Cis and trans actions of the cholinesterase-like domain within the thyroglobulin dimer.
Thyroglobulin (Tg, precursor for thyroid hormone synthesis) is a large secreted glycoprotein composed of upstream regions I-II-III, followed by the approximately 570 residue cholinesterase-like (ChEL) domain. ChEL has two identified functions: 1) homodimerization, and 2) binding to I-II-III that facilitates I-II-III oxidative maturation required for intracellular protein transport. Like its homologs in the acetylcholinesterase (AChE) family, ChEL possesses two carboxyl-terminal alpha-helices. We find that a Tg-AChE chimera (swapping AChE in place of ChEL) allows for dimerization with monomeric AChE, proving exposure of the carboxyl-terminal helices within the larger context of Tg. Further, we establish that perturbing trans-helical interaction blocks homodimerization of the Tg ChEL domain. Additionally, ChEL can associate with neuroligins (a related family of cholinesterase-like proteins), demonstrating potential for Tg cross-dimerization between non-identical partners. Indeed, when mutant rdw-Tg (Tg-G2298R, defective for protein secretion) is co-expressed with wild-type Tg, the two proteins cross-dimerize and secretion of rdw-Tg is partially restored. Moreover, we find that AChE and soluble neuroligins also can bind to the upstream Tg regions I-II-III; however, they cannot rescue secretion, because they cannot facilitate oxidative maturation of I-II-III. These data suggest that specific properties of distinct Tg ChEL mutants may result in distinct patterns of Tg monomer folding, cross-dimerization with wild-type Tg, and variable secretion behavior in heterozygous patients.


The effect of adding group-based counselling to individual lifestyle counselling on changes in dietary intake. The Inter99 study--a randomized controlled trial.
Few studies have investigated the specific effect of single intervention components in randomized controlled trials. The purpose was to investigate the effect of adding group-based diet and exercise counselling to individual life-style counselling on long-term changes in dietary habits. The study was a randomized controlled intervention study. From a general Danish population, aged 30 to 60 years (n = 61,301), two random sample were drawn (group A, n = 11,708; group B, n = 1,308). Subjects were invited for a health screening program. Participation rate was 52.5%. All participants received individual life-style counselling. Individuals at high risk of ischemic heart disease in group A were furthermore offered group-based life-style counselling. The intervention was repeated for high-risk individuals after one and three years. At five-year follow-up all participants were invited for a health examination. High risk individuals were included in this study (n = 2 356) and changes in dietary intake were analyzed using multilevel linear regression analyses. At one-year follow-up group A had significantly increased the unsaturated/saturated fat ratio compared to group B and in men a significantly greater decrease in saturated fat intake was found in group A compared to group B (net change: -1.13 E%; P = 0.003). No differences were found between group A and B at three-year follow-up. At five-year follow-up group A had significantly increased the unsaturated/saturated fat ratio (net change: 0.09; P = 0.01) and the fish intake compared to group B (net change: 5.4 g/day; P = 0.05). Further, in men a non-significant tendency of a greater decrease was found at five year follow-up in group A compared to group B (net change: -0.68 E%; P = 0.10). The intake of fibre and vegetables increased in both groups, however, no significant difference was found between the groups. No differences between groups were found for saturated fat intake in women. Offering group-based counselling in addition to individual counselling resulted in small, but significantly improved dietary habits at five-year follow-up and a tendency of better maintenance, compared to individual counselling alone. The Inter99 study was approved by the local Ethics Committee (KA 98 155) and is registered with ClinicalTrials.gov (registration number: NCT00289237).


The measurement of ambient air moisture stable isotope composition for the accurate estimation of evaporative losses.
Accurate estimation of evaporative losses from a water body, using the Craig-Gordon model and the stable hydrogen and oxygen isotope composition of water, requires knowledge of the stable isotope composition of ambient air moisture. This is rarely measured in the field, and it is usually estimated assuming that recent rainfall remains in isotopic equilibrium with atmospheric moisture. However, the ambient air moisture stable isotope composition may vary significantly at different heights above the water body. In this study, we set up outdoor pan evaporation experiments and simultaneously measured the stable isotope composition of ambient moisture in the atmosphere at three different heights. Using these measurements, we calculated evaporative losses, compared them with the observed losses in the pan, and assessed the uncertainty introduced by differences in ambient moisture measurements. Three main steps in the experimental method:•Daily water sampling from the evaporation pan for analysis of stable hydrogen and oxygen isotope compositions.•Recording the stable isotope composition of ambient air moisture at three different heights using the Picarro L2130-i system over a period of experiments.•Calculating evaporative losses from the pan using the Craig-Gordon model and ambient air stable isotope composition measured at three different levels and comparing to the observed losses.


Management of Accidental Succinylcholine Ingestion: Navigating Uncharted Waters.
Accidental oral ingestion of intravenous drugs is rare and under-reported, which may lead to serious morbidity and worsen the outcome for the patient. Though oral ingestion of sedatives and opioid drugs is reported, literature regarding the ingestion of muscle relaxants and subsequent management is limited. We report an interesting case of oral intake of 10 ml (500 mg) of injection Succinylcholine, a depolarizing muscle relaxant, by a psychiatric patient awaiting electroconvulsive therapy (ECT) in the pre-procedure room. We hereby report the subsequent sequence of events along with the suggested recommendations to be followed in the case of such an eventuality. To the best of our knowledge, this is the first case report of oral ingestion of a depolarizing neuromuscular blocking drug and its subsequent consequences.


Mice and rats achieve similar levels of performance in an adaptive decision-making task.
Two opposing constraints exist when choosing a model organism for studying the neural basis of adaptive decision-making: (1) experimental access and (2) behavioral complexity. Available molecular and genetic approaches for studying neural circuits in the mouse fulfill the first requirement. In contrast, it is still under debate if mice can perform cognitive tasks of sufficient complexity. Here we compare learning and performance of mice and rats, the preferred behavioral rodent model, during an acoustic flexible categorization two-alternative choice task. The task required animals to switch between two categorization definitions several times within a behavioral session. We found that both species achieved similarly high performance levels. On average, rats learned the task faster than mice, although some mice were as fast as the average rat. No major differences in subjective categorization boundaries or the speed of adaptation between the two species were found. Our results demonstrate that mice are an appropriate model for the study of the neural mechanisms underlying adaptive decision-making, and suggest they might be suitable for other cognitive tasks as well.


Characterization of a subtropical hawksbill sea turtle (Eretmocheyles imbricata) assemblage utilizing shallow water natural and artificial habitats in the Florida Keys.
In order to provide information to better inform management decisions and direct further research, vessel-based visual transects, snorkel transects, and in-water capture techniques were used to characterize hawksbill sea turtles in the shallow marine habitats of a Marine Protected Area (MPA), the Key West National Wildlife Refuge in the Florida Keys. Hawksbills were found in hardbottom and seagrass dominated habitats throughout the Refuge, and on man-made rubble structures in the Northwest Channel near Cottrell Key. Hawksbills captured (N = 82) were exclusively juveniles and subadults with a straight standard carapace length (SSCL) ranging from 21.4 to 69.0cm with a mean of 44.1 cm (SD = 10.8). Somatic growth rates were calculated from 15 recaptured turtles with periods at large ranging from 51 to 1188 days. Mean SSCL growth rate was 7.7 cm/year (SD = 4.6). Juvenile hawksbills (<50 cm SSCL) showed a significantly higher growth rate (9.2 cm/year, SD = 4.5, N = 11) than subadult hawksbills (50-70 cm SSCL, 3.6 cm/year, SD = 0.9, N = 4). Analysis of 740 base pair mitochondrial control region sequences from 50 sampled turtles yielded 12 haplotypes. Haplotype frequencies were significantly different compared to four other Caribbean juvenile foraging aggregations, including one off the Atlantic coast of Florida. Many-to-one mixed stock analysis indicated Mexico as the primary source of juveniles in the region and also suggested that the Refuge may serve as important developmental habitat for the Cuban nesting aggregation. Serum testosterone radioimmunoassay results from 33 individuals indicated a female biased sex ratio of 3.3 females: 1 male for hawksbills in the Refuge. This assemblage of hawksbills is near the northern limit of the species range, and is one of only two such assemblages described in the waters of the continental United States. Since this assemblage resides in an MPA with intensive human use, basic information on the assemblage is vital to resource managers charged with conservation and species protection in the MPA.


Effects of polyurethane matrices on fungal tannase and gallic acid production under solid state culture.
The influence of the physical structure of polyurethane matrix as a support in a solid state culture in tannase production and gallic acid accumulation by Aspergillus niger Aa-20 was evaluated. Three different polyurethane matrices were used as the support: continuous, semi-discontinuous and discontinuous. The highest tannase production at 2479.59 U/L during the first 12 h of culture was obtained using the discontinuous matrix. The gallic acid was accumulated at 7.64 g/L at the discontinuous matrix. The results show that the discontinuous matrix of polyurethane is better for tannase production and gallic acid accumulation in a solid state culture bioprocess than the continuous and semi-discontinuous matrices.


Incidence of G-Induced Loss of Consciousness and Almost Loss of Consciousness in the Royal Air Force.
Exposure to sustained +Gz acceleration with inadequate G protection can result in G-induced loss of consciousness (G-LOC) or almost loss of consciousness (A-LOC). The UK Royal Air Force (RAF) last conducted a survey of G-LOC within their military aircrew in 2005 with interventions subsequently introduced. The aim of this study was to repeat the 2005 survey in order to evaluate the impact of those interventions. An anonymous questionnaire requesting details of G-LOC and A-LOC events was mailed to all RAF pilots (N = 1878) and weapons systems operators (WSOs) (N = 473), irrespective of aircraft currently flown. The questionnaire was returned by 809 aircrew (34.4% response rate). There were 120 (14.8%) aircrew who reported at least one episode of G-LOC and 260 (32.2%) reported at least one episode of A-LOC. The reported prevalence of G-LOC in the previous 2005 survey was 20.1% (N = 454). There was an increased reporting of G-LOC in the Hawk, Tucano, and Grob Tutor aircraft, with 5 G-LOC and 19 A-LOC events reported in the Grob Tutor compared to none in 2005. The prevalence of reported G-LOC has decreased in the surveyed populations, which may be due to the introduction of centrifuge training, but also may be influenced by patterns of G exposure and other factors. Scope for further reduction remains through correct execution of the anti-G straining maneuver (AGSM) with centrifuge training early in flying training and use of a structured conditioning program to increase the general strength of muscles involved in the AGSM.Slungaard E, McLeod J, Green NDC, Kiran A, Newham DJ, Harridge SDR. Incidence of G-induced loss of consciousness and almost loss of consciousness in the Royal Air Force. Aerosp Med Hum Perform. 2017; 88(6):550-555.


Platinum accumulation in the kidney and changes in creatinine clearance following chemotherapy with cisplatin in humans.
To evaluate the effects of repeated administration of cisplatin on kidney platinum (Pt) accumulation and renal function, Pt content in the kidney was determined in 31 autopsy cases and changes in creatinine clearance (CCr) were retrospectively assessed in 26 of 31 autopsy cases. There was no significant correlation between the cumulative dose of cisplatin and Pt content in the kidney. However, the kidney Pt level was correlated with the dose of cisplatin injected within a 7-month period prior to the patient's death. We suggest that the dose of cisplatin injected during a relatively short period before a patient's death affects Pt content in the kidney. Neither the total dose of cisplatin nor the Pt content in the kidney was significantly correlated with the decrease in CCr following chemotherapy with cisplatin.


Breaking the cycle of violence among youth living in metropolitan Atlanta: a case history of kids alive and loved.
More teenagers in the United States die from gunshot wounds than from all natural causes of disease combined. Firearm-related mortality accounts for almost half of all deaths among African American teens. Residents of central cities have the highest probability of experiencing violent crimes. This article describes an innovative community-based intervention designed to break the cycle of violence among youth in metropolitan Atlanta. The intervention, Kids Alive and Loved (KAL), emerged from the African American community as one mother's response to the violent death of her 17-year-old son. The authors describe how her response to tragedy gave birth to a culturally appropriate intervention for youth exposed to violence. This article delineates the evolution of KAL, the role of community partners in the design of the intervention, and how diffusion of innovation theory has implications for understanding the KAL approach to breaking the cycle of violence.


Laboratory-confirmed dengue fever and chikungunya fever cases at the Narita Airport Quarantine Station in 2013.
Fourteen patients were laboratory-confirmed cases of imported infectious diseases at the Narita Airport Quarantine Station in 2013. Blood tests were performed on 283 subjects suspected of having imported infectious diseases. Of these, 11 were diagnosed as having dengue fever (dengue) and 3 as having chikungunya fever (chikungunya) using real-time RT-PCR. The possible countries from which dengue virus infections were contracted were Thailand, Laos, Sri Lanka, and some other countries in Southeast Asia and South Asia. The 3 chikungunya cases were also diagnosed in individuals that returned from Southeast Asia. Most of the patients with dengue had a fever of over 38℃. The other symptoms were generalized fatigue, dull headache, pain behind the eyes, arthralgia, and digestive symptoms. Four of the patients were unaware of any mosquito bites. The information obtained from the confirmed cases showed that it is important to consider both the destination to which individuals travelled and the clinical symptoms, regardless of whether the subjects were aware of mosquito bites. The detection rate of chikungunya at the Quarantine Station was higher than that of dengue in all reported cases in Japan.


Rotavirus antigen, cytokine, and neutralising antibody profiles in sera of children with and without HIV infection in Blantyre, Malawi.
Rotavirus and HIV infection are major causes of death among children in sub-Saharan Africa. A previous study reported no association between concomitant HIV infection and rotavirus disease severity among hospitalised children in Malawi. This study examined rotavirus antigenaemia and broader immune responses among HIV-infected and uninfected children. Stored (-80°C), paired sera from acute and convalescent phases of Malawian children less than 5 years old, hospitalised for acute gastroenteritis in the primary study, collected from July 1997 to June 1999, were utilised. Among children older than 15 months, HIV infection was defined as the presence of HIV antibody in the blood, when confirmed by at least 2 established methods. For those younger than 15 months, nested polymerase chain reaction (PCR) amplification of proviral DNA was used for verification. All were followed for up to 4 weeks after hospital discharge. Rotavirus antigen levels in sera were measured with Premier™ Rotaclone® rotavirus enzyme immunoassay (EIA) kit. Acute-phase sera were examined for 17 cytokines, using Luminex fluorescent bead human cytokine immunoassay kit. Rotavirus-specific IgA and neutralising activity were determined by EIA and microneutralisation (MN) assay, respectively. Human strains and bovine-human reassortants were propagated in MA104 cells with serum-free Iscove's Modified Dulbecco's Medium (IMDM). Differences in results, from specimens with and without HIV infection, were analysed for statistical significance using the chi-square test. We detected rotavirus antigen in 30% of the HIV-infected and 21% HIV-uninfected, in the acute-phase sera. HIV-infected children developed slightly prolonged rotavirus antigenaemia compared to HIV-uninfected children. Rotavirus-specific IgA seroconversion rates and neutralising titres were similar in HIV-infected and HIV-uninfected children, thus, HIV infection had no major effect on immune responses to rotavirus infection.


Evaluation of two public genome references for chinese hamster ovary cells in the context of rna-seq based gene expression analysis.
RNA-Seq is a powerful transcriptomics tool for mammalian cell culture process development. Successful RNA-Seq data analysis requires a high quality reference for read mapping and gene expression quantification. Currently, there are two public genome references for Chinese hamster ovary (CHO) cells, the predominant mammalian cell line in the biopharmaceutical industry. In this study, we compared these two references by analyzing 60 RNA-Seq samples from a variety of CHO cell culture conditions. Among the 20,891 common genes in both references, we observed that 31.5% have more than 7.1% quantification differences, implying gene definition differences in the two references. We propose a framework to quantify this difference using two metrics, Consistency and Stringency, which account for the average quantification difference between the two references over all samples, and the sample-specific effect on the quantification result, respectively. These two metrics can be used to identify potential genes for future gene model improvement and to understand the reliability of differentially expressed genes identified by RNA-Seq data analysis. Before a more comprehensive genome reference for CHO cells emerges, the strategy proposed in this study can enable more robust transcriptome analysis from CHO cell RNA-Seq data. Biotechnol. Bioeng. 2017;114: 1603-1613. © 2017 Wiley Periodicals, Inc.


Role of capsaicin-sensitive nerve fibers in uterine contractility in the rat.
The possible participation of capsaicin-sensitive sensory nerves in the modulation of neurogenic contractions was studied in nonpregnant and term pregnant rat uteri. Neurogenic contractions were elicited by electric field stimulation (40 V, 1-70 Hz, 0.6 msec) in intact uteri and uteri that were previously exposed to capsaicin in vitro. In capsaicin pretreated preparations obtained both from nonpregnant and term pregnant rats, a dose-dependent increase in the amplitude of uterine contractions was detected. Prior systemic treatment of the rats with capsaicin (130 mg/kg, s.c.) abolished the effect of in vitro capsaicin administration on the amplitude of neurogenic contractions. Use of a specific antagonist of calcitonin gene-related peptide revealed that depletion of this peptide, which normally elicits uterine smooth muscle relaxation, may be responsible for the increased responsiveness of the uterus to low-frequency stimulation. Experiments on the localization of calcitonin gene-related peptide in uterine tissue specimens exposed to capsaicin revealed dose-dependent depletion of calcitonin-gene related peptide-immunoreactive nerves innervating blood vessels and the myometrium. The findings indicate that capsaicin-sensitive afferent nerves, by the release of sensory neuropeptides, significantly contribute to the modulation of uterine contractility both in nonpregnant and term pregnant rats. It is suggested that uterine sensory nerve activation may be part of a trigger mechanism leading to preterm contractions evoked by, for example, inflammation.


[Advancement of the study in meibomian lipids].
Meibomian lipids are believed to compose the lipid layer of tear film and play important roles in maintaining the stability of tear film. Changes in composition of meibomian lipids contribute to the ocular surface disease, such as dry eye. The understanding of composition and role of meibomian lipids can help us understand the cause of these disease. This review is intended to summarize the current state of knowledge about meibomian lipids, lipidomic analysis of meibomian lipids and changes of medium regarding to diseases.


Melting glaciers: a probable source of DDT to the Antarctic marine ecosystem.
Persistent organic pollutants reach polar regions by long-range atmospheric transport and biomagnify through the food web accumulating in higher trophic level predators. We analyzed Adélie penguin (Pygoscelis adeliae) samples collected from 2004 to 2006 to evaluate current levels of sigmaDDT (p,p'-DDT + p,p'-DDE) in these birds, which are confined to Antarctica. Ratios of p,p'-DDT to p,p'-DDE in Adélie penguins have declined significantly since 1964 indicating current exposure to old rather than new sources of sigmaDDT. However, sigmaDDT has not declined in Adélie penguins from the Western Antarctic Peninsula for more than 30 years and the presence of p,p'-DDT in these birds indicates that there is a current source of DDT to the Antarctic marine food web. DDT has been banned or severely restricted since peak use in the 1970s, implicating glacier meltwater as a likely source for DDT contamination in coastal Antarctic seas. Our estimates indicate that 1-4 kg x y(-1) sigmaDDT are currently being released into coastal waters along the Western Antarctic Ice Sheet due to glacier ablation.


Preventing shortages of biologic medicines.
Shortages of small-molecule injectable drugs have captured the attention of patients, healthcare providers, regulators and policy makers in recent years. While these shortages have several causes, non-compliance with current good manufacturing practice and subsequent shutdowns of manufacturing facilities have played a central role. Sterile injectable drugs are particularly susceptible to manufacturing quality disruptions because of their sensitivity to contamination. Biologics are subject to the same fill-finish contamination risk as sterile injectables, but their active ingredients are also sensitive to subtle changes in the manufacturing process and to storage and handling of their final dosage forms. Originator biologics will lose market exclusivity in the years ahead as patents expire and as competitors develop biosimilar products. The availability of therapeutic alternatives may provide opportunities to reduce costs and increase patient access, but this should not come at the expense of critical investments in the manufacturing of these complex and sensitive products.


[Investigations on the utilisation of parenterally administered amino acids by premature and small-for-dates neonates. II. Investigations of elimination half life time, elimination constants, transfer and clearances of amino acids after short-time infusion of L-amino acid mixtures (author's transl)].
10 ml/kg body weight of a 5% standardized L-amino-acid-mixture was given within the first 25 days to 25 prematurely born infants of 32--37 weeks gestationale and 5 small-for-dates newborns. We studied the metabolic kinetics of these amino acids in the serum and calculated elimination half life time, elimination constants, clearance and transfer rates of the single amino acids. The results were correlated with sex, nutritional state and age. 5 additional experiments were performed with 3 prematurely born infants and 2 small-for-dates newborns. The following results were of special interests: 1. Nearly all amino-acids had half life times between 30 and 50 min. Ornithine and tryptophane were more (half life time: 70 and about 50 min, respectively) slowly eliminated, while arginine and glutamic acid had half life times of between 5 and 15 min. 2. There was no statistically significant differences between sexes. 3. Eutrophic prematures eliminated the amino acids much faster then the hypotrophic ones. 4. Hypotrophic premature infants showed slow elimination of nearly all amino acids compared with hypotrophic newborns. 5. The elimination of nearly all amino acids studied grew faster with increasing age. The most significant differences were registered between the 1st and 3rd weeks of life.


Left ventricular diastolic filling response to stationary bicycle exercise during pregnancy and the postpartum period.
The purpose of this study was to determine the effects of pregnancy and of maximal exercise on left ventricular diastolic filling response. Transmitral pulsed Doppler echocardiography was obtained in 10 healthy women during each trimester of pregnancy and at 12 weeks after delivery. Doppler studies were performed at rest and at each exercise workload. The P-R interval, the early and atrial peak flow velocities, the mitral early deceleration time, and the isovolumetric relaxation time were analyzed. Data are expressed as the mean and standard deviation of the mean. Values obtained during the last trimester of pregnancy were used as the pregnant value; values at the 12 weeks after delivery were used as the nonpregnant value. Paired t -test, analysis of variance, and mixed models were used to determine significance with a probability value of <.05. Pregnancy significantly increased the early and atrial peak flow velocities. Pregnancy decreased the P-R interval, the early deceleration time, and the isovolumetric relaxation time. Exercise significantly decreased these diastolic functions; but pregnancy, in any of the 3 trimesters, did not significantly affect this response. Pregnancy increased left ventricular diastolic camber stiffness at rest and shifted left ventricular diastolic filling during exercise from predominantly early to atrial filling. This finding suggests that there is an increase in left ventricular chamber stiffness during maximal upright bicycle exercise in pregnancy.


Cellobiose quinone oxidoreductase from the white rot fungus Phanerochaete chrysosporium is produced by intracellular proteolysis of cellobiose dehydrogenase.
The fungus Phanerochaete chrysosporium was grown in a 10-l automatic fermenter using cellobiose as carbon source to monitor the induction of cellobiose dehydrogenase (CDH) and cellobiose quinone oxidoreductase (CBQ) enzymes, and to search for tentative cbq and cdh genes and their transcriptional products. After 24 h of induction, CDH was detected in the culture supernatant and a protein was recognized by a specific anti-CDH polyclonal antibody in the sonicated biomass. Northern blot experiments performed with several fungal RNA samples showed, after 24 h of induction, only one single species of an mRNA transcript corresponding in size to the cdh gene (2.5 kb) The relative amount of this transcript decreased as a function of time. Southern blot experiments done with genomic DNA and database search in the recently available genome information also ruled out the presence in this strain of a separate cbq gene distinct from the cdh gene. Taken together, these results demonstrated that CBQ originates from the cdh gene. Furthermore, it is not produced by differential splicing but by a posttranslational, predominantly intracellular, proteolytic cleavage.


Neural dynamics underlying emotional transmissions between individuals.
Emotional experiences are frequently shaped by the emotional responses of co-present others. Research has shown that people constantly monitor and adapt to the incoming social-emotional signals, even without face-to-face interaction. And yet, the neural processes underlying such emotional transmissions have not been directly studied. Here, we investigated how the human brain processes emotional cues which arrive from another, co-attending individual. We presented continuous emotional feedback to participants who viewed a movie in the scanner. Participants in the social group (but not in the control group) believed that the feedback was coming from another person who was co-viewing the same movie. We found that social-emotional feedback significantly affected the neural dynamics both in the core affect and in the medial pre-frontal regions. Specifically, the response time-courses in those regions exhibited increased similarity across recipients and increased neural alignment with the timeline of the feedback in the social compared with control group. Taken in conjunction with previous research, this study suggests that emotional cues from others shape the neural dynamics across the whole neural continuum of emotional processing in the brain. Moreover, it demonstrates that interpersonal neural alignment can serve as a neural mechanism through which affective information is conveyed between individuals.


Effect of inseminator on reproductive performance in dairy cattle.
The reproductive efficiency of dairy cattle is critical to the economic success and sustainability of the enterprise. Due to its many advantages, artificial insemination has inevitably taken its place in herd management today, and different artificial insemination methods have been developed until today. At the same time, artificial insemination is carried out by commercial artificial insemination operators or do-it-yourself operators in cattle farms. One of the factors affecting the success of artificial insemination is inseminators. The aim of this study was to determine the effect of inseminators on conception rates in reproductive challenges and to reveal the differences between the success rates of inseminators. A total of 227,297 artificial insemination acts carried out by 35 inseminators for 4 years belonging to the Konya Cattle Breeders' Association in Turkey were assessed for this purpose. In the study, the insemination success rate was determined as 41.36%. In addition, the percentage of inseminators with a total insemination success rate of more than 50% is very low, such as 2.9%, and the percentage of those who achieved 35.9% and below was 9.3%. As a result, it can be stated that the inseminators having a lower success rate than the overall should be retrained to avoid costs or losses in the farms.


The genetics of colony form and function in Caribbean Acropora corals.
Colonial reef-building corals have evolved a broad spectrum of colony morphologies based on coordinated asexual reproduction of polyps on a secreted calcium carbonate skeleton. Though cnidarians have been shown to possess and use similar developmental genes to bilaterians during larval development and polyp formation, little is known about genetic regulation of colony morphology in hard corals. We used RNA-seq to evaluate transcriptomic differences between functionally distinct regions of the coral (apical branch tips and branch bases) in two species of Caribbean Acropora, the staghorn coral, A. cervicornis, and the elkhorn coral, A. palmata. Transcriptome-wide gene profiles differed significantly between different parts of the coral colony as well as between species. Genes showing differential expression between branch tips and bases were involved in developmental signaling pathways, such as Wnt, Notch, and BMP, as well as pH regulation, ion transport, extracellular matrix production and other processes. Differences both within colonies and between species identify a relatively small number of genes that may contribute to the distinct "staghorn" versus "elkhorn" morphologies of these two sister species. The large number of differentially expressed genes supports a strong division of labor between coral branch tips and branch bases. Genes involved in growth of mature Acropora colonies include the classical signaling pathways associated with development of cnidarian larvae and polyps as well as morphological determination in higher metazoans.


Clinical predictors for hepatocellular carcinoma in patients with primary biliary cirrhosis.
Although hepatocellular carcinoma (HCC) occurs with increased frequency in patients with primary biliary cirrhosis (PBC), the exact frequency is relatively low. Optimal selection of PBC patients for HCC screening needs to be determined for effective screening. In this study, we aimed to explore clinical predictors of HCC in PBC patients. We performed a case-control study using 17 PBC patients with HCC identified from 1976 to 2002 at the Mayo Clinic. Control PBC patients who had no evidence of HCC were selected for each case by matching the first year of their visit to the Mayo Clinic. All medical information was collected within 2 years from when the cases were diagnosed with HCC. Logistic regression models were used for the analyses. Age, sex, history of blood transfusion, current smoking, histologic stage at PBC diagnosis, any signs of portal hypertension, Mayo score, hemoglobin level, platelet count, aspartate aminotransferase level, and albumin level were associated with the presence of HCC (P < .05 for each). In multivariable analysis, older age (OR, 1.7; 95% confidence interval [CI], 1.1-2.5 for 5 years), male sex (OR, 9.7; 95% CI, 1.4-68.3), history of blood transfusion (OR, 5.0; 95% CI, 1.0-24.3), and any signs of portal hypertension (OR, 22.9; 95% CI, 3.4-155.3) were associated significantly with increased odds of HCC and yielded an excellent diagnostic performance (area under the receiver operating characteristics curve rate, 0.91). Older age, male sex, history of blood transfusion, and any signs of portal hypertension or cirrhosis indicate higher likelihood of HCC and should be considered for HCC screening. Further studies in larger patient cohorts are required to verify the diagnostic model.


Organic anion transport and reabsorptive fluid flux in diabetic retinal vessels: a microperfusion study.
Microperfusion techniques were used to characterize in vitro the fluorescein transport and fluid movement across isolated diabetic rabbit retinal vessels. Unbranched segments of arterioles of alloxan-induced diabetic rabbits were isolated and cannulated on a microperfusion apparatus. Movement of 3H-inulin, the reference tracer, and 14C-fluorescein, the test tracer, across the vascular walls was studied using microperfusion techniques adapted by us to study vascular permeability. The diabetic retinal vessels showed a fluid flux from bath to lumen of 3.6 +/- 0.6 nl min-1 mm-1 with perfusion rates of 7.1 +/- 0.3 nl min-1. This value was 43% (P < or = 0.05) lower when compared with normal values obtained in previous work. Bath to lumen fluorescein fluxes were 2.1 +/- 0.5 and 7.6 +/- 1.6 x 10(-12) mol min-1 mm-1, for fluorescein concentrations of 0.87 x 10(-4) and 1.5 x 10(-4) M, respectively. These values are 23% (P < or = 0.05) and 38.7% (P < or = 0.05), respectively, lower when compared to values obtained from normal retinal vessels. Probenecid, a competitive inhibitor of organic anion transport, added to the bath, had no effect. Increasing bath concentrations of fluorescein caused increasing fluid flux from bath to lumen, and again, probenecid added to the bath has no effect. These results show a significant decrease of fluorescein transport and of net fluid flux across diabetic retinal vessel walls from outside into the lumen, suggesting that in experimental alloxan-induced diabetes there is an early, probably functional, involvement of the retinal vessels in the initial alteration of the blood-retinal barrier (BRB).


Evolutionary Consequences of Male Driven Sexual Selection and Sex-Biased Fitness Modifications in Drosophila melanogaster and Members of the simulans Clade.
Males have evolved a variety of behavioral, morphological, and physiological traits to manipulate their mates in order to maximize their chances of success. These traits are bound to influence how females respond to male behaviors and influence the nature of sexual selection/conflict. A common consequence of aggressive male mating strategies in Drosophila melanogaster is the reduction of female lifespan. Our study shows that this is common across members of the simulans clade. Reduced life expectancy of females implies that female contribution to a population is less than that of males per generation. Fitness differences between the sexes in every generation will invariably affect overall population fitness. How natural selection responds to the female deaths and thereby the unequal fitness of the sexes has rarely been addressed. We shed light on this issue and provide evidence, which suggests that additional gains of fitness by males due to their longevity and continued mating may provide one explanation as to why the loss of female fitness may be "invisible" (effectively neutral) to natural selection. Male driven sexual selection and additional, transgenerational gains of male fitness can be an important force of evolutionary change and need to be tested with other organisms.


Teamwork and patient safety in dynamic domains of healthcare: a review of the literature.
This review examines current research on teamwork in highly dynamic domains of healthcare such as operating rooms, intensive care, emergency medicine, or trauma and resuscitation teams with a focus on aspects relevant to the quality and safety of patient care. Evidence from three main areas of research supports the relationship between teamwork and patient safety: (1) Studies investigating the factors contributing to critical incidents and adverse events have shown that teamwork plays an important role in the causation and prevention of adverse events. (2) Research focusing on healthcare providers' perceptions of teamwork demonstrated that (a) staff's perceptions of teamwork and attitudes toward safety-relevant team behavior were related to the quality and safety of patient care and (b) perceptions of teamwork and leadership style are associated with staff well-being, which may impact clinician' ability to provide safe patient care. (3) Observational studies on teamwork behaviors related to high clinical performance have identified patterns of communication, coordination, and leadership that support effective teamwork. In recent years, research using diverse methodological approaches has led to significant progress in team research in healthcare. The challenge for future research is to further develop and validate instruments for team performance assessment and to develop sound theoretical models of team performance in dynamic medical domains integrating evidence from all three areas of team research identified in this review. This will help to improve team training efforts and aid the design of clinical work systems supporting effective teamwork and safe patient care.


The Clinical Aspects and Prognostic Factors Concerning Survival in Patients With Recurrent Cervical Cancer After Radical Hysterectomy and Adjuvant Chemoradiotherapy.
To investigate the recurrence patterns and prognostic factors of patients with recurrent cervical cancer after radical hysterectomy with node dissection (RHND) followed by adjuvant radiotherapy (RT)/concurrent chemoradiotherapy (CCRT). The medical records of 153 patients with pre-operative International Federation of Gynecology and Obstetrics stage IB-IIA cervical cancer, who were treated with RHND followed by adjuvant RT/CCRT at the Liaoning Cancer Hospital between January 1, 2012 and May 31, 2018, were retrospectively analyzed. The median disease progression-free survival time was 16 months, and 75.2% (115/153) of patients had a relapse within two years. The survival of patients with multi-site relapse was significantly lower in comparison to those with relapse in a single site (p < 0.001). The survival rate of patients with distant metastasis (DM) and combined recurrence (DM with localregional recurrence [LR]) was significantly lower than that of patients with only LR (p = 0.006, p < 0.001). Furthermore, the survival rate of patients with combined recurrence was significantly lower than that of patients with only DM (p = 0.046). Multivariate analysis showed that resection margin involvement, para-aortic and common iliac lymph node metastasis, DM, no treatment after disease relapse, and early disease relapse were independent prognostic factors associated with poor survival. Most of the cervical cancer patients who received initial RHND followed by adjuvant RT/CCRT had a relapse within two years. Resection margin involvement, para-aortic and common iliac lymph node metastasis, DM, no treatment after recurrence, and early disease relapse were found to be prognostic factors in patients with recurrent cervical cancer after RHND followed by adjuvant RT/CCRT.


[The relationship between insulin resistance and endothelium-dependent vasodilatation in obese subjects].
To study the relationship between insulin resistance and endothelium-dependent vasodilatation function in obese subjects with normal blood glucose, serum cholesterol and blood pressure and to explore the non-traditional risk factors associated with atherosclerosis. 24 euglycemic obese males (OB) and 12 age and sex matched lean controls (Lean) underwent oral glucose tolerance test (OGTT), euglycemic hyperinsulinemia clamp study with an insulin infusion rate of 120 mU/m(2)/min to evaluate the peripheral glucose disposal rate (GDR) in steady-state, brachial artery ultrasound study to assess the endothelium-dependent vasodilation (EDV) and independent vasodilation (EIV), the combined intima-media thickness (IMT) of common carotid arteries examined by high resolution ultrasound, and fasting lipids profile and free fatty acids (FFAs) concentration. The blood glucose and serum cholesterol levels were all normal in the OB group but the serum FFA and triglyceride concentrations were elevated compared with those of the controls (FFA: 888 microM/l +/- 158 microM/l vs 508 microM/l +/- 137 microM/l, P < 0.05; TG: 1.8 mM/l +/- 0.6 mM/l vs 1.1 mM/l +/- 0.5 mM/l, P < 0.05) The GDR in steady-state was 6.7 mg x min(-1) x kg(-1) +/- 1.4 mg x min(-1) x kg(-1) in the OB group, significantly lower than that of the controls (12.2 mg x min(-1) x kg(-1) +/- 3.1 mg x min(-1) x kg(-1), P < 0.05). In the OB group the flow-mediated dilation of the brachial artery caused by reactive hyperemia (EDV) were impaired and the relaxation time was shorter than that of the controls (P < 0.05) but the EIV was not significantly different between these two groups. There was a significantly positive correlation between GDR and EDV (r = 0.438, P < 0.05). The IMT of common carotid arteries was thicker in the OB group than in the control group (0.50 mm +/- 0.02 mm vs 0.34 mm +/- 0.19 mm). The endothelium-dependent vasodilation was impaired in obese subjects even though without hyperglycemia, hypercholesterolemia, and hypertension. Traditional cardiovascular risk factors cannot fully account for the increased risk of cardiovascular disease in metabolic syndrome. Elevated circulating FFA concentration, is characteristic of metabolic syndrome, may play a role in endothelial dysfunction.


Assessing the Feasibility of a Multimodal Approach to Pain Evaluation in Early Stages after Spinal Cord Injury.
This research evaluates the feasibility of a multimodal pain assessment protocol during rehabilitation following spinal cord injury (SCI). The protocol amalgamates clinical workup (CW), quantitative sensory testing (QST), and psychosocial factors (PSF) administered at 4 (T1), 12 (T2), and 24 (T3) weeks post injury and at discharge (T4). Molecular blood biomarkers (BB) were evaluated via gene expression and proteomic assays at T1 and T4. Different pain trajectories and temporal changes were identified using QST, with inflammation and pain-related biomarkers recorded. Higher concentrations of osteopontin and cystatin-C were found in SCI patients compared to healthy controls, indicating their potential as biomarkers. We observed altered inflammatory responses and a slight increase in ICAM-1 and CCL3 were noted, pointing towards changes in cellular adhesion linked with spinal injury and a possible connection with neuropathic pain. Despite a small patient sample hindering the correlation of feasibility data, descriptive statistical analyses were conducted on stress, depression, anxiety, quality of life, and pain interferences. The SCI Pain Instrument (SCIPI) was efficient in distinguishing between nociceptive and neuropathic pain, showing a progressive increase in severity over time. The findings emphasize the need for the careful consideration of recruitment setting and protocol adjustments to enhance the feasibility of multimodal pain evaluation studies post SCI. They also shed light on potential early adaptive mechanisms in SCI pathophysiology, warranting the further exploration of prognostic and preventive strategies for chronic pain in the SCI population.


Amendments to the quick disability determination process. Final rule.
We are amending our regulations to extend the quick disability determination process (QDD), which is operating now in the Boston region, to all of the State disability determination services (DDSs). We also are removing from the QDD process the existing requirements that each State DDS maintain a separate QDD unit and that each case referred under QDD be adjudicated within 20 days. These actions stem from our continuing effort to improve our disability adjudication process.


Primary mediastinal seminoma.
Two new cases of primary mediastinal seminoma are presented and 126 cases from the literature are analysed. Radiotherapy, alone or combined with surgery, is thought by some authors to be the treatment of choice and the role of chemotherapy is uncertain. We propose a classification by which it is possible to compare various therapeutic approaches and to establish their value.


High-Performance Visible-Blind UV Phototransistors Based on n-Type Naphthalene Diimide Nanomaterials.
This study investigates the performance of single-crystalline nanomaterials of wide-band gap naphthalene diimide (NDI) derivatives with methylene-bridged aromatic side chains. Such materials are found to be easily used as high-performance, visible-blind near-UV light detectors. NDI single-crystalline nanoribbons are assembled using a simple solution-based process (without solvent-inclusion problems), which is then applied to organic phototransistors (OPTs). Such OPTs exhibit excellent n-channel transistor characteristics, including an average electron mobility of 1.7 cm2 V-1 s-1, sensitive UV detection properties with a detection limit of ∼1 μW cm-2, millisecond-level responses, and detectivity as high as 1015 Jones, demonstrating the highly sensitive organic visible-blind UV detectors. The high performance of our OPTs originates from the large face-to-face π-π stacking area between the NDI semiconducting cores, which is facilitated by methylene-bridged aromatic side chains. Interestingly, NDI-based nanoribbon OPTs exhibit a distinct visible-blind near-UV detection with an identical detection limit, even under intense visible light illumination (for example, 104 times higher intensity than UV light intensity). Our findings demonstrate that wide-band gap NDI-based nanomaterials are highly promising for developing high-performance visible-blind UV photodetectors. Such photodetectors could potentially be used for various applications including environmental and health-monitoring systems.


Pre- and postexposure prophylaxis of Ebola virus infection in an animal model by passive transfer of a neutralizing human antibody.
A neutralizing human monoclonal antibody, KZ52, protects guinea pigs from lethal Ebola Zaire virus challenge. Administration before or up to 1 h after challenge resulted in dose-dependent protection by the antibody. Interestingly, some antibody-treated animals survived despite developing high-level viremia, suggesting that the mechanism of protection by KZ52 may extend beyond reduction of viremia by virus neutralization. KZ52 is a promising candidate for immunoprophylaxis of Ebola virus infection.


Extracellular Matrix Proteome and Phosphoproteome of Potato Reveals Functionally Distinct and Diverse Canonical and Non-Canonical Proteoforms.
The extracellular matrix (ECM) has a molecular machinery composed of diverse proteins and proteoforms that combine properties of tensile strength with extensibility exhibiting growth-regulatory functions and self- and non-self-recognition. The identification of ECM proteoforms is the prerequisite towards a comprehensive understanding of biological functions accomplished by the outermost layer of the cell. Regulatory mechanisms of protein functions rely on post-translational modifications, phosphorylation in particular, affecting enzymatic activity, interaction, localization and stability. To investigate the ECM proteoforms, we have isolated the cell wall proteome and phosphoproteome of a tuberous crop, potato (Solanum tuberosum). LC-MS/MS analysis led to the identification of 38 proteins and 35 phosphoproteins of known and unknown functions. The findings may provide a better understanding of biochemical machinery and the integrated protein and phosphoprotein network of ECM for future functional studies of different developmental pathways and guidance cues in mechanosensing and integrity signaling.


CT detection and location of intraorbital foreign bodies. Experiments with wood and glass.
The series comprises 27 patients examined by CT to detect, locate or exclude a foreign body. 22 of them actually had an orbital foreign body. In three cases CT was the primary method and showed the foreign body correctly, while in 18 it was first detected in plain films and CT was performed to locate it. 20 metallic foreign bodies were hyperdense in appearance. Two cases had wooden foreign bodies, one with a density of +10 HU and the other hypodense with a value of about -434 to -446 HU. The latter piece of wood was first interpreted falsely as a bubble of gas. The results proved that the detection of metal is easy, but differentiation between wood and gas is problematical. Experiments conducted to determine the CT densities of different pieces of wood gave results varying from -618 HU to +23 HU. The highest densities obtained for glass varied from +522 HU to +2000 HU. The density of a plastic lens was -105 HU.


Antimicrobial Metabolites from Streptomyces sp. SN0280.
One new indole derivative, chloroindole (1), one new diketone, streptoone A (2), two new ketonic acids, streptoones B (3) and C (4), and one known macrolide antibiotic, X-14952B (5), were isolated from Streptomyces sp. SN0280. Extensive NMR, HRESIMS, and IR analysis was used to elucidate their structures. Streptoone A (2) displayed antibacterial activity (MIC value of 7.81 μg/mL) against Clavibater michiganensis, comparable with the positive control streptomycin (MIC value of 7.81 μg/mL). Streptoone B (3) showed antifungal activity (MIC value of 15.63 μg/mL) against Phytophthora capsici (positive control carbendazol MIC value of 7.81 μg/mL). These molecules provide new templates for the potential treatment and management of these phytopathogens.


ANCA-associated vasculitis with renal involvement.
Systemic vasculitis is a rare but severe group of diseases characterized by inflammation and necrosis of blood vessels. The size of the vessel affected varies among the different forms of vasculitis and there are three main subgroups: large, medium and small vessel vasculitis. Among small vessel vasculitis, the antineutrophil cytoplasmic antibody (ANCA)-associated forms are of particular importance. This subgroup includes: microscopic polyangiitis, granulomatosis with polyangiitis (Wegener's), eosinophilic granulomatosis with polyangiitis (Churg-Strauss) and the form limited to the kidney. ANCA are serum autoantibodies directed against proteins present in the cytoplasmic granules of neutrophils and represent the serological markers of small vessel vasculitis. Renal involvement is present in the majority of patients with ANCA-associated vasculitis (AAV) and the consequences of a missed or delayed diagnosis of renal vasculitis are potentially life threatening. Patient survival and the risk of end-stage renal disease are closely associated with renal function at presentation. The gold standard for diagnosis remains renal biopsy. In 2010, a new histopathological classification based on the percent of normal glomeruli, cellular crescent or global sclerotic glomeruli was proposed. The aim of this classification was to predict the renal prognosis. Nowadays, remission can be achieved and maintained in most cases with a combination of high-dose steroid and immunosuppressive drugs. This therapy has to be continued for at least 24 months after a substantial remission has been obtained because early cessation of treatment is associated with an increased risk of relapse. For this reason, patients should be regularly monitored in order to promptly diagnose and treat a possible recurrence of AAV. This review will focus on kidney involvement in AAV with an overview of the clinical-pathological characteristics and therapeutic strategy for these conditions.


[Comparative study of intravenous midazolam and diazepam used as sedative agents during gastroscopy].
Midazolam was compared with diazepam for the intravenous sedation of gastroscopy patients. 0.1 mg X kg-1 midazolam were given to patients less than 65 years old, and 0.085 mg X kg-1 after that age; diazepam doses were 0,2 mg X kg-1 and 0.15 mg X kg-1 respectively. The effect and duration of action of both drugs were similar. The main differences between them were as follows: the intravenous injection of midazolam was not painful, and virtually no venous complications were seen; amnesia and patient contentedness were more frequent with midazolam; gastric secretion was reduced with midazolam. Neither drug had clinically significant cardiovascular effects.


Postpartum follicular and luteal activity in Holstein-Friesian cows genetically selected for high or low mature bodyweight: relationships with follicle stimulating hormone , insulin , insulin-like growth factor-1 and growth hormone.
To investigate ovarian follicular and luteal activity during the postpartum period of cows genetically selected for high or low mature bodyweight, in relation to metabolic and reproductive endocrine parameters, to determine whether there are differences between strains that could affect fertility outcomes. The presence of follicles > or =5 mm diameter and luteal structures was mapped in the ovaries of 12 high (heavy-strain) and 12 low (light-strain) mature bodyweight cows by daily trans-rectal ultrasonography from Day 7 postpartum until the end of their first normal oestrous cycle. Blood samples were collected daily, for measurement of concentrations of follicle stimulating hormone (FSH), progesterone, growth hormone (GH), insulin-like growth factor-1 (IGF-1), and insulin. Intervals to first ovulation were calculated from ultrasonography data. Heavy-strain cows had shorter intervals than light-strain cows from calving to the emergence of the first (9.0 (SE 0.9) vs 12.4 (SE 1.3) days) and second (16.4 (SE 1.8) vs 20.6 (SE 1.6) days) dominant follicles (p<0.05). Concentrations of FSH in heavy-strain cows prior to the emergence of the second, third and fourth dominant follicles were higher than in light-strain cows (p<0.05). Heavy-strain cows were more likely to have a large (>15 mm diameter) follicle earlier than light-strain cows (p<0.01). Concentrations of insulin and IGF-1, but not those of GH, were higher in heavy- than light-strain cows during the postpartum period (p=0.01 and p=0.02, respectively), and concentrations of both on Day 6 were inversely related to the time of emergence of the first dominant follicle (p<0.01). Concentrations of progesterone were similar in both strains of cow until Day 10 of the first oestrous cycle, but thereafter were higher in light- than heavy-strain cows until Day 16. Progesterone concentrations in heavy-strain cows declined earlier and more rapidly than in their lighter counterparts. These results indicate that there is a rapid postpartum resumption of follicular activity in both heavy- and light-strain cows, but that there is an earlier emergence of dominant follicles and ovulation in the former. Differences in luteal function, in terms of lower dioestrus progesterone concentrations and an earlier onset of luteolysis, in heavy- than light-strain cows might be sufficient to impair the fertility of the former.


Circulating immune biomarkers as predictors of the response to pembrolizumab and weekly low dose carboplatin and paclitaxel in NSCLC and poor PS: An interim analysis.
The combination of standard-dose chemotherapy and immunotherapy has been shown to be beneficial for patients with non-small cell lung cancer (NSCLC) with good performance status (PS). However, treatment options for patients with poor PS are limited. In the present study, the feasibility and immunological effects of low-dose chemotherapy with carboplatin and paclitaxel combined with immunotherapy with pembrolizumab were examined in patients with metastatic NSCLC and a poor PS. Patients with advanced NSCLC and a PS of 2 were randomized to single-agent pembrolizumab at 200 mg every 3 weeks or pembrolizumab combined with weekly carboplatin area under the curve 1 and paclitaxel 25 mg/m2. Blood for circulating immune cell phenotyping, soluble program death ligand 1 (sPD-L1) and immune-modulatory microRNAs (miRNAs) was collected prior to treatment and at weeks 4 and 7. Ten patients were randomized to the combination arm and 10 to the single-agent arm. Therapy was well tolerated. Four patients discontinued carboplatin due to hypersensitivity reactions but continued pembrolizumab and paclitaxel treatments. Increases in activated CD4+ T cells and in immune-regulatory miRNA, and decreases in myeloid derived suppressor cells were observed in the blood of patients in the combination arm and not in the single-agent arm. Changes in circulating regulatory T cells and sPD-L1 were not observed. Seven patients in the combination arm manifested a partial response compared with only two in the single-agent arm. Weekly low-dose chemotherapy carboplatin and paclitaxel was well tolerated and immunologically active when combined with pembrolizumab in patients with advanced NSCLC and a PS of 2. This combination merits further study in this patient population.


Asymmetric Synthesis of (-)-Pterocarine and (-)-Galeon via Chiral Phase Transfer-Catalyzed Atropselective Formation of Diarylether Cyclophane Skeleton.
Pterocarine and galeon are typical examples of diarylether heptanoids (DAEHs) with planar chirality due to the strictly constrained conformations in their molecular skeletons. The characterized oxa[1,7]metapara-cyclophane motifs in DAEHs impose great challenges for their enantioselective synthesis. The asymmetric syntheses of (-)-pterocarine and (-)-galeon are demonstrated by employing a chiral phase transfer-catalyzed highly enantioselective SNAr cyclization as the key step for the formation of a diarylether cyclophane skeleton.


[Selection of Radiomic Features for the Classification of Triple-negative Breast Cancer Based on Radiogenomics].
Subtype classification of breast cancer by analyzing the gene expression profile of cancer cells is becoming a standard procedure. Breast cancer subtype classification is more useful than the conventional method because the characteristics of subtype classification is directly connected with the treatment method. However, genetic testing is invasive, and a part of cancer cells may not represent the overall nature of the cancer. In the computer-aided diagnosis (CAD) scheme for differentiation of triple-negative breast cancer (TNBC) by estimating the genetic properties of cancer based on Radiogenomics, principal component analysis (PCA) and least absolute shrinkage and selection operator (Lasso) were used for reducing the dimension of radiomic features, and we compared usefulness of both. We collected 81 magnetic resonance (MR) images, which included 30 TNBC and 51 others, from the public database. From the MR slice images, we selected the slice containing the largest area of the cancer and manually marked the cancer region. We subsequently calculated 294 radiomic features in the cancer region, and reduced the dimension of radiomic features. Finally, linear discriminant analysis, with the dimensionally compressed 10 image features, was used for distinguishing between TNBC and others. Area under the curve (AUC) was 0.60 when we used PCA, whereas AUC was 0.70 when we used Lasso (p=0.0058). Therefore, Lasso is useful for the determination of radiomic features in Radiogenomics.


Intracarotid 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine administration: biochemical and behavioral observations in a primate model of hemiparkinsonism.
Cynomolgus monkeys received intracarotid injections of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to produce a chronic unilateral model of parkinsonism. Extensive dopamine (DA) depletion was observed in the caudate nucleus and putamen on the side ipsilateral to the injection and this was associated with contralateral tremor, rigidity, and bradykinesia. A dose of 1.25 mg of MPTP caused ipsilateral DA loss of 99.4% in the caudate nucleus, 99.8% in the putamen, and 74.2% in the nucleus accumbens. A dose of 2.5 mg caused ipsilateral DA depletion of 99.3% in the caudate nucleus, 99.5% in putamen, and 90.1% in the nucleus accumbens. The unilateral aspect of the lesion was dose sensitive, with the 2.5-mg dose causing bilateral asymmetric DA depletion. Tissue concentrations of serotonin were not affected by the toxin. These findings confirm that intracarotid injection of MPTP may produce a useful primate model of hemiparkinsonism that can be associated with selective unilateral DA depletion when the appropriate dose of toxin is used.


Formal Total Synthesis of (±)-Lycojaponicumin C.
The formal total synthesis of (±)-lycojaponicumin C has been accomplished. Key transformations include a Rh-catalyzed formal [3 + 2] cycloaddition reaction to construct the bicyclic [3.3.0] scaffold bearing two vicinal quaternary carbon centers, a stereoselective γ-hydroxyl directed Michael addition to introduce the vinyl group at a bulky position, and a late-stage ring-closing metathesis reaction to form the cyclohexanone ring.


Pseudohyperkalemia: Look before you treat.
Pseudohyperkalemia is defined as a reported rise in serum potassium concentration along with a normal effective plasma potassium concentration. We present a case report of a 57-year-old gentleman with a history of chronic lymphocytic leukemia, who presented with an elevation in serum potassium along with a normal plasma potassium concentration. Through an exploration of the literature, we demonstrate that pseudohyperkalemia is an important phenomenon to watch for as it may sometimes lead to unnecessary and potentially dangerous treatment.


[Pulmonary embolism associated to HIV infection].
Antiphospholipid antibodies are frequently found in patients with HIV infection mainly in advanced forms of disease. Despite its elevated prevalence the association with thrombotic events is rare. The author present a clinical case of a HIV patient in which the first manifestation of the disease was a pulmonary embolism.


Bronchogenic cyst of the stomach: A case report.
Gastric bronchogenic cysts (BCs) are extremely rare cystic masses caused by abnormal development of the respiratory system during the embryonic period. Gastric bronchial cysts are rare lesions first reported in 1956; as of 2019, only 37 cases are available in the MEDLINE/PubMed online databases. BCs usually have no clinical symptoms in the early stage, and their imaging findings also lack specificity. Therefore, they are difficult to diagnose before histopathological examination. A 55-year-old woman presented at our hospital with intermittent epigastric pain. She had a slightly high level of serum carbohydrate antigen 72-4 (CA 72-4). Endoscopic ultrasound found that a cystic mass originated from the submucosa of the posterior gastric wall near the cardia, indicating a diagnosis of cystic hygroma of the stomach. Furthermore, a computed tomography scan demonstrated a quasi-circular cystic mass closely related to the lesser curvature of the gastric fundus with a low density. Because the imaging examinations did not suggest a malignancy and the patient required complete resection, she underwent laparoscopic surgery. As an intraoperative finding, this cystic lesion was located in the posterior wall of the fundus and contained some yellow viscous liquid. Finally, the pathologists verified that the cyst in the fundus was a gastric BC. The patient recovered well with normal CA 72-4 levels, and her course was uneventful at 10 mo. This is a valuable report as it describes an extremely rare case of gastric BC. Moreover, this is the first case of BC to present with elevated CA 72-4 levels.


Massively Parallel Sequencing of Peritoneal and Splenic B Cell Repertoires Highlights Unique Properties of B-1 Cell Antibodies.
B-1 cells are a unique subset of B cells that are positively selected for expressing autoreactive BCRs. We isolated RNA from peritoneal (B-1a, B-1b, B-2) and splenic (B-1a, marginal zone, follicular) B cells from C57BL/6 mice and used 5'-RACE to amplify the IgH V region using massively parallel sequencing. By analyzing 379,000 functional transcripts, we demonstrate that B-1a cells use a distinct and restricted repertoire. All B-1 cell subsets, especially peritoneal B-1a cells, had a high proportion of sequences without N additions, suggesting predominantly prenatal development. Their transcripts differed markedly and uniquely contained VH11 and VH12 genes, which were rearranged only with a restricted selection of D and J genes, unlike other V genes. Compared to peritoneal B-1a, the peritoneal B-1b repertoire was larger, had little overlap with B-1a, and most sequences contained N additions. Similarly, the splenic B-1a repertoire differed from peritoneal B-1a sequences, having more unique sequences and more frequent N additions, suggesting influx of B-1a cells into the spleen from nonperitoneal sites. Two CDR3s, previously described as Abs to bromelain-treated RBCs, comprised 43% of peritoneal B-1a sequences. We show that a single-chain variable fragment designed after the most prevalent B-1a sequence bound oxidation-specific epitopes such as the phosphocholine of oxidized phospholipids. In summary, we provide the IgH V region library of six murine B cell subsets, including, to our knowledge for the first time, a comparison between B-1a and B-1b cells, and we highlight qualities of B-1 cell Abs that indicate unique selection processes.


[Relationship between prenatal and postnatal echographic diagnosis of uropathy: is mass screening useful?].
Urinary tract malformations have a 1-2% incidence. An early diagnosis of these defects allows to realize as fast as possible the best medical and/or surgical treatment, preventing or at least slowing down the evolution toward chronic renal failure. Urinary tract malformations are particularly suitable for a "prevention" program due to their elevated incidence, to the "silent period" preceding complications and to the therapeutic possibilities strictly related to an early diagnosis. Ultrasound screening can easily identify congenital urinary tract abnormalities, especially obstructive ones, but unfortunately almost half of the cases escape even the most expert "eye". It is necessary a complete check-up right after birth. We tried to compare the method and the results of prenatal screening with the postnatal one, already used in many hospitals. We believe, even following our experience, that the screening for urinary tract malformations of all the neonatal population is a goal to pursue and achieve as soon as possible for its high sensitivity and specificity.


Metatranscriptomics as a tool to identify fungal species and subspecies in mixed communities - a proof of concept under laboratory conditions.
High-throughput sequencing (HTS) enables the generation of large amounts of genome sequence data at a reasonable cost. Organisms in mixed microbial communities can now be sequenced and identified in a culture-independent way, usually using amplicon sequencing of a DNA barcode. Bulk RNA-seq (metatranscriptomics) has several advantages over DNA-based amplicon sequencing: it is less susceptible to amplification biases, it captures only living organisms, and it enables a larger set of genes to be used for taxonomic identification. Using a model mock community comprising 17 fungal isolates, we evaluated whether metatranscriptomics can accurately identify fungal species and subspecies in mixed communities. Overall, 72.9% of the RNA transcripts were classified, from which the vast majority (99.5%) were correctly identified at the species level. Of the 15 species sequenced, 13 were retrieved and identified correctly. We also detected strain-level variation within the Cryptococcus species complexes: 99.3% of transcripts assigned to Cryptococcus were classified as one of the four strains used in the mock community. Laboratory contaminants and/or misclassifications were diverse, but represented only 0.44% of the transcripts. Hence, these results show that it is possible to obtain accurate species- and strain-level fungal identification from metatranscriptome data as long as taxa identified at low abundance are discarded to avoid false-positives derived from contamination or misclassifications. This study highlights both the advantages and current challenges in the application of metatranscriptomics in clinical mycology and ecological studies.


[An immune adjuvant activity of mycolic acid-containing glycolipid, trehalose-2,3,6'-trimycolate, derived from Gordona aurantiaca].
A mycolic acid-containing glycolipid, trehalose-2,3,6'-trimycolate (GaGM), derived from Gordona aurantiaca, an acid-fast bacterium closely related taxonomically to Mycobacterium, was investigated for its immune adjuvant activity on cell-mediated responses in the mouse. I.V. injection of liposomes containing GaGM enhanced the generation of cytotoxic T-lymphocyte (CTL) against syngeneic and allogeneic tumor cells. In addition, the injection of GaGM augmented the natural killer (NK) activity and the antibody-dependent cellular cytotoxicity (ADCC). These results suggest that the injection of GaGM induces the production of interleukin-2 (IL-2), since such effector cells as CTL, NK and K cells have been shown to require IL-2 for their development.


[Blink and masseter inhibitory reflexes in Parkinson's disease].
Idiopathic Parkinson's disease (PD) is a chronic, progressive, neurodegenerative disorder with prevalence from 60 to 187 per 100 000 persons in general population. The aim of the study was to determine the abnormalities of the blink reflex (BR) and the masseter inhibitory reflex (MIR) in parkinsonian patients, as indices of the functional status of brainstem neuronal network, and abnormality level dependence on disease progression. The investigation was conducted at the Clinic of Neurology, Clinical Center Nis, comprising a group of 60 subjects of both sexes, suffering from idiopathic Parkinson's disease in I-IV stages, according to the Hoehn and Yahr scale. The control group included 30 healthy subjects of both sexes and corresponding age. Testing of the patients was performed during the "on" phase by registering MIR and BR. Latency of polysynaptic R2 and R2' blink reflex responses and latency of polysynaptic S2 response, as well as a silent period of MIR, are linearly shortened in the subjects with PD, and more expressed in the subsequent stages of the disease compared to the control group. There is a positive correlation between the applied neurophysiological tests results and clinical stage of PD.


Development and application of hepatocellular carcinoma risk prediction model based on clinical characteristics and liver related indexes.
Hepatocellular carcinoma (HCC) is difficult to diagnose with poor therapeutic effect, high recurrence rate and has a low survival rate. The survival of patients with HCC is closely related to the stage of diagnosis. At present, no specific serological indicator or method to predict HCC, early diagnosis of HCC remains a challenge, especially in China, where the situation is more severe. To identify risk factors associated with HCC and establish a risk prediction model based on clinical characteristics and liver-related indicators. The clinical data of patients in the Affiliated Hospital of North Sichuan Medical College from 2016 to 2020 were collected, using a retrospective study method. The results of needle biopsy or surgical pathology were used as the grouping criteria for the experimental group and the control group in this study. Based on the time of admission, the cases were divided into training cohort (n = 1739) and validation cohort (n = 467). Using HCC as a dependent variable, the research indicators were incorporated into logistic univariate and multivariate analysis. An HCC risk prediction model, which was called NSMC-HCC model, was then established in training cohort and verified in validation cohort. Logistic univariate analysis showed that, gender, age, alpha-fetoprotein, and protein induced by vitamin K absence or antagonist-II, gamma-glutamyl transferase, aspartate aminotransferase and hepatitis B surface antigen were risk factors for HCC, alanine aminotransferase, total bilirubin and total bile acid were protective factors for HCC. When the cut-off value of the NSMC-HCC model joint prediction was 0.22, the area under receiver operating characteristic curve (AUC) of NSMC-HCC model in HCC diagnosis was 0.960, with sensitivity 94.40% and specificity 95.35% in training cohort, and AUC was 0.966, with sensitivity 90.00% and specificity 94.20% in validation cohort. In early-stage HCC diagnosis, the AUC of NSMC-HCC model was 0.946, with sensitivity 85.93% and specificity 93.62% in training cohort, and AUC was 0.947, with sensitivity 89.10% and specificity 98.49% in validation cohort. The newly NSMC-HCC model was an effective risk prediction model in HCC and early-stage HCC diagnosis.


IL-6 promotes PD-L1 expression in monocytes and macrophages by decreasing protein tyrosine phosphatase receptor type O expression in human hepatocellular carcinoma.
We have previously discovered a relationship between the low expression of protein tyrosine phosphatase, receptor type O (PTPRO) in tumor-infiltrating T cells and immunosuppression. The aim of the present study was to investigate the relationship between decreased PTPRO and increased programmed death ligand 1 (PD-L1) in both the peripheral monocytes and tumor-infiltrating macrophages of human hepatocellular carcinoma (HCC). The expression and correlation of all the indices were explored in monocytes and tumor-infiltrating macrophages within both human and mice HCC. The mechanic regulations were studied by using both in vitro and in vivo studies. We found a significant decrease in PTPRO in HCC peripheral monocytes that was associated with increased PD-L1 expression in peripheral monocytes and tumor-associated macrophages (TAMs) in HCC. Monocyte PD-L1 and PTPRO therefore could serve as valuable prognostic indicators for post-surgery patients with HCC and were associated with increased T-cell exhaustion (Tim3+T cells). A depletion of PTPRO promoted PD-L1 secretion in both monocytes and macrophages through the JAK2/STAT1 and JAK2/STAT3/c-MYC pathways. Increased IL-6 expression was associated with activation of JAK2/STAT3/c-MYC and with decreased PTPRO expression through the STAT3/c-MYC/miR-25-3 p axis. Monocytes and TAMs showed significantly increased miR-25-3 p expression, which could target the 3' untranslated region of PTPRO. The miR-25-3 p expression positively correlated with serum IL-6 levels, but inversely correlated with PTPRO in HCC monocytes. IL-6/STAT3/c-MYC activation enhanced in vitro miR-25-3 p transcription and decreased PTPRO, while further promoting PD-L1 secretion. Adoptive cell transfer of c-MYC/miR-25-3 p-modified monocytes promoted tumor growth by downregulating PTPRO and causing a PD-L1-induced immunosuppression in an orthotopic tumor transplantation model. Increased serum IL-6 downregulated PTPRO expression in HCC monocytes and macrophages by activating STAT3/c-MYC/miR-25-3 p and by further enhancing PD-L1 expression through JAK2/STAT1 and JAK2/STAT3/c-MYC signaling.


Guidelines for the Pharmacotherapy of Schizophrenia in Adults.
The present guidelines address the pharmacotherapy of schizophrenia in adults across different stages, phases, and symptom domains. Guidelines were developed using the ADAPTE process, which takes advantage of existing guidelines. Six guidelines were identified for adaptation, with recommendations extracted from each. For those specific to the pharmacotherapy of schizophrenia in adults, a working group selected between guidelines and recommendations to create an adapted guideline. Recommendations can be categorized into 6 areas that include 1) first-episode schizophrenia, 2) acute exacerbation, 3) relapse prevention and maintenance treatment, 4) treatment-resistant schizophrenia, 5) clozapine-resistant schizophrenia, and 6) specific symptom domains. For each category, recommendations are made based on the available evidence, which is discussed and linked to other established guidelines. In most cases, evidence-based recommendations are made that can be used to guide current clinical treatment and decision making. Notably, however, there is a paucity of established evidence to guide treatment decision making in the case of clozapine-resistant schizophrenia, a subsample that represents a sizable proportion of those with schizophrenia.


Viper fangs: functional limitations of extreme teeth.
The fangs of vipers are extremely long, rotating, hollow teeth. Analysis of video records of more than 750 strikes recorded at 60 or 250 frames per second for 285 individuals representing 86 species in 31 genera shows that vipers reposition fangs after initial contact with prey in more than a third of the strikes. Repositioning resulted when fangs missed prey entirely or hit prey regions that did not permit adequate penetration. The prevalence of repositioning, even among species that normally release prey, suggests strong selective pressure for rapid neuromotor response to fang placement error. The rapidity of repositioning suggests the existence of (a) fine-scale sensory detection of fang penetration depth, (b) rapid modulation of contraction of antagonistic muscles, and (c) possibly neurological modifications to shorten transmission time between sensory input and motor output. Extreme fang length has apparently coevolved with extreme functions.


Correlation between Immunohistochemical Markers in Hepatocellular Carcinoma Cells and In Vitro High-Throughput Drug Sensitivity Screening.
This study analyzed the correlation between immunohistochemical markers in hepatocellular carcinoma cells and the results of in vitro high-throughput drug sensitivity screening, to provide a reference for individualized drug treatment in patients with liver cancer. Seventy-four patients with hepatocellular carcinoma were included in this study from December 2019 to June 2021, and their liver cancer cells were used for in vitro high-throughput drug sensitivity screening. According to the screening results, the patients were divided into relatively sensitive and insensitive groups, and the correlations between sensitivity and immunohistochemistry results were analyzed statistically. Alpha-fetoprotein (AFP)-positive liver cancer cells were significantly more sensitive to gemcitabine than AFP-negative cells (χ 2 = 6.102, P=0.014). AFP was also positively correlated with sensitivity of liver cancer cells to three combined regimens containing oxaliplatin (L-OHP) and epirubicin (EPI) : L-OHP + EPI + irinotecan + 5-fluorouracil (5-FU), L-OHP + irinotecan + EPI, and L-OHP + EPI (χ 2 = 8.168, P=0.004, χ 2 = 5.705, P=0.017, and χ 2 = 8.275, P=0.004, respectively). Gemcitabine and L-OHP + EPI + irinotecan + 5-FU, L-OHP + EPI, and L-OHP + irinotecan + EPI were more effective against AFP-positive compared with AFP-negative liver cancer cells according to in vitro high-throughput drug sensitivity screening. These results may guide the selection of personalized drug treatments for patients with advanced liver cancer in the future but still need further clinical studies to confirm.


Structural Components for Amplification of Positive and Negative Strand VEEV Splitzicons.
RNA is a promising nucleic acid technology for both vaccines and therapeutics, and replicon RNA has gained traction as a next-generation RNA modality. Replicon RNA self-amplifies using a replicase complex derived from alphaviral non-structural proteins and yields higher protein expression than a similar dose of messenger RNA. Here, we debut RNA splitzicons; a split replicon system wherein the non-structural proteins (NSPs) and the gene of interest are encoded on separate RNA molecules, but still exhibit the self-amplification properties of replicon RNA. We designed both positive and negative strand splitzicons encoding firefly luciferase as a reporter protein to determine which structural components, including the 5' untranslated region (UTR), a 51-nucleotide conserved sequence element (CSE) from the first nonstructural protein, the subgenomic promoter (SGP) and corresponding untranslated region, and an internal ribosomal entry site (IRES) affect amplification. When paired with a NSP construct derived from the whole, wild type replicon, both the positive and negative strand splitzicons were amplified. The combination of the 51nt CSE, subgenomic promoter and untranslated region were imperative for the positive strand splitzicon, while the negative strand was amplified simply with inclusion of the subgenomic promoter. The splitzicons were amplified by NSPs in multiple cell types and show increasing protein expression with increasing doses of NSP. Furthermore, both the positive and negative strand splitzicons continued to amplify over the course of 72 h, up to >100,000-fold. This work demonstrates a system for screening the components required for amplification from the positive and negative strand intermediates of RNA replicons and presents a new approach to RNA replicon technology.


Predicting the risk of future diabetic foot ulcer occurrence: a prospective cohort study of patients with diabetes in Tanzania.
The aim of this study was to identify the parameters that predict the risk of future foot ulcer occurrence in patients with diabetes. 1810 (male (M)/female (F): 1012/798) patients, with no foot ulcer at baseline, participated in this study. Data from a set of 28 parameters were collected at baseline. During follow-up, 123 (M/F: 68/55) patients ulcerated. Survival analyses together with logistic regression were used to identify the parameters that could predict the risk of future diabetic foot ulcer occurrence. A number of parameters (HR (95% CI)) including neuropathy (2.525 (1.680 to 3.795)); history of ulceration (2.796 (1.029 to 7.598)); smoking history (1.686 (1.097 to 2.592)); presence of callus (1.474 (0.999 to 2.174)); nail ingrowth (5.653 (2.078 to 15.379)); foot swelling (3.345 (1.799 to 6.218)); dry skin (1.926 (1.273 to 2.914)); limited ankle (1.662 (1.365 to 2.022)) and metatarsophalangeal (MTP) joint (2.745 (1.853 to 4.067)) ranges of motion; and decreased (3.141 (2.102 to 4.693)), highly decreased (5.263 (1.266 to 21.878)), and absent (9.671 (5.179 to 18.059)) sensation to touch; age (1.026 (1.010 to 1.042)); vibration perception threshold (1.079 (1.060 to 1.099)); duration of diabetes (1.000 (1.000 to 1.000)); and plantar pressure at the first metatarsal head (1.003 (1.001 to 1.005)), temperature sensation (1.019 (1.004 to 1.035)) and temperature tolerance (1.523 (1.337 to 1.734)) thresholds to hot stimuli and blood sugar level (1.027 (1.006 to 1.048)) were all significantly associated with increased risk of ulceration. However, plantar pressure underneath the fifth toe (0.990 (0.983 to 0.998)) and temperature sensation (0.755 (0.688 to 0.829)) and temperature tolerance (0.668 (0.592 to 0.0754)) thresholds to cold stimuli showed to significantly decrease the risk of future ulcer occurrence. Multivariate survival model indicated that nail ingrowth (4.42 (1.38 to 14.07)); vibration perception threshold (1.07 (1.04 to 1.09)); dry skin status (4.48 (1.80 to 11.14)); and temperature tolerance threshold to warm stimuli (1.001 (1.000 to 1.002)) were significant predictors of foot ulceration risk in the final model. The mean time to ulceration was significantly (p<0.05) shorter for patients with: dry skin (χ2=11.015), nail ingrowth (χ2=14.688), neuropathy (χ2=21.284), or foot swelling (χ2=16.428). Nail ingrowth and dry skin were found to be strong indicators of vulnerability of patients to diabetic foot ulceration. Results highlight that assessments of neuropathy in relation to both small and larger fiber impairment need to be considered for predicting the risk of diabetic foot ulceration.


Evaluation of the buffy-coat smear for rapid detection of bacteremia.
Buffy-coat smears prepared from 599 blood samples, obtained simultaneously with blood for culture, were examined for bacteria. Each blood sample was quantitatively cultured on chocolate agar. Twenty-one blood cultures (3.6%) from 17 patients were positive. Two (11.7%) of the 17 patients had positive buffy-coat smears. One patient had two positive smears, for a total of three positive buffy-coat smears from 21 positive blood cultures. Chart reviews were performed for two patients with positive buffy-coat smears, three patients with positive blood cultures (containing the same organisms as the two study patients) but negative buffy-coat smears, and six additional patients who had had positive buffy-coat smears between 1974 and 1979. Of the eight patients with positive smears, six (75%) were infected with Gram-positive cocci, three of which were Streptococcus pneumoniae. Mortality was 100%.


Impact of metal ions on PCR inhibition and RT-PCR efficiency.
Inhibition of PCR by metal ions can pose a serious challenge in the process of forensic DNA analysis. Samples contaminated with various types of metal ions encountered at crime scenes include swabs from metal surfaces such as bullets, cartridge casings, weapons (including guns and knives), metal wires and surfaces as well as bone samples which contain calcium. The mechanism behind the impact of metal ions on DNA recovery, extraction and subsequent amplification is not fully understood. In this study, we assessed the inhibitory effects of commonly encountered metals on DNA amplification. Of the nine tested metals, zinc, tin, iron(II) and copper were shown to have the strongest inhibitory properties having IC50 values significantly below 1 mM. In the second part of the study, three commercially available DNA polymerases were tested for their susceptibility to metal inhibition. We found that KOD polymerase was the most resistant to metal inhibition when compared with Q5 and Taq polymerase. We also demonstrate how the calcium chelator ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) can be used as an easy and non-destructive method of reversing calcium-induced inhibition of PCR reactions.


Phase 1 Clinical Study of siRNA Targeting Carbohydrate Sulphotransferase 15 in Crohn's Disease Patients with Active Mucosal Lesions.
Carbohydrate sulphotransferase 15 [CHST15] is a specific enzyme biosynthesizing chondroitin sulphate E that binds various pathogenic mediators and is known to create local fibrotic lesions. We evaluated the safety of STNM01, a synthetic double-stranded RNA oligonucleotide directed against CHST15, in Crohn's disease [CD] patients whose mucosal lesions were refractory to conventional therapy. This was a randomized, double-blind, placebo-controlled, concentration-escalation study of STNM01 by a single-dose endoscopic submucosal injection in 18 CD patients. Cohorts of increasing concentration of STNM01 were enrolled sequentially as 2.5nM [n = 3], 25nM [n = 3], and 250nM [n = 3] were applied. A cohort of placebo [n = 3] was included in each concentration. Safety was monitored for 30 days. Pharmacokinetics was monitored for 24h. The changes from baseline in the segmental Simple Endoscopic Score for CD [SES-CD] as well as the histological fibrosis score were evaluated. STNM01 was well tolerated and showed no drug-related adverse effects in any cohort of treated patients. There were no detectable plasma concentrations of STNM01 at all measured time points in all treatment groups. Seven of nine subjects who received STNM01 showed reduction in segmental SES-CD at Day 30, when compared with those who received placebo. Histological analyses of biopsy specimens revealed that STNM01 reduced the extent of fibrosis. Local application of STNM01 is safe and well tolerated in CD patients with active mucosal lesions.


Three-dimensional distribution function theory for the prediction of protein-ligand binding sites and affinities: application to the binding of noble gases to hen egg-white lysozyme in aqueous solution.
The three-dimensional distribution function theory of molecular liquids is applied to lysozyme in mixtures of water and noble gases. The results indicate that the theory has the capability of predicting the protein-ligand binding sites and affinities. First, it is shown that the theory successfully reproduces the binding sites of xenon found by X-ray crystallography. Then, the ability of the theory to predict the size selectivity of noble gases is demonstrated. The effect of water on the selectivity is clarified by a theoretical analysis. Finally, it is demonstrated that the dose-response curve, which is employed in experiments for examining the binding affinity, is realized by the theory.


Impact of journal articles and grand rounds on practice: CT scanning in appendicitis.
The purpose of this study was to determine whether a report in a high-impact journal published in January 1998 changed practice patterns and to further explore the impact of a review of the subject in a department of surgery grand rounds (January 2000). Charts from all patients undergoing appendectomy at our institution during three time periods (January to December 1997, January to December 1999, and January to June 2000) were reviewed. Rates of CT scanning, negative appendectomy, and perforated/gangrenous appendicitis were compared for the three periods to determine the impact of the journal article and the subject review during grand rounds on practice patterns and outcomes. Charts from 230 (88%) of 262 patients who underwent appendectomy during the time periods were available for review. Age, percentage of male patients, temperature on admission, and white blood cell count did not differ among the groups. The rate of CT scanning increased significantly from 1997 to 1999 and again in 2000 (6.7%, 43%, and 70%, respectively; P < 0.001), whereas the proportion of perforated/gangrenous appendicitis decreased significantly from 33% in 1997 and 31% in 1999 to 13% in 2000 (P = 0.012). The use of CT scanning in appendicitis increased both after publication of a report in a high-impact journal and after review during grand rounds. A rate of CT scanning above 45% appeared to affect outcomes as well.


Chemical Constituents from Scindapsus officinalis (Roxb.) Schott. and Their Anti⁻Inflammatory Activities.
One new monoterpene glycoside (1), one new phenyl glycoside (2), one new caffeoyl derivative (3), were isolated from Scindapsus officinalis (Roxb.) Schott., along with four known compounds (4⁻7). Structures of the isolated compounds were elucidated by extensive analysis of spectroscopic data, especially 2D NMR data and comparison with literatures. All isolates were evaluated for anti-inflammatory activity against nitric oxide (NO) production in vitro. Compounds 3 and 7 exhibited moderate inhibitory effects on NO production with IC50 values of 12.2 ± 0.8 and 18.9 ± 0.3 μM, respectively.


Mortality following exenteration for malignant tumours of the orbit.
Orbital exenteration is a rare, but disfiguring procedure reserved for the treatment of locally invasive malignancy or potentially life threatening orbital neoplasms, when less destructive techniques are inadequate. The authors report their experience and analyses of 64 cases of orbital exenteration performed over a 13 year period, looking specifically at key factors affecting mortality associated with such a destructive surgical procedure. Records were reviewed retrospectively of all patients who had undergone exenteration of the orbit from 1 January 1991 to 1 April 2004 inclusive, at the Manchester Royal Eye Hospital. In all cases of deceased individuals, the cause of death was determined by liaison with the general practitioner and local health authority. Duplicate death certificates were requested for all deceased patients from the Registrar for Births, Deaths, and Marriages, Southport, UK. Kaplan-Meier analysis was used to estimate survival following exenteration. Overall, 1 year survival post-exenteration was high at 93%. After 3 years this had fallen to 67%, followed by 57% after 5 years, and 37% at 10 years. 13 patients died as a direct result of the orbital tumour. A further nine died of unrelated medical conditions, and two patients succumbed to malignant processes originating elsewhere in the body. There was no difference in survival rate at 3 years (p=0.99) and 5 years (p=0.454) between those with clear resection margins and those without. In this study it was found that there was an overall mortality rate of 38% over 12 years. The presence of clear surgical margins, although reassuring for the surgeon, should not be regarded as an indication of cure. However, an overall 1 year survival of 93% and a 10 year survival of 37% are reassuring in that a proportion of individuals achieve surgical cure following exenteration. 38% of patients died as a result of other medical causes over the 12 year follow up.


Candida accommodates non-culturable Helicobacter pylori in its vacuole - Koch's postulates aren't applicable.
The following are the responses to the "letter to the editor" ("Helicobacter is preserved in yeast vacuoles! Does Koch's postulates confirm it?") authored by Nader Alipour and Nasrin Gaeini that rejected the methods, results, discussions and conclusions summarized in the review article authored by Siavoshi F and Saniee P. In the article, 7 papers, published between 1998 and 2013, were reviewed. The 7 papers had been reviewed and judged very carefully by the assigned expertise of the journals involved, including the reviewers of the World Journal of Gastroenterology (WJG), before publication. In the review article, 121 references were used to verify the methods, results and discussions of these 7 papers. The review article was edited by the trustworthy British editor of the (WJG), and the final version was rechecked and finally accepted by the reviewers of (WJG). None of the reviewers made comments like those in this "letter to the editor", especially the humorous comments, which seem unprofessional and nonscientific. Above all, the authors' comments show a lack of understanding of basic and advanced microbiology, e.g. bacterial endosymbiosis in eukaryotic cells. Accordingly, their comments all through the letter contain misconceptions. The comments are mostly based on personal conclusions, without any scientific support. It would have been beneficial if the letter had been reviewed by the reviewers of the article by Siavoshi and Saniee.


The resistomes of Mycobacteroides abscessus complex and their possible acquisition from horizontal gene transfer.
Mycobacteroides abscessus complex (MABC), an emerging pathogen, causes human infections resistant to multiple antibiotics. In this study, the genome data of 1,581 MABC strains were downloaded from NCBI database for phylogenetic relatedness inference, resistance profile identification and the estimation of evolutionary pressure on resistance genes in silico. From genes associated with resistance to 28 antibiotic classes, 395 putative proteins (ARPs) were identified, based on the information in two antibiotic resistance databases (CARD and ARG-ANNOT). The ARPs most frequently identified in MABC were those associated with resistance to multiple antibiotic classes, beta-lactams and aminoglycosides. After excluding ARPs that had undergone recombination, two ARPs were predicted to be under diversifying selection and 202 under purifying selection. This wide occurrence of purifying selection suggested that the diversity of commonly shared ARPs in MABC have been reduced to achieve stability. The unequal distribution of ARPs in members of the MABC could be due to horizontal gene transfer or ARPs pseudogenization events. Most (81.5%) of the ARPs were observed in the accessory genome and 72.2% ARPs were highly homologous to proteins associated with mobile genetic elements such as plasmids, prophages and viruses. On the other hand, with TBLASTN search, only 18 of the ARPs were identified as pseudogenes. Altogether, our results suggested an important role of horizontal gene transfer in shaping the resistome of MABC.


Draft Genome Sequence of Ochrobactrum intermedium Strain SA148, a Plant Growth-Promoting Desert Rhizobacterium.
Ochrobactrum intermedium strain SA148 is a plant growth-promoting bacterium isolated from sandy soil in the Jizan area of Saudi Arabia. Here, we report the 4.9-Mb draft genome sequence of this strain, highlighting different pathways characteristic of plant growth promotion activity and environmental adaptation of SA148.