150 and received safety interventions. The FMEA was completed in one month in four 1-h meetings. A total of 55 staff hours were required and, additionally, 20 h by the facilitator. Streamlined FMEA provides a means of accomplishing a relatively large-scale analysis with modest effort. One potential value of FMEA is that it potentially provides a means of measuring the impact of quality improvement efforts through a reduction in risk scores. Future study of this possibility is needed.


In vitro flavon-3-ol oxidation mediated by a B ring hydroxylation pattern.
Flavonols are potent naturally occurring antioxidants. Chemical oxidation reactions in combination with modern spectroscopic techniques have been employed to identify oxidized flavonoid products. Although many oxidized derivatives have been generated from commercially available starting materials, few studies have developed a sequence of flavonoid substrates with a particular hydroxylation pattern to probe the mechanism of flavonoid oxidation. Here, we use AIBN (2,2'-azobisisobutyronitrile) in combination with a series of hydroxylated flavonols to probe the mechanism of flavonoid dimer formation and the role of singly or doubly oxidized species in generating and promoting oxidized flavonoid products. 3-Methoxyquercetin (3-hydroxyl group blocked) and luteolin (lack of 3-hydroxyl) were reacted with AIBN alone or with a second flavonol to serve as a C-3 hydroxyl donor to examine the mechanism of dimer formation. 3-Hydroxyflavones with increasing hydroxyl substitutions in the B ring were also reacted with AIBN in the presence or absence of an external nucleophile to examine the role of various hydroxyls in the formation of a carbocation intermediate via a doubly oxidized species. The presence of a free C-3 hydroxyl, coupled with a B ring ortho hydroxy unit, appears essential for dimer formation. An increase in the number of hydroxyls in the B ring facilitates products generated from a doubly oxidized species.


Upregulation of genes related to bone formation by γ-amino butyric acid and γ-oryzanol in germinated brown rice is via the activation of GABAB-receptors and reduction of serum IL-6 in rats.
Osteoporosis and other bone degenerative diseases are among the most challenging non-communicable diseases to treat. Previous works relate bone loss due to osteoporosis with oxidative stress generated by free radicals and inflammatory cytokines. Alternative therapy to hormone replacement has been an area of interest to researchers for almost three decades due to hormone therapy-associated side effects. In this study, we investigated the effects of gamma-amino butyric acid (GABA), gamma-oryzanol (ORZ), acylated steryl glucosides (ASG), and phenolic extracts from germinated brown rice (GBR) on the expression of genes related to bone metabolism, such as bone morphogenic protein-2 (BMP-2), secreted protein acidic and rich in cysteine (SPARC), runt-related transcription factor 2 (RUNX-2), osteoblast-specific transcription factor osterix (Osx), periostin, osteoblast specific factor (Postn), collagen 1&2 (Col1&2), calcitonin receptor gene (CGRP); body weight measurement and also serum interleukin-6 (IL-6) and osteocalcin, in serum and bone. Rats were treated with GBR, ORZ, GABA, and ASG at (100 and 200 mg/kg); estrogen (0.2 mg/kg), or remifemin (10 and 20 mg/kg), compared to ovariectomized non-treated group as well as non-ovariectomized non-treated (sham) group. Enzyme-linked immunosorbent assay was used to measure the IL-6 and osteocalcin levels at week 2, 4, and 8, while the gene expression in the bone tissue was determined using the Genetic Analysis System (Beckman Coulter Inc., Brea, CA, USA). The results indicate that groups treated with GABA (100 and 200 mg/kg) showed significant upregulation of SPARC, calcitonin receptor, and BMP-2 genes (P < 0.05), while the ORZ-treated group (100 and 200 mg/kg) revealed significant (P < 0.05) upregulation of Osx, Postn, RUNX-2, and Col1&2. Similarly, IL-6 concentration decreased, while osteocalcin levels increased significantly (P < 0.05) in the treated groups as compared to ovariectomized non-treated groups. GABA and ORZ from GBR stimulates osteoblastogenesis by upregulation of bone formation genes, possibly via the activation of GABAB receptors and by inhibiting the activity of inflammatory cytokines and reactive oxygen species. Therefore, it could be used effectively in the management of osteoporosis.


A temporal dimension to the influence of pollen rewards on bee behaviour and fecundity in Aloe tenuior.
The net effect of pollen production on fecundity in plants can range from negative--when self-pollen interferes with fecundity due to incompatibility mechanisms, to positive--when pollen availability is associated with increased pollinator visitation and fecundity due to its utilization as a reward. We investigated the responses of bees to pollen and nectar rewards, and the effects of these rewards on pollen deposition and fecundity in the hermaphroditic succulent shrub Aloe tenuior. Self-pollinated plants failed to set fruit, but their ovules were regularly penetrated by self-pollen tubes, which uniformly failed to develop into seeds as expected from ovarian self-incompatibility (or strong early inbreeding depression). Bees consistently foraged for pollen during the morning and early afternoon, but switched to nectar in the late afternoon. As a consequence of this differential foraging, we were able to test the relative contribution to fecundity of pollen- versus nectar-collecting flower visitors. We exposed emasculated and intact flowers in either the morning or late afternoon to foraging bees and showed that emasculation reduced pollen deposition by insects in the morning, but had little effect in the afternoon. Despite the potential for self-pollination to result in ovule discounting due to late-acting self-sterility, fecundity was severely reduced in artificially emasculated plants. Although there were temporal fluctuations in reward preference, most bee visits were for pollen rewards. Therefore the benefit of providing pollen that is accessible to bee foragers outweighs any potential costs to fitness in terms of gender interference in this species.


Native gel electrophoresis of human telomerase distinguishes active complexes with or without dyskerin.
Telomeres, the ends of linear chromosomes, safeguard against genome instability. The enzyme responsible for extension of the telomere 3' terminus is the ribonucleoprotein telomerase. Whereas telomerase activity can be reconstituted in vitro with only the telomerase RNA (hTR) and telomerase reverse transcriptase (TERT), additional components are required in vivo for enzyme assembly, stability and telomere extension activity. One such associated protein, dyskerin, promotes hTR stability in vivo and is the only component to co-purify with active, endogenous human telomerase. We used oligonucleotide-based affinity purification of hTR followed by native gel electrophoresis and in-gel telomerase activity detection to query the composition of telomerase at different purification stringencies. At low salt concentrations (0.1 M NaCl), affinity-purified telomerase was 'supershifted' with an anti-dyskerin antibody, however the association with dyskerin was lost after purification at 0.6 M NaCl, despite the retention of telomerase activity and a comparable yield of hTR. The interaction of purified hTR and dyskerin in vitro displayed a similar salt-sensitive interaction. These results demonstrate that endogenous human telomerase, once assembled and active, does not require dyskerin for catalytic activity. Native gel electrophoresis may prove useful in the characterization of telomerase complexes under various physiological conditions.


Chemosensory age discrimination in the snake Boa constrictor (Serpentes: Boidae).
Many snakes are able to use their chemosensory system to detect scent of conspecifics, which is important in many social contexts. Age discrimination based on chemical cues may be especially important to ensure access to sexually mature potential partners. In this study, we used 24 individual Boa constrictor snakes (12 adults mature and 12 non-mature individuals) that had been captured in different areas of Ecuador, and were maintained in captivity at the Vivarium of Quito. We used tongue-flick experiments to examine whether these snakes were able to discriminate between scents from mature and non-mature individuals. Results showed that B. constrictor snakes used chemical cues to recognize conspecifics and that the scent of individuals of different ages elicited chemosensory responses of different magnitudes. The scents from adult conspecifics elicited the quickest and highest chemosensory responses (i.e., short latency times and high tongue-flick rates), although we did not find differential responses to scent of males and females. The magnitude of the responses was lower to scent of sub adult individuals, and then even lower to scent of juvenile snakes, but in all cases the scent of snakes was discriminated from a blank control. We discuss the potential chemical mechanisms that may allow age recognition and its implications for social and sexual behavior of this snake species.


The ability of the acoustic energy/stone burden ratio to predict gallstone clearance in biliary lithotripsy.
The relationship between acoustic energy, stone burden, and stone clearance in biliary extracorporeal shock wave therapy (ESWL) is unclear. We hypothesized that there is a consistent relationship between these parameters that enables gallstone clearance to be predicted. Using a porcine model and various gallstone burdens, we performed biliary ESWL with varying energy levels and number of shock waves. The acoustic energy/stone burden ratio (AE/SB) was calculated that incorporated these variables. Stone clearance increased with higher energy settings and fewer stones present (p less than 0.005). The relationship between AE/SB ratio and stone clearance was logarithmic (r 2 = 0.82). The plateau of this curve showed optimal stone clearance at approximately 20 joules/g, beyond which point the benefit of further ESWL may not outweigh the risks. This relationship between AE/SB ratio and stone clearance may be able to predict the optimal energy required and probable outcome of biliary ESWL prior to the procedure.


[Walk training and drug therapy in peripheral arterial occlusive disease. A randomized, prospective, placebo-controlled double-blind study].
It was demonstrated in a prospective, randomized and placebo-controlled double-blind study of two groups of 15 patients each with peripheral arterial occlusive disease in stage IIb of Fontaine that the combination of intravenous and oral administration of Pentoxifylline with walk training was superior to walk training alone. Comparison of the two groups revealed a significant increase in pain-free walking capacity, as well as reduction in plasma viscosity and platelet aggregation in the patients of the combination group. A significant reduction in platelet aggregation and rigidity occurred with time in this group only, while there was no significant difference between the groups with regard to these two parameters at the end of the treatment period.


Colonization by Arbuscular Mycorrhizal Fungi of Sorghum Leads to Reduced Germination and Subsequent Attachment and Emergence of Striga hermonthica.
TWO SORGHUM CULTIVARS: the Striga-tolerant S-35 and the Striga-sensitive CK60-B were grown with or without arbuscular mycorrhizal (AM) fungi, and with or without phosphorus addition. At 24 and 45 days after sowing (DAS) of sorghum, root exudates were collected and tested for effects on germination of preconditioned Striga hermonthica seeds. Root exudates from AM sorghum plants induced lower germination of S. hermonthica seeds than exudates from non-mycorrhizal sorghum. The magnitude of this effect depended on the cultivar and harvest time. A significantly (88-97%) lower germination of S. hermonthica seeds upon exposure to root exudates from AM S-35 plants was observed at both harvest times whereas for AM inoculated CK60-B plants a significantly (41%) lower germination was observed only at 45 DAS. The number of S. hermonthica seedlings attached to and emerged on both sorghum cultivars were also lower in mycorrhizal than in non-mycorrhizal plants. Again, this reduction was more pronounced with S-35 than with CK60-B plants. There was no effect of phosphorus addition on Striga seed germination, attachment or emergence. We hypothesize that the negative effect of mycorrhizal colonization on Striga germination and on subsequent attachment and emergence is mediated through the production of signaling molecules (strigolactones) for AM fungi and parasitic plants.


Comparison of Rx-defined morbidity groups and diagnosis- based risk adjusters for predicting healthcare costs in Taiwan.
Medication claims are commonly used to calculate the risk adjustment for measuring healthcare cost. The Rx-defined Morbidity Groups (Rx-MG) which combine the use of medication to indicate morbidity have been incorporated into the Adjusted Clinical Groups (ACG) Case Mix System, developed by the Johns Hopkins University. This study aims to verify that the Rx-MG can be used for adjusting risk and for explaining the variations in the healthcare cost in Taiwan. The Longitudinal Health Insurance Database 2005 (LHID2005) was used in this study. The year 2006 was chosen as the baseline to predict healthcare cost (medication and total cost) in 2007. The final sample size amounted to 793,239 (81%) enrolees, and excluded any cases with discontinued enrolment. Two different kinds of models were built to predict cost: the concurrent model and the prospective model. The predictors used in the predictive models included age, gender, Aggregated Diagnosis Groups (ADG, diagnosis- defined morbidity groups), and Rx-defined Morbidity Groups. Multivariate OLS regression was used in the cost prediction modelling. The concurrent model adjusted for Rx-defined Morbidity Groups for total cost, and controlled for age and gender had a better predictive R-square = 0.618, compared to the model adjusted for ADGs (R2 = 0.411). The model combined with Rx-MGs and ADGs performed the best for concurrently predicting total cost (R2 = 0.650). For prospectively predicting total cost, the model combined Rx-MGs and ADGs (R2 = 0.382) performed better than the models adjusted by Rx-MGs (R2 = 0.360) or ADGs (R2 = 0.252) only. Similarly, the concurrent model adjusted for Rx-MGs predicting pharmacy cost had a better performance (R-square = 0.615), than the model adjusted for ADGs (R2 = 0.431). The model combined with Rx-MGs and ADGs performed the best in concurrently as well as prospectively predicting pharmacy cost (R2 = 0.638 and 0.505, respectively). The prospective models showed a remarkable improvement when adjusted by prior cost. The medication-based Rx-Defined Morbidity Groups was useful in predicting pharmacy cost as well as total cost in Taiwan. Combining the information on medication and diagnosis as adjusters could arguably be the best method for explaining variations in healthcare cost.


Adenylate cyclase is required for chemotaxis to phosphotransferase system sugars by Escherichia coli.
We report that in Escherichia coli, chemotaxis to sugars transported by the phosphotransferase system is mediated by adenylate cyclase, the nucleotide cyclase linked to the phosphotransferase system. We conclude that adenylate cyclase is required in this chemotaxis pathway because mutations in the cyclase gene (cya) eliminate or impair the response to phosphotransferase system sugars, even though other components of the phosphotransferase system known to be required for the detection of these sugars are relatively unaffected by such mutations. Moreover, merely supplying the mutant bacteria with the products of this enzyme, cyclic AMP and cyclic GMP, does not restore the chemotactic response. Because a residual chemotactic response is observed in certain strains with residual cyclic GMP synthesis but no cyclic AMP synthesis, it appears that the guanylate cyclase activity rather than the adenylate cyclase activity of the enzyme may be required for chemotaxis to sugars transported by the phosphotransferase system. Mutations in the cyclic nucleotide phosphodiesterase gene, which increase the level of both cyclic AMP and cyclic GMP, also reduce chemotaxis to these sugars. Therefore, it appears that control of the level of a cyclic nucleotide is critical for the chemotactic response to phosphotransferase system sugars.


The pupillary light reflex in diabetes mellitus: evaluation of a newly developed infrared light reflection method.
Pupillary light reflexes were measured in 18 diabetic patients without clinical signs of neuropathy and in 18 control subjects, using a newly developed infrared light reflection technique called IRIS. This method has some important advantages. Apart from being non traumatic, it enables the recording of dynamic pupillary light reflexes simultaneously in both eyes. Furthermore, it has a large amplitude and time resolution (1 ms), permitting accurate determination of latency values. It was found that 78% of the diabetic patients show a significantly prolonged latency of the constriction reaction, while 39% of the diabetic patients show an abnormally prolonged dilatation latency. All patients with an abnormally prolonged dilatation latency also show a prolonged constriction latency. This finding suggests that in diabetes mellitus the parasympathetic system is affected before abnormalities of the sympathetic nervous system can be demonstrated. Assessment of the pupillary light reflex, using the sensitive IRIS method, is an easily applicable and reliable method for detection of subclinical autonomic nervous system dysfunction associated with diabetes mellitus. The method may also provide an objective tool for the assessment of therapy results in diabetic neuropathy.


Status of bi- and multi-nuclear platinum anticancer drug development.
Cisplatin has become one of the most commonly used compounds for the treatment of a wide spectrum of human malignancies. Unfortunately, cisplatin has several major drawbacks. Driven by the impressive impact of cisplatin on cancer chemotherapy, great efforts have been made to develop new derivatives with improved pharmacological properties. Among the over 30 platinum agents which have entered clinical trials after the onset of clinical studies with cisplatin in the early 1970s, only carboplatin and oxaliplatin have received worldwide approval so far, nedaplatin, lobaplatin and heptaplatin have gained regionally limited approval. It has become quite evident that mere analogues of cisplatin or carboplatin will not probably offer any substantial clinical advantages over the existing drugs. Therefore, people turned to synthesize non-classical platinum anticancer drugs which were capable of forming a different range of DNA adducts which could display a different spectrum of anticancer activity compared to cisplatin. This review will summarize the structural types and structure-activity rules of non-classical bi- and multi-nuclear platinum anticancer drugs, and discuss their future potential as anticancer agents.


Practical rare event sampling for extreme mesoscale weather.
Extreme mesoscale weather, including tropical cyclones, squall lines, and floods, can be enormously damaging and yet challenging to simulate; hence, there is a pressing need for more efficient simulation strategies. Here, we present a new rare event sampling algorithm called quantile diffusion Monte Carlo (quantile DMC). Quantile DMC is a simple-to-use algorithm that can sample extreme tail behavior for a wide class of processes. We demonstrate the advantages of quantile DMC compared to other sampling methods and discuss practical aspects of implementing quantile DMC. To test the feasibility of quantile DMC for extreme mesoscale weather, we sample extremely intense realizations of two historical tropical cyclones, 2010 Hurricane Earl and 2015 Hurricane Joaquin. Our results demonstrate quantile DMC's potential to provide low-variance extreme weather statistics while highlighting the work that is necessary for quantile DMC to attain greater efficiency in future applications.


Novel application of imaging surface plasmon resonance for in situ studies of the surface exploration of marine organisms.
The surface interactions of exploring cyprids of the barnacle Semibalanus balanoides were studied in situ using imaging surface plasmon resonance. It was demonstrated how the deposition of a proteinaceous adhesive could be followed in real time as the cyprids explored and temporarily attached to a surface. Furthermore, the amount of protein left on the surface when the cyprids moved on could be quantified. Clear differences were demonstrated between an oligo(ethyleneglycol) coated surface and a bare gold substrate. It is anticipated that this technique will be a valuable tool in the development of novel surface chemistries that can prevent biofouling.


Are Filter-Tipped Cigarettes Still Less Harmful than Non-Filter Cigarettes?--A Laser Spectrometric Particulate Matter Analysis from the Non-Smokers Point of View.
Environmental tobacco smoke (ETS) is associated with human morbidity and mortality, particularly chronic obstructive pulmonary disease (COPD and lung cancer. Although direct DNA-damage is a leading pathomechanism in active smokers, passive smoking is enough to induce bronchial asthma, especially in children. Particulate matter (PM) demonstrably plays an important role in this ETS-associated human morbidity, constituting a surrogate parameter for ETS exposure. Using an Automatic Environmental Tobacco Smoke Emitter (AETSE) and an in-house developed, non-standard smoking regime, we tried to imitate the smoking process of human smokers to demonstrate the significance of passive smoking. Mean concentration (C(mean)) and area under the curve (AUC) of particulate matter (PM2.5) emitted by 3R4F reference cigarettes and the popular filter-tipped and non-filter brand cigarettes "Roth-Händle" were measured and compared. The cigarettes were not conditioned prior to smoking. The measurements were tested for Gaussian distribution and significant differences. C(mean) PM2.5 of the 3R4F reference cigarette: 3911 µg/m³; of the filter-tipped Roth-Händle: 3831 µg/m³; and of the non-filter Roth-Händle: 2053 µg/m³. AUC PM2.5 of the 3R4F reference cigarette: 1,647,006 µg/m³·s; of the filter-tipped Roth-Händle: 1,608,000 µg/m³·s; and of the non-filter Roth-Händle: 858,891 µg/m³·s. The filter-tipped cigarettes (the 3R4F reference cigarette and filter-tipped Roth-Händle) emitted significantly more PM2.5 than the non-filter Roth-Händle. Considering the harmful potential of PM, our findings note that the filter-tipped cigarettes are not a less harmful alternative for passive smokers. Tobacco taxation should be reconsidered and non-smoking legislation enforced.


Cancer hyperthermia using magnetic nanoparticles.
Magnetic-nanoparticle-mediated intracellular hyperthermia has the potential to achieve localized tumor heating without any side effects. The technique consists of targeting magnetic nanoparticles to tumor tissue followed by application of an external alternating magnetic field that induces heat through Néel relaxation loss of the magnetic nanoparticles. The temperature in tumor tissue is increased to above 43°C, which causes necrosis of cancer cells, but does not damage surrounding normal tissue. Among magnetic nanoparticles available, magnetite has been extensively studied. Recent years have seen remarkable advances in magnetite-nanoparticle-mediated hyperthermia; both functional magnetite nanoparticles and alternating-magnetic-field generators have been developed. In addition to the expected tumor cell death, hyperthermia treatment has also induced unexpected biological responses, such as tumor-specific immune responses as a result of heat-shock protein expression. These results suggest that hyperthermia is able to kill not only local tumors exposed to heat treatment, but also tumors at distant sites, including metastatic cancer cells. Currently, several research centers have begun clinical trials with promising results, suggesting that the time may have come for clinical applications. This review describes recent advances in magnetite nanoparticle-mediated hyperthermia.


Fine structure and evolution of the rat serum albumin gene.
The exons, their boundaries, and approximately half of the intronic deoxyribonucleic acid of the rat serum albumin gene were sequenced. In addition to the 14 exons identified earlier by R-loop analysis, a small exon was detected between the "leader" exon (Z) and exon B. The leader exon encoded the 5'-untranslated portion of albumin messenger ribonucleic acid and the "pre-pro" oligopeptide present on the nascent protein. The sites of initiation and termination of transcription were tentatively identified by comparison of the 5' and 3' gene-flanking sequences with those of other eucaryotic genes. All 28 intron/exon junctions conformed to the "GT-AG rule" (Breathnach et al., Proc. Natl. Acad. Sci. 75:4853-4857, 1978). The three homologous domains of albumin were encoded by three subgenes that consisted of four exons each and evolved by intragenic duplication of a common ancestor. The second and forth exons of each subgene appeared to be the result of an even earlier duplication event. We propose a model for the evolution of this gene that accounts for the observed patterns of exon size and homology.


Sleep deprivation reduces total plasma homocysteine levels in rats.
Hyperhomocysteinemia has been associated with pathological and stressful conditions and is a risk factor for cardiovascular disease. Since sleep deprivation is a stressful condition that is associated with disruption of various physiological processes, we investigated whether it would also be associated with increases in plasma homocysteine levels. Further, since hyperhomocysteinemia may promote oxidative stress, and we had previously found evidence of oxidative stress in brain following sleep deprivation, we also searched for evidence of systemic oxidative stress by measuring glutathione and thiobarbituric acid reactive substance levels. Rats were sleep deprived for 96 h using the platform technique. A group was killed after sleep deprivation and another two groups were allowed to undergo sleep recovery for 24 or 48 h. Contrary to expectation, plasma homocysteine was reduced in sleep-deprived rats as compared with the control group and did not revert to normal levels after 24 or 48 h of sleep recovery. A trend was observed towards decreased glutathione and increased thiobarbituric acid reactive substance levels in sleep-deprived rats. It is possible that the observed decreases in homocysteine levels may represent a self-correcting response to depleted glutathione in sleep-deprived animals, which would contribute to the attenuation of the deleterious effects of sleep deprivation.


Multinucleate cell angiohistiocytoma: a case report and review of the literature.
Multinucleate cell angiohistiocytoma (MCAH) is a rare, benign vascular proliferation. Fewer than 80 cases have been reported to date, which may relate to under-recognition of this entity. Lesions are commonly asymptomatic and appear as erythematous to violaceous papules on the lower extremities and dorsal hands of middle-aged to elderly women. The characteristic histopathologic and immunohistochemical features of MCAH are essential for definitive diagnosis of MCAH. Multinucleate cell angiohistiocytoma follows a slowly progressive course, although spontaneous regression has been reported in rare cases. We present a case of MCAH to increase awareness and elucidate the characteristic clinical and histopathologic features of this disorder.


The role of ribonucleases in regulating global mRNA levels in the model organism Thermus thermophilus HB8.
RNA metabolism, including RNA synthesis and RNA degradation, is one of the most conserved biological systems and has been intensively studied; however, the degradation network of ribonucleases (RNases) and RNA substrates is not fully understood. The genome of the extreme thermophile, Thermus thermophilus HB8 includes 15 genes that encode RNases or putative RNases. Using DNA microarray analyses, we examined the effects of disruption of each RNase on mRNA abundance. Disruption of the genes encoding RNase J, RecJ-like protein and RNase P could not be isolated, indicating that these RNases are essential for cell viability. Disruption of the TTHA0252 gene, which was not previously considered to be involved in mRNA degradation, affected mRNA abundance, as did disruption of the putative RNases, YbeY and PhoH-like proteins, suggesting that they have RNase activity. The effects on mRNA abundance of disruption of several RNase genes were dependent on the phase of cell growth. Disruption of the RNase Y and RNase HII genes affected mRNA levels only during the log phase, whereas disruption of the PhoH-like gene affected mRNA levels only during the stationary phase. Moreover, disruption of the RNase R and PNPase genes had a greater impact on mRNA abundance during the stationary phase than the log phase, whereas the opposite was true for the TTHA0252 gene disruptant. Similar changes in mRNA levels were observed after disruption of YbeY or PhoH-like genes. The changes in mRNA levels in the bacterial Argonaute disruptant were similar to those in the RNase HI and RNase HII gene disruptants, suggesting that bacterial Argonaute is a functional homolog of RNase H. This study suggests that T. thermophilus HB8 has 13 functional RNases and that each RNase has a different function in the cell. The putative RNases, TTHA0252, YbeY and PhoH-like proteins, are suggested to have RNase activity and to be involved in mRNA degradation. In addition, PhoH-like and YbeY proteins may act cooperatively in the stationary phase. This study also suggests that endo-RNases function mainly during the log phase, whereas exo-RNases function mainly during the stationary phase. RNase HI and RNase HII may have similar substrate selectivity.


In vitro inhibition of Pseudomonas aeruginosa elastase by metal-chelating peptide derivatives.
Pseudomonas aeruginosa elastase is a zinc metalloendopeptidase, probably responsible for the tissue destruction observed during infections with this organism. The elastase of a virulent Pseudomonas aeruginosa strain (Habs serotype 1) was isolated and found to have a molecular weight of 35,000; it readily degraded elastin and cartilage proteoglycans. A series of amino acid and peptide derivatives containing the metal-chelating moieties hydroxamate, phosphoryl, or thiol were synthesized and tested as potential inhibitors of the enzyme. Inhibition constants (K(i)s) for the compounds were determined with the chromophoric substrate furylacryloyl-glycyl-l-leucyl-l-alanine. The hydroxamic acid derivatives of benzyloxycarbonyl-glycine, benzyloxycarbonyl-l-leucine and benzyloxycarbonyl-l-phenylalanine had inhibition constants in the range of 11 to 28 muM. The 2-mercaptoacetyl derivatives of l-leucyl-d-phenylalanine and l-leucyl-l-phenylalanine had K(i) values of 34 and 1.5 muM, respectively, demonstrating the stereospecificity of the inhibition. The most potent inhibitors tested were 2- mercaptoacetyl-l-phenylalanyl-l-leucine and phosphoryl-l-leucyl-l-phenylala-nine (K(i) = 0.2 muM). Similar compounds lacking the metal-chelating moiety were about 3 orders of magnitude poorer inhibitors. When the inhibition of the enzyme activity towards azocasein, elastin, or cartilage was examined, inhibitor concentrations approximately 50-fold higher than the respective K(i)s were required to obtain 60 to 90% inhibition. Virtually complete inhibition was achieved with these substrates at inhibitor concentrations 500-fold higher than the respective K(i)s (0.1 to 14 mM). Although, 2-mercaptoacetyl-l-phenylalanyl-l-leucine and phosphoryl-l-leucyl-l-phenylalanine exhibited the same affinity to the enzyme, the latter was inferior in inhibiting cartilage proteoglycan degradation. 2-Mercaptoacetyl-l-phenylalanyl-l-leucine represents a class of potent elastase inhibitors that might prove useful in the management of P. aeruginosa infections.


Hydrogen formation and its regulation in Ruminococcus albus: involvement of an electron-bifurcating [FeFe]-hydrogenase, of a non-electron-bifurcating [FeFe]-hydrogenase, and of a putative hydrogen-sensing [FeFe]-hydrogenase.
Ruminococcus albus 7 has played a key role in the development of the concept of interspecies hydrogen transfer. The rumen bacterium ferments glucose to 1.3 acetate, 0.7 ethanol, 2 CO2, and 2.6 H2 when growing in batch culture and to 2 acetate, 2 CO2, and 4 H2 when growing in continuous culture in syntrophic association with H2-consuming microorganisms that keep the H2 partial pressure low. The organism uses NAD(+) and ferredoxin for glucose oxidation to acetyl coenzyme A (acetyl-CoA) and CO2, NADH for the reduction of acetyl-CoA to ethanol, and NADH and reduced ferredoxin for the reduction of protons to H2. Of all the enzymes involved, only the enzyme catalyzing the formation of H2 from NADH remained unknown. Here, we report that R. albus 7 grown in batch culture on glucose contained, besides a ferredoxin-dependent [FeFe]-hydrogenase (HydA2), a ferredoxin- and NAD-dependent electron-bifurcating [FeFe]-hydrogenase (HydABC) that couples the endergonic formation of H2 from NADH to the exergonic formation of H2 from reduced ferredoxin. Interestingly, hydA2 is adjacent to the hydS gene, which is predicted to encode an [FeFe]-hydrogenase with a C-terminal PAS domain. We showed that hydS and hydA2 are part of a larger transcriptional unit also harboring putative genes for a bifunctional acetaldehyde/ethanol dehydrogenase (Aad), serine/threonine protein kinase, serine/threonine protein phosphatase, and a redox-sensing transcriptional repressor. Since HydA2 and Aad are required only when R. albus grows at high H2 partial pressures, HydS could be a H2-sensing [FeFe]-hydrogenase involved in the regulation of their biosynthesis.


Heart Rate Variability Responses of Individuals With and Without Saline-Induced Obstructive Sleep Apnea.
Postoperative development of obstructive sleep apnea (OSA) has been attributed to the fluid overloaded state of patients during the postoperative period. In this context, alterations in cardiac autonomic regulation caused by OSA may explain the increased postoperative risk for adverse cardiovascular events. This study tests the hypothesis that individuals with fluid overload-induced OSA will experience autonomic dysregulation, compared to those without fluid overload-induced OSA. Twenty-one normotensive, nonobese (mean body mass index 24.5 kg/m2) males (mean age 37 years) underwent a sleep study. Participants were randomly assigned to infusion with saline during sleep either at the minimum rate (control) or as a bolus of 22 mL/kg body weight (intervention). Participants were blinded to the intervention and crossed over to the other study arm after 1 week. Measures of heart rate variability were calculated from electrocardiography recordings presaline and postsaline infusion in the intervention arm. Heart rate variability measures computed were: standard deviation of the RR interval; root mean square of successive differences; low-frequency, high-frequency, and total power; and the ratio of low-frequency to high-frequency power. Although presaline infusion values were similar, postsaline infusion values of the standard deviation of the RR interval and high-frequency power were lower in the group whose apnea-hypopnea index increased in response to saline infusion, compared to the group whose apnea-hypopnea index did not increase in response to saline infusion (P < .05 for both). Fluid overload-induced OSA is accompanied by a reduction in heart rate variability, consistent with vagal withdrawal. Future work should explore autonomic dysregulation in the postoperative period and its association with adverse events.


Vascular smooth muscle: quantitation of cell thickness in the wall of arterioles in the living animal in situ.
The wall of microarterial vessels, in the living animal in situ, was examined at magnification up to x 6500 on the television screen. The optical resolution af the cell components of the wall was sufficiently clear to permit image formation and splitting for permanent recording and quantitation. Thickness of single smooth muscle cells at rest was estimated to be 2.08 microns (S.D. +/- 0.24 micron) and 2.78 microns (S.D. +/- 0.59 micron) by two different approaches. Changes in cell thickness during activity were also recorded and quantitated.


Sweat but no gain: inhibiting proliferation of multidrug resistant cancer cells with "ersatzdroges".
ATP-binding cassette (ABC) drug transporters consuming ATPs for drug efflux is a common mechanism by which clinical cancers develop multidrug resistance (MDR). We hypothesized that MDR phenotypes could be suppressed by administration of "ersatzdroges," nonchemotherapy drugs that are, nevertheless, ABC substrates. We reasoned that, through prolonged activation of the ABC pumps, ersatzdroges will force MDR cells to divert limited resources from proliferation and invasion thus delaying disease progression. We evaluated ABC substrates as ersatzdroge by comparing their effects on proliferation and survival of MDR cell lines (MCF-7/Dox and 8226/Dox40) with the effects on the drug-sensitive parental lines (MCF-7 and 8226/s, respectively) in glucose-limited condition. The changes in glucose and energy demands were also examined in vitro and in vivo. MCF-7/Dox showed higher ATP demand and susceptibility to glucose resource limitation. Ersatzdroges significantly decreased proliferation of MCF-7/Dox when the culture media contained physiological glucose concentrations (1.0 g/L) or less, but had no effect on MCF-7. Similar evidence was obtained from 8226/Dox40 and 8226/s comparison. In vivo 18F-FDG-PET imaging demonstrated that glucose uptake was increased by systemic administration of an ersatzdroge in tumors composed of MDR. These results suggest that administration of ersatzdroges, by increasing the metabolic cost of resistance, can suppress proliferation of drug-resistance phenotypes. This provides a novel and relatively simple application model of evolution-based strategy, which can exploit the cost of resistance to delay proliferation of drug-resistant cancer phenotypes. Furthermore, suggested is the potential of ersatzdroges to identify tumors or regions of tumors that express the MDR phenotype.


Pharmacological modulation of bronchial hyperreactivity.
The airways of asthmatics are hyperreactive, not only to allergens but to a wide range of non-specific physical and chemical stimuli. Four mechanisms which have been proposed to be causes of hyperreactivity are (i) a decrease in baseline airway calibre (ii) an increase in the responsiveness of the bronchial smooth muscle (iii) an imbalance in the autonomic neurophysiological regulation of airway calibre and (iv) epithelial changes which lead to an increase in the accessibility of allergens and non-specific stimuli to the mast cells, sensory nerve endings or bronchial smooth muscle beneath the airway mucosa. We have studied the effects of some of the proposed mediators of asthma on lung mechanics and irritant receptor activity in dogs with a respiratory tract infection and in dogs following exposure to SO2 gas at a concentration (400 ppl) sufficient to damage the airway epithelium (Jackson and Richards, 1980). In dogs naturally infected with Bordetella bronchiseptica there was an increased airway reactivity to histamine which was almost completely abolished by vagotomy. The hyperreactivity to histamine was shown to be due to an increased responsiveness of lung irritant receptors. In dogs exposed to SO2 gas changes in RL induced by 5-hydroxytryptamine or electrical stimulation of the vagus nerves were significantly increased when compared to the responses in normal dogs. Altounyan (1970) has shown that continuous exposure to allergens in patients with seasonal allergic asthma also leads to an increase in non-specific bronchial reactivity. Pharmacological modulation of airway reactivity can be divided into the acute and long term effects of drugs. Acute effects of drugs have been studied by several groups of workers using the airway responses to inhalation of various non-specific stimuli (e.g. distilled water, cold air, SO2 gas). Beta-stimulants and sodium cromoglycate are usually effective inhibitors and anticholinergic drugs are effective when a reflex component contributes to the bronchoconstriction. Few controlled studies of the effect of long-term treatment with drugs on bronchial hyperreactivity have been carried out although as early as 1970 Altounyan showed that long-term treatment with sodium cromoglycate could inhibit the development of non-specific hyperreactivity during the pollen season in pollen-sensitive asthmatics. A number of further studies have confirmed these early observations.(ABSTRACT TRUNCATED AT 400 WORDS)


Topochemistry of blood cells of the Gallus domesticus (Aves, Galliforme).
Blood smears of both male and female chicken Gallus domesticus were analysed by using the following topochemical methods: a) Periodic acid-Schiff (PAS) for glycogen. b) Mercury-bromophenol blue for protein. c) O-Toluidine for myeloperoxidase. d) Sudan black B for lipid. The PAS reaction revealed glycogen in the cytoplasm of all thrombocytes and in a few heterophils. The presence of proteins was evidenced in all types of cells. However variation in the intensity of staining of protein granules was observed in the fusiform structures of the heterophils. A negative reaction for myeloperoxidase was found in all cells. Although some evidence of myeloperoxidase activity was show in the polymorphonuclears it was not enough to ascertain a positive reaction. Lipids were detected in the cytoplasm of few heterophils, eosinophils and monocytes.


[Reconstruction of the lower orbital wall in the remote posttraumatic period].
The authors share their experience gained in the treatment of 11 patients with different fractures of the lower orbital wall in remote periods after the injury (1 month to 4 years). A specific feature of late injury in all the patients we observed were disorders of binocular vision presenting as diplopia due to cicatricial changes in the orbit. X-Ray examinations showed deep fractures of the lower orbital wall involving the posterior half of the orbital space in all cases. An operation had to be performed in all the patients: inferior orbitotomy via a transcutaneous access combined with anterior highmorotomy and submucosal refracture of the upper wall of the maxillary sinus within the framework of improperly consolidated fracture along the borders of the orbital hernia. The choice of this operation was dictated by specific localization of the fractures (zones difficult to access), solidity of scar growth at the site of the fracture (because of long period elapsed after the injury), and by limited possibility of intervention via an isolated transorbital access. An accessory element: refracture of the lower deformed wall and its reposition in the proper state helped repair the defect in the wall with a homocartilage on the side of strips and the orbit. Binocular vision and mobility of the eyeball in the orbit were repaired in all the patients, the degree of posttraumatic enophtalmos was appreciably decreased.


Promoter elements of vav drive transgene expression in vivo throughout the hematopoietic compartment.
To develop a method for targeting expression of genes to the full hematopoietic system, we have used transgenic mice to explore the transcriptional regulation of the vav gene, which is expressed throughout this compartment but rarely outside it. Previously, we showed that a cluster of elements surrounding its promoter could drive hematopoietic-specific expression of a bacterial lacZ reporter gene, but the expression was confined to lymphocytes and was sporadically silenced. Those limitations are ascribed here to the prokaryotic reporter gene. With a human CD4 (hCD4) cell surface reporter, the vav promoter elements drove expression efficiently and stably in virtually all nucleated cells of adult hematopoietic tissues but not notably in nonhematopoietic cell types. In multiple lines, hCD4 appeared on most, if not all, B and T lymphocytes, granulocytes, monocytes, megakaryocytes, eosinophils, and nucleated erythroid cells. Moreover, high levels appeared on both lineage-committed progenitors and the more primitive preprogenitors. In the fetus, expression was evident in erythroid cells of the definitive but not the primitive type. These results indicate that a prokaryotic sequence can inactivate a transcription unit and that the vav promoter region constitutes a potent transgenic vector for the entire definitive hematopoietic compartment.


SOD1 and amyotrophic lateral sclerosis: mutations and oligomerization.
There are about 100 single point mutations of copper, zinc superoxide dismutase 1 (SOD1) which are reported (http://alsod.iop.kcl.ac.uk/Als/index.aspx) to be related to the familial form (fALS) of amyotrophic lateral sclerosis (ALS). These mutations are spread all over the protein. It is well documented that fALS produces protein aggregates in the motor neurons of fALS patients, which have been found to be associated to mitochondria. We selected eleven SOD1 mutants, most of them reported as pathological, and characterized them investigating their propensity to aggregation using different techniques, from circular dichroism spectra to ThT-binding fluorescence, size-exclusion chromatography and light scattering spectroscopy. We show here that these eleven SOD1 mutants, only when they are in the metal-free form, undergo the same general mechanism of oligomerization as found for the WT metal-free protein. The rates of oligomerization are different but eventually they give rise to the same type of soluble oligomeric species. These oligomers are formed through oxidation of the two free cysteines of SOD1 (6 and 111) and stabilized by hydrogen bonds, between beta strands, thus forming amyloid-like structures. SOD1 enters the mitochondria as demetallated and mitochondria are loci where oxidative stress may easily occur. The soluble oligomeric species, formed by the apo form of both WT SOD1 and its mutants through an oxidative process, might represent the precursor toxic species, whose existence would also suggest a common mechanism for ALS and fALS. The mechanism here proposed for SOD1 mutant oligomerization is absolutely general and it provides a common unique picture for the behaviors of the many SOD1 mutants, of different nature and distributed all over the protein.


A New Intraoperative Protocol for Reducing Perioperative Transfusions in Cardiac Surgery.
Perioperative anemia and blood product transfusion increases short-term and long-term morbidity and mortality during cardiac surgery. We hypothesized that streamlined cardiopulmonary bypass circuit and rotational thromboelastometry (ROTEM) would reduce blood product usage and improve outcomes. All patients with Society of Thoracic Surgeons risk scores at our institution from January 2013 to June 2015 were included. Individuals were chronologically stratified into 2 groups according to institutional changes to a streamlined bypass circuit and ROTEM-guided transfusion. Blood product transfusion, hematocrit, and observed to expected outcomes (O/E) were compared between the groups. Patients were defined as either control group (533 patients, 12 months) or intervention group (804 patients, 18 months). The intervention group was further subdivided into streamlined circuit (290 patients, 6 months) and ROTEM (514 patients, 12 months). Use of streamlined bypass circuit correlated with significantly reduced intraoperative transfusion of packed red blood cells (pRBCs) (23.8% versus 17.9%; p = 0.05) and platelets (28.0% versus 19.3; p = 0.01) with improvement in lowest intraoperative hematocrit (26.0 versus 26.9; p = 0.02). ROTEM was associated with a further reduction in intraoperative pRBCs (17.9% versus 11.28%; p = 0.01) and postoperative transfusion pRBCs (38.3% versus 23.5%; p = 0.02). The combination was associated with reduced intraoperative (44.6% versus 34.1; p < 0.001) and postoperative transfusions (45.6% versus 40.1; p < 0.001) in the intervention group, while maintaining a higher hematocrit at discharge (28.1 versus 29.1; p < 0.001). Finally, the intervention was associated with a statistically significant reduction in the O/E for reoperation (p = 0.003). Use of streamlined cardiopulmonary bypass circuit and ROTEM may reduce transfusion and reoperation rates and improve perioperative anemia in cardiac surgical patients. This study demonstrates reproducible intraoperative methods for reducing blood product usage and improving outcomes.


Knowledge and Awareness of Prostate Cancer Among Omani Men Attending a Teaching Hospital.
Prostate cancer is ranked as the fourth most prevalent cancer in the world and the second most common cancer affecting men. In Oman, prostate cancer is one of the most common cancers among men, with the majority of prostate cancer patients presenting in the more advanced stages of the disease. Public awareness of the risk factors, symptoms and emphasising the importance of seeking early medical attention could help to improve the outcomes and survival rates of prostate cancer patients. The aim of this study is to determine the awareness levels of the risk factors, symptoms and barriers to seeking early medical intervention among adult Omani men. A validated questionnaire measuring the knowledge of risk factors, symptoms and barriers to seeking early medical help was used to collect data from adult Omani men attending a teaching hospital in Muscat, Oman. Out of 720 men who were invited, 600 participated in the study (response rate = 83%). The most recognised risk factor was obesity (366; 61.0%), and the least was sexually transmitted diseases (204; 34.0%); the most recognised symptom was unexplained weight loss (26.5%), the least was changes in seminal fluid (13.3%); the most common barrier to seeking early intervention was "not trusting the medical knowledge of the doctors" (57.5%), the least was "difficulty in arranging transport" (19.5%). Age, education level, marital status and family history of cancer were significantly associated with the participants' knowledge of prostate cancer. Participants received a significant part of their prostate cancer awareness via the social media. National awareness campaigns using social media and information leaflets are needed to educate Omani men on prostate cancer awareness and prevention and to increase trust in the expertise of medical professionals. Further research exploring the barriers to seeking early medical intervention is needed.


Hierarchical Stereo Matching in Two-Scale Space for Cyber-Physical System.
Dense disparity map estimation from a high-resolution stereo image is a very difficult problem in terms of both matching accuracy and computation efficiency. Thus, an exhaustive disparity search at full resolution is required. In general, examining more pixels in the stereo view results in more ambiguous correspondences. When a high-resolution image is down-sampled, the high-frequency components of the fine-scaled image are at risk of disappearing in the coarse-resolution image. Furthermore, if erroneous disparity estimates caused by missing high-frequency components are propagated across scale space, ultimately, false disparity estimates are obtained. To solve these problems, we introduce an efficient hierarchical stereo matching method in two-scale space. This method applies disparity estimation to the reduced-resolution image, and the disparity result is then up-sampled to the original resolution. The disparity estimation values of the high-frequency (or edge component) regions of the full-resolution image are combined with the up-sampled disparity results. In this study, we extracted the high-frequency areas from the scale-space representation by using difference of Gaussian (DoG) or found edge components, using a Canny operator. Then, edge-aware disparity propagation was used to refine the disparity map. The experimental results show that the proposed algorithm outperforms previous methods.


Functional characteristics of reversibly immortalized hepatic progenitor cells derived from mouse embryonic liver.
Liver is a vital organ and retains its regeneration capability throughout adulthood, which requires contributions from different cell populations, including liver precursors and intrahepatic stem cells. To overcome the mortality of hepatic progenitors (iHPs) in vitro, we aim to establish reversibly immortalized hepatic progenitor cells from mouse embryonic liver. Using retroviral system to stably express SV40 T antigen flanked with Cre/LoxP sites, we establish a repertoire of iHP clones with varied differentiation potential. The iHP cells maintain long-term proliferative activity and express varied levels of progenitor markers (Pou5f1/Oct4 and Dlk) and hepatocyte markers (AFP, Alb and ApoB). Five representative iHP clones express hepatic/pancreatic transcription factors HNF3α/Foxa1, HNF3β/Foxa2, and HNF4α/MODY1. Dexamethasone is shown to promote the expression of hepatocyte markers AFP and TAT, along with ICG-uptake and glycogen storage functions in the iHP clones. Cre-mediated removal of SV40 T antigen reverses the proliferative activity of iHP cells. When iHP cells are subcutaneously implanted in athymic nude mice, no tumor formation is observed for up to 8 weeks. We demonstrate that the established iHP cells are stable, reversible, and non-tumorigenic hepatic progenitor-like cells, which should be valuable for studying liver organogenesis, metabolic regulations, and hepatic lineage-specific differentiation.


Thick and Diffuse Subarachnoid Blood as a Treatment Effect Modifier of Clazosentan After Subarachnoid Hemorrhage.
Background and Purpose- Clazosentan, an endothelin receptor antagonist, has been shown to reduce angiographic vasospasm and vasospasm-related morbidity after aneurysmal subarachnoid hemorrhage (SAH), although no effect on long-term functional outcome has been demonstrated. Thick clot on initial computed tomography is associated with an increased risk of vasospasm and delayed cerebral ischemia. In this post hoc analysis, we hypothesized that use of clazosentan in this subpopulation would provide stronger benefit. Methods- We analyzed SAH patients enrolled in the CONSCIOUS-2 and CONSCIOUS-3 studies (Clazosentan to Overcome Neurological Ischemia and Infarction Occurring After Subarachnoid Hemorrhage) and compared the effects of clazosentan 5 mg/h, 15 mg/h, and placebo starting the day after aneurysm repair. The analysis was performed separately based on the presence or absence of thick (≥4 mm) and diffuse (≥3 cisterns) SAH on admission computed tomography. The primary composite end point was all-cause mortality and vasospasm-related morbidity at 6 weeks, and the main secondary end point was the extended Glasgow Outcome Scale at 3 months, adjusted for admission clinical grade. Results- Of 1718 randomized patients, 919 (53%) had thick and diffuse SAH. The primary composite end point in this group occurred in 36% of placebo-treated patients (n=294), 30% patients treated with clazosentan 5 mg/h (n=514; relative risk, 0.82; 95% CI, 0.67-0.99), and 19% patients treated with clazosentan 15 mg/h (n=111; relative risk, 0.54; 95% CI, 0.36-0.80). Despite this, death or poor functional outcome (Glasgow Outcome Scale ≤4) occurred in 33% of placebo-treated patients, 34% of patients treated with clazosentan 5 mg/h (relative risk 1.02; 95% CI, 0.84-1.23), and 35% of patients treated with clazosentan 15 mg/h (relative risk 1.14; 95% CI, 0.88-1.48). Conclusions- In an enriched population with thick and diffuse SAH, clazosentan at a dose of 5 and 15 mg/h was able to significantly reduce vasospasm-related morbidity in a dose-dependent manner. The absence of an effect on long-term functional status likely reflects the complexity and multiplicity of factors that contribute to poor outcome after SAH. Clinical Trial Registration- URL: https://www.clinicaltrials.gov. Unique identifier: NCT00558311; NCT00940095.


An automated causality assessment algorithm to detect drug-induced liver injury in electronic medical record data.
The aim of this study was to develop an automated causality assessment algorithm to identify drug-induced liver injury. The Roussel Uclaf Causality Assessment Method (RUCAM) is an algorithm for determining the causal association between a drug and liver injury. In collaboration with hepatology experts, definitions were developed for the RUCAM criteria to operationalize an electronic RUCAM (eRUCAM). The eRUCAM was tested in a population of patients taking 14 drugs with a characteristic phenotype for liver injury. Quality assurance for programming specifications involved comparisons between scores generated by the eRUCAM, for probable and highly probable cases, and expert manual RUCAM (n = 20). Concordance between eRUCAM and manual RUCAM subscores and total score was tested using the Wilcoxon signed rank test. Causality scores were the same for 6 of 20 patients (30%) by manual and eRUCAM algorithms. Analysis of subscores revealed ≥80% concordance between manual and eRUCAM for five of the seven criteria. In general, the total scores tended to be higher for the eRUCAM compared with the manual RUCAM. Programming issues were identified for criterion 5 'non-drug causes of liver injury' where significant differences existed between manual and eRUCAM scoring (p = 0.001). For criterion 5, identical scores occurred in 9 of 20 patients (45%), and manual review identified additional codes, timing criteria, and laboratory results for improving subsequent eRUCAM revisions. The eRUCAM had generally good concordance with manual RUCAM scoring. These preliminary findings suggest that the eRUCAM algorithm is feasible and could have application in clinical practice and drug safety surveillance.


Monoclonal antibodies to choline acetyltransferase: production, specificity, and immunohistochemistry.
Immunohistochemical localization of choline acetyltransferase (ChAT) in cholinergic neurons has been difficult to achieve because of problems encountered in producing specific antisera. Here we describe the production and characterization of several distinct monoclonal antibodies to ChAT. Each of the monoclonal antibodies exhibits one of three general patterns of cross-species reactions; one pattern shows reactivity limited mainly to bovine ChAT, a second pattern shows reactivity only to ChAT from higher mammals including humans, and the third pattern shows reactivity to ChAT from all mammals tested. The antibodies bound specifically to two closely related bovine proteins of 68,000 and 70,000 daltons using the Western blotting technique. One of the antibodies was used to localize immunohistochemically known cholinergic structures in the rat brain, including motor neurons, basal forebrain neurons, and neostriatal neurons.


Interdental cleaning among persons with diabetes: relationships with individual characteristics.
Given the existence of many potential oral health complications for adults with diabetes (especially for those who do not practise regular oral self-care), and the specific importance of regular interdental cleaning, the research determined the proportion of U.S. adults with diabetes who practise daily interdental cleaning and their socio-demographic, economic and oral health characteristics related to this practice. Analyses were conducted using data collected from 573 dentulous adults with diabetes ≥ 30 years who participated in the U.S. 2009-2010 National Health and Nutrition Examination survey. Using complex sample survey software, findings were extrapolated to >15 million U.S. adults. Descriptive statistics were used to determine the frequency of interdental cleaning, and chi-square tests were used to identify salient individual characteristics related to this practice. 41.2% reported that they never used any interdental device, while 24.8% indicated that they practised interdental cleaning daily. Statistically significant relationships (P < 0.05) with daily interdental cleaning included female sex, ever having had treatment for gum disease and using mouthwash daily for a dental problem. Because so many adults with diabetes do not practise regular interdental cleaning, and in view of the important role that dental hygienists fulfil as oral healthcare educators, there is a great need for dental hygienists to teach and motivate adults with diabetes to practise regular interdental cleaning. This need is especially great for subgroups of these adults who are men, have not been treated for periodontitis and do not regularly use a mouthwash for dental problems.


Vanillin extracted from Proso and Barnyard millets induce apoptotic cell death in HT-29 human colon cancer cell line.
In the present study, we hypothesized that the active compound extracted from Proso and Barnyard millets inhibits cell proliferation and apoptosis induction in colon cancer cell line. The bioactive compounds from these millets were purified by supercritical fluid extraction and their structure was elucidated using spectroscopic methods. Extracted bioactive components from these millets were similar in chemical structure to the phenolic aldehyde-Vanillin [4-Hydroxy-3-methoxybenzaldehyde]. Cell proliferative effect was assessed by MTT assay using HT-29 cell line. Compound 1 significantly inhibited the proliferation of HT-29 cells when treated with concentrations of 250 µg/ml and 1,000 µg/ml for 48 h, while compound 2 moderately inhibited the proliferation of the HT-29 cell line at the same concentration and time period. Cytotoxic activity of extracted compounds by the release of lactate dehydrogenase confirms that these compounds were not toxic to the cells at 250 µg/ml of compounds 1 and 2. In addition, flow cytometry results show a significant cell arrest in the G0/G1 phase and increase in the apoptotic cells in sub G0 phase, in a dose-dependent manner when compared with the control. The conclusion of this study suggests that the anticancer property of these millets is mediated through the presence of vanillin.


Label-free Raman spectroscopic imaging to extract morphological and chemical information from a formalin-fixed, paraffin-embedded rat colon tissue section.
Animal models and archived human biobank tissues are useful resources for research in disease development, diagnostics and therapeutics. For the preservation of microscopic anatomical features and to facilitate long-term storage, a majority of tissue samples are denatured by the chemical treatments required for fixation, paraffin embedding and subsequent deparaffinization. These aggressive chemical processes are thought to modify the biochemical composition of the sample and potentially compromise reliable spectroscopic examination useful for the diagnosis or biomarking. As a result, spectroscopy is often conducted on fresh/frozen samples. In this study, we provide an extensive characterization of the biochemical signals remaining in processed samples (formalin fixation and paraffin embedding, FFPE) and especially those originating from the anatomical layers of a healthy rat colon. The application of chemometric analytical methods (unsupervised and supervised) was shown to eliminate the need for tissue staining and easily revealed microscopic features consistent with goblet cells and the dense populations of cells within the mucosa, principally via strong nucleic acid signals. We were also able to identify the collagenous submucosa- and serosa- as well as the muscle-associated signals from the muscular regions and blood vessels. Applying linear regression analysis to the data, we were able to corroborate this initial assignment of cell and tissue types by confirming the biological origin of each layer by reference to a subset of authentic biomolecular standards. Our results demonstrate the potential of using label-free Raman microspectroscopy to obtain superior imaging contrast in FFPE sections when compared directly to conventional haematoxylin and eosin (H&E) staining.


Block of squid axon K channels by internally and externally applied barium ions.
We have studied the interactions of Ba ion with K channels. Ba2+ blocks these channels when applied either internally or externally in millimolar concentrations. Periodic depolarizations enhance block with internal Ba2+, but diminish the block caused by external Ba2+. At rest, dissociation of Ba2+ from blocked channels is very slow, as ascertained by infrequent test pulses applied after washing Ba2+ form either inside or outside. The time constant for recovery from internal and external Ba2+ is the same. Frequent pulsing greatly shortens recovery time constant after washing away both Ba2+in and Ba2+out. Block by Ba2+ applied internally or externally is voltage dependent. Internal Ba2+ block behaves like a one-step reaction governed by a dissociation constant (Kd) that decreases e-fold/12 mV increase of pulse voltage: block deepens with more positive pulse voltage. For external Ba2+, Kd decreases e-fold/18 mV as holding potential is made more negative: block deepens with increasing negativity. Millimolar external concentrations of some cations can either lessen (K+) or enhance (NH+4, Cs+) block by external Ba2+. NH+4 apparently enhances block by slowing exist of Ba ions from the channels. Rb+ and Cs+ also slow clearing of Ba ions from channels. We think that (a) internally applied Ba2+ moves all the way through the channels, entering only when activation gates are open; (b) externally applied Ba2+ moves two-thirds of the way in, entering predominantly when activation gates are closed; (c) at a given voltage, Ba2+ occupies the same position in the channels whether it entered from inside or outside.


Plague in free-ranging mammals in western North Dakota.
From July through October of 1996, 48 blood samples were collected from coyotes (Canis latrans), badgers (Taxidea taxus), and raccoons (Procyon lotor) in western North Dakota (USA) for the purposes of determining antibody titers to the plague bacterium, Yersniia pestis. The passive hemagglutination paper-strip blood-sampling technique was utilized with hemagglutination inhibition controls. Two positive samples were obtained from McKenzie county, one from a coyote with a titer of 1:64 and one from a badger with a titer of 1:256. Considering coyote and badger population dynamics, this study documents plague in western North Dakota.


Conservative management of placenta accreta in a multiparous woman.
Placenta accreta refers to any abnormally invasive placental implantation. Diagnosis is suspected postpartum with failed delivery of a retained placenta. Massive obstetrical hemorrhage is a known complication, often requiring peripartum hysterectomy. We report a case of presumed placenta accreta in a patient following failed manual removal of a retained placenta. We describe an attempt at conservative management with methotrexate in a stable patient desiring future fertility. Treatment was unsuccessful and led to the development of a disseminated intrauterine infection complicated by a bowel obstruction, requiring both a hysterectomy and small bowel resection. In hemodynamically stable patients, conservative management of placenta accreta may involve leaving placental tissue in situ with subsequent administration of methotrexate. However, ongoing close observation is required to identify complications.


Laparoscopic-assisted vaginal myomectomy: a case report and literature review.
The purpose of this article is to present a case of laparoscopic myomectomy (LM) that led to the identification of a new minimally invasive technique [laparoscopic-assisted vaginal myomectomy (LAVM)] for removing multiple transmural uterine myomas and facilitating uterine suturing. In addition, we reviewed the literature to (1) describe the history leading up to LAVM, (2) relate the benefits of this technique to other more widely performed myomectomy procedures [LM and laparoscopic-assisted myomectomy (LAM)], and (3) identify criteria for LM and LAVM.


Genetic diversity in domesticated soybean (Glycine max) and its wild progenitor (Glycine soja) for simple sequence repeat and single-nucleotide polymorphism loci.
• The study of genetic diversity between a crop and its wild relatives may yield fundamental insights into evolutionary history and the process of domestication. • In this study, we genotyped a sample of 303 accessions of domesticated soybean (Glycine max) and its wild progenitor Glycine soja with 99 microsatellite markers and 554 single-nucleotide polymorphism (SNP) markers. • The simple sequence repeat (SSR) loci averaged 21.5 alleles per locus and overall Nei's gene diversity of 0.77. The SNPs had substantially lower genetic diversity (0.35) than SSRs. A SSR analyses indicated that G. soja exhibited higher diversity than G. max, but SNPs provided a slightly different snapshot of diversity between the two taxa. For both marker types, the primary division of genetic diversity was between the wild and domesticated accessions. Within taxa, G. max consisted of four geographic regions in China. G. soja formed six subgroups. Genealogical analyses indicated that cultivated soybean tended to form a monophyletic clade with respect to G. soja. • G. soja and G. max represent distinct germplasm pools. Limited evidence of admixture was discovered between these two species. Overall, our analyses are consistent with the origin of G. max from regions along the Yellow River of China.


QEVIS: Multi-grained Visualization of Distributed Query Execution.
Distributed query processing systems such as Apache Hive and Spark are widely-used in many organizations for large-scale data analytics. Analyzing and understanding the query execution process of these systems are daily routines for engineers and crucial for identifying performance problems, optimizing system configurations, and rectifying errors. However, existing visualization tools for distributed query execution are insufficient because (i) most of them (if not all) do not provide fine-grained visualization (i.e., the atomic task level), which can be crucial for understanding query performance and reasoning about the underlying execution anomalies, and (ii) they do not support proper linkages between system status and query execution, which makes it difficult to identify the causes of execution problems. To tackle these limitations, we propose QEVIS, which visualizes distributed query execution process with multiple views that focus on different granularities and complement each other. Specifically, we first devise a query logical plan layout algorithm to visualize the overall query execution progress compactly and clearly. We then propose two novel scoring methods to summarize the anomaly degrees of the jobs and machines during query execution, and visualize the anomaly scores intuitively, which allow users to easily identify the components that are worth paying attention to. Moreover, we devise a scatter plot-based task view to show a massive number of atomic tasks, where task distribution patterns are informative for execution problems. We also equip QEVIS with a suite of auxiliary views and interaction methods to support easy and effective cross-view exploration, which makes it convenient to track the causes of execution problems. QEVIS has been used in the production environment of our industry partner, and we present three use cases from real-world applications and user interview to demonstrate its effectiveness. QEVIS is open-source at https://github.com/DBGroup-SUSTech/QEVIS.


Diabetic nephropathy: where hemodynamics meets metabolism.
Diabetic nephropathy (DN), the most common cause of end stage renal disease in developed nations, is thought to result from interactions between metabolic and haemodynamic factors. Specific metabolically driven, glucose dependent pathways are activated within diabetic renal tissues. These pathways induce oxidative stress, polyol pathway flux, hexosamine flux and accumulation of advanced glycated end-products (AGEs). Haemodynamic factors are also implicated in the pathogenesis of DN and include elevations of systemic and intraglomerular pressure and activation of various vasoactive hormone pathways including the renin-angiotensin aldosterone system (RAAS), endothelin and urotensin. These altered hemodynamics act independently and in concert with metabolic pathways, to activate intracellular second messengers such as protein kinase C (PKC) and MAP kinase (MAPK), nuclear transcription factors such as nuclear factor-kappaB (NF-kappaB) and various growth factors such as the prosclerotic cytokines, transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF) and the angiogenic, permeability enhancing growth factor, vascular endothelial growth factor, VEGF. Ultimately these molecular mechanisms lead to increased renal albumin permeability, and extracellular matrix accumulation, which results in increasing proteinuria, glomerulosclerosis and tubulointerstitial fibrosis. In the past, the treatment of diabetic nephropathy has focused on control of hyperglycemia and the interruption of the RAAS with certain anti-hypertensive agents. Newer novel targets, some of which are linked to glucose dependent pathways, appear to be a major focus of new therapies directed against the development and progression of renal damage as a result of diabetes. It is likely that resolution of diabetic nephropathy will require synergistic therapies to target multiple mediators of this disease.


Role of complement in induction of antibody production in vivo. Effect of cobra factor and other C3-reactive agents on thymus-dependent and thymus-independent antibody responses.
In an in vivo study in mice, suppression by the C3-cleaving protein of cobra venom (CoF), and other C3-reactive agents (zymosan, aggregated IgG, anti-C3 antibodies, and type III pneumococcal polysaccharide) of the thymus-dependent antibody responses to sheep erythrocytes, ovalbumin, and human IgG was demonstrated. The thymus-independent antibody response to polyvinyl-pyrrolidone was however unaffected by CoF. These and other published observations suggest that there may be a requirement for functional C3 in induction of thymus-dependent but not thymus-independent antibody production. A model for the role of C3 in lymphocyte cooperation is proposed based on these data analyzed in the light of existing knowledge of this process. It is postulated that fixed C3 interacting with macrophage See PDF for Structure and B-cell C3 receptors might enhance or facilitate T-dependent presentation of antigen to B cells.


Ample Arsenite Bio-Oxidation Activity in Bangladesh Drinking Water Wells: A Bonanza for Bioremediation?
Millions of people worldwide are at risk of arsenic poisoning from their drinking water. In Bangladesh the problem extends to rural drinking water wells, where non-biological solutions are not feasible. In serial enrichment cultures of water from various Bangladesh drinking water wells, we found transfer-persistent arsenite oxidation activity under four conditions (aerobic/anaerobic; heterotrophic/autotrophic). This suggests that biological decontamination may help ameliorate the problem. The enriched microbial communities were phylogenetically at least as diverse as the unenriched communities: they contained a bonanza of 16S rRNA gene sequences. These related to Hydrogenophaga, Acinetobacter, Dechloromonas, Comamonas, and Rhizobium/Agrobacterium species. In addition, the enriched microbiomes contained genes highly similar to the arsenite oxidase (aioA) gene of chemolithoautotrophic (e.g., Paracoccus sp. SY) and heterotrophic arsenite-oxidizing strains. The enriched cultures also contained aioA phylotypes not detected in the previous survey of uncultivated samples from the same wells. Anaerobic enrichments disclosed a wider diversity of arsenite oxidizing aioA phylotypes than did aerobic enrichments. The cultivatable chemolithoautotrophic and heterotrophic arsenite oxidizers are of great interest for future in or ex-situ arsenic bioremediation technologies for the detoxification of drinking water by oxidizing arsenite to arsenate that should then precipitates with iron oxides. The microbial activities required for such a technology seem present, amplifiable, diverse and hence robust.


Efficient spatial separation of charge carriers over Sv-ZnIn2S4/NH2-MIL-88B(Fe) S-scheme heterojunctions for enhanced photocatalytic H2 evolution and antibiotics removal performance.
The exploration of highly efficient sunlight-assisted photocatalyst for photodegradation of organic contaminants or energy conversion is strongly encouraged. In this work, we designed a novel three-dimensional spindle-like Sv-ZIS@NMFe heterojunction made of amino functionalized NH2-MIL-88B(Fe) (NMFe) and ZnIn2S4 nanosheets with abundant sulfur vacancies (Sv-ZIS). The structural properties of NMFe materials, such as a clearly defined system of pores and cavities, were retained by the Sv-ZIS@NMFe composites. Additionally, the incorporation of sulfur vacancies, -NH2 functional groups, and well-matched energy level positions led to various synergistic effects that considerably enhanced internal electron transformation and migration, as well as improved adsorption performance. Consequently, under visible light irradiation, the optimized sample exhibited superior hydrogen production activity and tetracycline hydrochloride photodegradation performance. At last, density functional theory calculations was used to further elucidated the possible photoreactivity mechanism. This study demonstrates that the Sv-ZIS@NMFe heterojunction materials formed by ZnIn2S4 with suitable sulfur vacancies and amino functionalized Fe-MOFs have promising applications in photocatalysis.


Noninvasive Markers to Assess Liver Fibrosis.
Chronic liver disease represents a major public health problem, accounting for significant morbidity and mortality worldwide. Their prognosis and management greatly depends on the amount and progression of liver fibrosis with time and the risk of development of cirrhosis. Historically, liver biopsy was considered to be the gold standard for the detection of fibrosis. Nevertheless, liver biopsy is an invasive procedure that has limitations in terms of patient acceptance, risk-benefit ratio, cost-effectiveness, and its availability in various geographic regions. Moreover, it is a questionable gold standard due to significant sampling error and intraobserver and interobserver variability. These limitations have led to the development of noninvasive techniques for assessing the presence and the degree of liver fibrosis. This review aims to revise the most recent data from the literature about noninvasive methods useful in the evaluation of liver fibrosis.


Detection and partial characterization of a human mast cell carboxypeptidase.
Using a high performance liquid chromatography assay that detects the cleavage of the C-terminal leucine from angiotensin I, we have identified a carboxypeptidase activity in mast cells from human lung and in dispersed mast cell preparations from human skin. The enzyme activity was detected in a preparation of dispersed human mast cells from lung of greater than 99% purity and was released with histamine after stimulation with goat anti-human IgE. In nine preparations of dispersed human mast cells from lung of 10 to 99% purity, net percentage of release of carboxypeptidase correlated with the release of histamine, localizing carboxypeptidase to mast cell secretory granules. The enzyme activity was also detected in preparations of dispersed human mast cells from skin and in extracts of whole skin. The inhibitor profile and m.w. of carboxypeptidase activity from preparations of dispersed mast cells from skin was similar to that from dispersed mast cells from lung. Mast cell carboxypeptidase had a m.w. on gel filtration of 30,000 to 35,000. The enzyme in crude lysates of dispersed mast cell preparations had optimal activity between pH 8.5 and 9.5 and was inhibited by potato inhibitor, which distinguished it from carboxypeptidase in cultured human foreskin keratinocytes and adult fibroblasts, and from other proteolytic mast cell enzymes. The enzyme activity was also inhibited by EDTA, o-phenanthroline, and, to a small extent, by 8-OH quinoline, but not by Captopril, soybean trypsin inhibitor, or pepstatin. These findings demonstrate that human mast cell secretory granules contain carboxypeptidase in addition to tryptase and chymase. It appears that mast cells from skin may have a higher content of carboxypeptidase than do mast cells from lung.


[Electrophysiological study of the variations in innervation of the hand muscles].
An electrophysiological study was carried out searching for variations in the innervation of the muscles of the hand. For this purpose a simple and non time-consuming technique was designed for stimulating the ulnar and median nerves at the elbow and wrist. Recordings were performed with surface electrodes at the abductor pollicis brevis, adductor digiti minimi and first dorsal interosseus muscles. Several anomalous innervation patterns of the intrinsic muscles of the hand were found, and the innervation of the first dorsal interosseus by the median nerve was the most frequent.


[Effector peptides of the brain renin-angiotensin system in central mechanisms of learned and natural forms of drinking behavior in rats].
Central mechanisms of angiotensin involvement in initiation and realization of operant forms of drinking behavior were investigated. It was suggested that intracerebroventricular microinjection of angiotensin-II and angiotensin-IIl specifically affected the learned forms of drinking behavior. The experiments demonstrated that [des-Asp1]-angiotensin-I produced only the natural forms of drinking behavior. Angiotensins modulated specific forms of thirst-associated behavior such as exploring, grooming, and ingestive behavior. Injections of AT1 receptor antagonist losartan were associated with acute water intake decrease and sharp operant behavior inactivation.


Using eye tracking to identify faking attempts during penile plethysmography assessment.
Penile plethysmography (PPG) is considered the most rigorous method for sexual interest assessment. Nevertheless, it is subject to faking attempts by participants, which compromises the internal validity of the instrument. To date, various attempts have been made to limit voluntary control of sexual response during PPG assessments, without satisfactory results. This exploratory research examined eye-tracking technologies' ability to identify the presence of cognitive strategies responsible for erectile inhibition during PPG assessment. Eye movements and penile responses for 20 subjects were recorded while exploring animated human-like computer-generated stimuli in a virtual environment under three distinct viewing conditions: (a) the free visual exploration of a preferred sexual stimulus without erectile inhibition; (b) the viewing of a preferred sexual stimulus with erectile inhibition; and (c) the free visual exploration of a non-preferred sexual stimulus. Results suggest that attempts to control erectile responses generate specific eye-movement variations, characterized by a general deceleration of the exploration process and limited exploration of the erogenous zone. Findings indicate that recording eye movements can provide significant information on the presence of competing covert processes responsible for erectile inhibition. The use of eye-tracking technologies during PPG could therefore lead to improved internal validity of the plethysmographic procedure.


Community-based participatory research (CBPR) in South Africa: engaging multiple constituents to shape the research question.
Community engagement is an on-going, arduous, and necessary process for developing effective health promotion programs. The challenges are amplified when the particular health issue or research question is not prominent in the consciousness of the targeted community. In this paper, we explore the community-based participatory research (CBPR) model as a means to negotiate a mutual agenda between communities and researchers. The paper is focused on the (perceived) need for cervical cancer screening in an under-resourced community in Cape Town, South Africa. Cervical cancer is a significant health problem in this community and elsewhere in South Africa. Unlike HIV-AIDS, however, many Black South Africans have not been educated about cervical cancer and the importance of obtaining screening. Many may not consider screening a priority in their lives. Our research included extensive consultations and informal interviews with diverse community and regional stakeholders. Following these, we conducted 27 focus groups and 106 demographic surveys with randomly selected youth, parents, local health care personnel, educators and school staff. Focus group data were summarized and analyzed cross-sectionally. Community stakeholders were involved throughout this research. Our consultations, interviews, and focus group data were key in identifying the concerns and priorities of the community. By engaging community stakeholders, we developed a research framework that incorporated the community's concerns and priorities, and stressed the intersecting roles of poverty, violence, and other cultural forces in shaping community members' health and wellbeing. Community members helped to refocus our research from cervical cancer to 'cervical health,' a concept that acknowledged the impact on women's bodies and lives of HIV-AIDS and STDs, sexual violence, poverty, and multiple social problems. We conclude that the research agenda and questions in community-based health research should not be considered immutable. They need to be open to negotiation, creativity, and constant reinvention.


Morphological Characteristics of Biliary Strictures after Liver Transplantation Visualized Using SpyGlass™ Cholangioscopy.
Biliary complications following liver transplant are common. Endoscopic retrograde cholangiopancreatography (ERCP) and magnetic resonance cholangiopancreatography (MRCP) are the main techniques used to diagnose and treat biliary complications; however, these techniques have limits to the depth of visualization. In this report, we present five cases of orthotopic liver transplant patients with biliary complications that underwent ERCP- or MRCP-guided cholangioscopy with the SpyGlass™ DS Direct Visualization System (SDDVS). The SDDVS allowed for the visualization of the morphological characteristics of biliary strictures, and images collected using the SDDVS allowed for four of the cases to be treated endoscopically. Our findings suggest that cholangioscopy with the SDDVS is a promising method to guide the endoscopic treatment of biliary complications after liver transplantation.


A feasibility study of several 3D printing methods for applications in epilepsy surgery.
To describe the process of three-dimensional printing in epilepsy surgery using three different methods: low-force stereolithography (SLA), filament deposition modeling (FDM), and Polyjet Stratasys, while comparing them in terms of printing efficiency, cost, and clinical utility. MRI and CT images of patient anatomy have been limited to review in the two-dimensional plane, which provides only partial representation of intricate intracranial structures. There has been growing interest in 3D printing of physical models of this complex anatomy to be used as an educational tool and for surgical visualization. One specific application is in epilepsy surgery where there are challenges in visualizing complex intracranial anatomy in relation to implanted surgical tools. MRI and CT data from patients with refractory epilepsy from a single center that underwent surgery are converted into 3D volumes, or stereolithography files. These were then printed using three popular 3D printing methods: SLA, FDM, and Polyjet. Faculty were surveyed on the impact of 3D modeling on the surgical planning process. All three methods generated physical models with an increasing degree of resolution, transparency, and clinical utility directly related to cost of production and accurate representation of anatomy. Polyjet models were the most transparent and clearly represented intricate implanted electrodes but had the highest associated cost. FDM produced relatively inexpensive models that, however, were nearly completely opaque, limiting clinical utility. SLA produced economical and highly transparent models but was limited by single material capacity. Three-dimensional printing of patient-specific anatomy is feasible with a variety of printing methods. The clinical utility of lower-cost methods is limited by model transparency and lack of multi-material overlay respectively. Polyjet successfully generated transparent models with high resolution of internal structures but is cost-prohibitive. Further research needs to be done to explore cost-saving methods of modeling.


[Key components of pathogenesis of periodontal diseases in light of cytomorphological findings].
A new cytomorphometrical method for evaluation of periodontal status by cytograms of gingival impressions is based on estimation of destruction index and inflammatory destructive index. This method will be helpful in the diagnosis of periodontal diseases, monitoring of the pathological process after treatment, and in solution of theoretical problems, e.g. investigation of the key factors in the pathogenesis of periodontal diseases. The authors analyze the relationship between the two major components of pathogenesis of periodontal inflammation-inflammatory infiltrative process and destructive changes.


Effect of neodymium:YAG laser capsulotomy on retinal straylight values in patients with posterior capsule opacification.
To determine the effect of neodymium:YAG (Nd:YAG) laser capsulotomy on retinal straylight values in patients with posterior capsule opacification (PCO). Rotterdam Eye Hospital, Rotterdam, The Netherlands. This prospective study comprised 35 patients with PCO scheduled to have Nd:YAG capsulotomy. Before and after Nd:YAG capsulotomy, the best corrected visual acuity (BCVA) was determined, a slitlamp examination was performed, and retinal straylight was measured. The straylight measurements were performed using the C-Quant instrument utilizing the compensation-comparison method. Based on the median BCVA before Nd:YAG capsulotomy (20/50), patients were divided into 2 groups: those with visual acuity better than 20/50 (good VA group) and those with an acuity of 20/50 or worse (poor VA group). The BCVA and straylight values improved significantly after Nd:YAG capsulotomy. The improvement in the straylight value was statistically significant in the good VA group and poor VA group. The BCVA and straylight values behaved independently from each other. Before capsulotomy, the BCVA and the straylight value were moderately correlated. After Nd:YAG capsulotomy, there was no significant correlation between the 2 parameters. Also, the improvement in the 2 parameters after Nd:YAG capsulotomy was unrelated. After Nd:YAG capsulotomy, both BCVA and straylight values improved significantly. There was no uniform relation between BCVA and retinal straylight. Straylight must be considered an independent entry criterion for capsulotomy.


Optical measurement of the catalase-hydrogen peroxide intermediate (Compound I) in the liver of anaesthetized rats and its implication to hydrogen peroxide production in situ.
The spectrophotometric determination of the catalase-H2O2 intermediate (Compound I) was extended to the liver in situ in anaesthetized rats. The rate of H2O2 production was determined for the liver in situ with endogenous substrates, and in the presence of excess of glycollate. Glycollate infusion doubled H2O2 production rate in the liver of air-breathing rats, and caused a fourfold increase when rats breathed O2 at 1 times 10(5) Pa. Hyperbaric O2 up to 6 times 10(5) Pa did not increase H2O2 generation supported by endogenous substrates, nor did it increase H2O2 production above that produced by 1 times 10(5) Pa O2 in glycollate-supplemented rats. The rates of ethanol oxidation via hepatic catalase and via alcohol dehydrogenase in the whole body were separately measured. The contribution of hepatic catalase to ethanol oxidation was found to be approx. 10 percent in endogenous conditions and increased to 30 percent or more of the total ethanol oxidation in rats supplemented with glycolate.


The insulin receptor tyrosine kinase substrate p58/53 and the insulin receptor are components of CNS synapses.
The synapse is the primary locus of cell-cell communication in the nervous system. It is now clear that the synapse incorporates diverse cell signaling modalities in addition to classical neurotransmission. Here we show that two components of the insulin pathway are localized at CNS synapses, where they are components of the postsynaptic density (PSD). An immunochemical screen revealed that polypeptides of 58 and 53 kDa (p58/53) were highly enriched in PSD fractions from rat cerebral cortex, hippocampus, and cerebellum. These polypeptides were purified and microsequenced, revealing that p58/53 is identical to the insulin receptor tyrosine kinase substrate p58/53 (IRSp53). Our analysis of IRSp58/53 mRNA suggests that within rat brain there is one coding region for IRSp58 and IRSp53; we find no evidence of alternative splicing. We demonstrate that IRSp58/53 is expressed in the synapse-rich molecular layer of the cerebellum and is highly concentrated at the synapses of cultured hippocampal neurons, where it co-localizes with the insulin receptor. Together, these data suggest that insulin signaling may play a role at CNS synapses.