High-sensitivity troponin T and N-terminal pro-B-type natriuretic peptide (NT-proBNP) and risk of incident heart failure in patients with CKD: the Chronic Renal Insufficiency Cohort (CRIC) Study.
High-sensitivity troponin T (hsTnT) and N-terminal pro-B-type natriuretic peptide (NT-proBNP) strongly predict heart failure (HF) in the general population. However, the interpretation of levels of these biomarkers as predictors of HF is uncertain among patients with CKD. Here, we investigated whether hsTnT and NT-proBNP are associated with incident HF among patients with CKD. In a prospective cohort analysis, we studied 3483 people with CKD in the Chronic Renal Insufficiency Cohort (CRIC) Study recruited from June of 2003 to August of 2008 who were free of HF at baseline. We used Cox regression to examine the association of baseline levels of hsTnT and NT-proBNP with incident HF after adjustment for demographic factors, traditional cardiovascular risk factors, markers of kidney disease, pertinent medication use, and mineral metabolism markers. At baseline, hsTnT levels ranged from ≤5.0 to 738.7 pg/ml, and NT-proBNP levels ranged from ≤5 to 35,000 pg/ml. Compared with those who had undetectable hsTnT, participants in the highest quartile (>26.5 pg/ml) had a significantly higher rate of HF (hazard ratio, 4.77; 95% confidence interval, 2.49 to 9.14). Similarly, compared with those in the lowest NT-proBNP quintile (<47.6 pg/ml), participants in the highest quintile (>433.0 pg/ml) experienced a substantially higher rate of HF (hazard ratio, 9.57; 95% confidence interval, 4.40 to 20.83) [corrected]. In conclusion, hsTnT and NT-proBNP were strongly associated with incident HF among a diverse cohort of individuals with mild to severe CKD. Elevations in these biomarkers may indicate subclinical changes in volume and myocardial stress that subsequently contribute to clinical HF.


Management of low-grade squamous intraepithelial lesions during pregnancy.
Our purpose was to determine whether prenatal colposcopy is beneficial in pregnant women with squamous atypia, atypical squamous cells of undetermined significance, or low-grade squamous intraepithelial lesions on an initial screening Papanicolaou smear. A retrospective chart review identified a cohort of pregnant patients referred to the colposcopy clinic at Carolinas Medical Center between October 1991 and December 1994 with squamous atypia, atypical squamous cells of undetermined significance, or low-grade squamous intraepithelial lesions. Results of the colposcopic examination, cervical biopsy specimens, postpartum evaluation, and postpartum treatment were recorded. Descriptive statistics were used to tabulate numbers and percentages for all variables with 95% confidence intervals to determine disease progression. Prenatal colposcopy was performed on 253 women during the study period. The colposcopic impression was normal or consistent with low-grade squamous intraepithelial lesions in 235 (93%) of the women. Of the 67 women who had a cervical biopsy, 6 had a histologic diagnosis of high-grade squamous intraepithelial lesions. Postpartum Papanicolaou smears were obtained in 224 patients; 71 (32%) were normal, 145 (65%) were unchanged, and 8 (3%) showed high-grade squamous intraepithelial lesions. Of the 69 patients who had a postpartum cervical biopsy, 4 were found to have high-grade squamous intraepithelial lesions. Eight of the 10 women with biopsy-proved high-grade squamous intraepithelial lesions were compliant with treatment after delivery. Histologic examination of the cervix with tissue obtained by either loop conization or cold knife conization showed no evidence of invasive carcinoma. Squamous atypia, atypical squamous, cells of undetermined significance, or low-grade squamous intraepithelial lesions on a Papanicolaou smear in a pregnant patient does not require colposcopic evaluation during pregnancy. Progression of low-grade dysplasia to carcinoma during pregnancy is unusual, and no patient in this study was found to have invasive cancer.


Determination of ultra-trace mercury(II) by flow-injection/anodic stripping voltammetry using a track-etched microporous membrane electrode.
A new flow-injection/anodic stripping voltammetry has been demonstrated to assess ultra-trace mercury(II) using track-etched microporous membrane electrodes. The electrodes were prepared by the sputtering of gold or platinum onto both sides of a membrane filter with a smooth flat surface and with cylindrical pores having uniform diameter. The deposition of mercury from a mercury(II) solution was performed while the sample solution flowed through the membrane electrodes. After the deposition step, an anodic stripping voltammogram was obtained by sweeping the potential from 0 to +0.8 V vs. Ag/AgCl. In this case, the sample solution flowed through the pores of the 10-μm-thick membrane filters. Efficient electrolysis occurred during passage of the sample solution through the electrode, of which the pore size was 0.4 μm. In this study, the voltammetry described above was demonstrated using an FIA system. The continuous-flow mode showed a detection limit of 0.04 μg L(-1) when the experimental conditions of the flow rate and the deposition time were set at 0.5 mL min(-1) and 180 s. In the sample-injection mode equipped with a 1-mL sample loop, a linear relation was found for 0.5-4.0 μg L(-1) of a mercury(II) standard solution (r = 0.995). The detection limit was 0.05 μg L(-1). This method was applied to the ultra-trace determination of mercury(II) in river-water samples.


[Coherent singlet-triplet transitions at the initial step of tubulin assembly into microtubules].
Quantum chemistry calculations [DFT-B3LYP QM/MM method, 6-31G** basis set, + ab initio molecular dynamics] were used to study the action of Mg2+ on tubulin properties. It was shown that the hydration of the guanosine triphosphate-tubulin forms a protein zone structure, which includes a electron-occupied zone and a conductivity zone. The binding of Mg2+ to guanosine triphosphate-tubulin results in the unpairing of electrons in the occupied zone (triplet state formation) followed by their transition to the conductivity zone in which the inversion of spin occurs (singlet state formation). The formation of triplet state is the initial step in the subsequent protein dynamics in the picosecond range of time. The dynamics shows up as a coherent oscillating transition of tubulin between the triplet and singlet states, which is evidence of a simultaneous adjustment between nuclear and electron configurations of the protein (ab initio molecular dynamics calculations). The barrier between the triplet and singlet states does not exceed 0.60 kcal x mol(-1). The barrier overcome is considered as electron tunneling through the Fermi surface, which separates the occupied and conductivity zones. Zone formation occurs in the presence of the shell of biological water surrounding the protein.


Task-set switching and long-term memory retrieval.
The authors tested the hypothesis of a close relationship between the intentional component of task-set switching ("advance reconfiguration;" R. D. Rogers & S. Monsell, 1995) and long-term memory (LTM) retrieval. Consistent with this hypothesis, switch costs are reported to be larger when the switched-to task involves high retrieval demands (i.e., retrieval of episodic information) than when it involves low retrieval demands (i.e., retrieval of semantic information). In contrast, switch costs were not affected by a primary-task difficulty manipulation unrelated to intentional retrieval demands (Experiment 2). Also, the retrieval-demand effect on switch costs was eliminated when time for advanced preparation or task cues explicitly specifying the task rules were provided (Experiment 3). Overall, results were consistent with the hypothesis that the intentional switch-cost component reflects the time demands of retrieving appropriate task rules from LTM.


The differential release of basal ATPase, Ca2+-dependent ATPase, 5'-nucleotidase and cholesterol during homogenization of skeletal muscle.
The influence of homogenization times on the presence of constituents in the microsomal fraction of skeletal muscle was investigated. Membranes having Ca2+-activated ATPase activity have a fragmentation pattern distinct from that of membranes displaying Ca2+-independent or basal ATPase activity. These latter membranes were found in highest specific concentration in the microsomal fraction prepared from homogenates subjected to short periods of homogenization. 5'-Nucleotidase (EC 22.214.171.124) activity paralleled that of basal ATPase on short periods of homogenization, as also did the specific concentration of cholesterol. Longer periods of homogenization led to a decrease in the specific activity of basal atpase, which reached its lowest value at 120s of homogenization, whereas the specific activity of 5'-nucleotidase and the specific concentration of cholesterol decreased initially in a similar manner to basal ATPase, but both increased substantially after the longest period of homogenization.


Abortion of a sow caused by Pasteurella aerogenes.
Three strains of the Pasteurella aerogenes complex were isolated as sole pathogens from aborted fetuses of a sow aborted at the 12th week of gestation on a farm of 600 sows. Gross pathology showed no characteristic lesions. The isolates were biochemically identical and resembled P. pneumotropica on the basis of their strong indole and urease positivity but they produced gas, were ornithine decarboxylase negative and fermented mannitol but not trehalose. Only a few differences were apparent in biochemical characteristics between the isolated strains and P. aerogenes. They differed from the type strain of P. aerogenes in ornithine decarboxylase activity, indole production and lactose and mannitol fermentation; however, such strains do occur within this heterogeneous species. At the time of abortion the antibody titre of the aborted sow was 1 in 16 when examined with live bacterial suspension and 1 in 128 if boiled antigen was used. Similar strains could not be isolated from the vaginas of aborted sows or pregnant and newly farrowed sows in the same group. The bacteriological, serological and histological findings support the opinion of other workers on the occasional pathogenic nature of P. aerogenes.


In utero and in vitro exposure to beta-lactams impair kidney development in the rat.
beta-Lactam antibiotics are widely used because of their lack of toxicity in humans. However, during pregnancy, exposure of the fetus is likely to occur because beta-lactam antibiotics cross the placenta. The potential adverse effects of two penicillins (ampicillin, amoxicillin) and of one cephalosporin (ceftriaxone) were examined in rat kidney development. Two experimental approaches were used: metanephros organ cultures to analyze the direct effect of the drug and maternal treatment to assess the consequences of in utero exposure. For in vitro experiments, metanephroi were removed from 14-d-old fetuses and grown with or without the antibiotic at a concentration ranging from 10 to 1000 microg/ml for 6 d. For in vivo experiments, pregnant rats were treated with penicillin at 100 mg/kg per d for 5 d, a period overlapping early renal organogenesis. Both penicillins alter renal development in vitro in a dose-dependent manner, from a dose of 10 microg/ml for ampicillin and 100 microg/ml for amoxicillin. In young animals exposed to penicillins in utero, a mild oligonephronia was present and cystic tubule dilation was observed in newborn and in young animals as well. Ceftriaxone weakly impairs in vitro nephrogenesis except at the dose of 1000 microg/ml that blocks kidney development completely. No effect on nephron ontogeny was observed following in utero exposure, but an interstitial inflammation was present in the medulla of 2-wk-old rats. In conclusion, these data show that beta-lactams, at therapeutic doses, are harmful to fetal rat kidneys.


Non-participation in chlamydia screening in The Netherlands: determinants associated with young people's intention to participate in chlamydia screening.
In The Netherlands, a national chlamydia screening program started in 2008, but the participation was low and the screening was not cost-effective. This study aimed to explore unconscious and conscious associations with chlamydia screening (16-29 year-olds). In addition, we examined whether information presented in chlamydia screening invitation letters had an effect on the evaluation of these determinants compared to a no-letter group. An Internet survey was conducted that included self-report measures of attitude, susceptibility, severity, unrealistic optimism, subjective, moral, and descriptive norm, perceived behavioral control, outcome expectations, barriers, intention, and a response time measure to assess unconscious associations of chlamydia screening with annoyance, threat and reassurance. On the unconscious level, participants (N = 713) who received no information letter associated testing for chlamydia with annoyance and threat, but also with reassurance (all p's < .001). On the self-report measures, participants showed a low intention towards chlamydia screening (M = 1.42, range 1-5). Subjective norm, moral norm, perceived susceptibility and attitude were the most important predictors of the intention to screen (R2 = .56). Participants who rated their susceptibility as high also reported more risky behaviors (p < .001).In the groups that received a letter (N = 735), a weaker unconscious association of chlamydia screening with annoyance was found compared with the no-letter group (p < .001), but no differences were found in reassurance or threat. Furthermore, the letters caused a higher intention (p < .001), but intention remained low (M = 1.74). On a conscious level, giving information caused a more positive attitude, higher susceptibility, a higher subjective and moral norm, and more positive outcome expectations (all p's < .001). Subjective norm, moral norm, susceptibility, and attitude towards chlamydia might be crucial targets to increase chlamydia screening behavior among sexually active young people. This study shows that informational invitation letters increase the intention and the intention-predicting variables. More evidence is needed on whether screening behavior can be increased by the use of an alternative information letter adapted to the specific unconscious and conscious determinants revealed in this study, or that we need other, more interactive behavior change methods.


Dissociation of the proliferation and differentiation stimuli of granulocyte colony-stimulating factor (G-CSF).
One proposed therapeutic application of granulocyte colony-stimulating factor (G-CSF) is in differentiation induction therapy of myelodysplastic states (MDS) or acute myeloid leukemia (AML). G-CSF however has a substantial growth including effect which limits its potential as a differentiation inducing agent. We have therefore made a systematic search for agents which might restrain the proliferative effects of G-CSF whilst retaining the differentiation stimulus. Of all the agents we have tested on human bone marrow progenitor cells: (6-thioguanine, all-trans retinoic acid, vincristine, recombinant human alpha-2b and gamma-interferon) only the latter abolished the stimulation of cell growth and retained, or possibly increased, the differentiation effect of G-CSF. The antiproliferative drugs 6-thioguanine and vincristine both antagonized the neutrophil-granulocyte differentiation inducing action of G-CSF. Retinoic acid and alpha-2b interferon both had weak effects on proliferation and failed to enhance differentiation. These results suggest that it may be possible, by combining G-CSF with a suitable second agent, to utilize its substantial differentiation inducing effect without incurring the potentially hazardous effects of increased leukemic cell growth.


Electron paramagnetic resonance (EPR) studies on hydrogenase-1 (HYD1) purified from a mutant strain (AP6) of Escherichia coli enhanced in HYD1.
Hydrogenase-1 (HYD1), overexpressed by twofold, has been purified to homogeneity and to a high specific activity from a mutant strain (AP6) of Escherichia coli which lacks hydrogenase-2. Plasma emission spectroscopy indicated that 0.93 atom of nickel and 11.4 iron atoms were present in HYD1. EPR studies on the as isolated HYD1 detected a complex 3Fe-4S signal and a Ni(III) species. Reduction with hydrogen gas caused disappearance of both the 3Fe-4S cluster and initial Ni(III) signals. At the same time the EPR signature (small g = 2.19 signal) of the activated hydrogenase appeared. The detection of a 4Fe-4S cluster signal was noted. Reduction of HYD1 with sodium dithionite caused all nickel signals to disappear. The 4Fe-4S complex intensity was slightly increased. The EPR responses in the three oxidation-reduction states are consistent with other known (NiFe)-hydrogenases.


Effectiveness of Super-selective Embolization for Parasagittal Meningiomas and Its Effect on the Level of Inflammatory Factors.
To evaluate the effectiveness of super-selective embolization for parasagittal meningiomas and its effect on the level of inflammatory factors. A total of 48 patients with parasagittal meningiomas diagnosed and treated in our hospital from September 2018 to March 2020 were randomly included and assigned to receive meningioma resection (control group) or meningioma resection plus super-selective embolization (study group), with 24 patients in each group. Outcome measures included clinical indices, tumor resection outcome, inflammatory factor levels, and follow-up results. Patients in the study group had shorter operative time and less intraoperative bleeding volume than those in the control group (P < 0.05). The study group had more patients with Simpson I resection (87.50%) than the control group (62.50%) (P < 0.05). There was no statistically significant difference in the levels of inflammatory factors between the two groups of patients before treatment (P > 0.05). After treatment, the levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α, and hypersensitive C-reactive protein (hs-CRP) in patients in the study group were significantly lower than those of the control group. The results of the three-month follow-up showed that one patient in the study group was reoperated for tumor recurrence and there was no death record, while three patients in the control group were reoperated for tumor recurrence and one patient died. The difference in the recurrence and mortality between the two groups did not come up to the statistical standard (P > 0.05). Super-selective embolization for parasagittal meningiomas contributes to reducing intraoperative bleeding, effectively improving tumor resection and surgical safety, and lowering inflammatory factor levels. Further trials are, however, required before clinical promotion.


Rad52 deficiency decreases development of lung squamous cell carcinomas by enhancing immuno-surveillance.
RAD52 is involved in homologous recombination and DNA repair. This study focuses on lung cancer progression and how the DNA repair gene, Rad52, enables tumor cells to have sufficient genome integrity, i.e., the ability to repair lethal DNA damage, to avoid cell death. In this report, we analyze the phenotypic differences between wild type and Rad52-/- in inhibition of tumor phenotypes including cell growth, viability, cytolysis, and immune profiling. We demonstrated that loss of Rad52 not only increases the death of cells undergoing carcinogen-induced transformation in vivo, but that Rad52 loss also augments in vivo antitumor activity through an enhanced capacity for direct killing of LLC tumor cells by stimulated Rad52-/- NK and CD8+ T cells. We hypothesize that upon DNA damage, wild type cells attempt to repair DNA lesions, but those cells that survive will continue to divide with damage and a high likelihood of progressing to malignancy. Loss of Rad52, however, appears to increase genomic instability beyond a manageable threshold, acceding the damaged cells to death before they are able to become tumor cells. Our results suggest a key role for the complex interplay between the DNA damage response and host immunity in determining risk for Squamous Cell Lung Carcinoma.


Cytotoxic evaluation of biomechanically improved crosslinked ovine collagen on human dermal fibroblasts.
Earlier studies in our laboratory demonstrated that collagen extracted from ovine tendon is biocompatible towards human dermal fibroblast. To be able to use this collagen as a scaffold in skin tissue engineering, a mechanically stronger scaffold is required that can withstand manipulation before transplantation. This study was conducted to improve the mechanical strength of this collagen sponge using chemical crosslinkers, and evaluate their effect on physical, chemical and biocompatible properties. Collagen sponge was crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and glutaraldehyde (GA). Tensile test, FTIR study and mercury porosimetry were used to evaluate mechanical properties, chemical property and porosity, respectively. MTT assay was performed to evaluate the cytotoxic effect of crosslinked collagen sponge on human dermal fibroblasts. The FTIR study confirmed the successful crosslinking of collagen sponge. Crosslinking with EDC and GA significantly increased the mechanical strength of collagen sponge, with GA being more superior. Crosslinking of collagen sponge significantly reduced the porosity and the effect was predominant in GA-crosslinked collagen sponge. The GA-crosslinked collagen showed significantly lower, 60% cell viability towards human dermal fibroblasts compared to that of EDC-crosslinked collagen, 80% and non-crosslinked collagen, 100%. Although the mechanical strength was better when using GA but the more toxic effect on dermal fibroblast makes EDC a more suitable crosslinker for future skin tissue engineering.


Incremental multiple objective genetic algorithms.
This paper presents a new genetic algorithm approach to multiobjective optimization problems--incremental multiple objective genetic algorithms (IMOGA). Different from conventional MOGA methods, it takes each objective into consideration incrementally. The whole evolution is divided into as many phases as the number of objectives, and one more objective is considered in each phase. Each phase is composed of two stages. First, an independent population is evolved to optimize one specific objective. Second, the better-performing individuals from the single-objecive population evolved in the above stage and the multiobjective population evolved in the last phase are joined together by the operation of integration. The resulting population then becomes an initial multiobjective population, to which a multiobjective evolution based on the incremented objective set is applied. The experiment results show that, in most problems, the performance of IMOGA is better than that of three other MOGAs, NSGA-II, SPEA, and PAES. IMOGA can find more solutions during the same time span, and the quality of solutions is better.


[Evaluation of systemic involving of the connective tissue in children with different localisation of isolated abnormal chords of the left ventricle (ACLV)].
Evaluation of the systemic involving of the connective tissue (SICT) under the new Ghent nosology (2010) showed that in children with isolated ACLV born to parents exposed to the Chernobyl disaster, its expression is associated with their location and quantity. The degree of systemic involvement of connective tissue is confirmed by the results of the analysis of features echostructure of isolated ACLV (the presence of thickening and calcification), echomorphometry, assessment of systolic (hypokinetic organization of the central hemodynamics), and the relaxation functions of the heart (initiation of diastolic dysfunction). High level of SICT (score greater than 5) indicates systemic damage to the body and particularly the heart, which requires dynamic monitoring and preventive measures. Found that the diagnostic and monitoring of children with isolated ACLV may be based on registration of systemic involvement of connective tissue with the calculation of points under the new Ghent nosology of 2010.


Prosodic phonological representations early in visual word recognition.
Two experiments examined the nature of the phonological representations used during visual word recognition. We tested whether a minimality constraint (R. Frost, 1998) limits the complexity of early representations to a simple string of phonemes. Alternatively, readers might activate elaborated representations that include prosodic syllable information before lexical access. In a modified lexical decision task (Experiment 1), words were preceded by parafoveal previews that were congruent with a target's initial syllable as well as previews that contained 1 letter more or less than the initial syllable. Lexical decision times were faster in the syllable congruent conditions than in the incongruent conditions. In Experiment 2, we recorded brain electrical potentials (electroencephalograms) during single word reading in a masked priming paradigm. The event-related potential waveform elicited in the syllable congruent condition was more positive 250-350 ms posttarget compared with the waveform elicited in the syllable incongruent condition. In combination, these experiments demonstrate that readers process prosodic syllable information early in visual word recognition in English. They offer further evidence that skilled readers routinely activate elaborated, speechlike phonological representations during silent reading.


Preoperative Peroneal Artery Perforator Mapping Using Indocyanine Green Angiography: A Prospective Clinical Trial.
Perforator imaging is a prerequisite in preoperative planning of the peroneal perforator flap and the fibula skin island. Although reports indicate that indocyanine green angiography assessment method might be advantageous over conventional ultrasound-based techniques (i.e., Doppler and color duplex), in practice, clear evidence is lacking. Thus, a comparative assessment of the utility of indocyanine green angiography and ultrasound-based techniques in the identification of suitable lower leg skin perforators was performed. A prospective clinical cohort study with a series of 12 consecutive patients was conducted to assess indocyanine green angiography, Doppler ultrasound, and color duplex ultrasound techniques for preoperative perforator detection in the lower leg before free fibula flap harvest. Anatomical dissection served as a reference. Parameters measured were perforator spatial distance to the reference (precision), operative time expenditure, and ease of device usage for assessment/outcomes. This study included 12 patients, with a total of 27 perforators. Exhibition of technique sensitivity and positive predictive values were as follows: indocyanine green angiography, 93 percent and 100 percent; Doppler ultrasound, 82 percent and 82 percent; and color duplex ultrasound, 89 percent and 86 percent, respectively. With regard to the indocyanine green angiography technique, the distance to the actual perforator location was significantly shorter, which aided detection and lesser time expenditure during operation. The indocyanine green angiography technique proved to have high precision, sensitivity, positive predictive value, and easy-to-use capabilities because of its exceptional spatial and temporal information, compared to the conventional, ultrasound-based techniques. Therefore, indocyanine green angiography is superior for preoperative perforator imaging of the lateral lower leg. Diagnostic, II.


Optimizing the Prehospital-Hospital Emergency Care Path Application Value in Emergency Treatment of Patients with Cerebral Hemorrhage.
To explore the effect of optimizing the prehospital-hospital emergency nursing path on the rescue speed, rescue effect, and patient prognosis of patients with cerebral hemorrhage. A retrospective study was performed to select 227 patients with acute cerebral hemorrhage who visited our hospital from August 2018 to October 2019, and we compared the optimization of the prehospital-hospital emergency nursing pathway (research group) with traditional prehospital time spent in the rescue and the prognosis of patients in the emergency care pathway (control group) in the hospital. The GOS score, FMA score, and Barthel index were used to compare the prognosis of the two groups of patients. The prehospital-hospital emergency nursing route was optimized compared with the traditional nursing route. The rescue time of patients in each link was shorter (P < 0.05), and the incidence of complications was low (P < 0.05). The scales and scores all suggested that patients had a better prognosis (P < 0.05). Optimizing the prehospital-hospital emergency nursing path can significantly shorten the rescue time, improve the rescue effect, and improve the prognosis of patients.


Sternal fixation for isolated traumatic sternal fractures improves pain and upper extremity range of motion.
Sternal fractures are debilitating due to intractable pain, constant fracture movement and limited range of motion (ROM) of the upper extremities (UE). Traditional treatment comprises mainly of pain control, delaying return to daily activities. Recently, sternal fixation has gained popularity. There is, however, a lack of literature demonstrating efficacy. We report our experience of traumatically fractured sternal fixation. Following IRB approval, a retrospective chart review was completed for all patients undergoing sternal fixation by a single trauma surgeon at our Level I trauma center. Basic demographics were obtained. Primary outcomes included average cumulative pain scores, total cumulative narcotic amounts and total number of pain medication agents utilized prior to and after sternal fixation. Secondary outcome included physical therapy UE ROM before and after surgery. Paired t tests were used for comparison; significance set at p < 0.05. Thirteen patients underwent sternal fixation from 8/2016 to 2/2018. Average age was 54.4 ± 20.8 years; 54% were female. All patients experienced blunt trauma; average injury severity score was 15.8 ± 10.9 and abbreviated chest injury score was 2.5 ± 0.51. Average intensive care unit/hospital length of stay was 2.3/10.2 days. Average pain scores significantly improved by a score of 3.5 postoperatively (preoperative = 7.08 ± 2.3, postoperative = 3.54 ± 2.5; p = 0.001). Total pain medications required by sternal fixation patients significantly decreased by 1 medication postoperatively (preoperative = 4.2 medications, postoperative = 3.2 medications; p = 0.002). Average narcotic requirements significantly decreased by 7.59 morphine milligram milliequivalents (MME) after sternal fixation (preoperative amount = 71.78 MME, postoperative amount = 64.19 MME; p = 0.041). Every patient had limited UE ROM preoperatively; however, all but one patient resumed full UE ROM postoperatively (p < 0.001). There were no postoperative complications. Sternal fixation is a safe and effective procedure resulting in improved pain, decreased narcotic requirements, and faster recovery.


Smooth muscle Ca2+ sensitization causes hypercontractility of middle cerebral arteries in mice bearing the familial hemiplegic migraine type 2 associated mutation.
Familial hemiplegic migraine type 2 (FHM2) is associated with inherited point-mutations in the Na,K-ATPase α2 isoform, including G301R mutation. We hypothesized that this mutation affects specific aspects of vascular function, and thus compared cerebral and systemic arteries from heterozygote mice bearing the G301R mutation (Atp1a2+/-G301R) with wild type (WT). Middle cerebral (MCA) and mesenteric small artery (MSA) function was compared in an isometric myograph. Cerebral blood flow was assessed with Laser speckle analysis. Intracellular Ca2+ and membrane potential were measured simultaneously. Protein expression was semi-quantified by immunohistochemistry. Protein phosphorylation was analysed by Western blot. MSA from Atp1a2+/-G301R and WT showed similar contractile responses. The Atp1a2+/-G301R MCA constricted stronger to U46619, endothelin and potassium compared to WT. This was associated with an increased depolarization, although the Ca2+ change was smaller than in WT. The enhanced constriction of Atp1a2+/-G301R MCA was associated with increased cSrc activation, stronger sensitization to [Ca2+]i and increased MYPT1 phosphorylation. These differences were abolished by cSrc inhibition. Atp1a2+/-G301R mice had reduced resting blood flow through MCA in comparison with WT mice. FHM2-associated mutation leads to elevated contractility of MCA due to sensitization of the contractile machinery to Ca2+, which is mediated via Na,K-ATPase/Src-kinase/MYPT1 signalling.


A Single Transcription Factor (PDD1) Determines Development and Yield of Winter Mushroom (Flammulina velutipes).
Most of the edible mushrooms cannot be cultivated or have low yield under industrial conditions, partially due to the lack of knowledge on how basidioma (fruiting body) development is regulated. From winter mushroom (Flammulina velutipes), one of the most popular industrially cultivated mushrooms, a transcription factor, PDD1, with a high-mobility group (HMG)-box domain was identified based on its increased transcription during basidioma development. pdd1 knockdown by RNA interference affected vegetative growth and dramatically impaired basidioma development. A strain with an 89.9% reduction in the level of pdd1 transcription failed to produce primordia, while overexpression of pdd1 promoted basidioma development. When the transcriptional level of pdd1 was increased to 5 times the base level, the mushroom cultivation time was shortened by 9.8% and the yield was increased by at least 33%. RNA sequencing (RNA-seq) analysis revealed that pdd1 knockdown downregulated 331 genes and upregulated 463 genes. PDD1 positively regulated several genes related to fruiting, including 6 pheromone receptor-encoding genes, 3 jacalin-related lectin-encoding genes, FVFD16, and 2 FVFD16 homolog-encoding genes. PDD1 is a novel transcription factor with regulatory function in basidioma development found in industrially cultivated mushrooms. Since its orthologs are widely present in fungal species of the Basidiomycota phylum, PDD1 might have important application prospects in mushroom breeding.IMPORTANCE Mushrooms are sources of food and medicine and provide abundant nutrients and bioactive compounds. However, most of the edible mushrooms cannot be cultivated commercially due to the limited understanding of basidioma development. From winter mushroom (Flammulina velutipes; also known as Enokitake), one of the most commonly cultivated mushrooms, we identified a novel transcription factor, PDD1, positively regulating basidioma development. PDD1 increases expression during basidioma development. Artificially increasing its expression promoted basidioma formation and dramatically increased mushroom yield, while reducing its expression dramatically impaired its development. In its PDD1 overexpression mutants, mushroom number, height, yield, and biological efficiency were significantly increased. PDD1 regulates the expression of some genes that are important in or related to basidioma development. PDD1 is the first identified transcription factor with defined functions in mushroom development among commercially cultivated mushroom species, and it might be useful in mushroom breeding.


Staying good while playing god--the ethics of breeding farm animals.
Modern genetics has given us some very efficient tools with which to alter the characteristics of animals. To date, farm animal breeders have mainly used these tools to increase productivity. Thus, each new generation of farm animals matures faster, yields more milk, or produces more meat or eggs, than the previous one. Despite these apparent benefits, modern farm animal breeding has had severe negative consequences, including effects on the quality of the animals' lives and biodiversity. The aim of this paper is to discuss the goals and consequences of farm animal breeding within an ethical context. First, a description of what has happened to broilers and dairy cattle as breeders have pursued the goal of ever more efficient production is given. Second, the ethical values that ought to underlie future breeding schemes are discussed. It is suggested that there are in fact two very different ethical approaches: the 'quality of life-based' approach and the 'preservationist' approach. A view combining elements from both approaches is advanced. Finally, an example is given of how it is possible, in practice, to pursue an ethically defensible breeding goal without compromising production efficiency.


A large-scale genetic screen in Arabidopsis to identify genes involved in pollen exine production.
Exine, the outer plant pollen wall, has elaborate species-specific patterns, provides a protective barrier for male gametophytes, and serves as a mediator of strong and species-specific pollen-stigma adhesion. Exine is made of sporopollenin, a material remarkable for its strength, elasticity, and chemical durability. The chemical nature of sporopollenin, as well as the developmental mechanisms that govern its assembly into diverse patterns in different species, are poorly understood. Here, we describe a simple yet effective genetic screen in Arabidopsis (Arabidopsis thaliana) that was undertaken to advance our understanding of sporopollenin synthesis and exine assembly. This screen led to the recovery of mutants with a variety of defects in exine structure, including multiple mutants with novel phenotypes. Fifty-six mutants were selected for further characterization and are reported here. In 14 cases, we have mapped defects to specific genes, including four with previously demonstrated or suggested roles in exine development (MALE STERILITY2, CYP703A2, ANTHER-SPECIFIC PROTEIN6, TETRAKETIDE α-PYRONE REDUCTASE/DIHYDROFLAVONOL-4-REDUCTASE-LIKE1), and a number of genes that have not been implicated in exine production prior to this screen (among them, fatty acid ω-hydroxylase CYP704B1, putative glycosyl transferases At1g27600 and At1g33430, 4-coumarate-coenzyme A ligase 4CL3, polygalacturonase QUARTET3, novel gene At5g58100, and nucleotide-sugar transporter At5g65000). Our study illustrates that morphological screens of pollen can be extremely fruitful in identifying previously unknown exine genes and lays the foundation for biochemical, developmental, and evolutionary studies of exine production.


IC14, an anti-CD14 antibody, inhibits endotoxin-mediated symptoms and inflammatory responses in humans.
CD14 is a receptor for cell wall components of Gram-negative and Gram-positive bacteria that has been implicated in the initiation of the inflammatory response to sepsis. To determine the role of CD14 in LPS-induced effects in humans, 16 healthy subjects received an i.v. injection of LPS (4 ng/kg) preceded (-2 h) by i.v. IC14, a recombinant chimeric mAb against human CD14, at a dose of 1 mg/kg over 1 h, or placebo. In subjects receiving IC14, saturation of CD14 on circulating monocytes and granulocytes was >90% at the time of LPS injection. IC14 attenuated LPS-induced clinical symptoms and strongly inhibited LPS-induced proinflammatory cytokine release, while only delaying the release of the anti-inflammatory cytokines soluble TNF receptor type I and IL-1 receptor antagonist. IC14 also inhibited leukocyte activation, but more modestly reduced endothelial cell activation and the acute phase protein response. The capacity of circulating monocytes and granulocytes to phagocytose Escherichia coli was only marginally reduced after infusion of IC14. These data provide the first proof of principle that blockade of CD14 is associated with reduced LPS responsiveness in humans in vivo.


Breast cancer mortality among American Indian and Alaska Native women, 1990-2009.
We compared breast cancer death rates and mortality trends among American Indian/Alaska Native (AI/AN) and White women using data for which racial misclassification was minimized. We used breast cancer deaths and cases linked to Indian Health Service (IHS) data to calculate age-adjusted rates and 95% confidence intervals (CIs) by IHS-designated regions from 1990 to 2009 for AI/AN and White women; Hispanics were excluded. Mortality-to-incidence ratios (MIR) were calculated for 1999 to 2009 as a proxy for prognosis after diagnosis. Overall, the breast cancer death rate was lower in AI/AN women (21.6 per 100,000) than in White women (26.5). However, rates in AI/ANs were higher than rates in Whites for ages 40 to 49 years in the Alaska region, and ages 65 years and older in the Southern Plains region. White death rates significantly decreased (annual percent change [APC] = -2.1; 95% CI = -2.3, -2.0), but regional and overall AI/AN rates were unchanged (APC = 0.9; 95% CI = 0.1, 1.7). AI/AN women had higher MIRs than White women. There has been no improvement in death rates among AI/AN women. Targeted screening and timely, high-quality treatment are needed to reduce mortality from breast cancer in AI/AN women.


Advocating for equitable management of hereditary cancer syndromes.
The potential for preventive medicine to lead to more equitable health outcomes exists with the inclusion of genetic testing in medicine. Because of the medical implications of genetic testing for hereditary cancer syndromes and the financial cost attached to recommended management, ensuring equitable access to cancer screening and prevention must be made a priority. For patients with Hereditary Breast and Ovarian Cancer (HBOC) syndrome, the benefits of early detection and prevention are clear, significant, and create the opportunity to provide more equitable, personalized, preventive healthcare. Thus, for genetics providers who offer testing access for their patients, it is important to reflect on the ethical responsibility of advocating for access to appropriate management. Cancer genetic counselors can advocate for health equity by providing thorough pre-test genetic counseling, collaborating with other disciplines to coordinate care, lobbying state, and national representatives to pass legislation promoting health equity, and developing a management clinic that helps to ensure follow-up. Equitable access to and benefit from hereditary cancer risk management must be achieved in the pursuit of personalized preventive medicine.


Photocatalytic oxidation of triclosan.
In the spring of 2003, there was an outbreak of the severe respiratory syndrome (SARS) in Hong Kong. Health concerns have thus triggered an increased and predominant use of various types of household cleansing agents such as triclosan (5-chloro-2-(2,4-dichlorophenoxy)phenol). However, it has been reported recently that triclosan could be photochemically converted to toxic 2,8-dichlorodibenzo-p-dioxin (2,8-Cl(2)DD) in the environment. It is therefore necessary to develop environmentally friendly methods for the treatment of triclosan. To this end, photocatalytic degradation of triclosan in aqueous solution was conducted using TiO(2) (Degussa P25) under irradiation of UV light (lambda < 365 nm). It was found that triclosan could be degraded by this approach. Hydrogen peroxide was added to enhance the degradation process, and the optimal initial hydrogen peroxide concentration for triclosan degradation was 0.005% (w/v). Product identification indicated that triclosan oxidation occurred at its phenol moiety and yielded quinone and hydroquinone intermediates. The formation of a dichlorophenol intermediate in triclosan degradation suggested bond-breaking of the ether linkage occurred during the process. Moreover, no chlorinated dibenzo-p-dioxin congener was detected. These findings confirm that the photocatalytic degradation of triclosan is an environmentally friendly process.


Transient diabetes insipidus and preeclampsia.
A woman with triplet gestation had polydipsia and polyuria during pregnancy. She subsequently had preeclampsia for which she was delivered. The presumed diabetes insipidus was transient, resolving by postpartum day 5. The association of preeclampsia and diabetes insipidus may be confusing to the practitioner. Careful attention to fluid status and consideration of this abnormality will lead to the best outcome when hypertension is accompanied by polydipsia and polyuria.


The evaluation and choice of laboratory equipment and reagents.
Many new types of laboratory equipment are appearing on the market, as well as a large number of reagent sets or kits. The importance of adequate evaluation of the reliability of such new products cannot be overemphasized.Commercially prepared reagent sets or kits fall into two categories. In the one, a set of reagents is offered for use in performance of a particular conventional procedure; this type of prepared reagent should present no problem, provided that the reagents are stable. In the other category, a reagent kit or set is offered which has been designed to simplify the performance of a laboratory analysis, often by combining reagents and reducing the number of steps and the time required. Such a kit should be evaluated carefully in order to determine its efficiency and limitations. In general it is difficult to simplify greatly a laboratory procedure without sacrificing its accuracy.


Identifying chemical functionalization on individual carbon nanotubes and graphene by local vibrational fingerprinting.
Chemical functionalization of carbon nanotubes (CNTs) and graphene allows for fine-tuning their physical and chemical properties to realize fascinating new fundamental phenomena as well as exotic applications. A primary challenge in such endeavors is the need to identify the chemical nature of attached functionalities at a single-nano-object level in a spatially resolved manner. Here we report the vibrational fingerprinting of functional groups that are attached to individual CNTs and graphene flakes. In order to achieve this, we decorate noncovalently functionalized CNTs and graphene with nanoparticles, which leads to the appearance of Raman peaks that can be correlated with the vibrational modes characteristic of the functional groups with diffraction-limited spatial resolution. The presented strategy is generic enough to be extended to other chemical modification routes on a range of nanostructures and hence will allow for rapid characterization of chemical modification of individual (semi)conducting nanostructures.


Assessing health plan quality.
Health plan accreditation and report cards represent the beginning of a rapidly evolving information flow that impacts all stakeholders within the health care system. By understanding what information is available--and its limitations--stakeholders can work together to promote improved health plan quality.


The concept of the sexual reproduction cycle and its evolutionary significance.
The concept of a "sexual reproduction cycle (SRC)" was first proposed by Bai and Xu (2013) to describe the integration of meiosis, sex differentiation, and fertilization. This review discusses the evolutionary and scientific implications of considering these three events as part of a single process. Viewed in this way, the SRC is revealed to be a mechanism for efficiently increasing genetic variation, facilitating adaptation to environmental challenges. It also becomes clear that, in terms of cell proliferation, it is appropriate to contrast mitosis with the entire SRC, rather than with meiosis alone. Evolutionarily, it appears that the SRC was first established in unicellular eukaryotes and that all multicellular organisms evolved within that framework. This concept provides a new perspective into how sexual reproduction evolved, how generations should be defined, and how developmental processes of various multicellular organisms should properly be compared.


Microwave-assisted high-speed chemistry: a new technique in drug discovery.
In both lead identification and lead optimization processes there is an acute need for new organic small molecules. Traditional methods of organic synthesis are orders of magnitude too slow to satisfy the demand for these compounds. The fields of combinatorial and automated medicinal chemistry have been developed to meet the increasing requirement of new compounds for drug discovery; within these fields, speed is of the essence. The efficiency of microwave flash-heating chemistry in dramatically reducing reaction times (reduced from days and hours to minutes and seconds) has recently been proven in several different fields of organic chemistry. We believe that the time saved by using focused microwaves is potentially important in traditional organic synthesis but could be of even greater importance in high-speed combinatorial and medicinal chemistry.


Dependency of cerebral blood flow on mean arterial pressure in patients with acute bacterial meningitis.
Patients with acute bacterial meningitis are often treated with sympathomimetics to maintain an adequate mean arterial pressure (MAP). We studied the influence of such therapy on cerebral blood flow (CBF). Prospective physiologic trial. The Department of Infectious Diseases, Copenhagen University Hospital, Denmark. Sixteen adult patients with acute bacterial meningitis. Infusion of norepinephrine to increase MAP. During a rise in MAP induced by norepinephrine infusion, we measured relative changes in CBF by transcranial Doppler ultrasonography of the middle cerebral artery, recording mean flow velocity (Vmean), and by the arterial to jugular oxygen saturation difference. In 10 out of 16 patients, serial measurements were performed until recovery or death. Individual autoregulation curves were analyzed by a computer program. Autoregulation was classified as impaired if Vmean increased by >10% per 30 mm Hg increase in MAP and if no lower limit of autoregulation was identified by the computer program; otherwise, autoregulation was classified as preserved. Initially, Vmean increased from a median value of 46 cm/sec (range, 30-87 cm/sec) to 63 cm/sec (33-105 cm/sec) (p < .0001), and arterial to jugular oxygen saturation difference decreased from 0.28 (0.16-0.51) to 0.21 (0.08-0.39) (p < .001) when MAP was raised from 69 mm Hg (55-102 mm Hg) to 110 mm Hg (93-129 mm Hg). CBF autoregulation was restored in eight of ten patients undergoing serial examination after 7 (range, 2-10) days. Six of these patients had an uncomplicated course, one had a protracted recovery, and one died. Autoregulation was not restored in two patients; one died and one had a protracted recovery. In patients in the early phase of acute bacterial meningitis, CBF autoregulation is impaired. With recovery from meningitis, the cerebral vasculature regains the ability to maintain cerebral perfusion at a constant level despite variations in MAP.


Skeletal muscle stem cells in comfort and stress.
Investigations on developmental and regenerative myogenesis have led to major advances in decrypting stem cell properties and potential, as well as their interactions within the evolving niche. As a consequence, regenerative myogenesis has provided a forum to investigate intrinsic regulators of stem cell properties as well as extrinsic factors, including stromal cells, during normal growth and following injury and disease. Here we review some of the latest advances in the field that have exposed fundamental processes including regulation of stress following trauma and ageing, senescence, DNA damage control and modes of symmetric and asymmetric cell divisions. Recent studies have begun to explore the nature of the niche that is distinct in different muscle groups, and that is altered from prenatal to postnatal stages, and during ageing. We also discuss heterogeneities among muscle stem cells and how distinct properties within the quiescent and proliferating cell states might impact on homoeostasis and regeneration. Interestingly, cellular quiescence, which was thought to be a passive cell state, is regulated by multiple mechanisms, many of which are deregulated in various contexts including ageing. These and other factors including metabolic activity and genetic background can impact on the efficiency of muscle regeneration.


Structural coloring in large scale core-shell nanowires.
We demonstrated two complementary size-dependent structural coloring mechanisms, interference and scattering, in indefinitely long core-shell nanowire arrays. The unusual nanostructures are comprised of an amorphous semiconducting core and a polymer shell layer with disparate refractive indices but with similar thermomechanical properties. Core-shell nanowires are mass produced from a macroscopic semiconductor rod by using a new top-to-bottom fabrication approach based on thermal size reduction. Nanostructures with diameters from 30 to 200 nm result in coloration that spans the whole visible spectrum via resonant Mie scattering. Nanoshell coloration based on thin film interference is proposed as a structural coloration mechanism which becomes dominant for nanowires having 700-1200 nm diameter. Controlled color generation in any part of visible and infrared spectral regions can be achieved by the simple scaling down procedure. Spectral color generation in mass-produced uniform core-shell nanowire arrays paves the way for applications such as spectral authentication at nanoscale, light-scattering ingredients in paints and cosmetics, large-area devices, and infrared shielding.


Auxin dynamics: the dazzling complexity of a small molecule's message.
The phytohormone auxin is a potent regulator of plant development. Since its discovery in the beginning of the twentieth century many aspects of auxin biology have been extensively studied, ranging from biosynthesis and metabolism to the elucidation of molecular components of downstream signaling. With the identification of the F-box protein TIR1 as an auxin receptor a major breakthrough in understanding auxin signaling has been achieved and recent modeling approaches have shed light on the putative mechanisms underlying the establishment of auxin gradients and maxima essential for many auxin-regulated processes. Here, we review these and other recent advances in unraveling the entanglement of biosynthesis, polar transport and cellular signaling events that allow small auxinic molecules to facilitate their complex regulatory action.


Coordinated Management of COVID-19 Response: Lessons From Whole-of-Society and Whole-of-Health Strategies in Wuhan, China.
Background: The outbreak of novel coronavirus disease 2019 (COVID-19) has been challenging globally following the scarcity of medical resources after a surge in demand. As the pandemic continues, the question remains on how to accomplish more with the existing resources and improve the efficiency of existing health care delivery systems worldwide. In this study, we reviewed the experience from Wuhan - the first city to experience a COVID-19 outbreak - that has presently shown evidence for efficient and effective local control of the epidemic. Material and Methods: We performed a retrospective qualitative study based on the document analysis of COVID-19-related materials and interviews with first-line people in Wuhan. Results: We extracted two themes (the evolution of Wuhan's prevention and control strategies on COVID-19 and corresponding effectiveness) and four sub-themes (routine prevention and control period, exploration period of targeted prevention and control strategies, mature period of prevention and control strategies, and recovery period). How Wuhan combatted COVID-19 through multi-tiered and multi-sectoral collaboration, overcoming its fragmented, hospital-centered, and treatment-dominated healthcare system, was illustrated and summarized. Conclusion: Four lessons for COVID-19 prevention and control were summarized: (a) Engage the communities and primary care not only in supporting but also in screening and controlling, and retain community and primary care as among the first line of COVID-19 defense; (b) Extend and stratify the existing health care delivery system; (c) Integrate person-centered integrated care into the whole coordination; and (d) Delink the revenue relationship between doctors and patients and safeguard the free-will of physicians when treating patients.


N-(4-hydroxyphenyl)-retinamide increases lecithin:retinol acyltransferase activity in rat liver.
N-(4-Hydroxyphenyl)-retinamide (4-HPR; Fenretinide) is a synthetic retinoid which is undergoing investigation as a cancer chemopreventive agent. However, 4-HPR alters vitamin A kinetics and reduces the concentration of plasma retinol. We have conducted studies to examine the effects of 4-HPR on the activity of the enzyme lecithin:retinol acyltransferase (LRAT). This enzyme is implicated in the absorption and storage of vitamin A and is regulated, in liver, by vitamin A nutritional status. To determine whether 4-HPR, like retinoic acid, is able to induce liver LRAT activity, vitamin A-deficient rats having negligible liver LRAT activity were treated with single doses of 4-HPR (0.02-2.5 mg) and liver homogenates were assayed for LRAT activity using 3H-retinol bound to the cellular-retinol binding protein, CRBP, as substrate. Treatment with 4-HPR resulted in a dose- and time-dependent increase in liver LRAT activity which reached a maximum at 24 h. The activity of LRAT assayed in vitro and of hepatic 3H-retinyl ester content determined after an in vivo pulse of 3H-retinol were highly correlated (r = 0.802, P < 0.0002). When vitamin A-sufficient rats were fed a 4-HPR-supplemented diet for 30 d, LRAT activity differed significantly from control values in the liver (P < 0.0001) but not the small intestines. Changes in hepatic retinol metabolism which favor the esterification of vitamin A may be related to the mechanism by which 4-HPR alters vitamin A kinetics in vivo.


Plasma oxidized polyhydroxymethylsiloxane--a new smooth surface for supported lipid bilayer formation.
A novel substrate for preparation of supported lipid bilayers (SLBs), smooth at the subnanometer scale and of variable thickness from ten to several hundred nanometers, was developed by surface oxidation of spin-coated poly(hydroxymethylsiloxane) (PHMS) films. The deposited polymeric thin films were modified by a combination of oxygen plasma and thermal treatment (PHMS(ox)), in order to convert the outermost surface layer of the polymer film to a stable SiO(2) film, suitable for SLB formation. The hydrophilic, SiO(2)-like surfaces were characterized by XPS, wetting angle, ellipsometry, and AFM. Lipid bilayers were formed on this surface using the well-known vesicle adsorption-rupture-fusion process, usually performed on glass or vapor-deposited SiO(2). Reproducible formation of homogeneous SLBs of different compositions (POPC, DOEPC, and POPC/DOPS) was demonstrated on the new SiO(2) surface by quartz crystal microbalance with dissipation (QCM-D), surface plasmon resonance (SPR), and optical reflectometry measurements. The SLB formation kinetics on the PHMS(ox)-coated sensors showed very similar characteristics, for all investigated PHMS thicknesses, as on reference sensors coated with vapor-deposited SiO(2). The good adhesive properties of the PHMS to gold allows for the preparation of thin PHMS(ox) layers compatible with SPR. The much smaller roughness at the nanometer scale of the PHMS(ox) surfaces, compared to standard vapor-deposited SiO(2)-coated sensors, makes them advantageous for AFM and optical experiments and promising for patterning. To benefit optical experiments with the PHMS(ox) surfaces, it was also investigated how the PHMS film thickness influences the SPR and reflectometry responses upon SLB formation.


Addition of S-1 to the epidermal growth factor receptor inhibitor gefitinib overcomes gefitinib resistance in non-small cell lung cancer cell lines with MET amplification.
Most non-small cell lung cancer (NSCLC) tumors with activating mutations in the epidermal growth factor receptor (EGFR) are initially responsive to EGFR tyrosine kinase inhibitors (EGFR-TKI) such as gefitinib and erlotinib, but they almost invariably develop resistance to these drugs. A secondary mutation in EGFR (T790M) and amplification of the MET proto-oncogene have been identified as mechanisms of such acquired resistance to EGFR-TKIs. We have now investigated whether addition of the oral fluoropyrimidine derivative S-1 to gefitinib might overcome gefitinib resistance in NSCLC cell lines. The effects of gefitinib on EGFR signaling and on the expression both of thymidylate synthase and of the transcription factor E2F-1 in gefitinib-resistant NSCLC cells were examined by immunoblot analysis. The effects of S-1 (or 5-fluorouracil) and gefitinib on the growth of NSCLC cells were examined in vitro as well as in nude mice. Gefitinib induced down-regulation of thymidylate synthase and E2F-1 in gefitinib-resistant NSCLC cells with MET amplification but not in those harboring the T790M mutation of EGFR. The combination of 5-fluorouracil and gefitinib synergistically inhibited the proliferation of cells with MET amplification, but not that of those with the T790M mutation of EGFR, in vitro. Similarly, the combination of S-1 and gefitinib synergistically inhibited the growth only of NSCLC xenografts with MET amplification. Our results suggest that the addition of S-1 to EGFR-TKIs is a promising strategy to overcome EGFR-TKI resistance in NSCLC with MET amplification.


Fathers' views and experiences of their own mental health during pregnancy and the first postnatal year: a qualitative interview study of men participating in the UK Born and Bred in Yorkshire (BaBY) cohort.
The prevalence of fathers' depression and anxiety in the perinatal period (i.e. from conception to 1 year after birth) is approximately 5-10%, and 5-15%, respectively; their children face increased risk of adverse emotional and behavioural outcomes, independent of maternal mental health. Critically, fathers can be protective against the development of maternal perinatal mental health problems and their effects on child outcomes. Preventing and treating paternal mental health problems and promoting paternal psychological wellbeing may therefore benefit the family as a whole. This study examined fathers' views and direct experiences of paternal perinatal mental health. Men in the Born and Bred in Yorkshire (BaBY) epidemiological prospective cohort who met eligibility criteria (baby born <12 months; completed Mental Health and Wellbeing [MHWB] questionnaires) were invited to participate. Those expressing interest (n = 42) were purposively sampled to ensure diversity of MHWB scores. In-depth interviews were conducted at 5-10 months postpartum with 19 men aged 25-44 years. The majority were first-time fathers and UK born; all lived with their partner. Data were analysed using thematic analysis. Four themes were identified: 'legitimacy of paternal stress and entitlement to health professionals' support', 'protecting the partnership', 'navigating fatherhood', and, 'diversity of men's support networks'. Men largely described their 'stress' with reference to exhaustion, poor concentration and irritability. Despite feeling excluded by maternity services, fathers questioned their entitlement to support, noting that services are pressured and 'should' be focused on mothers. Men emphasised the need to support their partner and protect their partnership as central to the successfully navigation of fatherhood; they used existing support networks where available but noted the paucity of tailored support for fathers. Fathers experience psychological distress in the perinatal period but question the legitimacy of their experiences. Men may thus be reluctant to express their support needs or seek help amid concerns that to do so would detract from their partner's needs. Resources are needed that are tailored to men, framed around fatherhood, rather than mental health or mental illness, and align men's self-care with their role as supporter and protector. Further research is needed to inform how best to identify and manage both parents' mental health needs and promote their psychological wellbeing, in the context of achievable models of service delivery.


Antimicrobial susceptibility testing of cystic fibrosis and non-cystic fibrosis clinical isolates of Pseudomonas aeruginosa: a comparison of three methods.
Pseudomonas aeruginosa is an important pathogen in humans, particularly in the context of nosocomial infection and infections of the cystic fibrosis (CF) lung. In order to provide clinicians with information about the likely effectiveness of specific antimicrobial treatment for P. aeruginosa infections, clinical laboratories employ in vitro antimicrobial susceptibility testing. Two commonly employed methods are the CLSI disc-diffusion and Etest methods. The purpose of this study is to compare the accuracy of susceptibility results generated by these two methods against agar dilution as the reference method. Susceptible or nonsusceptible (resistant and intermediate) results of the Etest and CLSI disc-diffusion methods are compared with CLSI agar dilution results for a large cohort of clinical cystic fibrosis (n = 71) and non-cystic fibrosis (n = 83) isolates using CLSI interpretive criteria. An unacceptable number of major and very major errors were observed for various antimicrobials tested against both CF and non-CF isolates when using the Etest and CLSI disc-diffusion methods. The potential for error in standard laboratory antimicrobial susceptibility testing should be considered by clinicians when being guided by the results of such tests in the prescription of antimicrobial agents for P. aeruginosa infection.


A model of Periventricular Leukomalacia (PVL) in neonate mice with histopathological and neurodevelopmental outcomes mimicking human PVL in neonates.
Periventricular leukomalacia (PVL), a brain injury affecting premature infants is commonly associated with cerebral palsy. PVL results from hypoxia-ischemia (HI) with or without infection and is characterized by white matter necrotic lesions, hypomyelination, microglial activation, astrogliosis, and neuronal death. It is important to study a PVL mouse model that mimics human PVL in symptomatology, anatomic and molecular basis. In our neonate mice model, bilateral carotid arteries were temporary ligated at P5 followed by hypoxic exposure (FiO2 of 8% for 20 min.). At P5 in mice, the white matter is more vulnerable to HI injury than the grey matter. In our PVL model, mice suffer from significant hind limb paresis, incoordination and feeding difficulties. Histologically they present with ventriculomegally, white matter loss, microglial activation and neuronal apoptosis. HI injury increases proinflammtory cytokines, activates NF-kB, activates microglia and causes nitrative stress. All these inflammatory mediators lead to oligodendroglial injury and white matter loss. Neurobehavioral analysis in the PVL mice model at P60 showed that the HI group had a significant decrease in hind limb strength, worsening rotarod testing and worsening performance in the open field test. This new PVL model has great advantages far beyond just mimicking human PVL in clinical features and histopathology. Long term survival, the development of cerebral palsy and the ability of using this model in transgenic animals will increase our understanding of the mechanistic pathways underlying PVL and defining specific targets for the development of suitable therapeutics.


[Twilight consciousness disorder caused by overdose of indomethacin].
A case pertaining to legal psychiatric practice is described: the development of dusk clouding of consciousness in a patient with chronic alcoholism after the intake of indomethacin in high doses, foregoing insomnia and episodic alcohol abuse. Possible casuistry of the case forms no basis for ignoring the fact that in legal psychiatry, analogous episodes because of the intake of the high doses of certain drugs including indomethacin may be repeated.


MBD-Net: Multi-Branch Dilated Convolutional Network With Cyst Discriminator for Renal Multi-Structure Segmentation.
In surgery-based renal cancer treatment, one of the most essential tasks is the three-dimensional (3D) kidney parsing on computed tomography angiography (CTA) images. In this paper, we propose an end-to-end convolutional neural network-based framework to segment multiple renal structures, including kidneys, kidney tumors, arteries, and veins from arterial-phase CT images. Our method consists of two collaborative modules: First, we propose an encoding-decoding network, named Multi-Branch Dilated Convolutional Network (MBD-Net), consisting of residual, hybrid dilated convolutional, and reduced-dimensional convolutional structures, which improves the feature extraction ability with relatively fewer network parameters. Given that renal tumors and cysts have confusing geometric structures, we also design the Cyst Discriminator to effectively distinguish tumors from cysts without labeling information via gray-scale curves and radiographic features. We have quantitatively evaluated our approach on a publicly available dataset from MICCAI 2022 Kidney Parsing for Renal Cancer Treatment Challenge (KiPA2022), with mean Dice similarity coefficient (DSC) as 96.18%, 90.99%, 88.66% and 80.35% for the kidneys, kidney tumors, arteries, and veins respectively, winning the stable and top performance in the challenge.Clinical relevance-The proposed CNN-Based framework can automatically segment 3D kidneys, renal tumors, arteries, and veins for kidney parsing techniques, benefiting surgery-based renal cancer treatment.


Suicide and schizophrenia: a nationwide psychological autopsy study on age- and sex-specific clinical characteristics of 92 suicide victims with schizophrenia.
The authors examined the clinical characteristics of suicide victims with schizophrenia in the general population of Finland. As part of the nationwide National Suicide Prevention Project in Finland, all suicides over a 12-month period of persons with DSM-III-R schizophrenia were investigated by using the psychological autopsy method. Clinical characteristics and their variation with age, sex, and illness duration were examined. Among all suicide victims, 7% (N = 92) were identified as having suffered schizophrenia. Suicides occurred throughout the course of schizophrenia. Both active illness (78%) and depressive symptoms (64%) were highly prevalent immediately before suicide, and a history of suicide attempts (71%) was also common. Women were more likely than men to have committed suicide during an acute exacerbation of the illness. Marked variation in depressive symptoms, alcoholism, and suicide methods was found among sexes and age groups. Alcoholism was most common among middle-aged men (45%), whereas middle-aged women had a high rate of depressive symptoms (88%). Younger male subjects most often used violent suicide methods. Suicide may occur at any point during the course of schizophrenia. The results indicate clinically important variation in depression, alcoholism, and suicide methods among suicide victims with schizophrenia. This suggestion of age- and sex-specific risk factors for suicide in schizophrenia needs further investigation.


Effects of brain-derived neurotrophic factor and noise on transitions in temporal lobe epilepsy in a hippocampal network.
Brain-derived neurotrophic factor (BDNF) has recently been implicated in the modulation of receptor activation leading to dynamic state transitions in temporal lobe epilepsy (TLE). In addition, the crucial role of neuronal noise in these transitions has been studied in electrophysiological experiments. However, the precise role of these factors during seizure generation in TLE is not known. Building on a previously proposed model of an epileptogenic hippocampal network, we included the actions of BDNF-regulated receptors and intrinsic noise. We found that the effects of both BDNF and noise can increase the activation of N-methyl-D-aspartate receptors leading to excessive C a 2 + flux, which induces abnormal fast spiking and bursting. Our results indicate that the combined effects have a strong influence on the seizure-generating network, resulting in higher firing frequency and amplitude. As correlations between firing increase, the synchronization of the entire network increases, a marker of the ictogenic transitions from normal to seizures-like dynamics. Our work on the effects of BDNF dynamics in a noisy environment might lead to an improved model-based understanding of the pathological mechanisms in TLE.


P38 MAP kinase is involved in oleuropein-induced apoptosis in A549 cells by a mitochondrial apoptotic cascade.
Lung cancer is one of malignant tumors that cause great threats to human health, which causes the fastest growing morbidity and mortality. Oleuropein as natural production exerts anticancer effects in several cancer cells. In the study, we investigated apoptotic effect of oleuropein on A549 cells and the underlying mechanisms. Oleuropein markedly decreased cell viability in A549 cells by resulting in G2/M phase arrest, but failed to decreased cell viability in BEAS-2B cells significantly. Apoptosis by oleuropein was confirmed by apoptotic morphology, accumulation in a sub-G1 peak, nucleus fragmentation and cleavage of PARP. Dose-dependent elevation in p-p38MAPK and p-ATF-2 was observed whereas apparent changes could not be observed in p-JNK and p-c-Jun, showing activation of p38MAPK but not JNK. Interestingly, ERK1/2 appeared to be constant while p-ERK1/2 was reduced dose-dependently. Oleuropein caused decrease in mitochondrial membrane potential, increase in Bax/Bcl- 2 ratio, release of mitochondrial cytochrome c and activation of caspase-9 and caspase-3, implying that mitochondrial apoptotic pathway was activated. Additionally, oleuropein-induced apoptosis was dramatically attenuated by Z-VAD-FMK (caspase inhibitor). The p38MAPK inhibitor prevented production of apoptotic bodies and reduced expressions of cleaved-PARP, p-P38, p-ATF-2 and release of cytochrome c. Taken together, these results demonstrated p38MAPK signaling pathway mediated oleuropein-induced apoptosis via mitochondrial apoptotic cascade in A549 cells. Oleuropein has the potential to be a therapeutic drug for lung cancer treatment.


Cardiac Irradiation Predicts Activity Decline in Patients Receiving Concurrent Chemoradiation for Locally Advanced Lung Cancer.
Many patients with lung cancer are inactive due to their disease and underlying comorbidities, and activity levels can decline further during cancer therapy. Here we explore dosimetric predictors of activity decline in a cohort of patients who underwent continuous activity monitoring during definitive concurrent chemoradiotherapy (CRT) for locally advanced lung cancer. We identified patients who participated in prospective clinical trials involving the use of a commercial fitness tracker throughout the course of CRT. For each patient, we applied linear regression to log-transformed daily step counts to compute the weekly rate of activity change from 1 week before radiation therapy (RT) initiation to 2 weeks after RT completion. Clinical and dosimetric factors were tested as predictors of activity change using linear regressions. Forty-six patient met the eligibility criteria. Median age was 66 years (range, 38-90). Pretreatment Eastern Cooperative Oncology Group performance status was 0, 1, and 2 for 17%, 70%, and 13%, respectively. Mean lung dose ranged from 5.0 to 23.5 Gy, mean esophagus dose from 1.1 to 39.6 Gy, and mean heart dose from 0.6 to 31.5 Gy. Median daily step count average was 5861 (interquartile range, 3540-8282) before RT and 3422 (interquartile range, 2364-5395) 2 weeks after RT completion. Rate of activity change was not significantly associated with age, performance status, or mean RT dose received by lungs or esophagus. In multivariate analysis, mean heart dose was significantly associated with rate of activity decline, with a 3.1% reduction in step count per week for every 10 Gy increase in mean heart dose (95% confidence interval: 0.5-5.7, P = .023). Extent of cardiac irradiation is associated with the rate of physical activity decline during CRT for lung cancer. Our novel finding contributes to the growing body of evidence that adverse effects of cardiac irradiation may be manifested at early time points.


Mapping the Distribution of Anthrax in Mainland China, 2005-2013.
Anthrax, a global re-emerging zoonotic disease in recent years is enzootic in mainland China. Despite its significance to the public health, spatiotemporal distributions of the disease in human and livestock and its potential driving factors remain poorly understood. Using the national surveillance data of human and livestock anthrax from 2005 to 2013, we conducted a retrospective epidemiological study and risk assessment of anthrax in mainland China. The potential determinants for the temporal and spatial distributions of human anthrax were also explored. We found that the majority of human anthrax cases were located in six provinces in western and northeastern China, and five clustering areas with higher incidences were identified. The disease mostly peaked in July or August, and males aged 30-49 years had higher incidence than other subgroups. Monthly incidence of human anthrax was positively correlated with monthly average temperature, relative humidity and monthly accumulative rainfall with lags of 0-2 months. A boosted regression trees (BRT) model at the county level reveals that densities of cattle, sheep and human, coverage of meadow, coverage of typical grassland, elevation, coverage of topsoil with pH > 6.1, concentration of organic carbon in topsoil, and the meteorological factors have contributed substantially to the spatial distribution of the disease. The model-predicted probability of occurrence of human cases in mainland China was mapped at the county level. Anthrax in China was characterized by significant seasonality and spatial clustering. The spatial distribution of human anthrax was largely driven by livestock husbandry, human density, land cover, elevation, topsoil features and climate. Enhanced surveillance and intervention for livestock and human anthrax in the high-risk regions, particularly on the Qinghai-Tibetan Plateau, is the key to the prevention of human infections.


The nature of the sulphur atom liberated from xanthine oxidase by cyanide. Evidence from e.p.r. spectroscopy after 35S substitution.
Active xanthine oxidase was labelled specifically with 33S in the cyanide-labile site of the molybdenum centre. The Very Rapid molybdenum (V) e.p.r. signal, generated from this, shows strong coupling of 33S to molybdenum, providing unambiguous evidence that, at least in the signal-giving species, this sulphur atom is a ligand of molybdenum. The structure of the signal-giving species is discussed.


Correct microarray analysis approaches in 'Hsa-circRNA11783-2 in peripheral blood is correlated with coronary artery disease and type 2 diabetes mellitus'.
We read with great interest the article by Li et al. The results are illuminating for understanding the relationship among Hsa-circRNA11783-2, coronary artery disease (CAD) and type 2 diabetes mellitus (T2DM). However, from our perspective, the bioinformatics analyses need further context as the statistics for differential fold changes in expression data are not explained fully. The authors seem to use unadjusted p values for detecting differentially expressed circular RNA (differentially expressed genes (DEGs)) between the control, CAD and T2DM group. Due to the high false positives caused by a large number of probes and multiple comparisons, it seems essential to analyse microarray data properly to reach a reliable result by a statistical method. Only selecting circular RNA with greater than two fold change with unadjusted p values < 0.05 in expression is not reliable and suitable for high-level microarray analysis.


Prominent Spasticity and Hyperreflexia of the Legs in a Nepalese Patient with Friedreich Ataxia.
Friedreich ataxia (FRDA) is an autosomal recessive spinocerebellar ataxia caused by mutations of FXN. Hypotonus and hyporeflexia of the lower extremities are observed in most FRDA patients. Patients with hyperreflexia, called Friedreich ataxia with retained reflexes (FARR), have also been identified. We herein report the case of a 16-year-old Nepalese boy presenting with early-onset ataxia with prominent spasticity and hyperreflexia of the legs. Mutational analyses established the diagnosis of FRDA presenting as FARR. A haplotype analysis revealed that expanded alleles of the patient shared a common haplotype with Indian and European FRDA patients, suggesting that the mutation descended from a common founder.


Warming of the Eurasian landmass is making the Arabian Sea more productive.
The recent trend of declining winter and spring snow cover over Eurasia is causing a land-ocean thermal gradient that is particularly favorable to stronger southwest (summer) monsoon winds. Since 1997, sea surface winds have been strengthening over the western Arabian Sea. This escalation in the intensity of summer monsoon winds, accompanied by enhanced upwelling and an increase of more than 350% in average summertime phytoplankton biomass along the coast and over 300% offshore, raises the possibility that the current warming trend of the Eurasian landmass is making the Arabian Sea more productive.


Culturomics and pyrosequencing evidence of the reduction in gut microbiota diversity in patients with broad-spectrum antibiotics.
The human gut flora is currently widely characterised using molecular techniques. Microbial culturomics (large-scale culture conditions with identification of colonies using MALDI-TOF or 16S rRNA) is part of the rebirth of bacterial culture that was initiated by environmental microbiologists for the design of axenic culture for intracellular bacteria in clinical microbiology. Culturomics was performed on four stool samples from patients treated with large-scale antibiotics to assess the diversity of their gut flora in comparison with other culture-dependent studies. Pyrosequencing of the V6 region was also performed and was compared with a control group. Gut richness was also estimated by bacterial counting after microscopic observation. In total, 77 culture conditions were tested and 32,000 different colonies were generated; 190 bacterial species were identified, with 9 species that had not been isolated from the human gut before this study, 7 newly described in humans and 8 completely new species. A dramatic reduction in diversity was observed for two of the four stool samples for which antibiotic treatment was prolonged and uninterrupted. The total number of bacteria was generally preserved, suggesting that the original population was replaced but was sustained in size. Discordances between culture and pyrosequencing biodiversity biomarkers highlight the depth of bias of molecular studies. Stool samples studied showed a dramatic reduction in bacterial diversity. Considering the variable antibiotic concentration in the gut, this reduction in the number of species is possibly linked to the production of bacteriocin in the upper digestive tract by specific bacteria, such as Lactobacillus spp.


Restriction in V kappa gene use and antigen selection in anti-myeloperoxidase response in mice.
Anti-neutrophil cytoplasmic Abs, directed primarily toward myeloperoxidase (MPO) and proteinase 3, are detected in the majority of patients with distinct forms of small vessel vasculitides and pauci-immune necrotizing glomerulonephritis. However, the origin of these autoantibodies remains unknown. We studied the V region gene use in murine anti-MPO Abs derived from Spontaneous Crescentic Glomerulonephritis/Kinjoh mice. A total of 13 anti-MPO-producing hybridomas were generated from four unimmunized mice. Ten of the 13 hybridomas (corresponding to 3 of 4 clones) expressed Vkappa1C but differed in their use of VH genes. The remaining three hybridomas expressed a Vkappa5 gene. Anti-MPO hybridomas from individual mice were derived from single clones as deduced by sequence similarity and splice-site identity. We found a statistically significant bias of amino acid replacement mutations to the complementarity-determining regions (CDR) in the Vkappa1C-expressing hybridomas. Intriguingly, all 10 Vkappa1C hybridomas share a lysine to glutamate mutation in the CDR1. To determine the effects of somatic V gene mutations on binding to MPO, we generated an anti-MPO Ab with an unmutated Vkappa1C L chain and compared its ability to bind MPO with its mutated counterpart. The mutated hybridoma-derived Ab has a 4.75-fold higher avidity for MPO than the unmutated Ab. These results suggest that: 1) the L chain plays a dominant role in determining Ab specificity to MPO, 2) the anti-MPO Ab response is oligoclonal, consistent with Ag selection, and 3) MPO is a driving Ag in the murine anti-MPO Ab response.


Cancer mortality in Italy, 2008, and predictions for 2012.
This report provides up-to-date data and statistics for cancer mortality in Italy in 2008, and predicts the figures and rates for selected cancer sites for 2012. Cancer death certifications (for 30 sites) and resident population estimates in 2008 stratified by sex and age were obtained from the World Health Organization (WHO) database (WHOSIS). Mortality rates were age-standardized on the world standard population. Cancer deaths registered in Italy in 2008 were 172,783 (97,773 men and 75,010 women), corresponding to age-standardized death rates of 144.1/100,000 men and 84.3/100,000 women. The projected cancer deaths in 2012 are 178,000 (100,000 men, 78,000 women) and the corresponding rates 132,5/100,000 men and 80.5/100,000 women. The favorable trend in lung cancer mortality among men was confirmed, with rates of 37.7/100,000 in 2008 (all ages) and 33.3 for 2012. Other tobacco-related cancers also declined in men but not in women, including pancreatic cancer, whose rates tended to level off over the last 3 years. The fall in female cancer mortality rates continues to be led by favorable trends in breast cancer (16.1/100,000 in 2007 and 15.2 in 2012), intestinal cancer, stomach cancer and uterine cancer. However, the female lung cancer mortality was still rising with 7743 deaths in 2008 (9.5/100,000), and lung cancer is predicted to become the second cause of female cancer mortality by 2012 (8,500 deaths, 9.8/100,000). Reduced tobacco and alcohol consumption are largely responsible for the favorable trends in cancer mortality in men. Advances in treatment and management accounted for the reduced mortality from colorectal cancer, breast cancer, leukemias and a few other cancers, as well as improved diagnosis for colorectal, cervical and breast cancer. The rising epidemic of tobacco-related deaths in women indicates the need for targeted tobacco-control strategies.


P2X7 signaling promotes microsphere embolism-triggered microglia activation by maintaining elevation of Fas ligand.
The cerebral microvascular occlusion elicits microvascular injury which mimics the different degrees of stroke severity observed in patients, but the mechanisms underlying these embolic injuries are far from understood. The Fas ligand (FasL)-Fas system has been implicated in a number of pathogenic states. Here, we examined the contribution of microglia-derived FasL to brain inflammatory injury, with a focus on the potential to suppress the FasL increase by inhibition of the P2X(7)-FasL signaling with pharmacological or genetic approaches during ischemia. The cerebral microvascular occlusion was induced by microsphere injection in experimental animals. Morphological changes in microglial cells were studied immunohistochemically. The biochemical analyses were used to examine the intracellular changes of P2X(7)/FasL signaling. The BV-2 cells and primary microglia from mice genetically deficient in P2X(7) were used to further establish a linkage between microglia activation and FasL overproduction. The FasL expression was continuously elevated and was spatiotemporally related to microglia activation following microsphere embolism. Notably, P2X(7) expression concomitantly increased in microglia and presented a distribution pattern that was similar to that of FasL in ED1-positive cells at pathological process of microsphere embolism. Interestingly, FasL generation in cultured microglia cells subjected to oxygen-glucose deprivation-treated neuron-conditioned medium was prevented by the silencing of P2X(7). Furthermore, FasL induced the migration of BV-2 microglia, whereas the neutralization of FasL with a blocking antibody was highly effective in inhibiting ischemia-induced microglial mobility. Similar results were observed in primary microglia from wild-type mice or mice genetically deficient in P2X(7). Finally, the degrees of FasL overproduction and neuronal death were consistently reduced in P2X(7)(-/-) mice compared with wild-type littermates following microsphere embolism insult. FasL functions as a key component of an immunoreactive response loop by recruiting microglia to the lesion sites through a P2X(7)-dependent mechanism. The specific modulation of P2X(7)/FasL signaling and aberrant microglial activation could provide therapeutic benefits in acute and subacute phase of cerebral microembolic injury.


Polyamine and polyamine analog regulation of spermidine/spermine N1-acetyltransferase in MALME-3M human melanoma cells.
In MALME-3M human melanoma cells the polyamine analog N1,N12-bis(ethyl)spermine (BESPM) suppresses the key polyamine biosynthetic enzymes, ornithine and S-adenosylmethionine decarboxylase, and increases the polyamine catabolizing enzyme, spermidine/spermine N1-acetyl-transferase (SSAT) by more than 200-fold. In the present study increases in SSAT activity in MALME-3M cells treated with 10 microM BESPM were found to be accompanied by a substantial (up to 45-fold) accumulation of SSAT mRNA. By Northern blot analysis three RNA transcripts were found to hybridize with the coding region of human SSAT cDNA: a minor high molecular weight (approximately 3.5 kilobases) species designated form A and two lower molecular weight species designated forms B and C (approximately 1.5 and approximately 1.3 kilobases, respectively). Form A increased uniformly during BESPM treatment and was most obvious in nuclear RNA preparations. On the basis of size similarity to the transcribing region of the gene and hybridization with the coding region of SSAT cDNA and its prevalence in nuclear mRNA preparations, form A is thought to represent precursor SSAT RNA. Form C is present in control cells and increases steadily during treatment, whereas form B increases transiently during early treatment (1-3 h). By RNase H digestion assay, form B was found to have a 200-base pair longer poly(A) tract and as such may represent a precursor to form C. Accumulation of SSAT mRNA was found to be a result of increased gene transcription and stabilization of SSAT mRNA. Nuclear run-on studies indicated a 2-4-fold increase in the transcription rate of the SSAT gene. As indicated by actinomycin D studies, the SSAT mRNA half-life increased with BESPM treatment from 17 to 64 h. The natural polyamine, spermine, also increased SSAT mRNA (5.5-fold at 24 h) and behaved similarly to BESPM in inducing the appearance of the same three transcript forms. The polyamine was much less effective than the analog at increasing enzyme activity. Lowering intracellular polyamine pools with inhibitors of biosynthesis decreased basal SSAT mRNA levels by at least 70% indicating, that the gene can be down-regulated as well as up-regulated by polyamines. These findings indicate that SSAT represents a unique example of gene expression being positively influenced at the RNA level by polyamines and their analogs.


Nutritional status among hospitalized children with mixed diagnoses at a referral teaching hospital in Manizales, Colombia.
Despite advances in the treatment of malnutrition in pediatric hospitals, this remains a problem that must be recognized and to call the attention of academia and health authorities to be handled in time. To evaluate the nutritional status of hospitalized children at a referral teaching hospital and to describe the current prevalence of malnutrition. The study was conducted during the months of February, March and April 2010 at the University Hospital Rafael Henao Toro Colombian Red Cross in Manizales, Colombia, South America. We evaluated all patients admitted to a day of each month by calculating the Z score of the, weight for age, height or length for age, body mass index, mid-arm circumference, triceps and subscapular folds for children under 5 years and height for age and body mass index for school children and adolescents. A total of 174 children (age 1-216 mo) were evaluated during the 3-days survey. There were 52.8% children less than 60 months old, 17.2% between 61 and 120 months and 29.9% over 121 months. There were 44.3% females and 55.7% males. Children from the urban residence were 83.3% and 16.6% were rural. The overall prevalence of underweight was 27%, stunting 22.4%, wasting 16.6% and overweight and obesity 6.3%. Males less than five years old were more affected than older children. Rural children were more affected than urban children. The prevalence of overweight children was greater in children 61-120 months than other ages. Given the observed levels of malnutrition, it takes up a system for early identification of children hospitalized with nutritional risk in order to provide adequate and timely support and prevent hospital-acquired malnutrition. This requires the use of previously validated pediatric protocols.


Single enzyme RT-PCR of full-length ribosomal RNA.
The ribosome is a two-subunit, macromolecular machine composed of RNA and proteins that carries out the polymerization of α-amino acids into polypeptides. Efforts to engineer ribosomal RNA (rRNA) deepen our understanding of molecular translation and provide opportunities to expand the chemistry of life by creating ribosomes with altered properties. Toward these efforts, reverse transcription PCR (RT-PCR) of the entire 16S and 23S rRNAs, which make up the 30S small subunit and 50S large subunit, respectively, is important for isolating desired phenotypes. However, reverse transcription of rRNA is challenging due to extensive secondary structure and post-transcriptional modifications. One key challenge is that existing commercial kits for RT-PCR rely on reverse transcriptases that lack the extreme thermostability and processivity found in many commercial DNA polymerases, which can result in subpar performance on challenging templates. Here, we develop methods employing a synthetic thermostable reverse transcriptase (RTX) to enable and optimize RT-PCR of the complete Escherichia coli 16S and 23S rRNAs. We also characterize the error rate of RTX when traversing the various post-transcriptional modifications of the 23S rRNA. We anticipate that this work will facilitate efforts to study and characterize many naturally occurring long RNAs and to engineer the translation apparatus for synthetic biology.


Steamed broccoli sprouts alleviate DSS-induced inflammation and retain gut microbial biogeography in mice.
Inflammatory Bowel Diseases (IBD) are devastating conditions of the gastrointestinal tract with limited treatments, and dietary intervention may be effective, and affordable, for managing symptoms. Glucosinolate compounds are highly concentrated in broccoli sprouts, especially glucoraphanin, and can be metabolized by certain mammalian gut bacteria into anti inflammatory isothiocyanates, such as sulforaphane. Gut microbiota exhibit biogeographic patterns, but it is unknown if colitis alters these or whether the location of glucoraphanin metabolizing bacteria affects anti-inflammatory benefits. We fed specific pathogen free C57BL/6 mice either a control diet or a 10% steamed broccoli sprout diet, and gave a three-cycle regimen of 2.5% dextran sodium sulfate (DSS) in drinking water over a 34-day experiment to simulate chronic, relapsing ulcerative colitis. We monitored body weight, fecal characteristics, lipocalin, serum cytokines, and bacterial communities from the luminal and mucosa-associated populations in the jejunum, cecum, and colon. Mice fed the broccoli sprout diet with DSS treatment performed better than mice fed the control diet with DSS, including significantly more weight gain, lower Disease Activity Indexes, lower plasma lipocalin and proinflammatory cytokines, and higher bacterial richness in all gut locations. Bacterial communities were assorted by gut location, but were more homogenous across locations in the control diet + DSS mice. Importantly, our results showed that broccoli sprout feeding abrogated the effects of DSS on gut microbiota, as bacterial richness and biogeography were similar between mice receiving broccoli sprouts with and without DSS. Collectively, these results support the protective effect of steamed broccoli sprouts against dysbiosis and colitis induced by DSS. Evaluating bacterial communities across different locations in the gut provides a greater insight than fecal samples alone, and provides an additional metric by which to evaluate beneficial host-microbe interactions. Here, we show that 10% steamed broccoli sprouts in the diet protects mice from the negative effects of dextran sodium sulfate induced colitis, that colitis erases biogeographical patterns of bacterial communities in the gut, and that the cecum is not likely to be a significant contributor to colonic bacteria of interest in the DSS mouse model of ulcerative colitis. Mice fed the broccoli sprout diet during colitis performed better than mice fed the control diet while receiving DSS. The identification of accessible dietary components and concentrations that help maintain and correct the gut microbiome may provide universal and equitable approaches to IBD prevention and recovery, and broccoli sprouts represent a promising strategy.