Coxsackievirus B Escapes the Infected Cell in Ejected Mitophagosomes.
Coxsackievirus B (CVB) is a common enterovirus that can cause various systemic inflammatory diseases. Because CVB lacks an envelope, it has been thought to be inherently cytolytic, wherein CVB can escape from the infected host cell only by causing it to rupture. In recent years, however, we and others have observed that various naked viruses, such as CVB, can trigger the release of infectious extracellular microvesicles (EMVs) that contain viral material. This mode of cellular escape has been suggested to allow the virus to be masked from the adaptive immune system. Additionally, we have previously reported that these viral EMVs have LC3, suggesting that they originated from autophagosomes. We now report that CVB-infected cells trigger DRP1-mediated fragmentation of mitochondria, which is a precursor to autophagic mitochondrial elimination (mitophagy). However, rather than being degraded by lysosomes, mitochondrion-containing autophagosomes are released from the cell. We believe that CVB localizes to mitochondria, induces mitophagy, and subsequently disseminates from the cell in an autophagosome-bound mitochondrion-virus complex. Suppressing the mitophagy pathway in HL-1 cardiomyocytes with either small interfering RNA (siRNA) or Mdivi-1 caused marked reduction in virus production. The findings in this study suggest that CVB subverts mitophagy machinery to support viral dissemination in released EMVs.IMPORTANCE Coxsackievirus B (CVB) can cause a number of life-threatening inflammatory diseases. Though CVB is well known to disseminate via cytolysis, recent reports have revealed a second pathway in which CVB can become encapsulated in host membrane components to escape the cell in an exosome-like particle. Here we report that these membrane-bound structures derive from mitophagosomes. Blocking various steps in the mitophagy pathway reduced levels of intracellular and extracellular virus. Not only does this study reveal a novel mechanism of picornaviral dissemination, but also it sheds light on new therapeutic targets to treat CVB and potentially other picornaviral infections.


Epidemiology of acute coronary syndromes in Spain: estimation of the number of cases and trends from 2005 to 2049.
Acute coronary syndromes are a leading cause of mortality, morbidity, and health care cost in Spain. The aims of this report are to estimate the number of acute coronary syndromes cases in the Spanish population in 2013 and 2021, and the trend from 2005 to 2049. We estimated the number of acute coronary syndromes cases by sex and Spanish autonomous community using data from the most updated population and hospital registries. We present the estimated number of cases with an exact 95% confidence interval, assuming that the number of cases followed a Poisson distribution. There will be 115,752 acute coronary syndromes cases in Spain in 2013 (95% confidence interval, 114,822-116,687). Within 28 days, 39,086 of these patients will die and 85,326 will be hospitalized. Non-ST segment elevation acute coronary syndromes (56%) and acute myocardial infarction (81%) will be the most common admission and discharge diagnoses, respectively. We estimate approximately 109,772 acute coronary syndromes cases in 2021 (95% confidence interval, 108,868-110,635). The trend of acute coronary syndromes cases from 2005 to 2049 will stabilize in the population aged 25 to 74 years, but increase in those older than 74 years. Due to population aging, the number of acute coronary syndrome cases will increase overall until 2049, it may stabilize in the population aged <75 years. The acute coronary syndromes case-fatality has decreased in hospitalized patients but the proportion of sudden deaths remains unchanged.


Ten-year study of causes of moderate to severe angioedema seen by an inpatient allergy/immunology consult service.
The causes of angioedema are not well described, especially in the inpatient setting. The purpose of this study was to examine the causes of moderate to severe angioedema in patients requiring inpatient treatment. We performed a retrospective review in patients requiring inpatient consultation by the Division of Allergy and Immunology at our institution between 1995 and 2004. We focused on potential interactions among medications that elicited life-threatening angioedema requiring intubation. The allergy/immunology service was consulted on 69 patients with moderate to severe angioedema. Medications were the most common cause of angioedema (n = 64, 93%). In most cases (n = 46, 67%), the angioedema was attributed to two or more medications. Patients previously stable on ACE inhibitors (ACEI), aspirin (ASA), or non-steroidal anti-inflammatory drugs (NSAIDs) appeared more likely to develop angioedema soon after the addition of another drug (i.e., ACEI, ASA/NSAIDs, direct mast cell degranulators, and antibiotics). ACEI, ASA/NSAID, and direct mast cell degranulators were contributing causes in 36 patients (56%), 45 patients (70%), and 23 patients (36%), respectively. Twenty patients required intubation, 14 (70%) patients were on ACEI, 12 (60%) patients were on ASA/NSAID, and 7 (35%) patients were on direct mast cell degranulators. ACEI, ASA/NSAID, or direct mast cell degranulators were a cause in 95% (n = 19) of patients requiring intubation. The combination of ACEI and ASA/NSAID was the most frequent cause of angioedema among all patients (n = 17, 25%) and those requiring intubation (n = 8, 40%). Moderate to severe angioedema often is a result of interactions between two or more medications involved in different pathways causing angioedema. In particular, combinations of ACEI, ASA/NSAID, or direct mast cell degranulators may lead to life-threatening angioedema requiring intubation.


Sex difference in clinical and procedural outcomes in patients undergoing coronary atherectomy: a systematic review and meta-analysis.
Rotational and orbital coronary atherectomy (CA) are commonly utilized to treat complex calcified coronary lesions. We conducted a meta-analysis to evaluate sex differences in procedural complications and clinical outcomes after CA. PubMed, Google Scholar, and Cochrane databases were searched for all studies comparing sex differences in procedural and clinical outcomes following CA. The outcomes of interest were procedural complications (coronary dissection, stroke, major bleeding, coronary perforation, cardiac tamponade, and slow or no flow in target vessel) and the clinical outcomes (including early mortality, mid-term all-cause mortality, stroke, myocardial infarction, and target vessel revascularization). Pooled risk ratios (RRs) with their corresponding 95% confidence intervals (CIs) were calculated using the Mantel-Haenszel random-effects model. Six observational studies with 3517 patients (2420 men and 1035 women) were included in this meta-analysis. While there was no significant difference in the early mortality (RR, 1.14; 95% CI, 0.37-3.53; P = 0.83) between men and women, at a mean follow-up of 2.9 years, all-cause mortality was significantly higher in women (RR, 1.29; 95% CI, 1.11-1.49; P = 0.0009). Women had an increased risk of procedure-related stroke (RR, 3.98; 95% CI, 1.06-14.90; P = 0.04), coronary dissection (RR, 2.10; 95% CI, 1.23-3.58; P = 0.006), and bleeding (RR, 2.26; 95% CI, 1.30-3.93; P = 0.004), whereas the rates of coronary perforation, cardiac tamponade, and the risk of slow or no flow in the revascularized artery were similar in both. In our analysis, women undergoing CA are at increased risk of mid-term mortality and procedure-related complications including stroke, coronary dissection, and major bleeding.


Is the IS1-flanked r-determinant of the R plasmid NR1 a transposon?
The 23 kilobase multiple drug resistance r-determinant (r-det) of the R plasmid NR1 is an IS1-mediated transposon, Tn2671. Drug-resistant Escherichia coli transductants isolated after infection with bacteriophage P1::Tn2671 derivatives carry the intact r-det in their chromosomes. Independently isolated transductants carry the r-det at different locations on the chromosome. From the E. coli chromosome, Tn2671 can transpose to various locations on the phage P7 genome. Throughout these processes, r-det is maintained as a stable unit. Various possible molecular mechanisms, which all might contribute with characteristic frequencies to the transposition of Tn2671, are discussed. The results presented are relevant to the understanding of mechanisms for a wide spreading of drug resistance genes.


Can we trust projections of AMOC weakening based on climate models that cannot reproduce the past?
The Atlantic Meridional Overturning Circulation (AMOC), a crucial element of the Earth's climate system, is projected to weaken over the course of the twenty-first century which could have far reaching consequences for the occurrence of extreme weather events, regional sea level rise, monsoon regions and the marine ecosystem. The latest IPCC report puts the likelihood of such a weakening as 'very likely'. As our confidence in future climate projections depends largely on the ability to model the past climate, we take an in-depth look at the difference in the twentieth century evolution of the AMOC based on observational data (including direct observations and various proxy data) and model data from climate model ensembles. We show that both the magnitude of the trend in the AMOC over different time periods and often even the sign of the trend differs between observations and climate model ensemble mean, with the magnitude of the trend difference becoming even greater when looking at the CMIP6 ensemble compared to CMIP5. We discuss possible reasons for this observation-model discrepancy and question what it means to have higher confidence in future projections than historical reproductions. This article is part of a discussion meeting issue 'Atlantic overturning: new observations and challenges'.


Differences in dopamine beta-hydroxylase immunoreactivity between the brains of genetically epilepsy-prone and Sprague-Dawley rats.
Biochemical studies have indicated that norepinephrine is present in lower levels in certain brain regions of genetically epilepsy-prone rats (GEPR-9s) as compared to non-epileptic Sprague-Dawley (SD) rats. In this study, the immunocytochemical localization of dopamine beta-hydroxylase (DBH), the synthesizing enzyme for norepinephrine, was compared between GEPR-9s and SD rats. Brain regions caudal to the inferior colliculus, such as the cerebellum and locus coeruleus, showed no differences in the distribution of DBH-like immunoreactive (DBH-I) neurons and fibers. In contrast, differences in the distribution of DBH-I fibers were observed in more rostral brain regions including the central nucleus of the inferior colliculus, thalamus, piriform, orbital and somatosensory cortices and hippocampus. In these areas, the number, and often the staining intensity, of DBH-I processes was lower in GEPR-9s as compared to SD rats. It was interesting to note that other cortical regions displayed no differences in DBH immunoreactivity between GEPR-9s and SD rats. These results provide anatomical data that support previously described biochemical results. Furthermore, the reduced number of fibers and their decreased staining intensity in specific brain regions provide greater details to resolve the localization of deficiencies in the noradrenergic fiber plexus of GEPR-9s.


Changes in the deep subsurface microbial biosphere resulting from a field-scale CO2 geosequestration experiment.
Subsurface microorganisms may respond to increased CO2 levels in ways that significantly affect pore fluid chemistry. Changes in CO2 concentration or speciation may result from the injection of supercritical CO2 (scCO2) into deep aquifers. Therefore, understanding subsurface microbial responses to scCO2, or unnaturally high levels of dissolved CO2, will help to evaluate the use of geosequestration to reduce atmospheric CO2 emissions. This study characterized microbial community changes at the 16S rRNA gene level during a scCO2 geosequestration experiment in the 1.4 km-deep Paaratte Formation of the Otway Basin, Australia. One hundred and fifty tons of mixed scCO2 and groundwater was pumped into the sandstone Paaratte aquifer over 4 days. A novel U-tube sampling system was used to obtain groundwater samples under in situ pressure conditions for geochemical analyses and DNA extraction. Decreases in pH and temperature of 2.6 log units and 5.8°C, respectively, were observed. Polyethylene glycols (PEGs) were detected in the groundwater prior to scCO2 injection and were interpreted as residual from drilling fluid used during the emplacement of the CO2 injection well. Changes in microbial community structure prior to scCO2 injection revealed a general shift from Firmicutes to Proteobacteria concurrent with the disappearance of PEGs. However, the scCO2 injection event, including changes in response to the associated variables (e.g., pH, temperature and salinity), resulted in increases in the relative abundances of Comamonadaceae and Sphingomonadaceae suggesting the potential for enhanced scCO2 tolerance of these groups. This study demonstrates a successful new in situ sampling approach for detecting microbial community changes associated with an scCO2 geosequestration event.


Optical properties and mechanofluorochromism of new BODIPY dyes based on the pyridine-pyrimidine hybrid structure.
A new family of BODIPY dyes (PPB1-4) based on the pyridine-pyrimidine hybrid structure has been synthesized. These dyes show high cyan fluorescence in hexane with quantum yields (Φf) ranging from 0.33 to 0.90. Interestingly, moderate fluorescence of PPB1-4 is observed in the solid state (Φf, 0.11-0.26). X-ray structure analysis demonstrates that H-aggregation should be responsible for the red-shifted and quenched emission in the solid state. Moreover, PPB1-4 show high contrast and reversible mechanochromic fluorescence based on the proposed mechanism of mechanical stimuli triggered dimer dissolution and formation. In addition, rewritable mechanochromic fluorescence recording media fabricated with PPB4 have been successfully achieved.


Ku80 plays a role in non-homologous recombination but is not required for T-DNA integration in Arabidopsis.
Chromosomal breaks are repaired by homologous recombination (HR) or non-homologous end joining (NHEJ) mechanisms. The Ku70/Ku80 heterodimer binds DNA ends and plays roles in NHEJ and telomere maintenance in organisms ranging from yeast to humans. We have previously identified a ku80 mutant of the model plant Arabidopsis thaliana and shown the role of Ku80 in telomere homeostasis in plant cells. We show here that this mutant is hypersensitive to the DNA-damaging agent methyl methane sulphonate and has a reduced capacity to carry out NHEJ recombination. To understand the interplay between HR and NHEJ in plants, we measured HR in the absence of Ku80. We find that the frequency of intrachromosomal HR is not affected by the absence of Ku80. Previous work has clearly implicated the Ku heterodimer in Agrobacterium-mediated T-DNA transformation of yeast. Surprisingly, ku80 mutant plants show no defect in the efficiency of T-DNA transformation of plants with Agrobacterium, showing that an alternative pathway must exist in plants.


Multidrug resistance protein (MRP) expression in retinoblastoma correlates with the rare failure of chemotherapy despite cyclosporine for reversal of P-glycoprotein.
Failure of chemotherapy associated with expression of the multidrug resistance protein p170 frequently occurs in retinoblastoma (RB). Despite using cyclosporine, which inhibits p170 and improves our chemotherapy results, rare failures occur. In nonmetastatic primarily enucleated RBs, we show expression of p170 in 3 of 18 samples and expression of multidrug resistance protein (MRP), the second protein associated with resistance to chemotherapy, in 1 of 18 samples. All three RBs that failed chemotherapy without cyclosporine expressed MRP with p170. All three RBs that were enucleated immediately when chemotherapy failed despite the addition of cyclosporine expressed only MRP. One RB enucleated 2 years after failing chemotherapy with cyclosporine, despite radiation and salvage chemotherapy, expressed both p170 and MRP. Two metastatic RBs that expressed both p170 and MRP at diagnosis and at recurrence failed chemotherapy without cyclosporine, whereas one metastatic RB that expressed neither protein was cured by chemotherapy without cyclosporine. MRP may result in failure of chemotherapy despite the elimination of p170-expressing clones by cyclosporine.


22% Efficiency Inverted Perovskite Photovoltaic Cell Using Cation-Doped Brookite TiO2 Top Buffer.
Simultaneously achieving high efficiency and high durability in perovskite solar cells is a critical step toward the commercialization of this technology. Inverted perovskite photovoltaic (IP-PV) cells incorporating robust and low levelized-cost-of-energy (LCOE) buffer layers are supposed to be a promising solution to this target. However, insufficient inventory of materials for back-electrode buffers substantially limits the development of IP-PV. Herein, a composite consisting of 1D cation-doped TiO2 brookite nanorod (NR) embedded by 0D fullerene is investigated as a top modification buffer for IP-PV. The cathode buffer is constructed by introducing fullerene to fill the interstitial space of the TiO2 NR matrix. Meanwhile, cations of transition metal Co or Fe are doped into the TiO2 NR to further tune the electronic property. Such a top buffer exhibits multifold advantages, including improved film uniformity, enhanced electron extraction and transfer ability, better energy level matching with perovskite, and stronger moisture resistance. Correspondingly, the resultant IP-PV displays an efficiency exceeding 22% with a 22-fold prolonged working lifetime. The strategy not only provides an essential addition to the material inventory for top electron buffers by introducing the 0D:1D composite concept, but also opens a new avenue to optimize perovskite PVs with desirable properties.


An enhanced International Prognostic Index (NCCN-IPI) for patients with diffuse large B-cell lymphoma treated in the rituximab era.
The International Prognostic Index (IPI) has been the basis for determining prognosis in patients with aggressive non-Hodgkin lymphoma (NHL) for the past 20 years. Using raw clinical data from the National Comprehensive Cancer Network (NCCN) database collected during the rituximab era, we built an enhanced IPI with the goal of improving risk stratification. Clinical features from 1650 adults with de novo diffuse large B-cell lymphoma (DLBCL) diagnosed from 2000-2010 at 7 NCCN cancer centers were assessed for their prognostic significance, with statistical efforts to further refine the categorization of age and normalized LDH. Five predictors (age, lactate dehydrogenase (LDH), sites of involvement, Ann Arbor stage, ECOG performance status) were identified and a maximum of 8 points assigned. Four risk groups were formed: low (0-1), low-intermediate (2-3), high-intermediate (4-5), and high (6-8). Compared with the IPI, the NCCN-IPI better discriminated low- and high-risk subgroups (5-year overall survival [OS]: 96% vs 33%) than the IPI (5 year OS: 90% vs 54%), respectively. When validated using an independent cohort from the British Columbia Cancer Agency (n = 1138), it also demonstrated enhanced discrimination for both low- and high-risk patients. The NCCN-IPI is easy to apply and more powerful than the IPI for predicting survival in the rituximab era.


How electromagnetic fields can influence adult stem cells: positive and negative impacts.
The electromagnetic field (EMF) has a great impact on our body. It has been successfully used in physiotherapy for the treatment of bone disorders and osteoarthritis, as well as for cartilage regeneration or pain reduction. Recently, EMFs have also been applied in in vitro experiments on cell/stem cell cultures. Stem cells reside in almost all tissues within the human body, where they exhibit various potential. These cells are of great importance because they control homeostasis, regeneration, and healing. Nevertheless, stem cells when become cancer stem cells, may influence the pathological condition. In this article we review the current knowledge on the effects of EMFs on human adult stem cell biology, such as proliferation, the cell cycle, or differentiation. We present the characteristics of the EMFs used in miscellaneous assays. Most research has so far been performed during osteogenic and chondrogenic differentiation of mesenchymal stem cells. It has been demonstrated that the effects of EMF stimulation depend on the intensity and frequency of the EMF and the time of exposure to it. However, other factors may affect these processes, such as growth factors, reactive oxygen species, and so forth. Exploration of this research area may enhance the development of EMF-based technologies used in medical applications and thereby improve stem cell-based therapy and tissue engineering.


Unspecified gastroenteritis illness and deaths in the elderly associated with norovirus epidemics.
New variant strains of norovirus have emerged worldwide in recent years, evolving by mutation much like influenza viruses. These strains have been associated with a notable increase in the number of annual norovirus outbreaks. However, the impact of such increased norovirus activity on morbidity and mortality is not clear because norovirus infection is rarely specifically registered. We studied trends of gastroenteritis with unspecified cause in medical registrations (ie, general practitioner [GP] visits, hospitalizations, and deaths) and their association with known temporal trends in norovirus outbreaks in the Netherlands. Using weekly counts in the elderly (aged 65+ years) from 1999 through 2006, we applied Poisson regression analyses adjusted for additional pathogens and seasonal trends (linear, sine, and cosine terms). In the elderly, each notified norovirus outbreak was associated with an estimated 26 unspecified gastroenteritis GP visits (95% confidence interval = 17-34), 2.2 unspecified gastroenteritis hospitalizations (1.6-2.7), and 0.14 unspecified gastroenteritis deaths (0.08-0.21). For the heaviest norovirus season (2004-2005), these models attributed up to 3804 unspecified gastroenteritis GP visits, 318 unspecified gastroenteritis hospitalizations, and 21 unspecified gastroenteritis deaths to norovirus outbreaks among a total elderly population of 2.3 million. The recent increase in norovirus outbreak activity is associated with increases of unspecified gastroenteritis morbidity and even deaths in the elderly. Norovirus should not be regarded as an infection with trivial health risks.


Role of amino acids and peptides in the molecular signaling in skeletal muscle after resistance exercise.
Resistance exercise can effectively result in an increase in muscle mass, or hypertrophy, which generally becomes apparent after several weeks of training. Muscle hypertrophy requires muscle protein synthesis to exceed protein breakdown during an extended time period. It has been firmly established that the interaction between exercise and nutrition (i.e., protein intake) is necessary to attain net protein accretion in skeletal muscle. The stimulation of protein synthesis is caused in part by stimulation of mRNA translation initiation. There is relatively little information on the response of intracellular signaling controlling mRNA translation to exercise and nutrition, especially in humans, but the available data in humans seem to suggest that a single bout of resistance exercise does not substantially enhance PI-3 kinase/mTOR signaling during the first 2 h after exercise. Moreover, it is demonstrated that the ingestion of protein or amino acids after exercise is crucial to further stimulate molecular signaling that controls translation initiation. The aim of this review is to provide an overview of the intracellular signaling related to translational control and to provide a summary of the current knowledge about the response of the signaling pathways controlling the anabolic response to exercise and nutrient intake in vivo in humans.


Drug Use Evaluation of Ceftriaxone in Ras-Desta Memorial General Hospital, Ethiopia.
Ceftriaxone is one of the most commonly used antibiotics due to its availability, wide spectrum of activity and low toxicity. However, irrational use of ceftriaxone is one of the current issues in most countries, especially developing ones. The aim of this study was to evaluate ceftriaxone utilization in different wards of Ras-Desta Memorial General Hospital (RDMGH) in Ethiopia. An institution-based retrospective cross-sectional study was conducted on randomly selected 1,079 patients who were admitted to RDMGH from May 2017 to April 2018. A structured data abstraction format was used to collect data from patients' medical chart. Micromedex® drug interaction checker was used to identify significant drug-drug interactions, and national and international guidelines were used to evaluate the appropriateness of ceftriaxone use. The data were analyzed by using SPSS version 20.0. Among the 1,079 patients enrolled, ceftriaxone was utilized by 601 (55.7%) patients, and these patients were considered for evaluation of appropriateness and subsequent analysis. Among 601 patients, ceftriaxone was used for therapeutic purpose in 362 (60.2%) patients; of which, 359 (99.2%) were for empiric therapy and the rest 239 (39.8%) were for prophylaxis. In the majority of the patients, the dose of ceftriaxone was 2g/day 472 (78.6%) and for a duration of 2-7 days 409 (68.1%). Inappropriate use of ceftriaxone was observed among 237 (39.4%) patients with regard to indication, dose/frequency and duration. About half (49.3%) of the deviation from the guidelines was observed from surgical and gynecologic/obstetrics wards. Among the co-prescribed medications with ceftriaxone, ringer lactate, warfarin and heparin were found to have a significant drug-drug interaction. This study revealed that inappropriate use of ceftriaxone was high in RDMGH. This may increase the emergence of resistant pathogens which may lead to treatment failure and increase cost of therapy. Therefore, adherence to current evidence-based guidelines is recommended.


Localized hypertrophic neuropathy of the sciatic nerve in children: MRI findings.
Localized hypertrophic neuropathy (LHN) of the sciatic nerve in children is a rare condition characterized by a painless neurological deficit in the sciatic nerve territory. To demonstrate the role of MRI using a specific protocol and describe the primary findings in LHN. Imaging in four children (age 2 years to 12 years) is presented. All children presented with lower limb asymmetry. Three had a steppage gait. LHN was confirmed by electrophysiological studies and by MRI of the whole sciatic nerve with a dedicated protocol covering the lumbar spine and the lower limb. There were four direct MRI findings: (1) linear and focal hypertrophy with progressive enlargement of a peripheral nerve or plexus diameter, (2) abnormal hyperintensity of the nerve on T2-weighted images, (3) preserved fascicular configuration, and (4) variable enhancement after intravenous gadolinium administration. In addition there were atrophy and fatty infiltration of innervated muscles. MRI was helpful for determining the extent of lesions and in excluding peripheral nerve compression or tumour. MRI of the whole sciatic nerve is the method of choice for diagnosing LHN of the sciatic nerve.


Chromosome transfer in Proteus mirabilis mediated by hybrid plasmid.
A previously-described fused plasmid, P-lacRIdrd19, was found to mediate chromosomal transfer between cells of Proteus mirabilis strain PM5006; PM5006-(P-lacRIdrd19) was usually the donor and various auxotrophs of PM5006 resistant to nalidixic acid and/or streptomycin were recipients. The donor was usually counterselected with nalidixic acid and/or high concentrations of streptomycin. Recombination experiments with single markers indicated a 40-fold variation in recombination frequencies for different markers. Mapping double-auxotrophic markers by their gradient of transmission confirmed this variation and placed each of two independent isolates of eight markers in a linkage group his-ser-ura-pyrB-trp-cys-ade-ilv. Some donor markers did not register. Despite low recombination frequencies, interrupted mating experiments showed a polarity of early marker transfer. The segregation of unselected markers confirmed the order of some markers and showed that genetic material passed from the presumptive donor to the recipient. Recipients with two auxotrophic markers which could not be cotransduced by phage 5006M were converted to prototrophy by conjugation. The plasmid transferred to recipients at high frequency and all recombinants carried it. Recombinants could act as donors in further matings. Recombinants were fully susceptible to phage 5006M, unlike transductants of PM5006 by this phage. Direct involvement of the plasmid was indicated by drastically diminished recombination frequencies in crosses with recipients carrying P-lac as resident. P-lac had previously been shown to reduce the frequency of transfer of the hybrid plasmid to cells harbouring it. The histidine region was the first to register in recipients and recombined at the highest frequency of 5 times 10-6/donor cell. Some temporary association of plasmid and perhaps only the histidine region of the chromosome is favoured as the mechanism of chromosomal transfer. This could explain why not all donor markers could be mapped. Transduction and transformation were excluded as the cause of results.


Rv1288, a Two Domain, Cell Wall Anchored, Nutrient Stress Inducible Carboxyl-Esterase of Mycobacterium tuberculosis, Modulates Cell Wall Lipid.
Rv1288, a conserved hypothetical protein of M. tuberculosis (M.tb), was recently characterized as two-domain esterase enzyme by in silico study. In the present study, Rv1288 and its domains (Est and Lyt) were cloned individually from M.tb into E. coli for expression and purification. The purified rRv1288 and rEst proteins exhibited lipolytic activity with medium chain length esters as optimum substrates, while Lyt domain did not show enzymatic activity. However, presence of Lyt domain resulted in enhanced rate of protein aggregation at higher temperature. Both rRv1288 and rEst followed the similar patterns of substrate specificity, temperature and pH activity. Site directed mutagenesis confirmed the Ser-294, Asp-391 and His-425 as catalytic site residues. Rv1288 was found to be present in cell wall fraction of M.tb H37Ra. Peptidoglycan binding activity of Rv1288 and its domains demonstrated that the Lyt domain is essential for anchoring protein to the cell wall. Expression of rv1288 was up regulated in M.tb under nutrient starved condition. Over expression of rv1288 in surrogate host M. smegmatis led to change in colony morphology, enhanced pellicle and aggregate formation that might be linked with the changed lipid composition of bacterial cell wall. Cell wall of M. smegmatis expressing rv1288 had higher amount of lipids, with a significant increase in trehalose dimycolate content. Rv1288 also leads to increase in drug resistance of M. smegmatis. Rv1288 also enhanced the intracellular survival of M. smegmatis in Raw264.7 cell line. Overall, this study suggested that Rv1288, a cell wall localized carboxyl hydrolase with mycolyl-transferase activity, modulated the cell wall lipids to favor the survival of bacteria under stress condition.


Surface-coupled proton exchange of a membrane-bound proton acceptor.
Proton-transfer reactions across and at the surface of biological membranes are central for maintaining the transmembrane proton electrochemical gradients involved in cellular energy conversion. In this study, fluorescence correlation spectroscopy was used to measure the local protonation and deprotonation rates of single pH-sensitive fluorophores conjugated to liposome membranes, and the dependence of these rates on lipid composition and ion concentration. Measurements of proton exchange rates over a wide proton concentration range, using two different pH-sensitive fluorophores with different pK(a)s, revealed two distinct proton exchange regimes. At high pH (> 8), proton association increases rapidly with increasing proton concentrations, presumably because the whole membrane acts as a proton-collecting antenna for the fluorophore. In contrast, at low pH (< 7), the increase in the proton association rate is slower and comparable to that of direct protonation of the fluorophore from the bulk solution. In the latter case, the proton exchange rates of the two fluorophores are indistinguishable, indicating that their protonation rates are determined by the local membrane environment. Measurements on membranes of different surface charge and at different ion concentrations made it possible to determine surface potentials, as well as the distance between the surface and the fluorophore. The results from this study define the conditions under which biological membranes can act as proton-collecting antennae and provide fundamental information on the relation between the membrane surface charge density and the local proton exchange kinetics.


Nevirapine-associated early hepatotoxicity: incidence, risk factors, and associated mortality in a primary care ART programme in South Africa.
The majority of antiretroviral treatment programmes in sub-Saharan Africa are scaling up antiretroviral treatment using a fixed dose first-line antiretroviral regimen containing stavudine, lamivudine, and nevirapine. One of the primary concerns with the use of this regimen is nevirapine-associated hepatotoxicity. Study participants were 1809 HIV-infected, antiretroviral naïve adults initiating nevirapine-based antiretroviral therapy between November 2002 and December 2006. The primary outcome was early hepatotoxicity. Secondary outcomes were associations with hepatotoxicity and mortality at six months. The cumulative proportion of early hepatotoxicity ranged from 1.0-2.0% giving an incidence-rate at 102 days of 3.6-7.6 per 100 person-years. Median time to hepatotoxicity was 32 (IQR 28-58) days. At 12 weeks, only 8% of patients had alanine aminotransferase monitoring at all the time-points recommended by national guidelines. No association was found between age, gender, baseline CD4 count, concurrent tuberculosis infection, prior participation in a prevention of mother-to-child-transmission program, or baseline weight and early hepatotoxicity. There was no association between early hepatotoxicity and mortality. The cumulative proportion of early hepatotoxicity in nevirapine based antiretroviral therapy was low in this resource-constrained setting. Hepatotoxicity was not associated with mortality. Frequent routine monitoring of alanine aminotransferase proved difficult to implement in this public sector primary care programme. Focused monitoring in the first month may be a more cost-effective and pragmatic option in settings with limited resources. Correlation with clinical signs and symptoms may allow future alanine aminotransferase testing to be dictated by clinical criteria.


Correlation study of radiographic characteristics and operative difficulty in lateral-anterior lumbar interbody fusion (LaLIF) at the L4-5 level: a novel classification for case selection.
To analyze correlations between the realistic surgical difficulty of LaLIF and anatomic characteristics in radiographic images, in order to develop a simple classification to provide guiding information for case selection and evaluate the potential risks of the technique. Ninety-six consecutive cases who underwent LaLIF surgeries at the L4-5 level with MR T2-weighted images were analyzed. A novel classification based on the anatomic relationships among the disk, great vessels, and psoas muscle was used for grouping. Clinical outcomes and realistic surgical difficulty parameters were recorded, and comparisons were made among different types of classifications. Of the 96 analyzed cases, the time of surgical exposure was significantly longer for type C than for type B, and both of these were longer than that of type A. The VAS and ODI were significantly improved at a 1-year follow-up. There was no statistically significant difference among the three types. Type C had the highest incidence of complications, while Type A had the lowest. Analyses of another 304 MRI cases obtained in outpatient clinics showed that the distribution of the three types among these cases was consistent with that of the surgical cohort. Our novel and simple classification provides useful information for case selection. Type A provided the best indication and is most appropriate for a beginner in this technique. Type C includes the most challenging situations, which may have a high incidence of complications and require sophisticated surgical skills to achieve satisfactory outcomes and avoid approach-related complications.


Clinical and biologic characterization of T-cell neoplasias with rearrangements of chromosome 7 band q34.
In T cell malignancy, rearrangements of chromosome 14 have been observed with a break in the band that contains the alpha chain gene for the T cell receptor (TCR). Because the beta chain TCR gene is in chromosome band 7q34, we searched for and report finding specific rearrangements of 7q34 exclusively in T cell malignancies. The rearrangements were reciprocal translocations between 7q34 and other points: 1p34, 9q32, 9q34, 15q22, and 19p13. The malignancies containing a 7q34 translocation were either T cell acute lymphoblastic leukemias or T cell lymphoblastic lymphomas that had similarities in clinical, enzyme, immunologic, and cellular characteristics. Hybridization using a probe to the beta-TCR gene disclosed unique rearrangements consistent with clonality in every case. A common pattern with chromosome breakpoints involving TCR genes may be emerging in T cell neoplasia.


Maximizing Participant Engagement, Participation, and Retention in Cohort Studies Using Digital Methods: Rapid Review to Inform the Next Generation of Very Large Birth Cohorts.
Many current research needs can only be addressed using very large cohorts. In such studies, traditional one-on-one phone, face-to-face, or paper-based engagement may not be feasible. The only realistic mechanism for maintaining engagement and participation at this scale is via digital methods. Given the substantial investment being made into very large birth cohort studies, evidence for optimal methods of participant engagement, participation, and retention over sustained periods without in-person contact from researchers is paramount. This study aims to provide an overview of systematic reviews and meta-analyses evaluating alternative strategies for maximizing participant engagement and retention rates in large-scale studies using digital methods. We used a rapid review method by searching PubMed and Ovid MEDLINE databases from January 2012 to December 2019. Studies evaluating at least 1 e-engagement, participation, or retention strategy were eligible. Articles were screened for relevance based on preset inclusion and exclusion criteria. The methodological quality of the included reviews was assessed using the AMSTAR-2 (Assessing the Methodological Quality of Systematic Reviews 2) measurement tool, and a narrative synthesis of the data was conducted. The literature search yielded 19 eligible reviews. Overall, 63% (n=12) of these reviews reported on the effectiveness of e-engagement or participation promotion strategies. These evaluations were generally not conducted within very large observational digital cohorts. Most of the contributing reviews included multipurpose cohort studies (with both observational and interventional elements) conducted in clinical and research settings. Email or SMS text message reminders, SMS text messages or voice notifications, and incentives were the most commonly used design features to engage and retain participants. For parental outcomes, engagement-facilitation interventions influenced uptake and behavior change, including video feedback, goal setting, and intensive human facilitation and support. Participant-stated preferences for content included new knowledge, reminders, solutions, and suggestions about health issues presented in a clear, short, and personalized way. Perinatal and postpartum women valued self-monitoring and personalized feedback. Digital reminders and multiple SMS text messages were specific strategies that were found to increase adherence to medication and clinic attendance, respectively. This review adds to the growing literature evaluating methods to optimize engagement and participation that may apply to large-scale studies using digital methods; it is promising that most e-engagement and participation promotion strategies appear to be effective. However, these reviews canvassed relatively few strategies, suggesting that few alternative strategies have been experimentally evaluated. The reviews also revealed a dearth of experimental evidence generated within very large observational digital cohort studies, which may reflect the small number of such studies worldwide. Thus, very large studies may need to proactively build in experimental opportunities to test engagement and retention approaches to enhance the success of their own and other large digital contact studies.


Impression cytology study of epithelial phenotype of ocular surface reconstructed by preserved human amniotic membrane.
To determine the epithelial phenotype of the ocular surface reconstructed by preserved human amniotic membrane. Impression cytology was performed in 6 patients who received a large patch of amniotic membrane for conjunctival surface reconstruction during removal of acquired melanosis, conjunctival intraepithelial neoplasia, or bilateral inferior conjunctival chalasis, or for corneal surface reconstruction during removal of pannus associated with limbal deficiency caused by aniridia, toxic epidermal necrolysis, or chemical burn. The nongoblet epithelial cells covering the amniotic membrane were uniformly smaller and the cell density was almost twice that of age- and sex-matched normal control eyes at the corresponding site, and the goblet cell density was almost 10 times that of the control (both P < .05; Student paired t test) (N = 7 eyes). Furthermore, the conjunctival epithelial phenotype with goblet cells was found on corneal surfaces of all 3 patients with limbal deficiency. The success of conjunctival surface reconstruction correlated well with recovery of the conjunctival epithelial phenotype. The lack of corneal epithelial phenotype even on an avascular corneal stroma supports the concept that conjunctival transdifferentiation does not occur in vivo, and indicates that additional limbal stem cell transplantation is needed for effective corneal surface reconstruction in patients with limbal deficiency.


A randomized, controlled trial comparing two different continuous positive airway pressure systems for the successful extubation of extremely low birth weight infants.
To determine whether the use of the Infant Flow continuous positive airway pressure (IF CPAP) system reduces the rate of extubation failure among extremely low birth weight (ELBW) infants (infants with birth weight <1000 g) when compared with conventional CPAP delivered with a conventional ventilator and nasal prongs. A prospective, unmasked, randomized, controlled clinical trial was conducted in 162 eligible intubated ELBW infants who were hospitalized in 2 intensive care nurseries in Winston-Salem, North Carolina, between July 1997 and November 2000. Successful extubation was defined as no need for reintubation for any reason for at least 7 days after the first extubation attempt. The individual extubation success rates were 61.9% (52 of 84) in the conventional CPAP group and 61.5% (48 of 78) in the IF CPAP group. There were no significant differences in the extubation success rate in any birth weight subset between the 2 cohorts. The most common cause of extubation failure was apnea/bradycardia. Infants who were randomized to IF CPAP had fewer days on supplemental O(2) and shorter hospital stays. Extubation failure is a common problem, occurring in nearly 40% of ELBW infants who require mechanical ventilation. IF CPAP was as effective but no more effective than conventional CPAP in preventing extubation failure among ELBW infants. New strategies are needed to identify predictors of extubation success and to treat apnea/bradycardia, the most common cause of extubation failure, thereby reducing the likelihood of prolonged intubation in this high-risk cohort of premature infants.


Asperlicin antagonizes stimulatory effects of cholecystokinin on isolated islets.
Asperlicin, a product derived from the fungus Aspergillus alliaceus, antagonized the multiple stimulatory effects of cholecystokinin (CCK-8S) on isolated islets. At a level of 10 microM, asperlicin completely inhibited insulin release in response to 25 nM CCK-8S. Increasing the level of CCK-8S to 100 nM partially restored a secretory response, while an even greater insulin stimulatory effect was noted with 500 nM CCK-8S. The inhibitory effect of asperlicin on CCK-8S-induced release was reversible. Asperlicin exposure had no effect on glucose or glyceraldehyde-induced secretion. Asperlicin reduced, in parallel with secretion, the increase in 3H efflux from [3H]inositol prelabeled islets usually noted with CCK-8S addition. Asperlicin did not influence the small glucose-stimulated increase in 3H efflux. The results support the notion that asperlicin is a specific and potent antagonist of the multiple stimulatory effects of CCK-8S on islet tissue.


On the origin of open-circuit voltage losses in flexible n-i-p perovskite solar cells.
The possibility to manufacture perovskite solar cells (PSCs) at low temperatures paves the way to flexible and lightweight photovoltaic (PV) devices manufactured via high-throughput roll-to-roll processes. In order to achieve higher power conversion efficiencies, it is necessary to approach the radiative limit via suppression of non-radiative recombination losses. Herein, we performed a systematic voltage loss analysis for a typical low-temperature processed, flexible PSC in n-i-p configuration using vacuum deposited C60 as electron transport layer (ETL) and two-step hybrid vacuum-solution deposition for CH3NH3PbI3 perovskite absorber. We identified the ETL/absorber interface as a bottleneck in relation to non-radiative recombination losses, the quasi-Fermi level splitting (QFLS) decreases from ~1.23 eV for the bare absorber, just ~90 meV below the radiative limit, to ~1.10 eV when C60 is used as ETL. To effectively mitigate these voltage losses, we investigated different interfacial modifications via vacuum deposited interlayers (BCP, B4PyMPM, 3TPYMB, and LiF). An improvement in QFLS of ~30-40 meV is observed after interlayer deposition and confirmed by comparable improvements in the open-circuit voltage after implementation of these interfacial modifications in flexible PSCs. Further investigations on absorber/hole transport layer (HTL) interface point out the detrimental role of dopants in Spiro-OMeTAD film (widely employed HTL in the community) as recombination centers upon oxidation and light exposure.


Factors associated with unmet pediatric health care needs during the coronavirus pandemic.
To examine risk factors for unmet pediatric health care needs during the coronavirus pandemic in a national sample of US children under 17 years of age. In this cross-sectional study, we used data from the 2021 National Survey of Children's Health (n = 50,892). Unmet pediatric health care needs were assessed as follows: "during the past 12 months, was there any time when this child needed health care but it was not received?". Logistic regression with multiple imputations was used to evaluate bivariate and multivariable associations between predisposing, enabling, and need factors of health care services use and unmet pediatric health care needs. Approximately 3.6% of children had unmet health care needs, with significant differences by sample characteristics. Unmet health care needs were notably prevalent among select groups, including children with difficulty covering basic needs (10.6%) and those with ≥2 health conditions (8.9%). In multivariable analyses, factors associated with unmet pediatric health care needs included predisposing factors such as older age, ≥2 children with special health care needs in the household, primary language other than English or Spanish, and poor caregiver health; enabling factors such as difficulty covering basic needs, no insurance, and no personal doctor or nurse; and need factors such as poor perceived or evaluated child health. Reasons for unmet needs included financial, nonfinancial, and accessibility barriers. Several factors were associated with unmet health care needs. We identified children who would benefit from targeted interventions aimed at promoting health care services use.


Total Hip Arthroplasty in Teenagers: A Systematic Literature Review.
Total hip arthroplasty (THA) in teenagers is generally avoided. Nevertheless, recent THA procedures in a very young patient show improved functional outcomes and implant survival, resulting in lower revision rates. This review aims to present an overview of the available literature on THA in teenagers and to provide evidence to inform caregivers. In this systematic review, studies required a primary THA method and a teenage patient population. Studies must report at least one of the following outcome measures: functional outcomes, implant survival, and complications. In addition, demographic and surgical data were collected. Sixteen studies were analyzed, including 2040 patients and 2379 hips, with an average 7.7-year follow-up. The mean patient age was 18 years, with an average revision rate of 11.7%. The overall average relative improvement of the 2 most frequently used patient-reported (functional) outcome measures were 84.3 and 92.3% at the latest follow-up. Prosthesis, or liner loosening, was the cause of revision in 50.2% of the cases. Loosening was the most frequent complication (14.8%), together with prosthesis/liner wear (14.8%). Cementless fixation (70.7%), ceramic-on-ceramic articulation (34.7%), and the posterior surgical approach (82.3%) were the most applied techniques. The functional outcomes after THA in teenagers improved at follow-up. The average revision rate is relatively high, especially in the pre-1995 studies, with post-1995 studies reporting similar revision rates to the adult patient group. Research to further improve implant survival as well as the ease of revisions in teenagers is needed. Level III-systematic review.


The AIP Model of EMDR Therapy and Pathogenic Memories.
Eye Movement Desensitization and Reprocessing (EMDR) therapy has been widely recognized as an efficacious treatment for post-traumatic stress disorder (PTSD). In the last years more insight has been gained regarding the efficacy of EMDR therapy in a broad field of mental disorders beyond PTSD. The cornerstone of EMDR therapy is its unique model of pathogenesis and change: the adaptive information processing (AIP) model. The AIP model developed by F. Shapiro has found support and differentiation in recent studies on the importance of memories in the pathogenesis of a range of mental disorders beside PTSD. However, theoretical publications or research on the application of the AIP model are still rare. The increasing acceptance of ideas that relate the origin of many mental disorders to the formation and consolidation of implicit dysfunctional memory lead to formation of the theory of pathogenic memories. Within the theory of pathogenic memories these implicit dysfunctional memories are considered to form basis of a variety of mental disorders. The theory of pathogenic memories seems compatible to the AIP model of EMDR therapy, which offers strategies to effectively access and transmute these memories leading to amelioration or resolution of symptoms. Merging the AIP model with the theory of pathogenic memories may initiate research. In consequence, patients suffering from such memory-based disorders may be earlier diagnosed and treated more effectively.


[Results of pegylated interferon and ribavirin for the treatment of chronic hepatitis C in clinical practice: a 5-year experience].
Numerous clinical trials have demonstrated the efficacy of treatment with pegylated interferon and ribavirin but little is known about the results obtained in clinical practice. To evaluate treatment response and factors influencing the treatment of chronic hepatitis C in clinical practice. Between August 2001 and December 2005, we treated 219 patients with pegylated interferon (alpha 2a -fixed dose, or alpha 2b, according to weight) and ribavirin. Patients with genotype 1 or 4 received treatment with pegylated interferon alpha 2a (180 microg/week) and ribavirin (1000 mg/day if body weight was <75 kg or 1200 mg/day if body weight was >75 kg) or interferon alpha 2b (1.5 microg/kg/week) and ribavirin (10.6 mg/kg/day) for 48 weeks. Patients with genotype 2 or 3 were treated for 24 weeks with the same regimen of pegylated interferon alpha-2a or alpha-2b, but with 800 mg of ribavirin divided in two daily doses. Sustained viral response was defined as absence of HCV-RNA 6 months after the end of treatment. A total of 219 patients were included (69% men; mean age 44+/-10). As epidemiological antecedents, 22.4% of the treated patients had previously consumed drugs parenterally and 22.4% had received blood transfusions before 1992. Forty-seven percent of the patients with liver biopsy had fibrosis bridges or established liver cirrhosis. The genotype was distributed as follows: 69.8% genotype 1, 4.1% genotype 2, 17.8% genotype 3, and 8.2% genotype 4. Of the 219 patients, 76 (35%) were treated with pegylated interferon alpha 2a and 143 (65%) with interferon alpha 2b. Analysis of response by genotype revealed that sustained viral response was obtained in 46% genotype 1, 88.9% genotype 2, 78.9% genotype 3, and 33.3% genotype 4. Univariate analysis showed that the only variable influencing sustained viral response was genotype. Treatment with pegylated interferon and ribavirin in clinical practice shows a similar pattern of sustained viral response to that obtained in clinical research. The main variable correlated with sustained viral response continues to be viral genotype.


808-nm laser with exogenous chromophores for the treatment of benign oral lesions.
The aim of this study was to verify the effectiveness and safety of the 808-nm wavelength for use on benign lesions of the oral cavity that are stained with toluidine blue solution. The toluidine blue solution according to Mashberg is used in stomatology for the detection of leuko-erythroplasic lesions. In our experience, we have used it to stain even serous cysts, angiomas, and fibromas. We chose a 808-nm wavelength emitted from a power diode laser, because it is well absorbed by the blue color and barely by the healthy mucosa. The non-contact photocoagulation kept the surrounding borders intact when we used the pulsed-emission, 50 ms 7 W, without anesthesia. No sutures were needed. The leukoplasic borders of the lesions were histologically examined before and after the treatment. The vaporized lesions healed completely in about 5-7 days without any kind of treatment, leaving some slight elastic white scars. A mild edema-erythema followed the treatment for 5-7 days. The selective results confirm the effectiveness of this therapeutic procedure on pigmented benign lesions, pre-treated with exogenous chromophores and photocoagulated without bleeding, with a minimum energy 808-nm laser.


PCR-based pooling of dried blood spots for detection of malaria parasites: optimization and application to a cohort of Ugandan children.
Sensitive, high-throughput methods to detect malaria parasites in low-transmission settings are needed. PCR-based pooling strategies may offer a solution. We first used laboratory-prepared samples to compare 2 DNA extraction and 4 PCR detection methods across a range of pool sizes and parasite densities. Pooled Chelex extraction of DNA, followed by nested PCR of cytochrome b, was the optimal strategy, allowing reliable detection of a single low-parasitemic sample (100 parasites/μl) in pool sizes up to 50. This PCR-based pooling strategy was then compared with microscopy using 891 dried blood spots from a cohort of 77 Ugandan children followed for 2 years in an urban setting of low endemicity. Among 419 febrile episodes, 35 cases of malaria were detected using the PCR-based pooling strategy and 40 cases using microscopy. All five cases of malaria not detected by PCR were from samples stored for >2 years with parasitemia of <6,000/μl, highlighting the issue of possible DNA degradation with long-term storage of samples. Among 472 samples collected from asymptomatic children as part of routine surveillance, 15 (3.2%) were positive by PCR-based pooling compared to 4 (0.8%) by microscopy (P = 0.01). Thus, this PCR-based pooling strategy for detection of malaria parasites using dried blood spots offers a sensitive and efficient approach for malaria surveillance in low-transmission settings, enabling improved detection of asymptomatic submicroscopic infections and dramatic savings in labor and costs.


Post-electroconvulsive therapy recovery and reorientation time with methohexital and ketamine: a randomized, longitudinal, crossover design trial.
Methohexital, a barbiturate anesthetic commonly used for electroconvulsive therapy (ECT), possesses dose-dependent anticonvulsant properties, and its use can interfere with effective seizure therapy in patients with high seizure thresholds. Ketamine, an N-methyl-d-aspartate antagonist with epileptogenic properties not broadly used for ECT inductions, is a commonly used induction agent for general anesthesia. Recent studies suggest that the use of ketamine is effective in allowing successful ECT treatment in patients with high seizure thresholds without an increase in adverse effects. In this preliminary study, we directly compared the recovery and reorientation times of subjects receiving ketamine and methohexital for ECTs. Twenty patients were randomized in a crossover design to receive methohexital and ketamine for ECT inductions in alternating fashion in 6 trials. Primary outcome measures were recovery time (voluntary movement, respiratory effort, blood pressure, consciousness, and O2 saturation) and reorientation time. Secondary outcome measures were individual recovery variables, adverse effect occurrence, and seizure duration. Overall recovery time was not significantly different between the 2 treatment arms (F(1, 17) = 0.72; P = 0.41). Reorientation time was faster in the methohexital arm (F(1, 17) = 9.23; P = 0.007). Ketamine inductions resulted in higher number of adverse effects, higher subject dropout rates, and a longer reorientation time with respect to methohexital inductions. No significant difference in postanesthesia recovery time was found between the ketamine and methohexital arms. Intolerability to ketamine affected a significant proportion of subjects and suggests that ketamine should remain as an alternative or adjunctive agent for patients with high seizure thresholds.


Can Lipomatosis of the Nerve Occur or Extend Intradurally?
Diverse adipose lesions can affect peripheral nerves, including an intrinsic disorder known as lipomatosis of nerve (LN). This condition leads to massive nerve enlargement and has often been associated with nerve territory overgrowth. Although LN has been well documented as a peripheral lesion, it is uncertain whether LN can occur or extend intradurally. In the present 2-part study, we searched our institutional database and the world literature to identify any case of LN occurring or extending intradurally. Strict pathognomonic imaging and histopathologic features of LN were required to be present. We did not identify any case of LN that had occurred or extended intradurally in our institution. Specifically, in our database, we found no case of intradural LN, and an evaluation of the imaging studies of proximal examples of LN did not show any extension proximal to the spinal foramen. Our literature search identified 208 reports of potential interest, of which only 3 had reported on spinal LN. Although 2 of the 3 cases showed some similarities to LN, none had demonstrated features diagnostic for LN and none had demonstrated nerve territory overgrowth. A review of 16 cases of LN in proximal locations summarized in a recently reported systematic review did not reveal any cases with LN proximal to the foramen or in an intradural location. A review of our institutional cases and reported cases did not show any example of LN extending or occurring intradurally. It appears that LN is a benign tumor-like nerve lesion that is without a central location, unlike more well-known tumors such as schwannomas.


Common genetic variation in TP53 is associated with lung cancer risk and prognosis in African Americans and somatic mutations in lung tumors.
Lung cancer is primarily caused by tobacco smoking, but susceptibility is likely modified by common genetic variation. In response to many forms of cellular stress, including DNA damage, the p53 protein functions to induce cell cycle arrest, DNA repair, senescence, or apoptosis. We hypothesized that common TP53 haplotypes modulate pathways of lung carcinogenesis and lung cancer susceptibility or prognosis. To investigate our hypothesis, 14 polymorphisms in TP53, including haplotype tagging and coding single nucleotide polymorphisms, were genotyped in two studies from the greater Baltimore, Maryland area. One study is a case-control study and the second is a case-only study for which TP53 mutational spectra data are available. African Americans with Pro-T-A-G-G haplotypes of the combined TP53 polymorphisms TP53_01 (rs1042522), TP53_65 (rs9895829), TP53_66 (rs2909430), TP53_16 (rs1625895), and TP53_11 (rs12951053) had both an increased risk for lung cancer (odds ratio, 2.32; 95% confidence interval, 1.18-4.57) and a worsened lung cancer prognosis (hazards ratio, 2.38; 95% confidence interval, 1.38-4.10) compared with those with Arg-T-A-G-T haplotypes. No associations of TP53 polymorphisms with lung cancer were observed in Caucasians. In the case-only study, several polymorphisms in TP53 and TP53 haplotypes, overlapping regions of TP53 associated with risk and prognosis in African Americans, were associated with increased odds of somatic TP53 mutation in lung tumors in Caucasians. In conclusion, common genetic variation in TP53 could modulate lung cancer pathways, as suggested by the association with lung cancer in African Americans and somatic TP53 mutation frequency in lung tumors.


Influence of local anesthesia on the outcomes of non-surgical periodontal treatment.
There is limited evidence of the effects of local anesthesia (LA) on outcomes of non-surgical periodontal treatment (NSPT), in particular among the Chinese. This retrospective cohort study aimed to evaluate the effects of LA on short-term treatment outcomes of NSPT and to determine under what circumstances LA should be prescribed to improve these outcomes. Data from periodontal examinations of 3980 patients were used. The data were from 3-month re-evaluation records of an electronic periodontal charting record system in the Department of Periodontology of Peking University School and Hospital of Stomatology from June 2008 to January 2015. Descriptive analyses included changes in probing depth (PD) and the Mazza bleeding index (BI). Two-level (patient and tooth) logistic regression models and three-level (patient, tooth, and site) linear regression models were constructed to analyze the influence of LA on PD for all teeth/sites and teeth/sites with an initial PD ≥ 5 mm. Decreases in PD and BI at sites under LA using the initial PD were also compared. A significantly higher mean decrease in PD after NSPT was found in the LA group than in the no local anesthesia (NLA) group (0.98 vs. 0.54 mm, t = 24.12, P < 0.001). A significantly higher probability of decreases was found in the LA group in BI (percentages of teeth with BI > 1 and BI > 2) for all teeth (16.7% vs. 13.8%, t = 3.75, P < 0.001; 34.7% vs. 28.1%, t = 6.73, P < 0.001) and PD for teeth with PD ≥ 5 mm (32.3% vs. 17.3%, t = 28.48, P < 0.001). The difference in PD between the LA and NLA groups increased as the initial PD increased. The difference between the two groups was 0.12 to 0.22 mm for sites with a baseline PD < 7 mm; however, it increased to 0.41 to 1.37 mm for sites with a baseline PD ≥ 7 mm. LA improved the decrease in PD after NSPT. Root debridement at sites with initial PD ≥ 7 mm should be performed under routine LA.


Yersinia enterocolitica isolated from two cohorts of young children in Santiago, Chile: incidence of and lack of correlation between illness and proposed virulence factors.
Yersinia enterocolitica was isolated from children in two cohorts in Santiago, Chile. In a cohort containing a cross section of children aged 0 to 4 years, Y. enterocolitica was isolated from stool samples of 1.1% of children with diarrhea and 0.2% of age-matched control children. In a subgroup of this cohort from which weekly stool samples were obtained from all children irrespective of clinical status, 6% of children had asymptomatic Yersinia infections. In a birth cohort (with a greater representation of children less than 1 year of age and a significantly higher rate of diarrhea), Y. enterocolitica was isolated from 1.9% of children with diarrhea and 0.6% of controls (P = 0.05). Biogroup 1A strains (which lacked traditional phenotypic and molecular markers for pathogenicity) were isolated from seven children with diarrhea but from no control children in the birth cohort (P = 0.02). All other isolates, including all isolates from asymptomatic children, were "pathogenic" strains in biogroup 4, serogroup O3; no association between these isolates and occurrence of disease was found. Y. enterocolitica is found among young children in Santiago, with asymptomatic infections not uncommon occurrences. However, questions about the association between previously described virulence factors and diarrheal illness remain.


Resection of hepatocellular adenoma in patients with glycogen storage disease type Ia.
Because dietary modifications have prolonged the life expectancy of patients with glycogen storage disease type Ia (GSD Ia), the incidence of hepatocellular adenoma (HCA) to carcinoma (HCC) transformation is increasing. The objective of this retrospective study is to assess the safety and effectiveness of HCA resection in GSD Ia patients. Clinicopathologic, peri-operative, and long-term data were reviewed from patients who underwent HCA resection. Comparisons were made with Fisher's exact, Mann-Whitney U, and log-rank tests; survival was estimated with Kaplan-Meier analysis. From 1998 to 2006, 38 patients underwent HCA resection. Seven (22%) had GSD Ia. Post-operative mortality occurred in one GSD Ia patient. GSD Ia patients had greater morbidity (86% vs. 20%) and shorter time to adenoma progression (median 23 months vs. not yet reached) after partial hepatectomy compared to the general population (p<0.05). Six GSD Ia patients had no evidence of HCC and recovered after resection without long-term morbidity. Three GSD Ia patients underwent liver transplantation 77, 32, and 23 months after adenoma resection. Despite substantial morbidity, partial hepatectomy is feasible in GSD Ia patients and is an effective intermediate step in the prevention of HCC until definitive treatment with liver transplantation.


A discrete-time Multiple Event Process Survival Mixture (MEPSUM) model.
Traditional survival analysis was developed to investigate the occurrence and timing of a single event, but researchers have recently begun to ask questions about the order and timing of multiple events. A multiple event process survival mixture model is developed here to analyze nonrepeatable events measured in discrete-time that may occur at the same point in time. Building on both traditional univariate survival analysis and univariate survival mixture analysis, the model approximates the underlying multivariate distribution of hazard functions via a discrete-point finite mixture in which the mixing components represent prototypical patterns of event occurrence. The model is applied in an empirical analysis concerning transitions to adulthood, where the events under study include parenthood, marriage, beginning full-time work, and obtaining a college degree. Promising opportunities, as well as possible limitations of the model and future directions for research, are discussed.


Requirements for conversion of the Na(+)-driven flagellar motor of Vibrio cholerae to the H(+)-driven motor of Escherichia coli.
Bacterial flagella are powered by a motor that converts a transmembrane electrochemical potential of either H(+) or Na(+) into mechanical work. In Escherichia coli, the MotA and MotB proteins form the stator and function in proton translocation, whereas the FliG protein is located on the rotor and is involved in flagellar assembly and torque generation. The sodium-driven polar flagella of Vibrio species contain homologs of MotA and MotB, called PomA and PomB, and also contain two other membrane proteins called MotX and MotY, which are essential for motor rotation and that might also function in ion conduction. Deletions in pomA, pomB, motX, or motY in Vibrio cholerae resulted in a nonmotile phenotype, whereas deletion of fliG gave a nonflagellate phenotype. fliG genes on plasmids complemented fliG-null strains of the parent species but not fliG-null strains of the other species. FliG-null strains were complemented by chimeric FliG proteins in which the C-terminal domain came from the other species, however, implying that the C-terminal part of FliG can function in conjunction with the ion-translocating components of either species. A V. cholerae strain deleted of pomA, pomB, motX, and motY became weakly motile when the E. coli motA and motB genes were introduced on a plasmid. Like E. coli, but unlike wild-type V. cholerae, motility of some V. cholerae strains containing the hybrid motor was inhibited by the protonophore carbonyl cyanide m-chlorophenylhydrazone under neutral as well as alkaline conditions but not by the sodium motor-specific inhibitor phenamil. We conclude that the E. coli proton motor components MotA and MotB can function in place of the motor proteins of V. cholerae and that the hybrid motors are driven by the proton motive force.


Analysis of Danshen and twelve related Salvia species.
Danshen is a commonly used traditional Chinese herb, but over twenty Salvia species are used as Danshen by local herbalists. In this study, twelve Salvia species from the plateau of Sichuan and Yunnan Provinces were collected, analyzed and compared with Danshen by HPLC. The results showed that most of the Salvia species were good sources of rosmarinic acid and tanshinones. The highest amount of both rosmarinic acid and tanshinone IIA were found in S. przewalskii Maxim. These results pave the way for a better therapeutic exploitation of these plants.


Induction of apoptosis by ASK1, a mammalian MAPKKK that activates SAPK/JNK and p38 signaling pathways.
Mitogen-activated protein (MAP) kinase cascades are activated in response to various extracellular stimuli, including growth factors and environmental stresses. A MAP kinase kinase kinase (MAPKKK), termed ASK1, was identified that activated two different subgroups of MAP kinase kinases (MAPKK), SEK1 (or MKK4) and MKK3/MAPKK6 (or MKK6), which in turn activated stress-activated protein kinase (SAPK, also known as JNK; c-Jun amino-terminal kinase) and p38 subgroups of MAP kinases, respectively. Overexpression of ASK1 induced apoptotic cell death, and ASK1 was activated in cells treated with tumor necrosis factor-alpha (TNF-alpha). Moreover, TNF-alpha-induced apoptosis was inhibited by a catalytically inactive form of ASK1. ASK1 may be a key element in the mechanism of stress- and cytokine-induced apoptosis.


[Modern methods in analysis of microtubules dynamic in vivo].
Microtubules are involved in any vital cellular activities, including the maintenance of cell shape, division, migration and intracellular transport. Microtubule dynamics is regulated by the balance between their polymerization and depolymerization. Microtubule stability is dependent on their alpha and beta subunits composition, tubulin post-translational modifications and interaction of microtubules with microtubule-associated proteins (MAPs). Disruption of these processes can lead to a number of pathological conditions such as cancer, cardiovascular disease, or the fibrosis development. This review summarizes the current knowledge of the modern methods of microtubule polymerization analysis. This allows a better understanding of the structure and mechanisms played by microtubules in their physiological functions and the development of pathological conditions resulting from their disorder.


Characterization of the mouse proteasome regulator PA28b gene.
The proteasome regulator PA28, which can be upregulated by IFN, is important in the modulation of proteasome activity. Since the proteasome has been implicated in the processing of the major histocompatibility complex (MHC) class I antigens, it was of interest to determine the regulatory elements of PA28 at the genomic level. Although PA28 has been found in different species, the gene layout on the chromosome was not determined. In this study, the genetic organization of mouse PA28b was characterized. Two copies of the PA28b gene, namely b1 and b2, were found by restriction fragment mapping and Southern hybridization. By fluorescence in situ hybridization, the location of the two PA28b genes was determined on chromosomes 11 and 14. PA28b1 has 11 exons, whereas PA28b2 has no introns and appears to be a nonfunctional pseudogene. The 5' promoter region of PA28b1 contains several transcriptional factor binding sites including two IFN responsive elements. The expression levels of PA28 and other gene products involved in MHC class I antigen presentation appear to be correlated in various tissues. Notably, PA28 is expressed at high levels in immunological tissues such as spleen and peripheral blood leukocytes. Taken together, PA28 seems to be co-regulated with other molecules involved in MHC class I antigen presentation.


[Subclinical findings in the knees of taekwondo athletes: diagnostic ultrasound study].
Taekwondo is associated with an increased incidence of musculoskeletal injuries such as tendinopathy, synovitis, chondropathy, and ligament and meniscus injuries that may have an asymptomatic course in their initial stages, especially those located in the knee. To describe the presence of morphostructural abnormalities in asymptomatic taekwondo athletes' (TKD) knees through the use of diagnostic ultrasound (US). A cross-sectional, descriptive and comparative study. We evaluated 32 knees of 16 subjects (8 TKD and 8 recreational athletes). All subjects underwent sport-medical history and knee US. A variety of intra- and extra-articular morphostructural abnormalities were observed; the most frequent were synovitis, meniscal extrusion, and enthesopathy. The practice of Taekwondo abnormalities associated with an increased risk of knee injuries that may go unnoticed in the early stages. The use of US as an auxiliary tool in the diagnosis of these injuries and/or advisable since it can define in detail the anatomical structures subject to overuse, biomechanical stress, or repetitive trauma, and contribute to early detection of asymptomatic morphostructural alterations that may ensure timely preventive and therapeutic interventions.


Effect of green tea extract on obese women: a randomized, double-blind, placebo-controlled clinical trial.
To examine the effect of green tea extract (GTE) on obese women and to explore the relationship between GTE and obesity-related hormone peptides. A randomized, double-blind, placebo-controlled clinical trial was conducted from July 2006 to June 2007 in Taipei Hospital, Taiwan. Seventy-eight of 100 obese women aged between 16 and 60 years with BMI>27 kg/m(2) and who had not received any other weight control maneuvers within the last 3 months completed this study. The subjects were randomly divided into Groups A and B. Group A (n=41) received GTE while Group B (n=37) took cellulose as a placebo, one capsule (400mg) three times each day for 12 weeks. The body weight (BW), body mass index (BMI) and waist circumflex (WC) were measured at the beginning of the study and after 12 weeks of treatment with GTE. The data were compared and expressed as % reduction. There was only a 0.3% reduction in BW (0.15 kg) after 12 weeks of treatment with GTE. There was no statistical difference in % reduction in BW, BMI and WC between the GTE and placebo groups. Within group comparison revealed that the GTE group had significant reduction in LDL-cholesterol and triglyceride, and marked increase in the level of HDL-cholesterol, adiponectin and ghrelin. On the other hand, the placebo group showed significant reduction in triglyceride only, and a marked increase in the level of ghrelin alone. This study showed no statistical difference in % reduction in BW, BMI and WC between the GTE and placebo groups after 12 weeks of treatment. The intake of GTE (491 mg catechins containing 302 mg EGCG) for 12 weeks is considered safe as shown by the results.


[Castleman's disease in the cardiophrenic angle resected thoracoscopically; report of a case].
The patient was 55-years-old woman. She was referred to our hospital for an abnormal mass on a chest X-ray. She was pointed out it 12 years ago, but left it unattended. Therefore, we could observe the development of Castleman's disease for 12 years. An enhanced computed tomography showed a irregularly enhanced mass in the right cardiophrenic angle. We diagnosed as pleural tumor and performed the video assisted operation. The mass was growing from the anterior mediastinum. It was successfully resected thoracoscopically. The mass was elastic and hard, and 6.0 x 4.5 x 3.4 cm in size. The histopathological diagnosis was Castleman's disease, hyaline vascular type. We think that the tumor of Castleman's disease in the cadiophrenic angle may be resected by the thoracoscopic procedure with minimal thoracotomy.


[Selectivity of lysozyme oxidation by singlet oxygen].
Peptide analysis of tryptic hydrolysates of two lysozyme forms derived from oxidation of lysozyme with singlet oxygen shows that Trp-62, located at the active site, is destroyed. This is confirmed by the protective effect of the substrate (chitin), whose presense practically prevents the oxidation. A possibility of oxidating different tryptophan residues is discussed from the view-point of their availability to the reagent.


Neither ERK nor JNK/SAPK MAP kinase subtypes are essential for histone H3/HMG-14 phosphorylation or c-fos and c-jun induction.
The effects of EGF, TPA, UV radiation, okadaic acid and anisomycin on ERK and JNK/SAPK MAP kinase cascades have been compared with their ability to elicit histone H3/HMG-14 phosphorylation and induce c-fos and c-jun in C3H 10T1/2 cells. EGF and UV radiation activate both ERKs and JNK/SAPKs but to markedly different extents; EGF activates ERKs more strongly than JNK/SAPKs, whereas UV radiation activates JNK/SAPKs much more strongly than ERKs. Anisomycin and okadaic acid activate JNK/SAPKs but not ERKs, and conversely, TPA activates ERKs but not JNK/SAPKs. Nevertheless, all these agents elicit phosphorylation of ribosomal and pre-ribosomal S6, histone H3 and HMG-14, and the induction of c-fos and c-jun, showing that neither cascade is absolutely essential for these responses. We then analysed the relationship between ERKs, JNK/SAPKs and the transcription factors Elk-1 and c-Jun, implicated in controlling c-fos and c-jun, respectively. JNK/SAPKs bind to GST-cJun1-79, and ERKs, particularly ERK-2, to GST-Elk1(307-428); there is no cross-specificity of binding. Further, GST-Elk1(307-428) binds preferentially to active rather than inactive ERK-2. In vitro, JNK/SAPKs phosphorylate both GST-cJun1-79 and GST-Elk1(307-428), whereas ERKs phosphorylate GST-Elk1(307-428) but not GST-cJun1-79. Thus, neither ERKs nor JNK/SAPKs are absolutely essential for nuclear signalling and c-fos and c-jun induction. The data suggest either that activation of a single MAP kinase subtype is sufficient to elicit a complete nuclear response, or that other uncharacterised routes exist.


[Relations of melatonin level and insufficiency of T and B immune components in children with acute viral neuroinfections].
A study was made of the content of melatonin, cortisol and indicators of cellular (E-rosette formation, 3H-lymphocyte blast transformation) and humoral (E AC-rosette formation, IgA, IgM IgG) immunity in 36 children with acute viral neuroinfections (encephalitis, encephalomyelitis, chorio-ependymitis, polyradiculoneuritis). The children's age ranged from 7 to 14 years. A significant correlation was determined between the levels of melatonin, cortisol, 3H-lymphocyte blast transformation and IgG. Variants of immunomodulating therapy are provided, bearing in mind the disorders revealed.


TERT promoter mutation in adult granulosa cell tumor of the ovary.
The telomerase reverse transcriptase (TERT) gene is highly expressed in stem cells and silenced upon differentiation. Cancer cells can attain immortality by activating TERT to maintain telomere length and telomerase activity, which is a crucial step of tumorigenesis. Two somatic mutations in the TERT promoter (C228T; C250T) have been identified as gain-of-function mutations that promote transcriptional activation of TERT in multiple cancers, such as melanoma and glioblastoma. A recent study investigating TERT promoter mutations in ovarian carcinomas found C228T and C250T mutations in 15.9% of clear cell carcinomas. However, it is unknown whether these mutations are frequent in other ovarian cancer subtypes, in particular, sex cord-stromal tumors including adult granulosa cell tumors. We performed whole-genome sequencing on ten adult granulosa cell tumors with matched normal blood and identified a TERT C228T promoter mutation in 50% of tumors. We found that adult granulosa cell tumors with mutated TERT promoter have increased expression of TERT mRNA and exhibited significantly longer telomeres compared to those with wild-type TERT promoter. Extension cohort analysis using allelic discrimination revealed the TERT C228T mutation in 51 of 229 primary adult granulosa cell tumors (22%), 24 of 58 recurrent adult granulosa cell tumors (41%), and 1 of 22 other sex cord-stromal tumors (5%). There was a significant difference in overall survival between patients with TERT C228T promoter mutation in the primary tumors and those without it (p = 0.00253, log-rank test). In seven adult granulosa cell tumors, we found the TERT C228T mutation present in recurrent tumors and absent in the corresponding primary tumor. Our data suggest that TERT C228T promoter mutations may have an important role in progression of adult granulosa cell tumors.


Reviving red snapper.
Red snappers in the Gulf of Mexico once hovered on the brink of extinction, their population having dropped to 2 percent of what had historically swum in the Gulf. But thanks to a recently introduced plan that turns the conventional wisdom of fisheries management on its head, the picture has begun to change. Called Individual Fishing Quotas (IFQs), the new regulations, which give a guaranteed allotment of fish to each participant instead of applying industry-wide quotas, went into effect for Gulf of Mexico Red Snapper (Lutjanus campechanus) in early 2007. The results were immediate and so profound that the Gulf Fishery Management Council voted earlier this year to increase the annual limit on red snapper to nearly 7 million pounds from 5 million.


Nonpharmacological management of headaches during pregnancy.
Concerns about the effects of maternal medications on the growing baby limit the use of medication treatment for benign conditions, such as recurring headaches, during pregnancy and lactation. Nonpharmacological therapies hold particular promise for pregnant women due to the limited medication options. No controlled studies, however, have reported on the efficacy of nonpharmacological treatments for pregnant women. The first study evaluated the effectiveness of a combined nonpharmacological treatment (CT) consisting of relaxation, skin-warming biofeedback, and physical therapy for pregnant women with chronic headaches. In a second study, the CT protocol was compared with an attention control (AC) that received headache education and skin-cooling biofeedback. The first study resulted in significant symptom improvement in 79% of subjects, with an overall 72.9% reduction in headaches. In the second study, both groups improved with treatment; however the CT group was more likely to experience significant headache relief (72.7%) than the AC group (28.6%, chi 2(1) = 4.97, p < .03). Significant improvement was maintained at a 6-month follow-up for over 50% of patients. It is concluded that the combined nonpharmacological treatment was more effective than an attention control in reducing headaches during pregnancy. This treatment was effective regardless of predisposing variables.


Do community characteristics predict alcohol-related crime?
Alcohol-related crime is a substantial community problem. There is evidence to suggest that certain geographic areas experience higher rates of alcohol-related crime and that both individual and community factors are associated with alcohol-related crime. There is limited research at the community level despite communities being the target of interventions designed to reduce alcohol-related harm. This study aims to determine whether there are differences in alcohol-related crime at the community level and examines whether certain community characteristics are associated with increased alcohol-related crime. Routinely collected police data from 20 rural communities in New South Wales, Australia were analysed. The ratio of alcohol to non-alcohol-related criminal incidents was used as a proxy for alcohol-related crime. Predictor variables were population-adjusted community characteristics, including demographic and resource variables. Regression analyses suggest that there are differences between communities in alcohol-related crime. Less socioeconomic disadvantage and more GPs and licensed premises (pubs and clubs) are associated with greater alcohol-related crime at the community level. Decreasing the socioeconomic well-being of a community is not appropriate; however, introducing additional taxes to increase the cost of alcohol may decrease consumption and therefore alcohol-related crime. Reducing or capping the number of licensed premises, specifically the number of pubs and clubs, may be an appropriate strategy to reduce alcohol-related crime in rural communities.


Chronic treatment with N-acetyl-cystein delays cellular senescence in endothelial cells isolated from a subgroup of atherosclerotic patients.
Endothelial senescence may contribute to the pathogenesis of age-related vascular disorders. Furthermore, chronic exposure to risk factors for cardiovascular disease (CVD) accelerates the effects of chronological aging by generating stress-dependent damages, including oxidative stress, therefore promoting stress-induced premature senescence. Our objective was to determine whether a chronic treatment with an antioxidant (N-acetyl-cystein, NAC) could delay senescence of endothelial cells (EC) isolated and cultured from arterial segments of patients with severe coronary artery disease. If EC were considered as one population (n=26), chronic NAC treatment slightly shortened telomere attrition rate associated with senescence but did not significantly delay the onset of endothelial senescence. However, in a subgroup of NAC-treated EC (n=15) cellular senescence was significantly delayed, NAC decreased lipid peroxidation (HNE), activated the catalytic subunit of telomerase (hTERT) and inhibited telomere attrition. In contrast, in another subgroup of EC (n=11) characterized by initial short telomeres, no effect of NAC on HNE and high levels of DNA damages, the antioxidant was not beneficial on senescence, suggesting an irreversible stress-dependent damage. In conclusion, chronic exposure to NAC can delay senescence of diseased EC via hTERT activation and transient telomere stabilization, unless oxidative stress-associated cell damage has become irreversible.


Duplex Kidney in Adults: a Systematic Review of the Literature.
In this study, we describe a rare case of a male with a duplex renal collecting system with ectopic ureter insertion into the prostate who presented with sepsis and required urgent laparoscopic radical nephrectomy. This study offers urologists and emergency care practitioners a great overview of the variety of manifestations, anatomic differences, and therapeutic approaches for similar patients. modified reconstructive strategy could be a method of choice when addressing particular anatomies, such as incomplete duplex kidneys with ureteropelvic junction obstruction in both upper and lower poles when the obstruction is close to the confluence of the two ureters. There is a spectrum of acute presentations and management strategies in adult patients with duplex collecting systems. The majority of patients presenting with obstruction and infection are managed surgically with hemi-nephrectomy; stable patients who present with less extensive anomalies were managed conservatively or with ureteral dilatation or deroofing.


Overlapping-based optical freeform surface construction for extended lighting source.
The freeform optical system for an extended source was constructed by partially overlapping a few numbers of point-source freeform surfaces (PFSs) and extracting their contour. Each PFS redistributed the Lambertian emission of a point source into the prescribed light distribution or more frequently into a modified distribution. By adjusting the relative positions of the PFSs and the pattern of the modified light distribution, the optimized freeform surface could be obtained. As an example, an optical system with a height only four times the source radius is designed for achieving a uniform-illuminance distribution on the target. The optimized freeform surface was formed by two PFSs. The virtue-point-sources of the PFSs were located symmetrically on the extended source with a distance of a quarter of the source diameter from each other. Each PFS achieved an increasing-illuminance distribution. The illumination uniformity of this model can be improved by 55.4%, while the optical efficiency within the target area is maintained above 80%.


Multi-target spectral moment QSAR versus ANN for antiparasitic drugs against different parasite species.
There are many of pathogen parasite species with different susceptibility profile to antiparasitic drugs. Unfortunately, almost QSAR models predict the biological activity of drugs against only one parasite species. Consequently, predicting the probability with which a drug is active against different species with a single unify model is a goal of the major importance. In so doing, we use Markov Chains theory to calculate new multi-target spectral moments to fit a QSAR model that predict by the first time a mt-QSAR model for 500 drugs tested in the literature against 16 parasite species and other 207 drugs no tested in the literature using spectral moments. The data was processed by linear discriminant analysis (LDA) classifying drugs as active or non-active against the different tested parasite species. The model correctly classifies 311 out of 358 active compounds (86.9%) and 2328 out of 2577 non-active compounds (90.3%) in training series. Overall training performance was 89.9%. Validation of the model was carried out by means of external predicting series. In these series the model classified correctly 157 out 190, 82.6% of antiparasitic compounds and 1151 out of 1277 non-active compounds (90.1%). Overall predictability performance was 89.2%. In addition we developed four types of non Linear Artificial neural networks (ANN) and we compared with the mt-QSAR model. The improved ANN model had an overall training performance was 87%. The present work report the first attempts to calculate within a unify framework probabilities of antiparasitic action of drugs against different parasite species based on spectral moment analysis.


Enhanced affinity bifunctional bisphosphonates for targeted delivery of therapeutic agents to bone.
Skeletal diseases have a major impact on the worldwide population and economy. Although several therapeutic agents and treatments are available for addressing bone diseases, they are not being fully utilized because of their uptake in nontargeted sites and related side effects. Active targeting with controlled delivery is an ideal approach for treatment of such diseases. Because bisphosphonates are known to have high affinity to bone and are being widely used in treatment of osteoporosis, they are well-suited for drug targeting to bone. In this study, a targeted delivery of therapeutic agent to resorption sites and wound healing sites of bone was explored. Toward this goal, bifunctional hydrazine-bisphosphonates (HBPs), with spacers of various lengths, were synthesized and studied for their enhanced affinity to bone. Crystal growth inhibition studies showed that these HBPs have high affinity to hydroxyapatite, and HBPs with shorter spacers bind more strongly than alendronate to hydroxyapatite. The HBPs did not affect proliferation of MC3T3-E1 preosteoblasts, did not induce apoptosis, and were not cytotoxic at the concentration range tested (10(-6)-10(-4) M). Furthermore, drugs can be linked to the HBPs through a hydrazone linkage that is cleavable at the low pH of bone resorption and wound healing sites, leading to release of the drug. This was demonstrated using hydroxyapatite as a model material of bone and 4-nitrobenzaldehyde as a model drug. This study suggests that these HBPs could be used for targeted delivery of therapeutic agents to bone.


Protoplasts from yeast and mycelial forms of Candida albicans.
Protoplasts have been obtained in high yields from the yeast and mycelial forms of a variety of strains of Candida albicans by enzyme digestion of cells with commercially available lytic enzymes. The protoplast formation procedure was equally effective for exponential and stationary phase cells. Pretreatment with dithiothreitol and Pronase in the presence of EDTA and Tris was necessary. Other thiol reagents and conditions did not release protoplasts from all the strains of C. albicans tested. Treatment with digestive juice of the snail Helix pomatia required the addition of chitinase for the release of protoplasts from most strains tested. Conditions for maximizing the yield of protoplasts and the activities of beta-glucuronidase and chitinase were determined. Electron microscopy of C. albicans showed that the pretreatment conditions removed the outer layers and the treatment itself completely removed the inner layers of the cell wall. More than 90% of the protoplasts produced by this model were viable as assessed by vital staining with Janus Green B.


[Dark oxidation of unsaturated lipids and dihydroxy phenylalanine by photooxidized furocoumarins].
Well-known skin photosensitizers furocoumarins--psoralen, angelio in and 8-methoxypsoralen were shown to photosensitize oxidation of unsaturated lipids and dihydroxyphenylalanine via a two-step mechanism. At the first step (UV-irradiation at 366 nm) photooxidation of furocoumarins occurred, at the second step--the photooxidized furocoumarins oxidized unsaturated lipids or dihydroxyphenylalanine in darkness. The photochemical activity of furocoumarins in the two-step reaction decreased as follows: psoralen greater than angelicin greater than 8-methoxypsoralen. Similar photoreactions appear to participate in induction of phototoxic effects of furocoumarins.


Importance of bicarbonate ion in the vitreous space.
Intraocular irrigating solutions used during vitrectomy must be safe for all tissues surrounding the vitreous space. Bicarbonate and glucose were essential to the maintenance of retinal function. In this study the optimum concentration of bicarbonate in the irrigating solution for the retina was determined electrophysiologically with the use of the in vitro eyecup of the rabbit. In 15 to 25 mmole/L of bicarbonate solutions, the electroretinographic (ERG) b wave was well maintained. Higher concentrations of bicarbonate (35 to 45 mmole/L), which have been considered to be advantageous for the integrity of the corneal endothelium, were shown to suppress the retinal activity. These results were also found in in vivo eyecup, as determined with ERG and visual evoked response. Properly performed open-sky vitrectomy, itself, does not cause severe damage to the normal retinal function.


High-flux isobutanol production using engineered Escherichia coli: a bioreactor study with in situ product removal.
Promising approaches to produce higher alcohols, e.g., isobutanol, using Escherichia coli have been developed with successful results. Here, we translated the isobutanol process from shake flasks to a 1-L bioreactor in order to characterize three E. coli strains. With in situ isobutanol removal from the bioreactor using gas stripping, the engineered E. coli strain (JCL260) produced more than 50 g/L in 72 h. In addition, the isobutanol production by the parental strain (JCL16) and the high isobutanol-tolerant mutant (SA481) were compared with JCL260. Interestingly, we found that the isobutanol-tolerant strain in fact produced worse than either JCL16 or JCL260. This result suggests that in situ product removal can properly overcome isobutanol toxicity in E. coli cultures. The isobutanol productivity was approximately twofold and the titer was 9% higher than n-butanol produced by Clostridium in a similar integrated system.