New mechanistic insights into PAHs transport across wheat root cell membrane: Evidence for ABC transporter mediation.
Polycyclic aromatic hydrocarbons (PAHs) are a class of highly carcinogenic organic pollutants. Our previous results revealed that the active uptake of PAHs by plant roots is performed through H+/PAHs co-transport. However, the proteins and mechanisms of co-transport of PAHs remain unknown. We hypothesized that ABC transporters are involved in PAHs co-transport via the roots. We found a total of 47 ABC transporters with alkalinity and hydrophobicity which were up-regulated or newly expressed in the wheat roots after phenanthrene exposure. And the concentration of ABC transporters rose. There was a positive relationship between the concentration of phenanthrene and ABC transporter expression in the wheat roots. Additionally, the trend observed in the ABC transporters expression was also found in the gene expression. With energies below -6 kcal mol-1, a stable docking conformation formed between ABC transporters and PAHs. π-π stacking and van der Waals force bound PAHs to ABCB or ABCG. The binding strength of ABCB subfamily proteins with homodimers is stronger than that of ABCG subfamily proteins with single molecules. ABC transporters may transport PAHs by forming a dimer-shaped pocket, translocating it into cells, then opening it within the cells, to release the bound PAHs. These results contributed to our understanding of how ABC transporters aid plant root uptake of PAHs.


[Action of the microbial flora of the digestive tract on the biosynthesis of cholic acid in the rat (author's transl)].
Axenic and holoxenic (conventional) rats were fed a diet containing trace amounts of [2,4-3H]cholic and [24-14C]chemodeoxycholic acids. In the feces of both groups of rats, the percentage of labelled bile acids which were 3H-labelled was slightly different. In the experimental conditions used, the intestinal microflora only slightly modified the synthesis of 12alpha-hydroxylated bile acids.


E2F1 activates p53 transcription through its distal site and participates in apoptosis induction in HPV-positive cells.
The p53 tumor suppressor protein, one of the most extensively studied proteins, plays a pivotal role in cellular checkpoints that respond to DNA damage to prevent tumorigenesis. However, the transcriptional control of the p53 gene has not been fully characterized. We report that the transcription factor E2F1 binds only to the E2F1 distal site of the p53 promoter in the human papillomavirus positive carcinoma HeLa cell line. Moreover, we showed that etoposide, a DNA damaging agent, activates p53 transcription through the E2F1 pathway. This increase correlates with apoptosis induction as disruption of this pathway led to reduced apoptosis stimulation by the DNA damaging agent.


Field testing a complaints register proposal as a requirement of Australian general practice.
To investigate the feasibility, achievement and acceptance of indicators of quality general practice in the RACGP Standards for general practices (third edition), using complaints registers as a case study. A purposive sample of convenience of 200 general practices (stratified according to location and size) participated in a field test of quality and safety proposals during an accreditation survey visit between October 2004 and February 2005. Included was a test of the proposal for a complaints register (a document where complaints made to the practice are recorded). Achievement of the complaints register proposal, assessed by accreditation surveyors; questionnaire rating of the feasibility and acceptance of the proposal. Few practices used a formal complaints register (79/200; 39.5%), with large practices more likely (12/20; 60.0%) and very remote practices less likely (1/11; 9.1%) to use one. The proposal for complaints registers was rated feasible by 123 general practices (61.5%) and rated acceptable by 121 general practices (60.5%). The proposal for complaints registers in general practice, while popular with policymakers, gained limited support when tested in Australian general practice. This shows the need for a balance between the expectations of policymakers, the need to increase performance by setting standards, and the practicalities of every-day general practice.


Molecular heterogeneity and structural evolution during cerebellar ontogeny detected by monoclonal antibody of the mouse cell surface antigen BSP-2.
The monoclonal antibody anti-BSP-2 defines a set of glycoproteins present on the neuronal cell surface in dissociated mouse cerebellar cultures and on neurons and astrocytes in sections of the mouse cerebellum. This antibody was used in the present study to characterize the antigens recognized in cerebellar cultures and in the developing and adult mouse cerebellum in vivo. In extracts from cerebellar cultures and from late postnatal or adult cerebellum, the anti-BSP-2 antibody reacted with a triplet of glycosylated polypeptide chains of 180,000, 140,000 and 120,000 mol. wt. Early postnatal cerebellum contained a different form of BSP-2 antigen which migrated as one broad or several closely spaced diffuse bands in the 190,000-250,000 mol. wt. region of SDS polyacrylamide gels. During cerebellar ontogeny, the adult pattern emerged gradually between postnatal days 5 and 13. The cellular expression of the BSP-2 antigen was studied by immunohistochemistry on sections of the developing cerebellum. At postnatal day 3, the antigen was found mainly on cell bodies and fibers of the Bergmann glia and on astrocytes of the granular layer. Immature granule cells of the outer zone of the external granular layer lacked the antigen, but they appeared to acquire the antigen during their migration to the internal granular layer. At postnatal day 13, the immunofluorescence pattern was not different from the one seen in the adult. These results suggest that the neonatal 190,000-250,000 mol. wt. form of BSP-2 may at least in part be expressed by astroglial cells and they show a close correlation between the emergence of the adult forms of the antigen and the appearance of labeled granule cells in the internal granular layer. In vitro degradation implying cleavage of sialic acid residues, but probably also proteolysis and/or cleavage of different glycans converted the neonatal form of BSP-2 into the triplet pattern and ultimately into a p120 component. Neuraminidase digestion of the adult antigens produced small molecular weight shifts without converting one band into the other, but endogenous enzyme activities were capable of degrading the p180 and p140 bands by converting them into the p120 protein. Our findings support the idea that distinct, but structurally similar surface glycoproteins created by post-translational modifications from a common precursor molecule may be expressed by different cell types or during different developmental stages. As shown by sequential immunoprecipitation experiments, BSP-2 and the rat neuronal membrane protein D2 may belong to the same family of surface glycoproteins.


Glycosphingolipid synthesis is essential for MDCK cell differentiation.
Glycosphingolipids (GSLs), which are highly concentrated at the apical membrane of polarized epithelial cells, are key components of cell membranes and are involved in a large number of processes. Here, we investigated the ability of hypertonicity (high salt medium) to induce Madin-Darby Canine Kidney (MDCK) cell differentiation and found an increase in GSL synthesis under hypertonic conditions. Then, we investigated the role of GSLs in MDCK cell differentiation induced by hypertonicity by using two approaches. First, cultured cells were depleted of GSLs by exposure to D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP). Second, cells were transfected with an siRNA specific to glucosylceramide synthase, the key enzyme in GSL synthesis. Exposure of cells to both treatments resulted in the impairment of the development of the apical membrane domain and the formation of the primary cilium. Enzymatic inhibitions of the de novo and the salvage pathway of GSL synthesis were used to determine the source of ceramide responsible of the GSL increase involved in the development of the apical membrane domain induced by hypertonicity. The results from this study show that extracellular hypertonicity induces the development of a differentiated apical membrane in MDCK cells by performing a sphingolipid metabolic program that includes the formation of a specific pool of GSLs. The results suggest as precursor a specific pool of ceramides formed by activation of a Fumonisin B1-resistant ceramide synthase as a component of the salvage pathway.


Sonographically guided arterial blood sampling using a handheld Doppler device.
The purpose of this study was to evaluate the efficacy of a commercially available handheld Doppler device for sonographic guidance during arterial blood sampling in patients whose veins are inaccessible and whose arterial pulses are indistinct on palpation. From September 2000 through February 2001, blood samples were obtained from the brachial arteries of 20 patients ranging in age from 60 to 85 years. A commercially available continuous-wave, handheld Versadopp 10 Doppler device with a 10-MHz transducer, connected to a stethoscope, was aligned over each patient's brachial artery and used to determine the point of maximum flow. The blood-collection needle was inserted into the skin and advanced toward the point of maximum flow. Arterial blood samples were successfully obtained with no more than 2 passes in each patient. Sampling was unsuccessful in 2 patients; blood was obtained, however, from the contralateral brachial artery in both patients. This Doppler tool is convenient and easy to use and is helpful for obtaining arterial blood samples from obese or edematous patients whose veins cannot be accessed and who have weak, indistinct arterial pulses.


RNA-Seq transcriptome analysis to identify candidate genes involved in non-target site-based mesosulfuron-methyl resistance in Beckmannia syzigachne.
American sloughgrass (Beckmannia syzigachne Steud.) has become a dominant weed in fields with rice-wheat rotation. Moreover, herbicide resistance has rendered weed control difficult. We identified a biotype showing resistance to ALS inhibitor mesosulfuron-methyl with a resistant index 3.3, but without any ALS mutation. This study aims to identify and confirm the factors associated with non-target site resistance of this biotype to mesosulfuron-methyl using RNA-Seq. 118,111 unigenes were assembled, and 50.9% of these were annotated across seven databases. Eleven contigs related to metabolic resistance were identified based on differential expression via RNA-Seq which include a novel resistance-related transcription factor (MYC3) and two disease resistance proteins were also identified (At1g58602 and At1g15890). Fold changes in expression of these genes in comparison M-R vs. M-S ranged from 3.9 to 11.6, as confirmed by qPCR. The expression of a contig annotated as cytochrome P450 (CYP86B1) in resistant individuals was over 3 times higher than that in sensitive individuals at 0-72 h after mesosulfuron-methyl treatment. A similar trend was noted for three other genes annotated as glutathione S-transferase (GST), namely GST-T3, GST-U6, and GST-U14; the expression of GST-U6 in resistant individuals was up to 142.3 times higher than that in sensitive individuals at 24 h after mesosulfuron-methyl treatment. In addition, GST activity in resistant individuals was 2.1 to 5.3 times higher than that in sensitive individuals. The GR50 of resistant biotype decreased from 24.4 to 11.3 g a.i. ha-1 after P450 inhibitor malathion treatment. This study identified a cytochrome P450 gene CYP86B1 and three GST genes GST-T3, GST-U6, and GST-U14 that have higher expression in mesosulfuron-methyl resistant B. syzigachne, suggesting that both P450- and GST-based activities could be involved in resistance.


Cardiac characterization of mdx mice using high-resolution doppler echocardiography.
Duchenne muscular dystrophy is an X-linked neuromuscular disorder. The heart is traditionally involved, leading to heart failure. The mdx mouse is a natural animal model of the disease. We conducted a prospective study to analyze left ventricular (LV) function in mdx mice at different ages using high-resolution Doppler echocardiography. Echocardiography was performed with a 30-MHz cardiac probe. Wild-type and mdx mice were scanned at 10 and 12 months. We measured the interventricular septal wall thickness, posterior wall thickness, and LV diameter in systole and diastole. The LV shortening fraction, LV ejection fraction, and LV mass were then calculated. At 10 months, the shortening fractions in mdx and wild-type mice were relatively close (29% ± 5% versus 25% ± 4%). We found a significant difference in the posterior wall thickness change (40% ± 12% in mdx versus 28% ± 10% in wild-type; P = .048). The LV mass/body weight ratio was higher in mdx than wild-type mice (3.67 ± 0.25 versus 3.39 ± 0.26; P = .05). At 12 months, the LV mass was elevated in mdx mice compared to wild-type (152 ±16 versus 135 ± 3.7 mg; P = .04). The diastolic posterior wall thickness change was decreased in mdx mice at 12 months compared to wild-type (21% ± 7% versus 33% ± 4%; P = .01). The LV ejection fraction was not statistically different between mdx and wild-type mice (50% ± 6% versus 54% ± 2%). Ten-month-old mdx mice had a significantly higher posterior wall thickness than wild-type mice, but it was not significant at 12 months. In 12-month-old mdx mice, the posterior wall thickness change was significantly lower, and the LV mass was significantly higher. These findings indicate the role of LV function in the early stages of Duchenne muscular dystrophy.


The pro-inflammatory biomarker soluble urokinase plasminogen activator receptor (suPAR) is associated with incident type 2 diabetes among overweight but not obese individuals with impaired glucose regulation: effect modification by smoking and body weight status.
Recent evidence links the soluble urokinase plasminogen activator receptor (suPAR), a stable biomarker of systemic immune activation, to several chronic diseases, including type 2 diabetes. suPAR is also associated with adiposity and smoking. We hypothesised that this biomarker would be linked to incident type 2 diabetes in individuals with impaired glucose regulation and that this association would be modified by smoking and body weight status. The study included 1,933 participants with impaired glucose regulation, who were drawn from the Danish arm of the Anglo-Danish-Dutch Study of Intensive Treatment in People with Screen-Detected Diabetes in Primary Care (ADDITION) and for whom data on suPAR, BMI and smoking were available. Logistic regression analysis was used to estimate the odds for incident type 2 diabetes per twofold increase in suPAR levels. Interactions between both smoking and body weight status and suPAR were tested. During a 3-year follow-up (599 incident diabetes cases), there was a 48% overall increase in the odds of developing type 2 diabetes per twofold increase in suPAR (p = 0.006). This association was modified by body weight status in overweight, but not in obese individuals (OR 2.36, 95% CI 1.48, 3.76 in overweight group), and by smoking status (OR 2.05, 95% CI 1.20, 3.51 in non-smokers). After adjustment for other diabetes risk factors, the association between suPAR and type 2 diabetes was attenuated in the whole sample and among non-smokers, but remained robust among overweight participants. suPAR may be a good novel biomarker for systemic sub-clinical inflammation and immune activation linked to incident type 2 diabetes risk in overweight individuals and non-smokers. The observed interactions with adiposity and smoking should be investigated further.


Association between physician turnover and survival outcome after in-hospital cardiopulmonary resuscitation: A nationwide cohort study in South Korea.
We investigated the association between physician turnover and survival outcomes after in-hospital cardiopulmonary resuscitation (ICPR) in South Korea. This population-based cohort study used the South Korean national registration database as the data source. All adult patients admitted to the hospital and who underwent ICPR between 1 January 2010 and 31 December 2019, were included. Patients who underwent ICPR in March were included in the turnover group, while those who underwent ICPR in the other months were included in the non-turnover group. Propensity score (PS) matching was performed. Overall, 298,676 adult patients who underwent ICPR in 2,553 South Korean hospitals were included in the analysis. Among them, 26,342 (8.8%) and 272,334 (91.2%) were included in the turnover and non-turnover groups, respectively. In total, 7,009 (26.6%) and 6,903 (26.2%) of the 26,342 patients each in the turnover and non-turnover groups, respectively, were discharged alive after ICPR. Using logistic regression analysis in the PS-matched cohort, the two groups did not show any significant association in the live discharge rate after ICPR (odds ratio: 1.02, 95% confidence interval: 0.98, 1.06; P = 0.295). This non-significant association was also observed in patients who underwent ICPR in tertiary general hospitals that had cardiopulmonary resuscitation teams for ICPR (P = 0.136). Moreover, the median survival time in the turnover and non-turnover groups was 4.0 days (95% confidence interval: 3.8 days, 4.2 days; log-rank test, P = 0.796). Significant association between physician turnover and survival outcomes was not observed after ICPR in South Korea.


LncRNA THRIL aggravates sepsis-induced acute lung injury by regulating miR-424/ROCK2 axis.
Here, we aimed to investigate the role of long noncoding RNA (lncRNA) THRIL in septic-induced acute lung injury. C57BL/6 mice were injected with Adenoviruses (Ad)-shTHRIL or negative control (NC) before caecal ligation and puncture (CLP) operation. MPVECs were transfected with Ad-shTHRIL or NC, followed by lipopolysaccharide (LPS) treatment. MiR-424 and Rho-associated kinase 2 (ROCK2) were predicted and verified as direct targets of THRIL and miR-424, respectively, by using dual-luciferase reporter assay. ROCK2 overexpression vector and shTHRIL were co-transfected into mouse pulmonary microvascular endothelial cells for 24 h before LPS treatment. Our results showed that THRIL was highly expressed in the lung of sepsis mice. CLP triggered severe lung injury and apoptosis in mice, which was abolished by THRIL knockdown. Moreover, CLP treatment visibly increased protein concentration, the number of total cell of neutrophils, and macrophages in bronchoalveolar lavage fluid (BALF). Besides, elevated protein levels of tumor necrosis factor-α, interleukin-1β, and interleukin-6 were observed in both lung and BALF. However, inhibition of THRIL reduced the number of inflammatory cells and the production of pro-inflammatory cytokines in sepsis mouse model. The effect of THRIL on inflammatory response and apoptosis in the lung was confirmed in sepsis cell model. Moreover, mechanistic studies have shown that THRIL up-regulated ROCK2 level through sponging miR-424. Furthermore, ROCK2 overexpression reversed the inhibitory effects of THRIL knockdown on LPS-induced inflammatory response and apoptosis. Overall, in vivo and in vitro results suggested that THRIL accelerates sepsis-induced lung injury by sponging miR-424 and further restoring ROCK2.


Assessing the cellular and humoral immune response in Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) infected with Leishmania infantum (Nicolle, 1908).
The aim of this study was to evaluate aspects of the innate cellular and humoral immune response by evaluating hemocyte dynamics, phagocytosis, phenoloxidase (PO) activity and nitric oxide (NO) production in Rhipicephalus sanguineus sensu lato (s.l.) (Acari: Ixodidae) infected with Leishmania infantum and to assess the persistence of parasites at time 0 and 1, 2, 5, and 7 days post-infection (dpi). The total and differential count of the five types of hemocytes circulating in the hemolymph of R. sanguineus s.l. females showed the average total number of hemocytes in the group infected with L. infantum to be significantly higher (p < 0.05) on the 1st and 2nd dpi compared to the control group. The hemocyte differential count showed that the average number of plasmatocytes and granulocytes increased significantly on the 1st, 2nd, and 5th dpi with L. infantum compared to the control group (p < 0.001). Phagocytosis assays revealed that plasmatocytes and granulocytes were able to perform phagocytosis of latex beads and L. infantum on the 1st and 2nd dpi, respectively. NO production was significantly increased (p < 0.001) on the 1st, 2nd, and 5th dpi with L. infantum and PO activity increased significantly (p < 0.05) only on the 5th dpi. L. infantum DNA was significantly increased (p < 0.001) on the 5th and 7th dpi compared to time 0. Although there are no studies describing the response of R. sanguineus s.l. to an infection with L. infantum, these results suggest that R. sanguineus s.l. activates the cellular and humoral immune response after infection with L. infantum. Further studies are however, needed to assess the impact of such a response on fighting infection.


Hemangioma - A Rare Cause of Laryngeal Growth in an Adult and its Management with Sclerotherapy and Laser Surgery: A Case Report.
Hemangiomas are benign tumors of vascular origin commonly affecting head and neck region. Laryngeal Hemangiomas are however rare and have no well-defined management protocols. We present an interesting case of a 42-year-old adult who was diagnosed as a case of Laryngeal Hemangioma based on clinical and radiological findings and was managed with local injection of sclerosing agent followed by CO2 Laser fulguration. The patient had a satisfactory outcome and remains asymptomatic.


Autologous peripheral blood stem cell mobilization following dose-adjusted cyclophosphamide, doxorubicin, vincristine, and prednisolone chemotherapy alone or in combination with rituximab in treating high-risk non-Hodgkin's lymphoma.
The regimen of cyclophosphamide, doxorubicin, vincristine, and prednisolone (CHOP) is an efficient treatment of non-Hodgkin's lymphoma (NHL). This study aimed to assess the efficacy and toxicity of dose-adjusted CHOP alone or in combination with rituximab (R-CHOP) by examining the stem cell mobilization in NHL patients. Factors affecting the collection of CD34+ cells were also explored. Our retrospective study included 39 patients eligible for autologous stem cell transplantation: 14 patients who expressed CD20 and were financially eligible received R-CHOP for autologous peripheral blood stem cell (APBSC) mobilization; the remaining 25 patients received CHOP. The median CD34+ cell yield was 7.01×10(6) cells/kg body weight (range 1.49-28.39×10(6) cells/kg body weight), with only two patients failing to meet the target CD34+ cell harvest of ≥2.0×10(6) cells/kg body weight. The median number of apheresis procedures per patient was 1 (range 1-3). The APBSC mobilization yield of the CHOP group appeared to be higher than that of the R-CHOP group (P=0.005), whereas the success rate was similar between groups. R-CHOP elevated the complete response (CR) rate in B cell lymphoma patients as compared with CHOP (P=0.01). No significant differences in toxicity or engraftment were observed between the two groups. The present study demonstrated that dose-adjusted CHOP chemotherapy effectively mobilized APBSCs in NHL patients and that the addition of rituximab to dose-adjusted CHOP chemotherapy elevated the CR rate for patients with B-cell lymphoma.


Detection of hepatitis E virus genotypes 3 and 4 in donkeys in northern China.
Hepatitis E virus (HEV) is the causative agent of acute self-limiting hepatitis in humans in developing countries. Hepatitis E virus RNA was first detected in donkeys in Spain, but little is known about the possible presence of HEV in donkeys in China. To investigate the prevalence of HEV in donkeys in northern China. Investigation of the prevalence of HEV in donkeys using serological, molecular and phylogenetic approaches. A total of 401 donkey serum specimens were tested for serological and molecular detection of HEV via enzyme-linked immunosorbent assay and quantitative reverse transcription polymerase chain reaction. The amplified products were cloned in pMD18-T vector and sequenced. The alignment and phylogenetic analysis of partial HEV ORF2 genes were compared with the corresponding sequences of the obtained HEV representative strains. Serological results showed that 49 donkeys (12.22%, 95% CI: 9.18-15.83%) were positive for anti-HEV-specific antibodies, and 17 donkeys (4.24%, 95% CI: 2.49-6.70%) were positive for HEV viral RNA. On the basis of sequence alignment and phylogenetic analysis, all isolated HEV strains belonged to genotype 3 (HEV-3) or HEV-4, sharing more than 76.2-96.3% identities with 67 other HEV representative strains of HEV-1 to HEV-8. Further studies about the prevalence of HEV in organs or faecal samples from donkeys are needed to evaluate the possible role of HEV reservoir and to determine the risk factors associated with the transmission of this zoonotic virus in donkeys in China. This is the first report documenting the molecular analysis of donkey HEV strains worldwide and the serological evidence of HEV infection in donkeys in northern China. The results suggest that young donkeys are more susceptible to HEV infection compared with older donkeys. Further investigation is required to determine whether donkeys should be considered reservoirs for zoonotic HEV. The Summary is available in Chinese - see Supporting information.


Advance in glycosyltransferases, the important bioparts for production of diversified ginsenosides.
Ginsenosides are a series of glycosylated triterpenoids predominantly originated from Panax species with multiple pharmacological activities such as anti-aging, mediatory effect on the immune system and the nervous system. During the biosynthesis of ginsenosides, glycosyltransferases play essential roles by transferring various sugar moieties to the sapogenins in contributing to form structure and bioactivity diversified ginsenosides, which makes them important bioparts for synthetic biology-based production of these valuable ginsenosides. In this review, we summarized the functional elucidated glycosyltransferases responsible for ginsenoside biosynthesis, the advance in the protein engineering of UDP-glycosyltransferases (UGTs) and their application with the aim to provide in-depth understanding on ginsenoside-related UGTs for the production of rare ginsenosides applying synthetic biology-based microbial cell factories in the future.


Carbon dioxide assimilation and growth of red spruce (Picea rubens Sarg.) seedlings in response to ozone, precipitation chemistry, and soil type.
The influence of ozone, mist chemistry, rain chemistry, and soil type on CO2 assimilation and growth of red spruce (Picea rubens Sarg.) seedlings was investigated over a 4-month period under controlled laboratory and glasshouse conditions. Growth was evaluated through interval estimates of aboveground relative growth rates (RGR) and the partitioning of biomass components at harvest to root, stem, and needle fractions. Precipitation chemistry treatments and O3 exposure dynamics were based on reported characteristics of air chemistry and/or deposition in high-elevation forests of eastern North America. The two soils were collected from Camels Hump in the Green Mountains of Vermont and Acadia National Park on the Maine coast. Soil from Acadia had higher organic content, higher levels of extractable base cations, and lower levels of extractable aluminum and heavy metals. The only treatment variables that consistently influenced the growth of P. rubens were soil type and rain chemistry. In comparison with seedlings grown in soil from Acadia National Park, those grown in Camels Hump soil had significantly less needle (27%), stem (33%), and root (26%) biomass at harvest and statistically lower aboveground RGR within 2 months after initiation of the treatments. Seedlings grown in Camels Hump soil had significantly higher levels of aluminum (6.5X), copper (1.4X), and nickel (2.7X) in new needle tissue. The only influence of precipitation chemistry on the growth of P. rubens was a pattern of greater root and shoot biomass in seedlings experiencing the more acidic rain treatments. Interactive effects among the main treatment variables (e.g., acidic mist and O3, acidic rain and soil type) on seedling growth were not notable. Rates of CO2 assimilation and transpiration on a per gram needle dry weight basis [mol·g-1·s-1] were not influenced by any of the main treatment variables or their interaction. Because neither soil type nor precipitation chemistry influenced the efficiency of CO2 assimilation per gram dry weight of needle tissue, the physiological mechanism underlying the growth response of P. rubens is attributed to a change in either whole-plant allocation of carbon resources or a direct toxic effect in the rhizosphere on root growth.


Transretinal feeder vessel ligature in von Hippel-Lindau disease.
To present a new technique called transretinal feeder vessel ligature for the treatment of retinal angiomas. Case report of a patient with peripheral retinal angiomas previously treated unsuccessfully with photocoagulation who responded to this new, alternative surgical treatment. The retinal angiomas decreased in size although two new feeder vessels appeared and the lesions showed a regression pattern after additional laser therapy over the vascular tumors. A transretinal feeder vessel ligature in association with vitrectomy and photocoagulation may be useful for some advanced or non-responsive cases of retinal angiomas.


Engineered solubility tag for solution NMR of proteins.
The low solubility of many proteins hinders large scale expression and purification as well as biophysical measurements. Here, we devised a general strategy to solubilize a protein by conjugating it at a solvent-exposed position to a 6 kDa protein that was re-engineered to be highly soluble. We applied this method to the CARD domain of Apoptosis-associated speck-like protein containing a CARD (ASC), which represents one member of a class of proteins that are notoriously prone to aggregation. Attachment of the tag to a cysteine residue, introduced by site-directed mutagenesis at its self-association interface, improved the solubility of the ASC CARD over 50-fold under physiological conditions. Although it is not possible to use nuclear magnetic resonance (NMR) to obtain a high quality 2D correlation spectrum of the wild type domain under physiological conditions, we demonstrate that NMR relaxation parameters of the solubilized variant are sufficiently improved to facilitate virtually any demanding measurement. The method shown here represents a straightforward approach for dramatically increasing protein solubility, enabled by ease of labeling as well as flexibility in tag placement with minimal perturbation to the target.


Comparative analysis of visual outcomes following small-incision lenticule extraction with or without cyclotorsion compensation in eyes with high astigmatism: Contralateral eye study.
To compare the visual outcomes in patients undergoing small-incision lenticule extraction (SMILE) for correction of myopic astigmatism (≥-1.50 D) with or without manual cyclotorsion compensation. A prospective, double-blinded, randomized, contralateral study was conducted in the refractive services of a tertiary eye care center. Eligible patients with bilateral high myopic astigmatism (≥1.5 D) and intraoperative cyclotorsion (≥5°) undergoing SMILE between June 2018 and May 2019 were included. Cyclotorsion compensation was performed using triple centration method before femtosecond laser delivery. Uncorrected and corrected distance visual acuity (UDVA and CDVA, respectively) measurement, manifest refraction, slit-lamp biomicroscopy, and corneal tomography were performed preoperatively and at 1 and 3 months' postoperative visit. Astigmatic outcomes were analyzed using Alpins criteria. A total of 30 patients (60 eyes) were included in this study. The patients underwent bilateral SMILE surgery, with manual cyclotorsion compensation in one eye (CC group, n = 30 eyes) and no cyclotorsion compensation in the other eye (NCC group, n = 30 eyes). Preoperative astigmatism and intraoperative cyclotorsion were -2.0 D and 7.03° ± 1.06° (CC) and -1.75 D and 7.24° ± 0.98° (NCC) (P = 0.472 and 0.240, respectively). No significant differences were noted in mean refractive spherical equivalent (MRSE), UDVA, CDVA, and refractive error between the two groups at 3 months' postoperative visit. Astigmatic outcomes measured using Alpins criteria demonstrated no significant difference between the two cohorts. The cyclotorsion compensation technique provided no additional advantage in terms of astigmatic outcomes or postoperative visual quality, in eyes with high preoperative astigmatism and intraoperative cyclotorsion.


Asthma Care Protocol Implementation in the Pediatric Intensive Care Unit.
Asthma is a complex chronic disease characterized by inflammatory disorder causing airflow obstruction due to inflammation, bronchospasms, and mucus plugging. Children who fail to respond to initial first-line therapies often require hospitalization, and many with severe exacerbations and near-fatal asthma require admission to the pediatric intensive care unit (PICU). Nursing care of these PICU patients requires close monitoring and excellent assessment of their respiratory status. Administration of medications, such as albuterol, methylprednisolone, magnesium sulfate, and sedatives, is needed. Close communication with the care team is vital, including providers and respiratory therapy, to coordinate care and to communicate assessment findings.


Effect of therapeutic hyperoxia on maximal oxygen consumption and perioperative risk stratification in chronic obstructive pulmonary disease.
Maximal consumption of oxygen ( ̇VO(2)max) during exercise is used in patients with chronic obstructive pulmonary disease (COPD) to stratify perioperative risk. However, the impact of therapeutic hyperoxia (i.e., use of supplemental oxygen to prevent hypoxemia during exercise) on ( ̇VO(2)max and other ventilatory parameters during maximal exercise in the resting normoxic COPD population is poorly defined. A randomized, double-blind crossover study was performed in which resting normoxic subjects (n=16) with COPD underwent two standard symptom-limited, ramped-protocol bicycle ergometry cardiopulmonary exercise tests >5 days apart with FiO(2) of 0.21 (control) and ~0.28 (therapeutic hyperoxia). ̇VO(2)max and other ventilatory parameters were compared using a paired two-sample t-test. Therapeutic hyperoxia significantly increased ̇VO(2)max (12.2 ± 2.9 vs. 13.6 ± 3.8 ml/kg/min, P = 0.03), partial pressure of end-tidal carbon dioxide, and oxygen saturation and significantly decreased ̇VE-̇VCO(2) slope, but it did not affect exercise time, maximum watts achieved, maximum minute ventilation, or change in end-expiratory lung volume. Three of four subjects with ̇VO(2)max <10 ml/kg/min without supplemental oxygen increased ̇VO(2)max to ≥10 ml/kg/min on therapeutic hyperoxia and potentially changed perioperative risk category. Therapeutic hyperoxia in a resting normoxic COPD population significantly improves ̇VO(2)max and may change perioperative risk stratification by conventional criteria. Further studies are needed to determine if this change in stratification is appropriate.


Telomere shortening with aging in human esophageal mucosa.
Progressive telomere shortening with aging was studied using normal esophageal mucosal specimens from 177 human subjects aged between 0 and 102 years (yrs). We observed age-related shortening of the telomere, at a rate of 60 base pairs (bp) per year (yr). The mean telomere length of 12 neonates was 15.2 kilobase pairs (kbp) and that of 2 centenarians was 9.3 kbp. Mean (±SD) telomere lengths were 14.9±1.3, 14.0±1.8, 10.1±3.7, 10.4±3.3 and 9.5±3.1 kbp for the age groups less than 2 yrs, 2-20 yrs, 21-60 yrs, 61-80 yrs and 81-102 yrs, respectively. The variation in telomere length among individuals in the same age group was greater for the 3 older groups than for the 2 younger groups, as shown by the SDs. Furthermore, older individuals had greater telomere length variation than younger individuals, based on the lengths of DNA digested smears. Although the telomere length decreased significantly with aging at the rate of 60 bp per yr, differences in the mean telomere lengths between the 3 older age groups were not significant. Rapid shortening occurred in the young generations and there was no further substantial decrease in the esophageal mucosa after 60 yrs of age. Compared to the very rapid renewal rate of the esophageal epithelial cells, the annual reduction rate in telomere length was very low. These findings support the hypothesis that germ cells in the esophageal epithelium have a mechanism to lengthen telomeres.


Kinetics of collagen crosslinking in adult bovine articular cartilage.
Determine the kinetics of collagen crosslinking in adult bovine articular cartilage explants using radiolabel pulse-chase studies. Explant cultures of adult bovine articular cartilage were radiolabeled with [14C]lysine in medium including fetal bovine serum and ascorbate, and then maintained for chase periods up to 28 days. In some samples, beta-aminopropionitrile (BAPN) was included during chase to inhibit lysyl oxidase-mediated collagen crosslinking. Tissue was hydrolyzed and analyzed for [14C]metabolites in the forms of lysine, hydroxylysine, dehydrodihydroxylysinonorleucine (DeltaDHLNL), and hydroxylysyl pyridinoline (HP). Explant cultures of adult bovine articular cartilage metabolized lysine into hydroxylysine and the collagen crosslinks, DeltaDHLNL and HP. During chase, [14C]hydroxylysine maintained steady-state levels, [14C]DHLNL rose to a plateau, and [14C]HP increased gradually. Addition of BAPN inhibited formation of [14C]DHLNL. Analysis of raw data and that normalized to [14C]hydroxylysine gave characteristic time constants for formation of DeltaDHLNL and HP crosslinks of 1-2 and 7-30 days, respectively. The distribution of [14C]lysine metabolites in collagen crosslinks was described by peak values in [14C]DHLNL/[14C]hydroxylysine of 0.047-0.064 and in [14C]HP/[14C]hydroxylysine of 0.03. Collagen crosslinks form in cartilage explants in vitro according to the classical lysyl oxidase-mediated pathway.


[Emergency Triage. An Overview].
In emergency departments, patients present with different severities of diseases and traumatic injuries. However, patients with severe and life-threatening conditions compete for the same resources such as personal and structure. As a general rule, each patient should receive immediate diagnostic and treatment, independent of his or her severity of disease or traumatic injury. However, an unexpected number of patients presenting to the emergency department at the same time may exceed available resources. Thus, waiting times will occur and management of patients may be impeded. As a consequence, patients with diseases or traumatic injuries with a need for time-critical management, have to be detected at the time of presentation. After categorization, patients have to be prioritized and guided to the correct place of treatment ("triage"). Starting in Australia and the United States, nurse-driven triage systems have been introduced in the emergency departments. Aim of triage is to correctly identify at increased risk of death and guide them to rapid and correct treatment. In Germany, two five-level triage systems have been introduced: Manchester Triage System (MTS) and Emergency Severity Index (ESI). We give an overview of these risk assessment tools and discuss pros and cons. In addition, new options such as "team triage" and a combination with "Early Warning Scores" are reported. In summary, nurse-driven triage is an instrument to improve patient safety in emergency medicine. A structured and systematic triage of patients using validated triage assessment tools are recommended from national and international societies of emergency medicine. Therefore, nurse-driven triage is also a must in Germany.


Taking a rational approach in the treatment of avian mycobacteriosis.
Treatment for avian mycobacteriosis is still in its infancy and based on extrapolations from human medicine. The optimum drug choice, dose, or length of treatment has yet to be determined for most exotic animal species. Treatment should include multiple drugs for extended periods of time with appropriate monitoring of both drug levels and overall animal health. Risk to owners and handlers needs to be minimized through appropriate identification of the species of mycobacteri causing disease. More research is necessary on the pharmacokinetics of these drugs in other animal species and antibiotic resistance. Currently, euthanasia remains the most common action in the face of active mycobacteriosis.


Effects of forest harvesting nutrient removals on soil nutrient reserves.
Intensive harvesting of native eucalypt forests is carried out in the Eden area in the south east coastal region of New South Wales, Australia. Soil nutrient capital and nutrient removals in forest harvesting were estimated together with potential impacts of these removals on the nutrient capital balance. Soils were anlysed from eighty sites for phosphorus fractions, including organic phosphorus fractions, and total and exchangeable cations. Based on typical forest harvesting systems, it was found that 3-4 kg phosphorus would be removed per hectare. Due to equilibrium between the various soil phosphorus components, depletion would not be solely from the more available pools. It is expected that at least four forest rotations (320 years) would be required before any detectable change would occur, within forest communities. A similar depletion estimate was calculated for the potentially most vulnerable cation, calcium. The other nutrient cations, magnesium and potassium had considerably greater reserves.


Identity of angiotensinogen precursors of rat brain and liver.
Angiotensin modifies a number of functions of the central nervous system (CNS) including blood pressure regulation, thirst, and the secretion of the antidiuretic and adrenocorticotropic hormones. Whereas peripheral administration of angiotensin produces these effects, accumulating evidence implicates a brain angiotensin system which operates independently of the circulating renin-angiotensin system (reviewed in refs 1-3). All components of the renin-angiotensin system have been identified in the brain, but there is considerable controversy over whether these components are synthesized in the brain or derived from plasma and the extent to which these components interact to generate angiotensin in the brain in vivo. Whereas many different enzymes are able to cleave angiotensinogen (renin substrate) to release angiotensin I (AI) or angiotensin II (AII), angiotensinogen is the only known precursor of AI and AII. Definitive evidence for an independent brain angiotensin system requires the demonstration that the brain synthesizes an angiotensin precursor. Here we report the identification of angiotensinogen mRNA in rat brain by cell-free translation and show that the brain angiotensinogen precursors are identical to those previously identified for liver, the source of plasma angiotensinogen.


The management of female infertility by tubal microsurgical reconstruction: a ten year review.
The outcome of 368 consecutive tubal microsurgical operations excluding reversal of sterilizations, over a ten year period at the Rotunda Hospital, is presented. One hundred and ten conceptions resulted giving a pregnancy rate of 29.9% per procedure. The 'take home baby rate' was 24.2%, with miscarriage and ectopic pregnancy rates of 2.7% and 3% respectively.


KMUP-1 activates BKCa channels in basilar artery myocytes via cyclic nucleotide-dependent protein kinases.
This study investigated whether KMUP-1, a synthetic xanthine-based derivative, augments the delayed-rectifier potassium (K(DR))- or large-conductance Ca2+-activated potassium (BKCa) channel activity in rat basilar arteries through protein kinase-dependent and -independent mechanisms. Cerebral smooth muscle cells were enzymatically dissociated from rat basilar arteries. Conventional whole cell, perforated and inside-out patch-clamp electrophysiology was used to monitor K+- and Ca2+ channel activities. KMUP-1 (1 microM) had no effect on the K(DR) current but dramatically enhanced BKCa channel activity. This increased BKCa current activity was abolished by charybdotoxin (100 nM) and iberiotoxin (100 nM). Like KMUP-1, the membrane-permeable analogs of cGMP (8-Br-cGMP) and cAMP (8-Br-cAMP) enhanced the BKCa current. BKCa current activation by KMUP-1 was markedly inhibited by a soluble guanylate cyclase inhibitor (ODQ 10 microM), an adenylate cyclase inhibitor (SQ 22536 10 microM), competitive antagonists of cGMP and cAMP (Rp-cGMP, 100 microM and Rp-cAMP, 100 microM), and cGMP- and cAMP-dependent protein kinase inhibitors (KT5823, 300 nM and KT5720, 300 nM). Voltage-dependent L-type Ca2+ current was significantly suppressed by KMUP-1 (1 microM), and nearly abolished by a calcium channel blocker (nifedipine, 1 microM). In conclusion, KMUP-1 stimulates BKCa currents by enhancing the activity of cGMP-dependent protein kinase, and in part this is due to increasing cAMP-dependent protein kinase. Physiologically, this activation would result in the closure of voltage-dependent calcium channels and the relaxation of cerebral arteries.


Group I introns from Zygomycota: evolutionary implications for the fungal IC1 intron subgroup.
The origins of fungal group I introns within nuclear small-subunit (nSSU) rDNA are enigmatic. This is partly because they have never been reported in basal fungal phyla (Zygomycota and Chytridiomycota), which are hypothesized to be ancestral to derived phyla (Ascomycota and Basidiomycota). Here we report group I introns from the nSSU rDNA of two zygomycete fungi, Zoophagus insidians (Zoopagales) and Coemansia mojavensis (Kickxellales). Secondary structure analyses predicted that both introns belong to the IC1 subgroup and that they are distantly related to each other, which is also suggested by different insertion sites. Molecular phylogenetic analyses indicated that the IC1 intron of Z. insidians is closely related to the IC1 intron inserted in the LSU rDNA of the basidiomycete fungus Clavicorona taxophila, which strongly suggests interphylum horizontal transfer. The IC1 intron of C. mojavensis has a low phylogenetic affinity to other fungal IC1 introns inserted into site 943 of nSSU rDNA (relative to E. coli 16S rDNA). It is noteworthy that this intron contains a putative ORF containing a His-Cys box motif in the antisense strand, a hallmark for nuclear-encoded homing endonucleases. Overall, molecular phylogenetic analyses do not support the placement of these two introns in basal fungal IC1 intron lineages. This result leads to the suggestion that fungal IC1 introns might have invaded or been transferred laterally after the divergence of the four major fungal phyla.


TFG mutation induces haploinsufficiency and drives axonal Charcot-Marie-Tooth disease by causing neurite degeneration.
TFG-related axonal Charcot-Marie-Tooth (CMT) disease is a late-onset, autosomal dominant, hereditary motor, and sensory neuropathy characterized by slowly progressive weakness and atrophy of the distal muscles. The objective of this study was to determine the common pathogenic mechanism of TFG-related CMT type 2 (CMT2) caused by different mutations and establish a direct association between TFG haploinsufficiency and neurodegeneration. Three individuals carrying the TFG p.G269V mutation but with varying disease durations were studied. The effect of the p.G269V mutation was confirmed by analyzing protein samples extracted from the blood of two individuals. The functional consequences of both CMT2 mutant gene products were evaluated in vitro. The effect of TFG deficiency in the nervous system was examined using zebrafish models and cultured mouse neurons. Overexpression of p.G269V TFG failed to enhance soluble TFG levels by generating insoluble TFG aggregates. TFG deficiency disrupted neurite outgrowth and induced neuronal apoptosis both in vivo and in vitro and further impaired locomotor capacity in zebrafish, which was consistent with the phenotype in patients. Wnt signaling was activated as a protective factor in response to TFG deficiency. CMT2-related TFG mutation induces TFG haploinsufficiency within cells and drives disease by causing progressive neurite degeneration.


Focused Neuromuscular Ultrasound Approach for the Diagnosis of Chronic Inflammatory Demyelinating Polyneuropathy.
Previous ultrasonographic studies of individuals with chronic inflammatory demyelinating polyneuropathy (CIDP) have shown nerve enlargement at several sites. This prospective study compares only the bilateral median and ulnar nerves of individuals with CIDP with reference values to determine the clinical usefulness of this focused approach as a diagnostic tool. The cross-sectional area, echogenicity, and vascularity of the bilateral median and ulnar nerves of 25 subjects with CIDP were measured using ultrasound. Nineteen had typical CIDP based on the European Federation of Neurological Societies and the Peripheral Nerve Society guidelines, whereas six had atypical CIDP and were diagnosed based on clinical impression. Focal nerve enlargement was found in at least one segment in all subjects. Subjects with typical CIDP had larger cross-sectional areas compared with subjects with atypical CIDP. A focused ultrasound study, involving only the median and ulnar nerves, is sensitive for the detection of nerve enlargement in CIDP. Measuring the cross-sectional area of the median and ulnar nerves is clinically feasible and may help establish the diagnosis of CIDP.


Critical evaluation of biodegradation studies on synthetic plastics through a systematic literature review.
Increasing amounts of plastic waste in the environment and their fragmentation into smaller particles known as microplastics (particles, <5mm) have raised global concerns due to their persistency in the environment and their potential to act as vectors for harmful substances or pathogenic microorganisms. One possible solution to this problem is biodegradation of plastics by microorganisms. However, the scientific information on plastic-degrading microorganisms is scattered across different scientific publications. We conducted a systematic literature review (SLR) with predefined criteria using the online databases of Scopus and Web of Science to find papers on bacterial biodegradation of synthetic petroleum-based polymers. The aims of this SLR were to provide an updated list of all of the currently known bacteria claimed to biodegrade synthetic plastics, to determine and define the best methods to assess biodegradation, to critically evaluate the existing studies, and to propose directions for future research on polymer biodegradation in support of more rapid development of biodegradation technologies. Most of the bacteria identified here from the 145 reviewed papers belong to the phyla Proteobacteria, Firmicutes and Actinobacteria, and most were isolated from contaminated sites, such as landfill sites. Just under a half of the studies (44%) investigated the biodegradability of polyethylenes and derivates, particularly low-density polyethylenes. The methods used to monitor the biodegradation were mainly scanning electron microscopy and Fourier-transform infrared spectroscopy. We propose that: (1) future research should focus on biodegradation of microplastics arising from the most common pollutants (e.g. polyethylenes); (2) bacteria should be isolated from environments that are permanently contaminated with plastics; and (3) a combination of different observational methods should be used to confirm bacterial biodegradation of these plastics. Finally, when reporting, researchers need to follow standard protocols and include all essential information needed for repetition of the experiments by other research groups.


Use of covered self-expandable stents for benign colorectal disorders in children.
There is a lack of experience with covered self-expandable stents for benign colorectal disorders in children. Five children (4M, 1F) with a median age of 5years (range, 6months-9years) who underwent treatment with covered self-expandable plastic (SEPSs) or self-expandable metal stents (SEMSs) for a benign colorectal condition between April 2005 and November 2013 were recruited to this retrospective study. Etiologies included: anastomotic stricture with (n=1) or without (n=3) simultaneous enterocutaneous fistula, as well as an anastomotic leak associated with enterocutaneous fistula (n=1). All children suffered from either Hirschsprung's disease (n=3) or total colonic aganglionosis (Zuelzer-Wilson syndrome) (n=2). Median duration of individual stent placement was 23days (range, 1-87days). In all cases up to five different stents were placed over time. At follow-up two patients were successfully treated without further intervention. In another patient the anastomotic stricture resolved fully, but a coexisting enterocutaneous fistula persisted. Overall, three patients did not improve completely following stenting and required definite surgery. Stent-related problems were noted in all cases. There was one perforation of the colon at stent insertion. Further complications consisted of stent dislocation (n=4), obstruction (n=1), formation of granulation tissue (n=1), ulceration (n=1) and discomfort (n=3). Covered self-expandable stents enrich the armamentarium of interventions for benign colorectal disorders in children including anastomotic strictures and intestinal leaks. A stent can be applied either as an emergency procedure (bridge to surgery) or as an adjuvant treatment further to endoscopy and dilatation. Postinterventional problems are frequent but there is a potential for temporary or definite improvement following stent insertion.


GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.
We have used the Escherichia coli beta-glucuronidase gene (GUS) as a gene fusion marker for analysis of gene expression in transformed plants. Higher plants tested lack intrinsic beta-glucuronidase activity, thus enhancing the sensitivity with which measurements can be made. We have constructed gene fusions using the cauliflower mosaic virus (CaMV) 35S promoter or the promoter from a gene encoding the small subunit of ribulose bisphosphate carboxylase (rbcS) to direct the expression of beta-glucuronidase in transformed plants. Expression of GUS can be measured accurately using fluorometric assays of very small amounts of transformed plant tissue. Plants expressing GUS are normal, healthy and fertile. GUS is very stable, and tissue extracts continue to show high levels of GUS activity after prolonged storage. Histochemical analysis has been used to demonstrate the localization of gene activity in cells and tissues of transformed plants.


Continuous intercostal nerve block for pain relief after lumbar incision.
To relieve postoperative pain along a lumbar incision in 9 patients the intercostal nerves were blocked with catheters for continuous epidural anesthesia. The catheters were inserted near the intercostal nerves, above and beneath the incision, just before the wound was closed and 0.25% bupivacaine hydrochloride solution was infused periodically through the catheters. With this technique 5 of 9 patients had a satisfactory analgesic effect and could breathe deeply or cough without pain. The other 4 patients did not have satisfactory results and this was believed to be owing to inadequate insertion of the catheters. None of the patients had any complications. The technique is simple and can produce an analgesic effect repeatedly without causing pain for the patient.


Long-term renal function in heart transplant children on cyclosporine treatment.
Renal function deterioration is a reason of concern in heart transplantation. Our aim was to evaluate long-term renal function in heart transplant children on cyclosporine (CsA) treatment and to investigate the effect of several variables possibly involved in renal function deterioration. Creatinine clearances were retrospectively reviewed in 50 children (median follow 99.7 months after heart transplant). Gender, age, and body weight at transplant, rejection episodes, CsA cumulative dose, and trough levels were analyzed. After an initial increase of the glomerular filtration rate (GFR), renal function worsened in most patients; 28% of the children developed renal insufficiency (defined as GFR <80 ml/min per 1.73 m2), which was already evident in the first 3 years. Neither CsA dose, trough levels, nor other patient characteristics were found to be associated with renal function deterioration. In this study renal failure occurred in one-third of the patients. The lack of association of CsA with renal insufficiency may be explained by several reasons, including the limitations of the retrospective design of the study. However, it is possible that the nephrotoxic effect of CsA is more likely to occur in a set of predisposed patients. These must be soon identified to evaluate early a calcineurin inhibitor-sparing strategy.


All in a scent - naphthalene dependence confined to pregnancy: a case report.
Inhalant or volatile substance use is a health issue with significant medical and psychiatric sequelae. Inhalants or volatile substances are volatile organic substances found in domestic and commercial products which are inhaled to obtain pleasurable effects. They are easily accessible, cheap, and legal. Common inhalants are spray, paints, glue and shoe polish whilst naphthalene or 'mothball' abuse is reported less commonly. We report a case of a 29-year-old female who inhaled and ingested naphthalene during her pregnancy. This case report is unique because the dependence on naphthalene was confined to pregnancy and resolved as soon as she delivered. This brings up the question whether pregnancy in general increases the risk of substance dependence in vulnerable populations or whether the dependence in this patient during pregnancy is due to individual factors. The patient we report is a 29-year-old female who developed a strong desire to inhale mothballs during her third pregnancy. The pattern of use started in the first trimester meeting the criteria for dependence syndrome and resolved completely by the second day following delivery. She had features suggestive of harmful use in her second pregnancy as well. The case report emphasizes that pregnant women should be screened for psychoactive substance use. Equally important is the need for adequate psychoeducation about the myths and cultural beliefs associated with pregnancy-related cravings and the potentially devastating consequences of harmful cravings on the neonate and the mother. The case highlights how chemicals used in day-to-day activities can lead to dependence.


Challenges encountered in comparing international policy responses to COVID-19 and their effects.
A variety of policies have been implemented around the world in response to the COVID-19 pandemic. This study originally aimed to identify and compare policy responses of different countries and their effects on the pandemic. It quickly evolved into an identification of the heterogeneity among existing policies and the challenges in making meaningful comparisons of the impact of these policies. The process of collecting and comparing data from different sources was analysed through inductive thematic analysis to understand the obstacles that impede research designed to compare COVID-19 data and related policies. We identified the following obstacles: (1) no single reputable source of information and too much noise; (2) a lack of standards for how to measure and report data across countries; (3) variations in the content, implementation and enforcement of policies; and (4) politics, instead of science, leading the efforts in pandemic management. Heterogeneity in existing policies makes it challenging to compare the effects of various policies worldwide on the COVID-19 pandemic. Our findings call for an automatically updated informatics infrastructure across the globe for collecting and maintaining standardized data from multiple sources. There is a strong need for steadfast utilization of scientific and technical experts to inform and influence health policy. Increased investment in public health and emergency planning is essential to overcome the current pandemic, as well as future public health emergencies. Focused leadership and collaboration from world leaders in a unified mission to decrease the mortality and morbidity of the COVID-19 pandemic is imperative.


A Teething Problem: Artefactual X-Ray Appearances of Odontoid Fracture due to Superimposed Incisor.
We report a case of artefactual C2 fracture caused by a superimposed incisor tooth in a seven-year-old boy. CT refuted the diagnosis. Trauma clinicians should be aware of this entity to guide correct interpretation of trauma X-rays.


The interaction of p-nitrophenyl carbamate with urease.
1. p-Nitrophenyl carbamate and thiourea have been shown to be substrates for urease (urea amidohydrolase, EC 18.104.22.168) 2. Urease has been shown to have a lower Km, 0.67 mM, with p-nitrophenyl carbamate than with urea, 2.0 mM. 3. The V of urease for the hydrolysis of urea, p-nitrophenyl carbamate and thiourea has been shown to be the same, indicating a common rate-limiting step. 4. A mechanism has been proposed for urease where the initial rate-limiting step is the release of a molecule of ammonia from the substrates.


First record of Glabellula Bezzi (Diptera, Mythicomyiidae: Glabellulinae) in South America, with description of three new species from Brazil.
Studying material recently collected in Brazil, the authors found three new species of Glabellula (G. catiapatiuae sp. nov., G. rafaelae sp. nov., G. sarahae sp. nov.) that are described and illustrated herein. An identification key to the Brazilian species is also presented. This paper presents the first records of Glabellula in South America and the presence, for the first time, of the subfamily Glabellulinae in Brazil.


Theta synchronization in the human EEG and episodic retrieval.
Event-related desynchronization (ERD) and synchronization (ERS) was measured during episodic retrieval in a specially designed recognition task which forces subjects to avoid semantic search strategies. ERD represents the percentage of a decrease, ERS an increase in band power. The results show that only the theta band differentiates between good and bad episodic memory performers and that good performance is related to a large degree of theta synchronization. The delta and alpha bands did not yield significant effects. Topographical differences in theta ERS reveal that good performers use primarily their right hemisphere to retrieve episodic information. This finding agrees with respective results from PET studies.


DNA damage induced by a chromium(III) Schiff base complex is reversible under physiological condition.
[Cr(naphen)(H2O)(2)]+, where naphen is 1,2-bis(naphthylideneamino)ethane having the basic salen moiety, has been characterized structurally. [Cr(naphen)(H2O)(2)]+, which has an extended aromatic system and binds with calf thymus DNA (CT DNA) intercalatively, has been found to promote DNA cleavage in the presence of biological reductant such as ascorbate and oxidant like hydrogen peroxide. Results of electron paramagnetic resonance (EPR) experiments suggest involvement of hydroxyl radicals in the oxidative cleavage of DNA in the presence of the Cr(III) complex and hydrogen peroxide. The cell viability study on nicked DNA by [Cr(naphen)(H2O)(2)]+ has shown that the damage brought about to DNA could be repaired by Escherichia coli DNA repair enzymes.


Xenon enhancement for computed tomography of the spinal cord.
The spinal cord is not consistently shown by plain computed tomography or with iodide enhancement. Xenon enhancement increases the attenuation of the spinal cord and visualization is improved considerably; fluid-filled cavities, including syringohydromyelia, do not enhance and are clearly defined.


Maternal sepsis: a Scottish population-based case-control study.
To describe the risk of maternal sepsis associated with obesity and other understudied risk factors such as operative vaginal delivery. Population-based, case-control study. North NHS region of Scotland. All cases of pregnant, intrapartum and postpartum women with International Classification of Disease-9 codes for sepsis or severe sepsis recorded in the Aberdeen Maternal and Neonatal Databank (AMND) from 1986 to 2009. Four controls per case selected from the AMND were frequency matched on year-of-delivery. Cases and controls were compared; significant variables from univariable regression were adjusted in a multivariable logistic regression model. Dependent variables were uncomplicated sepsis or severe ('near-miss') sepsis. Independent variables were demographic, medical and clinical delivery characteristics. Unadjusted and adjusted odds ratios (OR) with 95% confidence intervals (95% CI) are reported. Controlling for mode of delivery and demographic and clinical factors, obese women had twice the odds of uncomplicated sepsis (OR 2.12; 95% CI 1.14-3.89) compared with women of normal weight. Age <25 years (OR 5.15; 95% CI 2.43-10.90) and operative vaginal delivery (OR 2.20; 95% CI 1.02-4.87) were also significant predictors of sepsis. Known risk factors for maternal sepsis were also significant in this study (OR for uncomplicated and severe sepsis respectively): multiparity (OR 6.29, 12.04), anaemia (OR 3.43, 18.49), labour induction (OR 3.92 severe only), caesarean section (OR 3.23, 13.35), and preterm birth (OR 2.46 uncomplicated only). Obesity, operative vaginal delivery and age <25 years are significant risk factors for sepsis and should be considered in clinical obstetric care.


Glycolytic Profiling of Mouse Embryonic Stem Cells (mESCs).
Mouse embryonic stem cells (mESCs) can be captured in vitro in different pluripotency states through media modulation, mimicking their natural environment during early embryo development. As highly proliferative cells, mESCs prefer to use glycolysis to support the energetic and biosynthetic demands, even in the presence of oxygen. Indeed, glycolysis can not only supply ATP at a much faster rate, when compared to other catabolic pathways, but also provides biosynthetic substrates to meet anabolic requirements. Considering that ESCs cultured in different media conditions display distinct metabolic requirements, it is of utmost importance to have a robust metabolic characterization methodology to understand how subtle metabolic variations may be coupled to ESC identity. Here we describe how to profile the glycolytic activity of naive mouse ESC, using the established Seahorse XFe24 Live-cell Metabolic Assay. This may be a useful protocol for understanding how the glycolytic function of mESCs changes in certain circumstances and how is it coupled to diverse pluripotency/differentiation phenotypes.


Pharmacologic denervation of frown muscles enhances baseline expression of happiness and decreases baseline expression of anger, sadness, and fear.
Individuals with intense frowning are commonly perceived as expressing negative emotions. Anger, fear, and sadness are associated with corrugator ("frown") muscle activity. We sought to study how faces were perceived by others after denervation of frown muscles with localized botulinum toxin injections for treatment of facial frown lines. Facial photographs were taken from volunteers before and after botulinum toxin injection. These photographs were shown to viewers who were naive to the procedure and asked to rate the expressed intensity of anger, sadness, fear, and happiness. As reference for this task we used a standard set of pictures of facial affect displaying different intensity levels for each emotion tested. Of 40 viewers, 39 were able to discriminate different intensity levels (0%, 25%, 50%, 75%, and 100%) of emotional affect in the control task. According to their ratings faces with denervated frown muscle activity expressed relatively less anger (-40%), fear (-49%), sadness (-10%), and more happiness (+71%). Frown muscle activity is essential for both negative and positive emotional expressions. Temporary denervation using botulinum toxin enhances the facial expression of positive emotion resulting in a shift rather than a loss of facial affect.


Heuristic approach uncovering biological significance of gene-lifestyle interactions in cardiometabolic traits.
Gene-lifestyle interaction studies using GWAS data contribute to a better understanding of individual responses to environmental exposures. Herein, we aimed at assessing the biological significance of overlapping genes reported in gene-lifestyle interaction studies in cardiometabolic health. A heuristic analysis of genes reporting significant interactions related to cardiometabolic traits was performed to determine the biological pathways common to the different traits. A total of 873 genes were analysed. Fine and condensed phenotypic solutions were obtained from overlapping genes common to more than one trait. This study revealed significant metabolic pathways associated with the impact of gene-environment interactions on cardiometabolic risk.


Temporal spacing of breeding activity in the natterjack toad, Bufo calamita.
The reproductive behaviour of the toad Bufo calamita was studied in a large population in the northern Rhineland, Germany. Toads bred at four spatially separated but neighbouring areas including 34 breeding ponds. Three methods were applied to monitor quantitatively the intensity of breeding and migratory activity: capture-mark-recapture, a mechanical tracking device, and radio-telemetry. In 1987 a twofold temporal spacing of breeding activity was observed: (1) division of the reproductive period into early, main, and late breeding periods; (2) subdivision of each breeding period into several short calling periods. Since weather conditions were favourable during most of the reproductive period, environmental factors contributed little (23.4%-41.0%) to the temporal variation in breeding intensity as shown by multivariate statistics. The period of vocalization (April to August) included the activity of three distinct male groups. The first group called from April to mid-May (2 calling periods), the second from May to early July (5 calling periods) and the third from mid-July to August (2 calling periods). Within a calling period most males stayed near their calling sites and migratory activity was low. Extensive movements were detected when a new group of males immigrated from the feeding regions to the breeding areas and males that had been calling previously emigrated, at least partly. However, exchange of males among the neighbouring breeding choruses were clearly audible. It is suggested that the energetic costs of vocalization impeded continuous calling activity in individuals and accounted for the periodical variations in chorus size. The short-time variations of breeding activity therefore probably reflect the physiological limits of toads in a situation where the weather permits continuous reproductive activity.


Substitution of Ala-251 of the D1 reaction centre polypeptide with a charged residue results in impaired function of photosystem II.
Ala-251 in the membrane-parallel helix in the D-E loop of the D1 polypeptide close to the Q(B) pocket of photosystem II (PS II), was mutated to aspartate (D), lysine (K), leucine (L) or serine (S) in Synechocystis 6803. O2 evolution rates (H2O-->DCBQ; 2,6-dichloro-p-benzoquinone) of A251D, A251L and A251S were lower, being 38, 16, 62 and 70%, respectively, of that of the control, and there was an even more drastic impairment of O2 evolution when measured from H2O to DMBQ (2,5-dimethyl-p-benzoquinone), demonstrating modifications in the Q(B) pocket. However, in all other mutants but A251K, the Q(B) function could sustain O2 evolution at a level high enough to support photosynthetic growth. The mutant A251S, carrying a substitution of alanine for a chemically quite similar residue serine, was less severely affected. Substitution by a positively charged residue drastically delayed chlorophyll a fluorescence relaxation in the non-photosynthetic strain A251K, implying strong impairment of Q(A)-to-Q(B) electron transfer. Delay of fluorescence relaxation was clear in A251D as well, carrying a substitution of alanine for a negatively charged residue. The effects of the substitutions of A251 demonstrate the importance of this residue of the D1 polypeptide in the conformation of the acceptor side of PS II and, accordingly, the effect on the acceptor-side function of PS II was very clear. Nevertheless, the tolerance of PS II activity to high-light-induced photoinhibition in vivo and the subsequent D1 degradation were not much impaired in any of the photosynthetic mutant strains as compared to the control.


Primary structure of PDC-109, a major protein constituent of bovine seminal plasma.
The two major protein components of bovine seminal plasma, PDC-109 and BSP I, have been purified by gel filtration, partition chromatography and reverse-phase high performance liquid chromatography from an 86% ethanol precipitate of bovine seminal plasma ejaculate. The complete 109-residue amino acid sequence of PDC-109 has been established by automated Edman degradation of the intact peptide as well as its proteolytic digestion and cyanogen bromide cleavage fragments. The 12,774 dalton structure has two structurally similar domains of 38 and 41 amino acids, each containing two disulfide bonds.


Influence of vertical transport on chemical evolution of dicarboxylic acids and related secondary organic aerosol from surface emission to the top of Mount Hua, Northwest China.
Dicarboxylic acids are strong hygroscopic organic compounds in the atmosphere, and thus significantly affect the cloud formation process and radiative forcing on a regional scale. So far, the evolution of dicarboxylic acids during vertical transport from the surface to the mountaintop has yet to be explicitly understood. In this study, the molecular distribution and stable carbon isotopic (δ13C) compositions of dicarboxylic acids and related organic compounds (DCRCs) in PM2.5 were measured simultaneously at the top (c. 2060 m a.s.l.) and foot (c. 400 m a.s.l.) of Mount (Mt.) Hua during the summer of 2020. Due to the strong anthropogenic emissions at ground level, the concentrations of DCRCs at foot of Mt. Hua were generally higher than those at the top. Oxalic acid (C2) was the predominant diacid in both sites, whose concentrations at foot and top of Mt. Hua were 87-852 and 40-398 ng m-3, respectively. Ratios of adipic acid to azelaic acid (C6/C9), phthalic aid to azelaic acid (pH/C9), glyoxal to methylglyoxal (Gly/mGly), and lower δ13C values (-21.0 ± 2.3 ‰ and - 21.9 ± 2.7 ‰) of C2 indicated that the contributions of anthropogenic sources to DCRCs in PM2.5 in the mountain region are more significant than biogenic sources. Aerosols from the foot of Mt. Hua could affect the atmosphere on the top of the mountain via vertical transport under the influence of daytime valley wind, even though the altitude of Mt. Hua is beyond the boundary layer most of time. The value δ13C of C2 is linearly correlated with C2/mGly, C2/pyruvic acid (Pyr), C2/glyoxylic acid (ωC2) at the top of the mountain, and C2/Gly, C2/ωC2 at the foot of the mountain, indicating that the formation pathway of C2 is mGly-Pyr-ωC2-C2 at the top of Mt. Hua and Gly-ωC2-C2 at the foot of Mt. Hua.


Biomechanical and transcriptional evidence that smooth muscle cell death drives an osteochondrogenic phenotype and severe proximal vascular disease in progeria.
Hutchinson-Gilford Progeria Syndrome results in rapid aging and severe cardiovascular sequelae that accelerate near end-of-life. We found a progressive disease process in proximal elastic arteries that was less evident in distal muscular arteries. Changes in aortic structure and function were then associated with changes in transcriptomics assessed via both bulk and single cell RNA sequencing, which suggested a novel sequence of progressive aortic disease: adverse extracellular matrix remodeling followed by mechanical stress-induced smooth muscle cell death, leading a subset of remnant smooth muscle cells to an osteochondrogenic phenotype that results in an accumulation of proteoglycans that thickens the aortic wall and increases pulse wave velocity, with late calcification exacerbating these effects. Increased central artery pulse wave velocity is known to drive left ventricular diastolic dysfunction, the primary diagnosis in progeria children. It appears that mechanical stresses above ~ 80 kPa initiate this progressive aortic disease process, explaining why elastic lamellar structures that are organized early in development under low wall stresses appear to be nearly normal whereas other medial constituents worsen progressively in adulthood. Mitigating early mechanical stress-driven smooth muscle cell loss/phenotypic modulation promises to have important cardiovascular implications in progeria patients.


Hyperspectral phenotyping on the microscopic scale: towards automated characterization of plant-pathogen interactions.
The detection and characterization of resistance reactions of crop plants against fungal pathogens are essential to select resistant genotypes. In breeding practice phenotyping of plant genotypes is realized by time consuming and expensive visual rating. In this context hyperspectral imaging (HSI) is a promising non-invasive sensor technique in order to accelerate and to automate classical phenotyping methods. A hyperspectral microscope was established to determine spectral changes on the leaf and cellular level of barley (Hordeum vulgare) during resistance reactions against powdery mildew (Blumeria graminis f.sp. hordei, isolate K1). Experiments were conducted with near isogenic barley lines of cv. Ingrid, including the susceptible wild type (WT), mildew locus a 12 (Mla12 based resistance) and the resistant mildew locus o 3 (mlo3 based resistance), respectively. The reflection of inoculated and non-inoculated leaves was recorded daily with a hyperspectral linescanner in the visual (400 - 700 nm) and near infrared (700 - 1000 nm) range 3 to 14 days after inoculation. Data analysis showed no significant differences in spectral signatures between non-inoculated genotypes. Barley leaves of the near-isogenic genotypes, inoculated with B. graminis f.sp. hordei differed in the spectral reflectance over time, respectively. The susceptible genotypes (WT, Mla12) showed an increase in reflectance in the visible range according to symptom development. However, the spectral signature of the resistant mlo-genotype did not show significant changes over the experimental period. In addition, a recent data driven approach for automated discovery of disease specific signatures, which is based on a new representation of the data using Simplex Volume Maximization (SiVM) was applied. The automated approach - evaluated in only a fraction of time revealed results similar to the time and labor intensive manually assessed hyperspectral signatures. The new representation determined by SiVM was also used to generate intuitive and easy to interpretable summaries, e.g. fingerprints or traces of hyperspectral dynamics of the different genotypes. With this HSI based and data driven phenotyping approach an evaluation of host-pathogen interactions over time and a discrimination of barley genotypes differing in susceptibility to powdery mildew is possible.


Synthesis, anti-amoebic activity and molecular docking simulation of eugenol derivatives against Acanthamoeba sp.
Amoebae of the genus Acanthamoeba can cause diseases such as amoebic keratitis and granulomatous amoebic encephalitis. Until now, treatment options for these diseases have not been fully effective and have several drawbacks. Therefore, research into new drugs is needed for more effective treatment of Acanthamoeba infections. Eugenol, a phenolic aromatic compound mainly derived from cloves, has a variety of pharmaceutical properties. In this study, nine eugenol derivatives (K1-K9), consisting of five new and four known compounds, were synthesized and screened for their antiamoebic properties against Acanthamoeba sp. The structure of these compounds was characterized spectroscopically by Fourier transform infrared (FTIR), Ultraviolet-Visible (UV-Vis), 1H and 13C Nuclear Magnetic Resonance (NMR) and mass spectrometer (MS). The derived molecules were screened for antiamoebic activity by determining IC50 values based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and observation of amoeba morphological changes by light and fluorescence microscopy. Most of the tested compounds possessed strong to moderate cytotoxic effects against trophozoite cells with IC50 values ranging from 0.61 to 24.83 μg/mL. Observation of amoebae morphology by light microscopy showed that the compounds caused the transformed cells to be roundish and reduced in size. Furthermore, fluorescence microscopy observation using acridine orange (AO) and propidium iodide (PI) (AO/PI) staining showed that the cells have damaged membranes by displaying a green cytoplasm with orange-stained lysosomes. Acidification of the lysosomal structure indicated disruption of the internal structure of Acanthamoeba cells when treated with eugenol derivatives. The observed biological results were also confirmed by interaction simulations based on molecular docking between eugenol derivatives and Acanthamoeba profilin. These interactions could affect the actin-binding ability of the protein, disrupting the shape and mobility of Acanthamoeba. The overall results of this study demonstrate that eugenol derivatives can be considered as potential drugs against infections caused by Acanthamoeba.


A blind reanalysis of a random subset of NCI bioassay studies: agreement between rats and mice.
There are many problems in the statistical analysis of long-term rodent studies. This paper discusses the alternative approaches to these problems suggested by the previous three papers and compares the results of the different analyses on 25 rat and mouse studies. It is concluded that many problems remain unresolved and that alternative, seemingly reasonable decision rules can give rise to dramatically different results.


Emergency separation of a xipho-omphalopagus twin in a developing country.
Female conjoined twins (thoraco-omphalopagus) were delivered by emergency caesarean section in a private nursing home. On examination, one of the twins was dead and was threatening the survival of the surviving twin (twin A). An emergency separation was performed to salvage the surviving twin.


Comparative study of viscerotropic pathogenicity of Leishmania major amastigotes and promastigotes based on identification of mitochondrial and nucleus sequences.
Experimental viscerotropic leishmaniasis is regularly caused by Leishmania tropica promastigotes. In the current investigation, the viscerotropic pathogenicities of Leishmania major amastigotes and promastigotes were compared and evaluated based on their heterogeneity traits and number of inoculated parasites in experimental mammalian hosts. Serous exudate from 50 patients was infected, 44 with L. major and 6 with L. tropica; only BALB/c mice inoculated with 1-2 × 10(4-6) L. major amastigotes manifested cutaneous lesions at the base of their tails. Five out of the 44 BALB/c mice inoculated with L. major died of sequela of viscerotropic adverse effect, while 2 × 10(6) L. major promastigotes showed viscerotropic signs in four BALB/c mice. The sequencing of the Cyt b gene showed a strain of L. major (GenBank accession number KM393221: haplotype diversity 0.9) containing two codon mutations, 86 and 126 in dead mice, whereas no significant mutant was observed in internal transcribed spacer (ITS)-ribosomal DNA (rDNA) sequences (haplotype diversity 0). Findings show that a lower dose of L. major amastigotes than promastigotes has more potential viscerotropic intensity in susceptible hosts. It illustrates that testing Cyt b as an evolutionary mitognome marker because of having its semi-conserved structure and low copy number is able to be utilized in the discrimination of new mutants.


Pax3 and Pax7 are expressed in commissural neurons and restrict ventral neuronal identity in the spinal cord.
Pax3 and Pax7 are transcription factors sharing high sequence identity and overlapping patterns of expression in particular in the dorsal spinal cord. Analysis of Pax3 and Pax7 double mutant mice demonstrates that both genes share redundant functions to restrict ventral neuronal identity in the spinal cord. In their absence, the En1 expression domain is expanded dorsally but that of Evx1 is not affected. In addition, Pax3 and Pax7 are expressed in commissural neurons and double mutant embryos exhibit highly reduced ventral commissure. Our findings reveal two distinct regulatory pathways for spinal cord neurogenesis, only one of which is dependent on Pax3/7 and 6.


Incorporation of [2-3H] ethanolamine into rat muscle phosphoglycerides.
The contribution of phosphatidylethanolamine methylation to phosphatidylcholine biosynthesis in rat muscle was investigated by studying the incorporation of [2-3H] ethanolamine. The specific radioactivities of individual molecular species of muscle phosphoglycerides were measured by a combination of argentation thin-layer chromatography and countercurrent distribution. The specific radioactivity of phosphatidylethanolamine was approximately one thousand times that of phosphatidylcholine. Amongst individual phosphatidylethanolamines, hexaenoic species possessed the highest specific radioactivities and tetraenoic the lowest. Because of the very low incorporation into phosphatidylcholine, the specific radioactivities of combined rather than of individual fractions were measured. The results indicate that the contribution of phosphatidylethanolamine methylation to the overall biosynthesis of phosphatidylcholine in muscle is of minor importance.


The in vitro effect of leptin on basal and growth hormone-releasing hormone-stimulated growth hormone secretion from the ovine pituitary gland.
We have studied the direct effect of leptin on the secretion of GH from the anterior pituitary gland of the sheep. Using primary cultures of ovine pituitary cells, leptin (10(-9)-10(-7) M) treatment for 30 min did not affect basal or GHRH (10(-7) M)-stimulated GH secretion. Following treatment for 24 h, a dose of 10(-7) M leptin stimulated basal GH secretion. In contrast, doses of 10(-7) and 10(-8) M leptin inhibited GHRH-stimulated GH secretion after 24 h of treatment. These results suggest that leptin can have long-term effects on the somatotropes, but no acute effect. Furthermore, leptin appears to have opposite effects on basal and GHRH-stimulated GH secretion.


Effects of somatostatin-14 and its analogs on the (Ca,Mg)ATPase in the rat anterior pituitary.
The effects of somatostatin-14 and its biologically active analogs, RC-160 and SMS 201-995, on (Ca,Mg)ATPase activity in vitro were studied in homogenates of anterior pituitary cells. It was found that somatostatin inhibited the (Ca,Mg)ATPase activity in the anterior pituitary and somatostatin analogs exerted slight biphasic effects on the calcium pump activity. The effects of specific brain (Ca,Mg)ATPase inhibitors, VOSO4 and LaCl3, were also studied in vitro. Neither VOSO4 nor LaCl3 enhance the inhibition of calcium pump activity caused by somatostatin. It is suggested that somatostatin may mediate its action in pituitary cells, among others, by the regulation of (Ca,Mg)ATPase activity.


Prognostic role of heart rate variability in patients with ST-segment elevation acute myocardial infarction treated by primary angioplasty.
The aim of our study was to assess the prognostic value of heart rate variability (HRV) in ST-segment elevation acute myocardial infarction (STEMI) patients treated by percutaneous transluminal coronary angioplasty (PTCA) and optimal medical therapy. We enrolled 182 consecutive patients with a first STEMI (59.1 ± 11 years; 82.4% men) treated by primary PTCA. HRV was assessed on 24-hour Holter ECG recordings before discharge and 1 and 6 months after discharge. The primary end point was the occurrence of major clinical events (MCE), defined as death or new acute myocardial infarction (AMI). At a follow-up of 42 ± 23 months, MCE occurred in 14 patients (7.6%; 3 deaths and 11 re-AMIs). HRV parameters before discharge were significantly lower in patients with MCE, with standard deviation of all RR intervals (SDNN) and very low frequency and low frequency (LF) amplitude being the most predictive variables. HRV assessed at follow-up instead did not significantly predict MCE. At multivariate analysis, only SDNN (HR 0.97; p = 0.02) and LF (HR 0.90; p = 0.04) remained significantly associated with MCE. Lower tertile SDNN and LF values were associated with a multivariate HR of 3.91 (p = 0.015) and of 2.92 (p = 0.048), respectively. Similar results were observed considering re-AMI only as the end point. In STEMI patients treated by PTCA, HRV assessed before discharge was an independent predictor of MCE and re-AMI.


Health and survival of red abalone, Haliotis rufescens, under varying temperature, food supply, and exposure to the agent of withering syndrome.
Withering syndrome (WS) is a disease of wild and cultured abalone caused by a Rickettsiales-like prokaryote (WS-RLP). This study compared the pathologic changes that occur during the progression of WS in red abalone to those caused by environmental stresses consisting of elevated temperature and food limitation and determined the impact of these stressors on WS prevalence and intensity. Farmed red abalone were administered a feed-based oxytetracycline therapeutic treatment to assure WS-RLP-free status prior to initiation of the experiment. Groups were then held in each of eight combinations of exposed vs. unexposed to WS-RLP, elevated vs. ambient temperature, and high vs. low food supply, for 447 days. Mortality was associated with starvation and disease but not elevated temperature alone. Elevated temperature significantly affected WS-RLP transmission: only 1.7% of WS-RLP- exposed abalone held at ambient temperature (12.3 degrees C) became infected compared to at least 72% of those held at elevated temperature (18.7 degrees C). Among exposed abalone at elevated temperature, fed animals exhibited greater infection prevalence but not greater infection intensity or digestive gland changes than starved animals, suggesting that abalone acquire infections by ingesting contaminated food. Food, temperature, WS-RLP exposure, and most of their interactions had significant effects on body condition and foot atrophy. Immunohistochemical detection of cell proliferation and apoptosis revealed no differences between normal digestive gland and that infected with WS-RLP. Body mass shrinkage, foot atrophy, elevated mortality, and decreased foot and digestive gland glycogen were observed in both WS-affected and starved, unexposed abalone, with the WS-RLP-exposed, starved group held at elevated temperature faring worst. Among exposed and unexposed animals, food supply but not temperature affected body mass and growth. These data demonstrate that the high morbidity and mortality exhibited by WS-RLP-infected abalone is a consequence of disease and not direct thermal stress. Drug residue analysis indicated oxytetracycline concentrations of up to 600 ppm in the digestive gland at 38 days post-treatment, an unusual degree of tissue retention of this therapeutant.


Little effect of insurance status or socioeconomic condition on disparities in minority appendicitis perforation rates.
To estimate how much of the gap in appendicitis perforation rates between minority and white children is explained by differences in socioeconomic and insurance factors. Observational analysis of hospital discharge information. The Healthcare Cost and Utilization Project database. Appendicitis perforation rates determined from the Healthcare Cost and Utilization Project database of hospital discharges from 2001 to 2008. The proportion of the gap between perforation rates explained by various patient- and hospital-level variables. There were no disparities observed in adult appendicitis perforation rates. The perforation rate for white children was 26.7%; black children, 35.5%; and Latino children, 36.5%. Gap analysis showed that only 12.0% of the difference in perforation rates between black and white children was explained by insurance status and only 12.7% of the difference between Latino and white children was explained. Income level only accounted for 7.2% of the gap for black children and 6.1% for Latino children. Age explained one-third of the gap for Latino children and one-third was not accounted for by measurable variables. Two-thirds of the difference between appendicitis perforation rates between black and white children was not explained by measurable factors. A very small amount of the gap between minority and white children's appendicitis rates is explained by the proxy factors for health insurance and poverty status that might relate to health care access. Appendicitis perforation rates are not an appropriate indicator of health care access.


The nature of prolonged word search.
Two alternative hypotheses were considered concerning the process of prolonged search for an uncommon word in response to hearing its definition. One alternative is that a conscious retrieval effort brings the target progressively closer to threshold. The second is that the retrieval process is a random neural exploration outside of conscious control. A tachistoscopic probe was devised to compare word recognition thresholds after 10 sec versus 30 sec of presumed search time. Results failed to show a difference between the two delay conditions and were interpreted as inconsistent with the first alternative but consistent with the second.


Child rearing styles, dental anxiety and disruptive behaviour; an exploratory study.
This was to explore the relation between children's dental anxiety, their behaviour during treatment and their parent's rearing style. Also the parents' preparation of the child for dental treatment was related to behaviour and parental rearing style. The parents of 100 children, referred to a secondary dental care clinic were asked to fill out the Child Rearing Practices Report (CRPR), the Child Fear Survey Schedule (CFSS) on behalf of their children and an additional questionnaire on child preparation prior to dental treatment. Four rearing styles were constructed by using the results of the Nurturance and the Restrictiveness domain of the CRPR. Dentists were asked to assess the behaviour of the child during dental habituation and dental treatment using the Venham scale. The dentists were unaware of the child's CFSS score. Parents were not present during actual dental treatment. Each child's dental anxiety was related to their behaviour displayed during dental treatment. Highly anxious children showed more disruptive behaviour than their low anxious counterparts did, especially during familiarisation. Parental rearing style was neither related to child dental anxiety prior to or to behaviour during dental treatment. Parents who use a permissive rearing style were less likely to tell their children that the dentist will not hurt them compared with authoritarian parents. Parents with an authoritative rearing style were more convinced that the behaviour of their child could be managed by the dentist than parents with a permissive and neglectful rearing style. After their children's dental rehabilitation parents were less inclined to accompany their child during treatment. Dental anxiety correlated positively with the behaviour displayed during treatment. No relation was found between parenting style and dental anxiety and behaviour during treatment. Parents showed more confidence in the child-dentist dyad after the rehabilitation of their child's teeth.


Modification of improved-genome editing via oviductal nucleic acids delivery (i-GONAD)-mediated knock-in in rats.
Improved genome-editing via oviductal nucleic acids delivery (i-GONAD) is a new technology that facilitates in situ genome-editing of mammalian zygotes exiting the oviductal lumen. The i-GONAD technology has been developed for use in mice, rats, and hamsters; however, oligonucleotide (ODN)-based knock-in (KI) is more inefficient in rats than mice. To improve the efficiency of i-GONAD in rats we examined KI efficiency using three guide RNAs (gRNA), crRNA1, crRNA2 and crRNA3. These gRNAs recognize different portions of the target locus, but also overlap each other in the target locus. We also examined the effects of commercially available KI -enhancing drugs (including SCR7, L755,507, RS-1, and HDR enhancer) on i-GONAD-mediated KI efficiency. The KI efficiency in rat fetuses generated after i-GONAD with crRNA2 and single-stranded ODN was significantly higher (24%) than crRNA1 (5%; p < 0.05) or crRNA3 (0%; p < 0.01). The KI efficiency of i-GONAD with triple gRNAs was 11%. These findings suggest that KI efficiency largely depends on the type of gRNA used. Furthermore, the KI efficiency drugs, SCR7, L755,507 and HDR enhancer, all of which are known to enhance KI efficiency, increased KI efficiency using the i-GONAD with crRNA1 protocol. In contrast, only L755,507 (15 μM) increased KI efficiency using the i-GONAD with crRNA2 protocol. None of them were significantly different. We attempted to improve the KI efficiency of i-GONAD in rats. We demonstrated that the choice of gRNA is important for determining KI efficiency and insertion and deletion rates. Some drugs (e.g. SCR7, L755,507 and HDR enhancer) that are known to increase KI efficiency in culture cells were found to be effective in i-GONAD in rats, but their effects were limited.