Characteristics and outcomes of antiretroviral-treated HIV-HBV co-infected patients in Canada?
Hepatitis B (HBV) and Human Immunodeficiency Virus (HIV) share common risk factors for exposure. Co-infected patients have an increased liver-related mortality risk and may have accelerated HIV progression. The epidemiology and demographic characteristics of HIV-HBV co-infection in Canada remain poorly defined. We compared the demographic and clinical characteristics and factors associated with advanced hepatic fibrosis between HIV and HIV-HBV co-infected patients. A retrospective cohort analysis was conducted using data from the Canadian Observational Cohort (CANOC) Collaboration, including eight sites from British Columbia, Quebec, and Ontario. Eligible participants were HIV-infected patients who initiated combination ARV between January 1, 2000 and December 14, 2014. Demographic and clinical characteristics were compared between HIV-HBV co-infected and HIV-infected groups using chi-square or Fisher exact tests for categorical variables, and Wilcoxon's Rank Sum test for continuous variables. Liver fibrosis was estimated by the AST to Platelet Ratio Index (APRI). HBV status and APRI values were available for 2419 cohort participants. 199 (8%) were HBV co-infected. Compared to HIV-infected participants, HIV-HBV co-infected participants were more likely to use injection drugs (28% vs. 21%, p = 0.03) and be HCV-positive (31%, vs. 23%, p = 0.02). HIV-HBV co-infected participants had lower baseline CD4 T cell counts (188 cells/mm3, IQR: 120-360) compared to 235 cells/mm3 in HIV-infected participants (IQR: 85-294) (p = 0.0002) and higher baseline median APRI scores (0.50 vs. 0.37, p < 0.0001). This difference in APRI was no longer clinically significant at follow-up (0.32 vs. 0.30, p = 0.03). HIV-HBV co-infected participants had a higher mortality rate compared to HIV-infected participants (11% vs. 7%, p = 0.02). The prevalence, demographic and clinical characteristics of the HIV-HBV co-infected population in Canada is described. HIV-HBV co-infected patients have higher mortality, more advanced CD4 T cell depletion, and liver fibrosis that improves in conjunction with ARV therapy. The high prevalence of unknown HBV status demonstrates a need for increased screening among HIV-infected patients in Canada.


Probing the seismic cycle timing with coseismic twisting of subduction margins.
Assessing the timing of great megathrust earthquakes is together crucial for seismic hazard analysis and deemed impossible. Geodetic instrumentation of subduction zones has revealed unexpected deformation patterns at subduction segments adjacent to those that hosted recent mega-earthquakes: coastal sites move landward with faster velocities than before the earthquake. Here, we show observations from the largest and best-monitored megathrust earthquakes, and from a scaled analog model, to reveal that these events create coseismic and postseismic deformation patterns typical of a complete gear-like rotation about a vertical axis, hereafter called twisting. We find that such twisting alters the interseismic velocity field of adjacent subduction segments depending on the time since the last earthquake. Early interactions accelerate while late interactions decelerate local kinematics. This finding opens the possibility of using megathrust earthquakes, the characteristics of the twisting pattern, and the ensuing geodetic velocity changes, as a proxy for estimating the timing of the seismic cycle at unruptured segments along the margin.


Characteristics and etiology of infection in febrile infants aged ≤90 days.
Infants ≤90 days old can exhibit non-specific signs of infection, even in cases of serious bacterial infection (SBI). This prospective study included infants aged ≤90 days hospitalized for fever from June 2017 to August 2019. Nasopharyngeal swabs were tested using multiplex real-time polymerase chain reaction (PCR) tests and 16S ribosomal RNA analysis of whole blood to determine causative microorganisms. Data pertaining to inflammatory markers, maximum body temperature (BT), and respiratory symptoms of infants and their cohabiting families were collected at admission. A total of 110 infants were enrolled (age range, 9-90 days), 17 (15.5%) of whom presented with SBIs. White blood cell (WBC) count and absolute neutrophil count (ANC) were significantly higher in patients with SBIs than in those without, although maximum BT did not significantly differ between the SBI and non-SBI groups (n = 93). One or more viruses were detected in 82 infants (74.5%). Viruses were detected more frequently in infants with respiratory symptoms than in those without respiratory symptoms (P = 0.038), and patients with SBIs experienced significantly less respiratory symptoms than those without SBIs (P = 0.049). Moreover, viruses were more often detected in infants from cohabiting families with respiratory symptoms than in those whose family members did not exhibit respiratory symptoms (P = 0.0018). White blood cell count, and ANC were significantly higher, and respiratory symptoms were less in infants ≤90 days old with SBIs than in those without SBIs. Microorganisms from nasopharyngeal by multiplex real-time PCR swabs could not be judged as SBI or non-SBI.


Safety of bronchoscopy in patients with malignant hematologic disorders.
Factors affecting the safety of bronchoscopy in patients with malignant hematologic disorders have not been well described. We evaluated the safety of bronchoscopy and describe factors affecting its complication rate in such patients. Between January 2009 and December 2018, 316 bronchoscopies in 282 patients with malignant hematologic disorders and pulmonary infiltrates were performed at our institution. The bronchoscopic procedure used and its complications were evaluated. The most common underlying disease was acute myeloid leukemia (134/282 patients, 47.5%). Platelet transfusion was performed the day before or the day of bronchoscopy in 42.4%, supplemental oxygen was administered before the procedure in 23.1%, and midazolam was used in 74.4%. Thirty-five bronchoscopies (11.1%) were complicated by hemoptysis and 7 patients developed pneumothorax, 4 of whom required thoracic drainage. Two patients (0.6%) were intubated within 48 h of the procedure and prolonged oxygen desaturation (> 48 h) occurred in 3.8%. Multivariate analysis showed that only use of midazolam significantly reduced the risk of prolonged oxygen desaturation (hazard ratio 0.28, 95% confidence interval 0.09-0.85, p = 0.03). Transbronchial lung biopsy significantly increased the risk of hemoptysis (hazard ratio 10.40, 95% confidence interval 4.18-25.90, p = 0.00), while use of midazolam significantly reduced the risk (hazard ratio 0.31, 95% confidence interval 0.14-0.73, p = 0.01). Bronchoscopy is relatively safe in patients with malignant hematologic disorders. Caution and judicious use of sedatives may improve the patient's procedural tolerance and lower complications.


Effects of smoking on platelets and on plasma thromboxane-prostacyclin balance in man.
To determine the influence of brief period of smoking on platelet release and platelet-vessel wall prostaglandins, we studied 13 non-smokers and 15 habitual smokers. Blood samples were collected before and immediately after smoking. Before smoking plasma beta-thromboglobulin levels were similar in the two groups. Plasma levels of thromboxane B2 (TXB2), stable metabolite of TXA2 were slightly higher (124 +/- 43 vs 101 +/- 36 pg/ml) and of 6-keto-PGF1 alpha (stable metabolite of PGI2) lower (39 +/- 23 vs 63 +/- 23 pg/ml) in smokers compared to non-smokers. Immediately after smoking, plasma TXB2 levels increased 197% (P less than 0.05) and plasma 6-keto-PGF1 alpha 44% (P-NS) in non-smokers. Similar increases were not observed in the smoker population, although plasma beta-thromboglobulin levels increased 40%. Platelet TXA2 generations induced by arachidonic acid and thrombin were similar in the two groups. These data show that brief period of smoking stimulates both TXA2 and PGI2 release in the non-smokers. Increase in TXA2 is more than in PGI2. Episodic increase in TXA2 may be a mechanism of vascular injury. Absence of increase in TXA2 and PGI2 with smoking in habitual smokers may reflect tolerance to effects of smoking.


A fully degradable transcatheter ventricular septal defect occluder: Towards rapid occlusion and post-regeneration absorption.
Degradable heart occluders are a promising replacement for currently clinically used non-degradable ones without concerns about the complications caused by the persistent residue of a foreign implant. However, the inherent mechanical properties of degradable occluders are poor and decline with material degradation, leading to a preference for a long degradation period upon designing a degradable heart occluder. This configuration can lower the risk of occluder dislodgement but reduce the benefits of degradable implants over their non-degradable counterparts due to a longer retention of foreign materials in the human body. Here, we fabricated a fully degradable ventricular septum defect (VSD) occluder consisting of polydioxanone (PDO) fiber and poly-L-lactic acid (PLLA) membrane featuring an auto-locking function. The degradable occluder showed an excellent shape recovery effect after transcatheter delivery and anchored securely to a heart defect as evidenced by in vitro and in vivo experiments. The degradable occluder could warrant robust fixation ability during the first 3-months of implantation within which tissue reconstruction was accomplished and be completely absorbed within 12 months. Benefitting from these merits, the degradable occluder displayed desired occlusion and no complications after being implanted in the VSD sites of canines during a 24-months follow-up. Compared with traditional non-degradable occluders, our degradable occluder could provide a potentially superior approach for rapidly repairing the congenital VSD without interfering with the normal development and physiological function of the heart.


Rapid and sensitive determination of the intermediates of advanced glycation end products in the human nail by ultra-performance liquid chromatography with electrospray ionization time-of-flight mass spectrometry.
The resolution of the intermediate advanced glycation end products (AGEs) in the human nail was carried out by the combination of 4,5-dimethyl-1,2-phenylenediamine (DMPD) derivatives and ultra-performance liquid chromatography with electrospray ionization time-of-flight mass spectrometry (UPLC-ESI-TOF-MS). The reaction of the reagent with 3-deoxyglucosone (3-DG), methylglyoxal (MG), and glyoxal (GO) effectively proceeds at 60°C for 2h. The resulting derivatives were efficiently separated by a gradient program (a mixture of water and acetonitrile containing 0.1% formic acid) using a reversed-phase ACQUITY UPLC BEH C(18) column (1.7 μm, 50×2.1 mm i.d.) and sensitively detected by TOF-MS. The detection limits (signal-to-noise ratio=5) of the TOF-MS were 10 to 50 fmol. A good linearity was achieved from the calibration curve, which was obtained by plotting the peak area ratios of the analytes relative to the internal standard (IS) (i.e., 2,3-hexanedione) versus the injected amounts of 3-DG, MG, and GO (r(2)>0.999), and the intra- and interday assay precisions were less than 6.89%. The derivatives of the compounds in the human nail were successfully identified by the proposed procedure. As we know, these three kinds of dicarbonyl intermediates in the formation of AGEs-3-DG, MG, and GO-were first found in human nail samples. Using these methods, the amounts of compound in the nails of healthy volunteers and diabetic patients were determined. When comparing the index from the diabetic patients with that from healthy volunteers, there is no significant difference in the content of the MG and GO in the nails. However, a statistically significant (P<0.001) correlation was observed between the 3-DG concentrations. Because the proposed method provides a good mass accuracy and the trace detection of the dicarbonyl intermediates of AGEs in the human nail, this analytical technique could be a noninvasive technique to assist in the diagnosis and assessment of disease activity in diabetic patients. Here we present a novel, sensitive, and simple method for the simultaneous determination of dicarbonyl compounds in the human nail.


Impact of a constant current electrical stimulation (CCES) system and hormonal growth-promoting (HGP) implants on meat quality and palatability of finished steers.
This study evaluated the effects of a constant current electrical stimulation (CCES) system and hormonal growth-promoting (HGP) implants on the quality and palatability of the longissimus thoracis et lumborum (LTL) from yearling-finished steers. The experiment used a total of 46 Angus cross steers, which were either non-implanted (n = 20) or implanted with trenbolone acetate and estradiol benzoate (n = 26). The CCES was applied to one side of each carcass during the slaughter process, whereas the other side remained unstimulated. Regardless of the application of HGP implants, the CCES reduced pH at 3 and 72 h post-mortem and shear force at all ageing times (P < 0.05), improved colour at 72 h post-mortem and during the retail display (P < 0.05), increased initial and overall tenderness (P < 0.01), and decreased the amount of perceived connective tissue and the proportion of trained panelists detecting spongy texture (P < 0.05) compared to meat from unstimulated carcass sides. Although CCES increased meat purge losses and reduced moisture content (P < 0.05), this did not affect meat juiciness (P > 0.10). CCES interacted with HGP to prevent increase in drip loss (P > 0.10), increase frequency of panelists detecting bloody/serumy flavour and typical texture, and reduce the proportion of panelists detecting rubbery texture in meat (P < 0.05). Regardless of stimulation treatment, meat from implanted animals had a more pronounced pH decline at 72 h post-mortem (P < 0.05) and a higher proportion of panelists finding no off-flavours (P < 0.05) or bloody/serumy flavour (P < 0.01) than non-implanted cattle. The CCES system tested in this study improved LTL quality and palatability of heavier beef carcasses.


Clinical characterization of autosomal dominant retinitis pigmentosa with NRL mutation in a three-generation Japanese family.
Retinitis pigmentosa (RP) is a heterogeneous group of inherited retinal disorders. NRL-associated autosomal dominant (AD)-RP is a rare form of AD-RP in the Japanese population. This study aimed to report a clinical characterization of NRL-associated retinopathy in a three-generation Japanese family. A total of 4 patients from a Japanese family were referred to The Jikei University School of Medicine for clinical and genetic examination. The patients included a male proband (41 years old), his daughters (5 and 6 years old), and his mother (71 years old); they underwent ophthalmic examinations, and genetic testing was performed using whole exome sequencing analysis, revealing a known variant [c.152C > T (p.Pro51Leu)] heterozygously in exon 2 of the NRL gene. Fundus photograph showed that retinal degeneration expanded to the macular and peripheral retina in an age-dependent manner. Fundus autofluorescence imaging showed hyper-autofluorescence (AF) within the macular with slightly hypo-AF in younger patients and obvious hypo-AF in older patients. Optical coherence tomography showed that the length of the ellipsoid zone tended to be longer in younger patients than in older patients. Goldmann perimetry showed an age-dependent decrease in the visual field. Furthermore, full-field electroretinographic findings revealed non-recordable rod and cone function in older patients and non-recordable rod function with preserved cone function in younger patients. Our results indicated that retinal construction and function were aggravated in an age-dependent manner, and retinal degeneration, especially in the macular region, revealed milder findings than in previous cases with NRL-associated AD-RP.


Fracture Dislocations of the Glenohumeral Joint.
Proximal humerus fracture dislocations typically result from high-energy mechanisms and carry specific risks, technical challenges, and management considerations. It is vital for treating surgeons to understand the various indications, procedures, and complications involved with their treatment. While these injuries are relatively rare in comparison with other categories of proximal humerus fractures, fracture dislocations of the proximal humerus require treating surgeons to consider patient age, activity level, injury pattern, and occasionally intra-operative findings to select the ideal treatment strategy for each injury. Proximal humerus fracture dislocations are complex injuries that require special considerations. This review summarizes recent literature regarding the evaluation and management of these injuries as well as the indications and surgical techniques for each treatment strategy. Thorough pre-operative patient evaluation and shared decision-making should be employed in all cases. While nonoperative management is uncommonly considered, open reduction and internal fixation (ORIF), hemiarthroplasty, and reverse total shoulder replacement are at the surgeon's disposal, each with their own indications and complication profile.


The influence of sex and diet on the characteristics of hibernation in Syrian hamsters.
Research on deep hibernators almost exclusively uses species captured from the wild or from local breeding. An exception is Syrian hamster (Mesocricetus auratus), the only standard laboratory animal showing deep hibernation. In deep hibernators, several factors influence hibernation quality, including body mass, sex and diet. We examined hibernation quality in commercially obtained Syrian hamsters in relation to body mass, sex and a diet enriched in polyunsaturated fatty acids. Animals (M/F:30/30, 12 weeks of age) were obtained from Harlan (IN, USA) and individually housed at 21 °C and L:D 14:10 until 20 weeks of age, followed by L:D 8:16 until 27 weeks. Then conditions were changed to 5 °C and L:D 0:24 for 9 weeks to induce hibernation. Movement was continuously monitored with passive infrared detectors. Hamsters were randomized to control diet or a diet 3× enriched in linoleic acid from 16 weeks of age. Hamsters showed a high rate of premature death (n = 24, 40%), both in animals that did and did not initiate torpor, which was unrelated to body weight, sex and diet. Time to death (31.7 ± 3.1 days, n = 12) or time to first torpor bout (36.6 ± 1.6 days, n = 12) was similar in prematurely deceased hamsters. Timing of induction of hibernation and duration of torpor and arousal was unaffected by body weight, sex or diet. Thus, commercially obtained Syrian hamsters subjected to winter conditions showed poor survival, irrespective of body weight, sex and diet. These factors also did not affect hibernation parameters. Possibly, long-term commercial breeding from a confined genetic background has selected against the hibernation trait.


Comparison of false discovery rate methods in identifying genes with differential expression.
Current high-throughput techniques such as microarray in genomics or mass spectrometry in proteomics usually generate thousands of hypotheses to be tested simultaneously. The usual purpose of these techniques is to identify a subset of interesting cases that deserve further investigation. As a consequence, the control of false positives among the tests called "significant" becomes a critical issue for researchers. Over the past few years, several false discovery rate (FDR)-controlling methods have been proposed; each method favors certain scenarios and is introduced with the purpose of improving the control of FDR at the targeted level. In this paper, we compare the performance of the five FDR-controlling methods proposed by Benjamini et al., the qvalue method proposed by Storey, and the traditional Bonferroni method. The purpose is to investigate the "observed" sensitivity of each method on typical microarray experiments in which the majority (or all) of the truth is unknown. Based on two well-studied microarray datasets, it is found that in terms of the "apparent" test power, the ranking of the FDR methods is given as Step-down<Step-up: dependent<Step-up: one-stage (BH95)<Step-up adaptive<qvalue. The BH95 method shows the best control of FDR at the target level. It is our hope that the observed results could provide some insight into the application of different FDR methods in microarray data analysis.


Tuberculous radiculomyelitis presenting in a toddler with lower extremity weakness and seizure.
Tuberculous radiculomyelitis is an uncommon but serious complication of tuberculosis that can lead to considerable morbidity and mortality. We present the case of a 21-month-old male Congolese refugee diagnosed with tuberculous radiculomyelitis who presented with gradual motor and speech regression, and likely an infection-related seizure 2 months before diagnosis.


Chromosome damage induced by nanomolar concentrations of bleomycin in porated mammalian cells.
We have examined chromosome damage caused by a wide range of bleomycin (BLM) concentrations in Chinese hamster ovary (CHO-K1) cells reversibly porated by the bacterial cytotoxin streptolysin-O (SLO). Chromosome damage was measured using the micronucleus cytokinesis block technique (employing cytochalasin-B). Treatment of exponentially growing cells with 0.045 IU/mL SLO for 5 min resulted in up to a thousand-fold and a million-fold increase in biological effectiveness, compared to treatment in the absence of SLO for 24 hr and 5 min, respectively. Increases in micronuclei of 4-5 times background level were observed after only 5 min exposure to the drug in the presence of SLO at doses as low as 100 pg/mL (approximately 70 pmol/L). These results indicate that the use of SLO may facilitate the treatment of cells with BLM for periods of time resembling acute exposure to ionizing radiations.


The Consequentialist Scale: Translation and empirical investigation in a Greek sample.
The Consequentialist Scale (Robinson, 2012) [89] assesses the endorsement of consequentialist and deontological moral beliefs. This study empirically investigated the application of the Greek translation of the Consequentialist Scale in a sample of native Greek speakers. Specifically, 415 native Greek speakers completed the questionnaire. To uncover the underlying structure of the 10 items in the Consequentialist Scale, an Exploratory Factor Analysis (EFA) was conducted. The results revealed a three-factor solution, where the deontology factor exhibited the same structure as the original work by Robinson (2012) [89], while the original consequentialism factor split into two separate factors. Significant Pearson's r correlations were observed between age and responses to the Consequentialist Scale. Separate EFAs were conducted for two age groups based on a medial split: younger (36 years old or less) and older (more than 36 years old). Interestingly, the younger group exhibited a two-factor solution with the same structure as the original work, while the older group showed a three-factor solution. A hierarchical k-means cluster analysis revealed that the cluster of participants who scored higher in deontology compared to consequentialism primarily consisted of older participants, whereas the two other clusters comprised of younger participants exhibited the reverse pattern. Neither gender nor previous experience with philosophy significantly affected scores on the Consequentialist Scale. Overall, our study provides evidence that the Consequentialist Scale is suitable for use in the Greek population.


Ganaplacide (KAF156) plus lumefantrine solid dispersion formulation combination for uncomplicated Plasmodium falciparum malaria: an open-label, multicentre, parallel-group, randomised, controlled, phase 2 trial.
Emergence of drug resistance demands novel antimalarial drugs with new mechanisms of action. We aimed to identify effective and well tolerated doses of ganaplacide plus lumefantrine solid dispersion formulation (SDF) in patients with uncomplicated Plasmodium falciparum malaria. This open-label, multicentre, parallel-group, randomised, controlled, phase 2 trial was conducted at 13 research clinics and general hospitals in ten African and Asian countries. Patients had microscopically-confirmed uncomplicated P falciparum malaria (>1000 and <150 000 parasites per μL). Part A identified the optimal dose regimens in adults and adolescents (aged ≥12 years) and in part B, the selected doses were assessed in children (≥2 years and <12 years). In part A, patients were randomly assigned to one of seven groups (once a day ganaplacide 400 mg plus lumefantrine-SDF 960 mg for 1, 2, or 3 days; ganaplacide 800 mg plus lumefantrine-SDF 960 mg as a single dose; once a day ganaplacide 200 mg plus lumefantrine-SDF 480 mg for 3 days; once a day ganaplacide 400 mg plus lumefantrine-SDF 480 mg for 3 days; or twice a day artemether plus lumefantrine for 3 days [control]), with stratification by country (2:2:2:2:2:2:1) using randomisation blocks of 13. In part B, patients were randomly assigned to one of four groups (once a day ganaplacide 400 mg plus lumefantrine-SDF 960 mg for 1, 2, or 3 days, or twice a day artemether plus lumefantrine for 3 days) with stratification by country and age (2 to <6 years and 6 to <12 years; 2:2:2:1) using randomisation blocks of seven. The primary efficacy endpoint was PCR-corrected adequate clinical and parasitological response at day 29, analysed in the per protocol set. The null hypothesis was that the response was 80% or lower, rejected when the lower limit of two-sided 95% CI was higher than 80%. This study is registered with EudraCT (2020-003284-25) and ClinicalTrials.gov (NCT03167242). Between Aug 2, 2017, and May 17, 2021, 1220 patients were screened and of those, 12 were included in the run-in cohort, 337 in part A, and 175 in part B. In part A, 337 adult or adolescent patients were randomly assigned, 326 completed the study, and 305 were included in the per protocol set. The lower limit of the 95% CI for PCR-corrected adequate clinical and parasitological response on day 29 was more than 80% for all treatment regimens in part A (46 of 50 patients [92%, 95% CI 81-98] with 1 day, 47 of 48 [98%, 89-100] with 2 days, and 42 of 43 [98%, 88-100] with 3 days of ganaplacide 400 mg plus lumefantrine-SDF 960 mg; 45 of 48 [94%, 83-99] with ganaplacide 800 mg plus lumefantrine-SDF 960 mg for 1 day; 47 of 47 [100%, 93-100] with ganaplacide 200 mg plus lumefantrine-SDF 480 mg for 3 days; 44 of 44 [100%, 92-100] with ganaplacide 400 mg plus lumefantrine-SDF 480 mg for 3 days; and 25 of 25 [100%, 86-100] with artemether plus lumefantrine). In part B, 351 children were screened, 175 randomly assigned (ganaplacide 400 mg plus lumefantrine-SDF 960 mg once a day for 1, 2, or 3 days), and 171 completed the study. Only the 3-day regimen met the prespecified primary endpoint in paediatric patients (38 of 40 patients [95%, 95% CI 83-99] vs 21 of 22 [96%, 77-100] with artemether plus lumefantrine). The most common adverse events were headache (in seven [14%] of 51 to 15 [28%] of 54 in the ganaplacide plus lumefantrine-SDF groups and five [19%] of 27 in the artemether plus lumefantrine group) in part A, and malaria (in 12 [27%] of 45 to 23 [44%] of 52 in the ganaplacide plus lumefantrine-SDF groups and 12 [50%] of 24 in the artemether plus lumefantrine group) in part B. No patients died during the study. Ganaplacide plus lumefantrine-SDF was effective and well tolerated in patients, especially adults and adolescents, with uncomplicated P falciparum malaria. Ganaplacide 400 mg plus lumefantrine-SDF 960 mg once daily for 3 days was identified as the optimal treatment regimen for adults, adolescents, and children. This combination is being evaluated further in a phase 2 trial (NCT04546633). Novartis and Medicines for Malaria Venture.


The Bak core dimer focuses triacylglycerides in the membrane.
Apoptosis, the intrinsic programmed cell death process, is mediated by the Bcl-2 family members Bak and Bax. Activation via formation of symmetric core dimers and oligomerization on the mitochondrial outer membrane (MOM) leads to permeabilization and cell death. Although this process is linked to the MOM, the role of the membrane in facilitating such pores is poorly understood. We recently described Bak core domain dimers, revealing lipid binding sites and an initial role of lipids in oligomerization. Here we describe simulations that identified localized clustering and interaction of triacylglycerides (TAGs) with a minimized Bak dimer construct. Coalescence of TAGs occurred beneath this Bak dimer, mitigating dimer-induced local membrane thinning and curvature in representative coarse-grain MOM and model membrane systems. Furthermore, the effects observed as a result of coarse-grain TAG cluster formation was concentration dependent, scaling from low physiological MOM concentrations to those found in other organelles. We find that increasing the TAG concentration in liposomes mimicking the MOM decreased the ability of activated Bak to permeabilize these liposomes. These results suggest that the presence of TAGs within a Bak-lipid membrane preserves membrane integrity and is associated with reduced membrane stress, suggesting a possible role of TAGs in Bak-mediated apoptosis.


Quantification methods for Bacillus cereus vegetative cells and spores in the gastrointestinal environment.
There is an interest to understand the fate and behaviour of the food-borne pathogen Bacillus cereus in the gut, a challenging environment with a high bacterial background. We evaluated the current detection methods to select an appropriate strategy for B. cereus monitoring during gastrointestinal experiments. Application of quantitative real-time PCR (qPCR) in a gastrointestinal matrix required careful selection of the qPCR reaction and elaborate optimization of the DNA extraction protocol. Primer competition and depletion problems associated with qPCR reactions targeting general 16S rRNA gene can be avoided by the selection of a target sequence that is unique for and widespread among the target bacteria, such as the toxin gene nheB in the case of pathogenic B. cereus. Enumeration of B. cereus during the ileum phase was impossible by plating due to overgrowth by intestinal bacteria, while a carefully optimized qPCR enabled specific detection and quantification of B. cereus. On the other hand, plating allowed the distinction of viable, injured and dead bacteria and the germination of spores, which was not possible with qPCR. In conclusion, both plating and qPCR were necessary to yield the maximal information regarding the viability and physiology of the B. cereus population in various gastrointestinal compartments.


Prevalence of essential tremor: a door-to-door survey in Sile, Istanbul, Turkey.
The aim of this study was to determine the prevalence of essential tremor (ET) in a Turkish population living in Sile, a rural town located on Black Sea coast of Istanbul. This study was designed as a descriptive cross-sectional field study. People aged 18 years and over were selected for the study. Tremor surveillance questionnaire was applied by door-to-door visits. Subsequently, all subjects who answered the questionnaire had a standard tremor examination. Diagnosis was made based on Washington Heights-Inwood Genetic Study of Essential Tremor (WHIGET) diagnosis and clinical evaluation scale. A total of 2227 people was screened. The prevalence of ET was found to be 3.09% (95% Cl=2.42-3.91%) in Turkish population aged over 18 years. The prevalence of ET increased by age in both genders.


Sinapic acid ameliorates bleomycin-induced lung fibrosis in rats.
Pulmonary fibrosis is a multifaceted disease with high mortality and morbidity, and it is commonly nonresponsive to conventional therapy. We explore the possible discourse of sinapic acid (SA) against the prevention of bleomycin (BLM)-instigated lung fibrosis in rats through modulation of Nrf2/HO-1 and NF-κB signaling pathways. Lung fibrosis was persuaded in Sprague-Dawley rats by a single intratracheal BLM (6.5 U/kg) injection. Then, these rats were treated with SA (10 and 20 mg/kg, p.o.) for 28 days. The normal control rats provided saline as a substitute of BLM. The lung function and biochemical, histopathological, and molecular alterations were studied in serum, bronchoalveolar lavage fluid (BALF), and the lungs tissues. SA treatment significantly restored BLM-induced alterations in body weight index and serum biomarkers [lactate dehydrogenase (LDH) and alkaline phosphatase (ALP)]. SA (10 and 20 mg/kg) treatment appeared to show a pneumoprotective effect through upregulation of antioxidant status, downregulation of inflammatory cytokines and MMP-7 expression, and reduction of collagen accumulation (hydroxyproline). Nrf2, HO-1, and TGF-β expression was downregulated in BLM-induced fibrosis model, while the reduced expression levels were significantly and dose-dependently upregulated by SA (10 and 20 mg/kg) treatment. We demonstrated that SA ameliorates BLM-induced lung injuries through inhibition of apoptosis and induction of Nrf2/HO-1-mediated antioxidant enzymes via NF-κB inhibition. The histopathological findings also revealed that SA treatment (10 and 20 mg/kg) significantly ameliorated BLM-induced lung injury. The present results showed the ability of SA to restore the antioxidant system and to inhibit oxidative stress, proinflammatory cytokines, extracellular matrix, and TGF-β. This is first report demonstrating that SA amoleriates BLM induced lung injuries through inhibition of apoptosis and induction of Nrf2 and HO-1 mediated antioxidant enzyme via NF-κB inhibition. The histopathological finding reveals that SA treatment (10 and 20 mg/kg) significantly ameliorates BLM induced lung injuries.


Evaluation of extraction methods for untargeted metabolomic studies for future applications in zebrafish larvae infection models.
Sample preparation in untargeted metabolomics should allow reproducible extractions of as many molecules as possible. Thus, optimizing sample preparation is crucial. This study compared six different extraction procedures to find the most suitable for extracting zebrafish larvae in the context of an infection model. Two one-phase extractions employing methanol (I) and a single miscible phase of methanol/acetonitrile/water (II) and two two-phase methods using phase separation between chloroform and methanol/water combinations (III and IV) were tested. Additional bead homogenization was used for methods III and IV (III_B and IV_B). Nine internal standards and 59 molecules of interest (MoInt) related to mycobacterial infection were used for method evaluation. Two-phase methods (III and IV) led to a lower feature count, higher peak areas of MoInt, especially amino acids, and higher coefficients of variation in comparison to one-phase extractions. Adding bead homogenization increased feature count, peak areas, and CVs. Extraction I showed higher peak areas and lower CVs than extraction II, thus being the most suited one-phase method. Extraction III and IV showed similar results, with III being easier to execute and less prone to imprecisions. Thus, for future applications in zebrafish larvae metabolomics and infection models, extractions I and III might be chosen.


Exploring Opportunities to Enhance the Screening and Surveillance of Hepatocellular Carcinoma in Non-Alcoholic Fatty Liver Disease (NAFLD) through Risk Stratification Algorithms Incorporating Ultrasound Elastography.
Non-alcoholic fatty liver disease (NAFLD), with its progressive form, non-alcoholic steatohepatitis (NASH), has emerged as a significant public health concern, affecting over 30% of the global population. Hepatocellular carcinoma (HCC), a complication associated with both cirrhotic and non-cirrhotic NAFLD, has shown a significant increase in incidence. A substantial proportion of NAFLD-related HCC occurs in non-cirrhotic livers, highlighting the need for improved risk stratification and surveillance strategies. This comprehensive review explores the potential role of liver ultrasound elastography as a risk assessment tool for HCC development in NAFLD and highlights the importance of effective screening tools for early, cost-effective detection and improved management of NAFLD-related HCC. The integration of non-invasive tools and algorithms into risk stratification strategies could have the capacity to enhance NAFLD-related HCC screening and surveillance effectiveness. Alongside exploring the potential advancement of non-invasive tools and algorithms for effectively stratifying HCC risk in NAFLD, we offer essential perspectives that could enable readers to improve the personalized assessment of NAFLD-related HCC risk through a more methodical screening approach.


Characterization of the dimeric CMG/pre-initiation complex and its transition into DNA replication forks.
The pre-initiation complex (pre-IC) has been proposed for two decades as an intermediate right before the maturation of the eukaryotic DNA replication fork. However, its existence and biochemical nature remain enigmatic. Here, through combining several enrichment strategies, we are able to isolate an endogenous dimeric CMG-containing complex (designated as d-CMG) distinct from traditional single CMG (s-CMG) and in vitro reconstituted dimeric CMG. D-CMG is assembled upon entry into the S phase and shortly matures into s-CMG/replisome, leading to the fact that only ~ 5% of the total CMG-containing complexes can be detected as d-CMG in vivo. Mass spectra reveal that RPA and DNA Pol α/primase co-purify with s-CMG, but not with d-CMG. Consistently, the former fraction is able to catalyze DNA unwinding and de novo synthesis, while the latter catalyzes neither. The two CMGs in d-CMG display flexibly orientated conformations under an electronic microscope. When DNA Pol α-primase is inactivated, d-CMG % rose up to 29%, indicating an incomplete pre-IC/fork transition. These findings reveal biochemical properties of the d-CMG/pre-IC and provide in vivo evidence to support the pre-IC/fork transition as a bona fide step in replication initiation.


Cost-effectiveness of gene-expression profiling for tumor-site origin.
Gene-expression profiling (GEP) reliably supplements traditional clinicopathological information on the tissue of origin (TOO) in metastatic or poorly differentiated cancer. A cost-effectiveness analysis of GEP TOO testing versus usual care was conducted from a US third-party payer perspective. Data on recommendation changes for chemotherapy, surgery, radiation therapy, blood tests, imaging investigations, and hospice care were obtained from a retrospective, observational study of patients whose physicians received GEP TOO test results. The effects of chemotherapy recommendation changes on survival were based on the results of trials cited in National Comprehensive Cancer Network and UpToDate guidelines. Drug and administration costs were based on average doses reported in National Comprehensive Cancer Network guidelines. Other unit costs came from Centers for Medicare & Medicaid Services fee schedules. Quality-of-life weights were obtained from literature. Bootstrap analysis estimated sample variability; probabilistic sensitivity analysis addressed parameter uncertainty. Chemotherapy regimen recommendations consistent with guidelines for final tumor-site diagnoses increased significantly from 42% to 65% (net difference 23%; P<0.001). Projected overall survival increased from 15.9 to 19.5 months (mean difference 3.6 months; two-sided 95% confidence interval [CI] 3.2-3.9). The average increase in quality-adjusted life-months was 2.7 months (95% CI 1.5-4.3), and average third-party payer costs per patient increased by $10,360 (95% CI $2,982-$19,192). The cost per quality-adjusted life-year gained was $46,858 (95% CI $13,351-$104,269). GEP TOO testing significantly altered clinical practice patterns and is projected to increase overall survival, quality-adjusted life-years, and costs, resulting in an expected cost per quality-adjusted life-year of less than $50,000.


Microlens formation by thin-film deposition with mesh-shaped masks.
Thin-film microlens arrays with high fill factors have been fabricated in a one-step process by shading a vapor beam in a vacuum-deposition apparatus with metallic meshes placed at defined distances to the substrate surface. To generate three-dimensional mask structures with the necessary depth profiles, microgalvanic technology has been applied. Profiles and optical properties of the microlenses have been studied theoretically and experimentally.


The developmental toxicity of PM2.5 on the early stages of fetal lung with human lung bud tip progenitor organoids.
Exposure to air pollution has been proven to be associated with impaired fetal lung development. However, due to the lack of reliable human source models, it is still challenging to deeply understand the human fetal lung development under PM2.5 exposure. Here, we utilized human embryonic stem cell (hESC) line H9 to generate lung bud tip progenitor organoids (LPOs), a process that mimics early stages of fetal lung development including definitive endoderm (DE) formation, anterior foregut endoderm (AFE) differentiation and lung progenitor cell specification, to evaluate potential pulmonary developmental toxicity of PM2.5. We demonstrated that PM2.5 exposure the entire LPOs induction from hESCs significantly affected cellular proliferation of LPOs, and altered the expression of lung progenitor cell markers NKX2.1, SOX2 and SOX9, which are canonically defined subsequently proximal-distal airways specification. To explore the dynamic influences of PM2.5 exposure at different stages of LPOs specification, we also found that PM2.5 exposure significantly affected the expression of several transcriptional factors that are important for the differentiation of DE and AFE. Mechanistically, we suggested PM2.5-induced developmental toxicity to LPOs was partially linked with the Wnt/β-catenin signaling pathway. Therefore, our findings further emphasize the substantial health risks in the development of respiratory system associated with prenatal exposure to PM2.5.


What floats a float nurse's boat?
Few studies have examined the personality traits of nurses, and none identified the personality traits of float pool nurses. Float pool nurses have specific personality traits that differ from unit-based nurses. Independence, Tough-Mindedness, Rule Consciousness, Social Boldness, Openness to Change, and Tension were six personality factors that were found to be statistically significantly different. As hospitals implement float pools or resource teams to meet staffing needs, gleaning insight into the specific personality traits of these individuals could assist in nurse recruitment and retention.


Isoflurane suppresses the self-renewal of normal mouse neural stem cells in a p53-dependent manner by activating the Lkb1-p53-p21 signalling pathway.
Isoflurane is widely used in anaesthesia for surgical operations. However, whether it elicits unwanted side effects, particularly in neuronal cells, remains to be fully elucidated. The Lkb1-p53-p21 signalling pathway is able to modulate neuronal self‑renewal and proliferation. Furthermore, the suppression of Lkb1‑dependent p21 induction leads to apoptosis. In the present study, whether Lkb1‑p53‑p21 signalling is involved in the response to isoflurane was investigated. A comparison of mouse primary, wild‑type neural stem cells (WT NSCs) with the p53‑/‑ NSC cell line, NE‑4C, revealed that isoflurane inhibited proliferation in a dose‑, a time‑ and a p53‑dependent manner. However, flow cytometric analysis revealed that the concentration of isoflurane which caused 50% inhibition (the IC50 value) induced cell cycle arrest in WT NSCs. Furthermore, the protein expression levels of LKB1, p53 and p21 were increased, although those of nestin and survivin decreased, following treatment of WT NSCs with isoflurane. On the other hand, isoflurane induced the phosphorylation of Ser15 in p53 in WT NSCs, which was associated with p53 binding to the p21 promoter, and consequentially, the transcriptional activation of p21. All these events were abrogated in NE‑4C cells. Taken together, the present study has demonstrated that isoflurane suppresses the self-renewal of normal mouse NSCs by activating the Lkb1-p53-p21 signalling pathway.


Orphan Drugs In Development For The Treatment Of Friedreich's Ataxia: Focus On Omaveloxolone.
Friedreich's Ataxia (FRDA) is a devastating and progressive ataxia, marked by mitochondrial dysfunction and oxidative stress. Nrf2 activators such as omaveloxolone (Omav) modulate antioxidative mechanisms, and thus may be viable therapeutic agents in FRDA.


Enantioselective determination of metoprolol and its metabolites in human urine high-performance liquid chromatography with fluorescence detection (HPLC-FLD) and tandem mass spectrometry (MS/MS).
A sensitive, stereoselective assay using solid phase extraction and high-performance liquid chromatography (HPLC) with fluorescence detection (FLD) was developed and validated for the analysis of enantiomers of metoprolol and its metabolites (α-hydroxymetoprolol, O-desmethylmetoprolol). Chiral separation was achieved using a CHIRALCEL OD-RH column, packed with cellulose tris-(3,5-dimethylphenyl-carbamate) stationary phase, employing a mobile phase composed by a mixture of 0.2% diethylamine in water and acetonitrile in gradient elution mode. Linear calibration curves were obtained over the range of 0.025-2.0μg/mL (R(2)>0.994) in urine for both enantiomers of metoprolol and its metabolites with quantitation limit of 0.025μg/mL. Intra and inter-day precision and accuracy were below 15% for both metoprolol and metabolites enantiomers. The recovery of enantiomer of metoprolol and its metabolite was greater than 68.0%, utilizing a SPE procedure. The method was tested with urine quality control samples and human urine fractions after administration of 50mg rac-metoprolol.


Changes in area and thickness of expanded and unexpanded axial pattern skin flaps in pigs.
The pig buttock flap model was used firstly, to compare changes in expanded axial skin flap area with control flaps prior to flap elevation and 3 or 4 months post-flap inset and, secondly, to compare the thickness of expanded and control flaps at elevation and for 3 to 6 months post-flap inset. Following 5 weeks' expansion and prior to elevation, the expanded tissue had gained a significant 63.3% mean increase in area compared with the control tissue (p less than 0.01). Immediately post elevation and inset, the expanded flaps were still significantly larger than the control flaps by a mean 29.8% (p less than 0.01) but had lost 56% of the original area gained. Little change in area occurred in the 3 months post-flap inset as the expanded flaps were still a mean 23.4% larger than the control (p less than 0.01). Dermal and cellular non-keratinised epidermal layers thickened markedly in expanded skin compared to control skin. Following elevation and inset of the flaps, both dermis and epidermis thickened in expanded and control flaps.


Impact of childhood epilepsy on reading and phonological processing abilities.
Although children with epilepsy tend to exhibit more reading difficulties than their classmates, no systematic studies have investigated the relationship between these difficulties and epilepsy. As functional neuroimaging studies have implicated both temporal and frontal lobes in the phonological aspect of reading [K.R. Pugh, B.A. Shaywitz, S.E. Shaywitz, et al. Brain 1996;119:1221-38], seizure activity originating in either region could interfere with phonological processing, whereas generalized seizures would not disturb this function as much. To explore this hypothesis, we compared the metaphonological skills of school-aged children with either temporal lobe epilepsy (TLE), frontal lobe epilepsy (FLE), or generalized absence seizures (ABS) with those of healthy controls. While the reading ability of all epileptic children was close to 2 years behind expectations, children with TLE did not differ from the controls on phonological tasks. In contrast, children with FLE exhibited significant deficits, whereas children with ABS showed difficulties restricted to phonemic segmentation. The results suggest that FLE and, to a lesser extent, generalized seizures may interfere with phonological processing, whereas TLE may affect other aspects of reading.


p21WAF1/CIP1 is more effective than p53 in growth suppression of mouse renal carcinoma cell line Renca in vitro and in vivo.
Although there are many controversial reports about the effect of p53 and p21(WAF1/CIP1) overexpression in different human tumor cells, the p53 gene is shown to be a more effective candidate for cancer gene therapy because of its more pronounced ability to induce apoptosis. In the present study, we present the effect of p53 and p21(WAF1/CIP1) overexpression on mouse renal carcinoma cells in vitro and in vivo. p53 and p21(WAF1/CIP1) genes were introduced into Renca cells using adenoviral vectors (Ad5CMV-p53 and Ad5CMV-p21). The induction of apoptosis was measured using Annexin V assay and DNA fragmentation analysis. The expression of proteins was examined using immunocytochemistry and Western blot methods. The ability of adenoviral vectors to inhibit tumorigenicity of Renca cells, as well as the growth of pre-established tumors was measured. In vitro growth assays revealed higher growth suppression after Ad5CMV-p21 infection. Although both vectors induced apoptosis, Ad5CMV-p53 was slightly more efficient. In vivo studies in Balb/c mice, demonstrated that tumorigenicity was completely suppressed by Ad5CMV-p21. Besides this, Ad5CMV-p21 significantly inhibited the growth of established tumors, while Ad5CMV-p53 did not. These data suggest that p21(WAF1/CIP1) is a more potent growth suppressor than p53 of mouse tumor cells Renca. The divergent responses of tumor cells to p21(WAF1/CIP1) overexpression could be due to various networks that differ between species.


Tensor fasciae latae perforator flap.
The tensor fasciae latae perforator flap and a new technique, named microdissection, for one-stage accurate formation of a thin flap were presented. Microdissection enables the perforator in the adipose tissue to be used as a lengthened pedicle, and the flap can be transferred without adding and reducing excess fat tissue to the recipient and donor sites. This microdissected thin tensor fasciae latae perforator flap can be used conveniently in many aspects of reconstructive surgery, especially in cases of severe bum scar contracture of the extremities.


The cingulate cortex in neurologic diseases: History, Structure, Overview.
A brief review of the history of key observations of cingulate organization, particularly in the limbic system framework, is presented from 1907 to the present. The reasons why it has not played a significant role in neurology reflect the fact that its arterial supply is complex and no cingulate syndrome has been identified. A flat map of its eight subregions and areas is presented as a basis for many subsequent chapters in this volume. Along with this is a summary of current functional attributes of many of its divisions including cognitive, emotion, mental (Theory of Mind), skeletomotor, and autonomic output and various models of its function based on connectivity studies.


Internalized homophobia in men: wanting in the first person singular, hating in the first person plural.
This paper focuses on the expression of internalized homophobia in men, arguing that the most powerful clinical use of this term depends upon its applicability to any man, without limitation to those whose primary object choice is homosexual. A number of dynamic situations are described to which the term might apply, elaborated by examples from contemporary culture and clinical practice. A central dynamic elucidated here is the move from an anxiety-ridden, first person singular voice to the promised safety of a first person plural voice--that is, from the dangerous position of "I want" to the more protected "we hate."


Intersite reliability of vertebral bone marrow lipidomics-derived lipid composition among children with varying degrees of bone fragility undergoing routine orthopedic surgery.
Lipidomics, a branch of metabolomics, is an attractive technique to characterize bone marrow lipid composition, which may be associated with skeletal acquisition and homeostasis. However, the reliability of lipidomics-derived lipid composition of the bone marrow is unknown, especially for pediatric populations with bone fragility. The purpose of this study was to evaluate the intersite reliability and standard error of measurement (SEM) of vertebral bone marrow lipid composition at the thoracic (T11/T12) and lumbar (L1/L2) spine determined by targeted lipidomics among children with varying degrees of bone fragility undergoing routine orthopedic surgery. Children aged between 12 and 19 years of age, with a confirmed diagnosis of adolescent idiopathic scoliosis or neuromuscular scoliosis and cerebral palsy, and undergoing routine posterior spinal fusion surgery at our institution were initially included in this study. Transpedicular vertebral body bone marrow samples were taken from thoracic (T) or lumbar (L) vertebrae. Further inclusion criteria involved having bone marrow extracted from both T11 and T12 (n = 24) or L1 and L2 (n = 19). Lipid composition was measured using a targeted lipidomics technique and examined as the saturated, monounsaturated, and polyunsaturated index and as individual fatty acids. Relative and absolute test-retest reliability was assessed using the intraclass correlation coefficient (ICC) and SEM. For the T11/T12 analysis: the ICC and SEM were 0.59 and 1.7% for the saturated index, 0.31 and 6.2% for the monounsaturated index, and 0.44 and 6.1% for the polyunsaturated index; the ICC showed a considerable range for individual fatty acids from 0.07 (fatty acid 20:2) to 0.82 (15:0) with 62.1% of the fatty acids having poor reliability (i.e., ICC < 0.50). For the L1/L2 analysis: the ICC and SEM were 0.50 and 2.4% for the saturated index, -0.12 and 6.0% for the monounsaturated index, and 0.00 and 4.9% for the polyunsaturated index; the ICC showed a considerable range for individual fatty acids from -0.34 (18:1_n-9) to 0.88 (15:0 and 18:3_n-3) with 79.3% of the fatty acids having poor reliability. The intersite test-retest reliability was poor-to-moderate for index measures and generally poor for individual fatty acids for the thoracic and lumbar spine. At this time, it is not recommended to pool bone marrow adipose tissue across vertebral sites for bone marrow adiposity research or clinical monitoring for pediatric populations with bone fragility.


DNA variations within the sea urchin Otx gene enhancer.
We performed both intra- and interspecific comparisons of the Otx gene in the sea urchin to investigate DNA variations within the enhancer elements. Intraspecific comparisons within Hemicentrotus pulcherrimus showed that indel variations were rare within the Otx enhancer, whereas SNP variations were found uniformly within the whole test region. A similar pattern of DNA variation was observed in comparisons between closely related species. On the other hand, both nucleotide substitution and indel variations were at high levels between distant species. Additionally, the regions corresponding to the Otx enhancer in two related species showed substantial activities during development. Our results suggest the possibility that a stabilizing selection has occurred during the evolution of the Otx gene enhancer in the sea urchin that maintains its expression pattern.


Interdomain cleavage of plasma fibronectin by zinc-metalloproteinase from Serratia marcescens.
Limited proteolysis of porcine plasma fibronectin by the 56 kDa proteinase (56K proteinase) (EC 126.96.36.199) from Serratia marcescens released six polypeptides: a 27 kDa peptide, the heparin-binding domain which comprises the NH2-terminal end; a 50 kDa peptide, a mid-molecule that mediates binding to gelatin or collagen; a 160 kDa peptide, that contained the heparin-binding domain with cell-spreading activity; and a 140 and a 20 kDa peptide which released from the 160 kDa peptide. Each fragment was purified and characterized by its chemical and biological properties, and it was found that they were respectively different domains. Both the 160 and the 140 kDa peptide contained one cysteine per mole of peptide. The 160 kDa peptides were connected by a 6 kDa peptide, which was present at the COOH-terminal end of the molecule and was biologically inactive. Only 6 kDa peptide contained a disulfide bond and produced 3 kDa peptide after reduction, whereas other fragments did not change with or without reduction on SDS-polyacrylamide gel electrophoresis. NH2-terminal sequence analyses of the released peptides showed that the 56K proteinase cleaved the fibronectin between the Arg-Thr (located at two different sites), Leu-Ser and Gln-Glu bonds. Out of 118 Arg residues, there are nine sequences containing Arg-Thr, and two of them near or at an interdomain location (at Arg 259 and 2239) were cleaved. Out of 124 Leu residues, there are 11 Leu-Ser sequences and only one, at 687, was cleaved. The above fragments with functional domain activity could be aligned according to the previously reported amino-acid sequence of human or bovine plasma fibronectin. The treatment of fibroblast cells by the 56K proteinase resulted in loss of morphological integrity and extracellular matrix.


May Thurner syndrome: description of a case with unusual clinical onset.
May-Thurner syndrome (MTS) is a congenital vascular alteration that is part of a restricted category of very rare vascular syndromes that have in common the compression of an arterial or venous vessel. MTS, first described in 1957, is due to compression of the left common iliac vein against the lumbar spine by the adjacent common iliac artery. It can cause chronic thrombosis of the left lower limb and can give edema, pain, claudication, thrombophlebitis, and, in severe cases, pulmonary embolism. Color Doppler and duplex Doppler ultrasound allow us to easily locate the deep vein thrombosis, to measure its extension, and to highlight the vascular changes typical of MTS: compression and consequent hypertension of the left common iliac vein. The therapy depends on the degree of venous stasis and on the presence of venous thrombosis; generally, it consists of the administration of short- or long-term anticoagulant and thrombolytic drugs. In cases of severe stenosis of the left common iliac vein, the first-choice treatment consists of positioning a vascular stent, which resolves compression and significantly reduces chronic thrombotic episodes. We describe a case of MTS with an unusual clinical onset with pulmonary embolism.


A review of the treatment of cancer of the male breast with a case follow-up.
The author presents a four-year follow-up of a male patient with cancer of the breast. The lesion was large, fungating, and ulcerating. It was treated effectively with cobalt without mastectomy.


Polymorphisms in methionine synthase (A2756G) and cystathionine beta-synthase (844ins68) and susceptibility to carcinomas of the upper gastrointestinal tract.
Folate deficiency is considered to increase the risk for the development of malignant tumors such as prostate and colorectal cancer. Methionine synthase (MTR) and cystathionine ss-synthase (CBS) are enzymes that play a central role in folate metabolism, thereby affecting DNA methylation and synthesis. A single A-->G substitution at nucleotide 2756 of the MTR and a 68 bp CBS insertion polymorphism in exon 8 have been associated with decreased enzyme activity. The purpose of this study is to compare the association of the MTR A2756G polymorphism and CBS insertion polymorphism with susceptibility to carcinomas of the upper gastrointestinal tract. Using the restriction fragment length polymorphism (RFLP)-PCR, the prevalence of MTR A2756G and CBS insertion polymorphism was determined in healthy controls (n = 257) and in patients with esophageal squamous cell carcinoma (ESCC) (n = 263), Barrett's esophagus-associated esophageal adenocarcinoma (BC) (n = 89), cardiac carcinoma (CC) (n = 144), or gastric carcinoma (GC) (n = 221) from German Caucasian subjects. No significant difference in MTR A2756G genotype distribution was observed between controls (A/A 66.9%, A/G 29.8%, G/G 3.3%) and patients with ESCC (A/A 61.7%, A/G 36.3%, G/G 2.1%), BC (A/A 69.2%, A/G 26.9%, G/G 3.9%), CC (A/A 51.8%, A/G 44.6%, G/G 3.6%), or GC (A/A 73.4%, A/G 20.9%, G/G 5.7%). Similarly, the CBS genotype (I: allele with 68 bp insertion; N: allele without insertion) distribution among German patients with ESCC (N/N 86.8%, I/N 13.2%), BC (N/N 90.2%, I/N 9.8%), CC (N/N 90.1%, I/N 9.9%) or GC (N/N 91.3%, I/N 8.7%) was not different from healthy controls (N/N 90.4%, I/N 9.6%). The gene allele constellation I/I was not present. The current study suggests that there is no association between MTR A2756G polymorphism and the CBS (844ins68) insertion polymorphism and cancer of the upper gastrointestinal tract.


Role of antioxidant defenses in the tolerance of severe dehydration by anurans. The case of the leopard frog Rana pipiens.
Many anurans have excellent dehydration tolerance that allows endurance of the loss of up to 50-60% of total body water. One of the effects of severe dehydration is circulatory impairment due the reduced volume and increased viscosity of blood, which leads to organ hypoxia. The rehydration situation, therefore, involves a reoxygenation of tissues that may include elements of oxidative stress that resemble the injury in post-ischemic reperfusion of mammalian organs. The role of endogenous defenses against oxygen radicals in the tolerance of severe dehydration by leopard frogs, Rana pipiens, was investigated by monitoring the activities of antioxidant enzymes and glutathione levels (reduced GSH and oxidized GSSG) in leg muscle and liver of control, 50%-dehydrated, and fully rehydrated frogs. The maximal activities of muscle catalase and liver glutathione peroxidase, measured per mg soluble protein, increased significantly by 52 and 74%, respectively, after dehydration whereas muscle superoxide dismutase and glutathione reductase activities responded oppositely, decreasing by 32 and 35%, respectively. Enzyme activities returned to control levels after full rehydration. Hepatic GSH and GSSG increased early in the rehydration process (30% recovery of total body water), but returned to control levels after full recovery. A similar trend was observed for liver GSSG. The elevation of antioxidant defenses against peroxides during dehydration could provide protection against post-hypoxic oxyradical stress during rehydration. Indeed, analysis of one product of lipid peroxidation, thiobarbituric acid reactive substances, in frog tissues gave no indication of oxidative stress during the dehydration/rehydration cycle.


Massively parallel tumor multigene sequencing to evaluate response to panitumumab in a randomized phase III study of metastatic colorectal cancer.
To investigate whether EGF receptor (EGFR) pathway mutations predicted response to monotherapy with panitumumab, an anti-EGFR monoclonal antibody, in a randomized phase III study of metastatic colorectal cancer. Using massively parallel multigene sequencing, we analyzed 320 samples for 9 genes, with multigene sequence data from 288 (90%) samples. Mutation rates were: KRAS (45%), NRAS (5%), BRAF (7%), PIK3CA (9%), PTEN (6%), TP53 (60%), EGFR (1%), AKT1 (<1%), and CTNNB1 (2%). In the randomized study and open-label extension, 22 of 138 (16%) wild-type KRAS (codons 12/13/61) patients versus 0 of 103 mutant KRAS (codons 12/13) patients had objective responses. Of 6 mutant KRAS (codon 61) patients, 1 with a Q61H mutation achieved partial response during the extension. Among wild-type KRAS (codons 12/13/61) patients, 0 of 9 patients with NRAS mutations, 0 of 13 with BRAF mutations, 2 of 10 with PIK3CA mutations, 1 of 9 with PTEN mutations, and 1 of 2 with CTNNB1 mutations responded to panitumumab. No patients responded to best supportive care alone. Panitumumab treatment was associated with longer progression-free survival (PFS) among wild-type KRAS (codons 12/13/61) patients [HR, 0.39; 95% confidence interval (CI), 0.28-0.56]. Among wild-type KRAS patients, a treatment effect for PFS favoring panitumumab occurred in patients with wild-type NRAS (HR, 0.39; 95% CI, 0.27-0.56) and wild-type BRAF (HR, 0.37; 95% CI, 0.24-0.55) but not mutant NRAS (HR, 1.94; 95% CI, 0.44-8.44). These results show the feasibility and potential clinical use of next-generation sequencing for evaluating predictive biomarkers.


TGF-beta-associated enhanced susceptibility to leishmaniasis following intramuscular vaccination of mice with Leishmania amazonensis antigens.
Leishmania amazonensis and Leishmania braziliensis are the main causal agents of anergic diffuse cutaneous leishmaniasis and hyperergic mucosal leishmaniasis in man, respectively. In this work we demonstrate that intramuscular vaccination of BALB/c mice with whole antigens of L. amazonensis (LaAg) but not L. braziliensis (LbAg) results in increased susceptibility to cutaneous leishmaniasis. LaAg vaccination resulted in an increased capacity of the draining lymph nodes to produce IL-10 and TGF-beta during antigen recall responses. In vitro cultivation with LaAg but not LbAg induced increased apoptosis of CD8+ T cells. Following infection with L. amazonensis, LaAg-vaccinated mice produced significantly more TGF-beta and a higher serum IgG1/IgG2a antibody ratio compared with LbAg-vaccinated and non-vaccinated animals. The association of TGF-beta with enhanced susceptibility to infection was confirmed in mice co-vaccinated with LaAg and neutralizing anti-TGF-beta antibodies. Upon parasite challenge, these animals developed much smaller lesion sizes and parasite burdens, comparable with non-vaccinated controls. The disease-promoting effect of LaAg vaccination is not a general event, as in contrast to BALB/c, the disease outcome in C57Bl/6 mice was unaltered. Together, these findings indicate that species-specific components of L. amazonensis activate overt TGF-beta production that predisposes more susceptible individuals to aggravated disease following vaccination.


Q fever during pregnancy: a narrative review.
Coxiella burnetii, the causative agent of Q fever, causes abortions in animals. Its effects on pregnancy in humans and the management of Q fever in pregnancy are uncertain. To summarize data on the effects of Q fever on pregnancy in women, the effects of pregnancy on Q fever complications and the optimal screening and management of Q fever during pregnancy. We searched for studies reporting on Q fever during pregnancy in women. We included randomized and observational studies, seroprevalence studies, case series and case reports, including clinical and histopathological studies. The accumulating data seems convincing that Q fever increases the risk of abortions in early pregnancy and prematurity or intrauterine fetal demise in late pregnancy. Data are based on sero-epidemiological associations of Q fever and adverse pregnancy outcomes and case reports showing the presence and effects of C. burnetii on the placenta and the fetus. Based on observational studies, acquisition of Q fever during pregnancy also increases the risk for maternal chronic Q fever. Treatment of recently infected women seems to improve these outcomes, based on case series only, but the optimal duration of treatment has not been studied. The efficacy of active surveillance during pregnancy, timing and frequency have not been determined in high-endemicity settings. Obstetricians should be aware of the risk for transmission of the disease during delivery. Currently available data are based mostly on case series and case reports, with some discrepancy between the French experience in chronic endemicity settings and Dutch experience in outbreak settings. Since infection with Q fever is largely asymptomatic, we believe that the accumulating information linking Q fever to adverse pregnancy outcomes justifies screening in the high-endemicity setting and treatment of infected women. High-quality research addressing the questions raised by this review is needed to determine the optimal public health policy.


Use of naproxen versus intracervical block for pain control during the 52-mg levonorgestrel-releasing intrauterine system insertion in young women: a multivariate analysis of a randomized controlled trial.
To compare the effectiveness of 550 mg naproxen sodium versus 6 mL 2%-lidocaine intracervical block in pain lowering at the 52-mg levonorgestrel-releasing intrauterine system (LNG-IUS) placement in young women. In this randomized controlled trial, 100 women aged 15-24 years were block-randomized to receive either 6 mL 2%-lidocaine intracervical block 5 min before the LNG-IUS insertion or 550 mg naproxen 30 min before the procedure. Forty-nine women received 550 mg naproxen and 51 received intracervical block. The primary outcome was pain at LNG-IUS insertion. Secondary outcomes were ease of insertion, insertion failures, and correct IUS positioning. Neither participants nor doctors were blinded. Pain at insertion was assessed by using a Visual Analog Scale (VAS). Women randomized to lidocaine intracervical block presented lower mean pain score at insertion, when compared to women who received naproxen (5.4 vs. 7.3, respectively; p < 0.001). Parous women had a 90.1% lower chance of experiencing severe pain (p = 0.004). There was a 49.8% reduction in the chance of severe pain for every 1-cm increase in the hysterometry (p = 0.002). The only complication observed during insertion was vasovagal-like reactions (7%). The insertion was performed without difficulty in 82% of the women. Participants in the intracervical block group presented higher proportion of malpositioned IUS on transvaginal ultrasound examination compared to women in naproxen group. Nevertheless, all the malpositioned IUS were inserted by resident physicians. Lidocaine intracervical block was found to be more effective than naproxen in reducing LNG-IUS insertion pain. RBR-68mmbp, Brazilian Registry of Clinical Trials, Retrospectively registered (August 4, 2020), URL of trial registry record: https://ensaiosclinicos.gov.br/rg/RBR-68mmbp/ .


T2-weighted and diffusion-weighted MRI for discriminating benign from malignant focal liver lesions: diagnostic abilities of single versus combined interpretations.
To compare the diagnostic accuracies of diffusion-weighted imaging (DWI), T2-weighted imaging (T2WI), and the combination of both sequences in discriminating benign from malignant focal liver lesions (FLLs). In all, 166 patients with 269 FLLs (153 benign and 116 malignant) were retrospectively evaluated. Two abdominal readers visually assessed the DWI, T2WI, and the combined (DWI+T2WI) image sets in an independent and blinded manner. The diagnostic abilities of each image set in discriminating the benign from the malignant FLLs set were compared using a binary logistic regression model. Pathologic results, consensus reading, and follow-up imaging were used as the reference standard. The overall characterization accuracy in all lesions of the combined set (80.3%) was significantly higher than those of the T2WI set (68.8%) and DWI set (73.2%) (combined vs. T2WI, P < 0.001; combined vs. DWI, P = 0.001), while there was no significant difference between the T2WI and DWI sets (P = 0.058). All image sets were more accurate in the characterization of malignant FLLs than of benign FLLs (P < 0.001). T2WI and DWI are complementary in discriminating benign from malignant FLLs; their combination improves diagnostic confidence.


The enhancing effect of a cation-π interaction on the cooperativity of halogen bonds: A computational study.
In this work, we investigate the effect of a cation-π interaction on the cooperativity of X⋯N halogen bonds in PhX⋯NCX⋯NH3 complexes, where Ph=phenyl and X=Cl, Br, I. Molecular geometries and interaction energies of the resulting complexes are studied at the MP2/aug-cc-pVDZ(-PP) computational level. The mechanism of the cooperativity between halogen bonds is analyzed using parameters derived from the noncovalent index, quantum theory of atoms in molecules and natural bond orbital methodologies. It is found that the divalent cations (Be2+, Mg2+) have a larger influence on the cooperativity of halogen bonds than monovalent ones (Li+, Na+). The formation of a cation-π interaction leads to strengthening of the halogen bonds, hence increases their cooperativity.


A novel emaravirus is associated with redbud yellow ringspot disease.
Yellow ringspot is the only virus-like disease reported in redbud (Cercis spp.) with symptoms including vein clearing, chlorotic ringspots and oak-leaf pattern. A putative new emaravirus was present in all trees displaying typical yellow ringspot symptoms and the name redbud yellow ringspot associated virus is proposed. The virus genome is composed of at least five RNA segments. Two coding regions were studied to determine isolate diversity with results pointing to a homogeneous virus population. Host range was evaluated using graft transmission and by testing species found in close proximity to infected trees. Mite transmission with Aculops cercidis, the predominant species found in redbud trees in the epicenter of the disease, was evaluated but was not found to be a vector of the virus. Based on this study and the accumulated knowledge on emaravirus evolution we propose that speciation is allopatric, with vectors being a major component of the process.


How to use outside collection agencies. Don't simply charge off accounts and forget them.
Collection agencies may be the key to receiving payment on overdue bills. Learn how they work and what to look for in a reputable collection agency.


Case management: the allocation of hours and influencing factors in the management of survivors of traumatic brain injury.
Case management is a service frequently provided to clients following a traumatic brain injury (TBI); however, the time required for this service is uncertain. To identify factors that influence case management for clients with TBI, a descriptive analysis of 64 client files were provided with case management services of a rehabilitation provider based in Sydney, Australia. These files were then further examined by using qualitative measures to determine the relationship among case management factors, time required and the severity of the injury. This information was statistically analyzed to identify what factors predicted the greatest use of case management time. The analysis identified inappropriate behavior, family dynamics, accommodation, and formal relationships as primary affectors on case management time. Other issues identified as less demanding of case management time were sustaining employment and provision of equipment. In terms of case management hours, the mean was 2.5 hr/ month. Where there was a combination of three factors, the time for case management increased to a mean of 3.97 hr, and up to 4.34 hr/month (range: 1.5-14.0 hr) of case management time where 4 factors were identified. A positive relationship between severity of posttraumatic amnesia (PTA) and care and needs score was also noted to impact on amount of case management time and duration of service.


Popliteal vein entrapment.
A case of popliteal vein entrapment by an abnormal 'third' head of the gastrocnemius muscle is reported. Complete relief of disabling symptoms was obtained by dividing the abnormal muscle band.


Fluorescence histochemical observations on the distribution of exogenous dihydroisoquinoline in the rat brain.
The distribution of fluorescence in the rat brain after i.p. or intracerebral (i.c.) injections of a fluorescent dihydroisoquinoline derivate of dopamine was studied. After low i.p. doses (50 mg/kg of body weight) the fluorescence was totally confined to the capillary endothelial cells in the cerebral cortex, neostriatum, and substantia nigra (Sn). After large i.p. doses (500 mg/kg) fluorescent material was also present in the neuropil of all the regions studied and some cells of the cerebral cortex and SN. Ater injections to the neostriatum or SN fluorescence was observed in the endothelial cells and some small to medium-sized rounded cells in both regions. A conspicious dark area contrasting with the background fluorescence was constantly present around capillaries, and this area was in contact with nonfluorescent multibranched cells of astrocytic type. In fluorescent cells the fluorescence was present both in the cytoplasm and the nucleus.


2,4-dithiouracil: the reproducible H-bonded structural motifs in the complexes with 18-membered crown ethers.
2,4-dithiouracil (DTU) forms in the crystals the H-bonded monohydrates of a 1:1:1 ratio with 18-crown-6 (18C6) 1, cis,syn,cis-isomer of dicyclohexano-18-crown-6 (DCH6A) 2, and benzo-18-crown-6 (B18C6) 3, while the anhydrous adduct with cis,anti,cis-isomer of dicyclohexano-18-crown-6 (DCH6B) 4 is of a 2:1 ratio. In 1-3 the components reproducibly alternate in the chains, while in 4 the chains are built of the alternative centrosymmetric dimers of 2,4-dithiouracil and the molecules of the cis,anti,cis-isomer of dicyclohexano-18-crown-6.


Susceptibility of erythrocyte lipids to oxidation and erythrocyte antioxidant status in myocardial infarction.
We evaluated the erythrocyte lipid susceptibility to oxidation and erythrocyte antioxidant status in patients with myocardial infarction (MI). Patients with MI were divided into two group according to the severity of the disease as severe (n = 30) or milder (n = 25). Malondialdehyde as a marker of lipid peroxidation was measured to show the lipid susceptibility to oxidation. Erythrocytes were stressed in vitro by hydrogen peroxide acting as the oxidative agents for 120 min. After designated time, erythrocyte MDA production was significantly higher in patients with severe MI than in controls and in patients with milder MI (p < 0.001, p < 0.001, respectively). Antioxidant status was determined by measuring the reduced glutathione levels and glutathione peroxidase activity of erythrocyte. In patients with MI, antioxidant status was significantly lower than in controls, and there was no significant difference between the patient groups. Determination of erythrocyte lipid susceptibility to oxidation may be a useful in vitro test to evaluate the degree of oxidative stress in myocardial infarction.


Receptors for luteinizing hormone releasing hormone (LHRH) expressed in human non-Hodgkin's lymphomas can be targeted for therapy with the cytotoxic LHRH analogue AN-207.
We determined by immunohistochemistry the presence of LHRH receptors in surgical specimens of human non-Hodgkin's lymphomas (NHL) and investigated the expression of LHRH receptors in two human NHL cell lines, RL and HT by RT-PCR, Western blot and radioligand binding studies. In in vivo experiments with nude mice bearing tumours of these NHL cell lines, the efficacy of cytotoxic LHRH analogue AN-207 and its cytotoxic radical AN-201 was examined. LHRH receptors were detected in 94.1% of the human NHL specimens and in both NHL cell lines. AN-207 significantly (P < 0.01) inhibited the growth of RL and HT tumours, while the non-targeted AN-201 had no effects. Blockade of the LHRH receptors with an excess of LHRH agonist Decapeptyl suppressed the antitumour effects of AN-207. Our findings indicate that LHRH receptors expressed in a high percentage of human NHL specimens can be used for effective targeted therapy with the cytotoxic LHRH analogue AN-207.


Neuropsychological features of benign partial epilepsy in children.
Rolandic paroxysmal epilepsy (RPE) is a useful model for investigating the complex links between epilepsy and cognitive dysfunction. 44 children with RPE who met the following (among other) criteria: negative CT scan, freedom from drug treatment, and IQ greater than or equal to 80, were assigned to three subgroups by side of EEG focus: left, right and bilateral. A neuropsychological battery elicited small differences in cognitive performance between the whole group and the controls and among the subgroups, only partially correlated with EEG side. A follow-up assessment showed that the short falls had disappeared along with the seizures and EEG anomalies, thus confirming the benign nature of RPE. Our findings suggest too that the mere presence of paroxysmal cortical activity is enough to trigger cognitive dysfunction.


A unified matrix hypothesis of DNA-directed morphogenesis, protodynamism and growth control.
A theoretical concept is proposed, in order to explain some enigmatic aspects of cellular and molecular biology of eukaryotic organisms. Among these are the C-value paradox of DNA redundancy, the correlation of DNA content and cell size, the disruption of genes at DNA level, the "Chromosome field" data of Lima de Faria (Hereditas 93:1, 1980), the "quantal mitosis" proposition of Holtzer et al. (Curr. Top. Dev. Biol. 7:229 1972), the inheritance of morphological patterns, the relations of DNA and chromosome organisation to cellular structure and function, the molecular basis of speciation, etc. The basic proposition of the "Unified Matrix Hypothesis" is that the nuclear DNA has a direct morphogenic function, in addition to its coding function in protein synthesis. This additional genetic information is thought to be largely contained in the non-protein coding transcribed DNA, and in the untranscribed part of the genome.


Endourologic management of upper and mid ureteral calculi: percutaneous antegrade extraction vs transurethral ureteroscopy.
Fifty patients requiring intervention for upper or mid ureteral calculi were managed initially with a percutaneous antegrade approach (25 patients) or a transurethral ureteroscopic approach (25 patients). In both groups, a crossover to the other modality was at times utilized to accomplish stone removal. Success for either group was defined as complete stone removal without the need for open intervention.


Gun-shot injuries in UK military casualties - Features associated with wound severity.
Surgical treatment of high-energy gun-shot wounds (GSWs) to the extremities is challenging. Recent surgical doctrine states that wound tracts from high-energy GSWs should be laid open, however the experience from previous conflicts suggests that some of these injuries can be managed more conservatively. The aim of this study is to firstly characterise the GSW injuries sustained by UK forces, and secondly test the hypothesis that the likely severity of GSWs can be predicted by features of the wound. The UK Military trauma registry was searched for cases injured by GSW in the five years between 01 January 2009 and 31 December 2013: only UK personnel were included. Clinical notes and radiographs were then reviewed. Features associated with energy transfer in extremity wounds in survivors were further examined with number of wound debridements used as a surrogate marker of wound severity. There were 450 cases who met the inclusion criteria. 96 (21%) were fatally injured, with 354 (79%) surviving their injuries. Casualties in the fatality group had a median New Injury Severity Score (NISS) of 75 (IQR 75-75), while the median NISS of the survivors was 12 (IQR 4-48) with 10 survivors having a NISS of 75. In survivors the limbs were most commonly injured (56%). 'Through and through' wounds, where the bullet passes intact through the body, were strongly associated with less requirement for debridement (p<0.0001). When a bullet fragmented there was a significant association with a requirement for a greater number of wound debridements (p=0.0002), as there was if a bullet fractured a bone (p=0.0006). More complex wounds, as indicated by the requirement for repeated debridements, are associated with injuries where the bullet does not pass straight through the body, or where a bone is fractured. Gunshot wounds should be assessed according to the likely energy transferred, extremity wounds without features of high energy transfer do not require extensive exploration.


Applying new clinicopathological characteristics to prognostication in advanced thyroid carcinoma.
In the February 2011 issue of Endocrine-Related Cancer, Deandreis et al. reported that increased FDG uptake was prognostic in patients with metastatic thyroid cancer. Fludeoxyglucose-positron emission tomography (FDG-PET) is routinely used in the staging and follow-up of patients with cancer. This study gives further evidence for the role of FDG-PET scanning in metastatic thyroid cancer, especially to identify patients with aggressive disease requiring systemic therapy.


13C direct detected COCO-TOCSY: a tool for sequence specific assignment and structure determination in protonless NMR experiments.
A novel experiment is proposed to provide inter-residue sequential correlations among carbonyl spins in (13)C detected, protonless NMR experiments. The COCO-TOCSY experiment connects, in proteins, two carbonyls separated from each other by three, four or even five bonds. The quantitative analysis provides structural information on backbone dihedral angles phi as well as on the side chain dihedral angles of Asx and Glx residues. This is the first dihedral angle constraint that can be obtained via a protonless approach. About 75% of backbone carbonyls in Calbindin D(9K), a 75 amino acid dicalcium protein, could be sequentially connected via a COCO-TOCSY spectrum. 49(3)J(C')(C') values were measured and related to backbone phi angles. Structural information can be extended to the side chain orientation of aminoacids containing carbonyl groups. Additionally, long range homonuclear coupling constants, (4)J(CC) and (5)J(CC), could be measured. This constitutes an unprecedented case for proteins of medium and small size.


Mutations of LRTOMT, a fusion gene with alternative reading frames, cause nonsyndromic deafness in humans.
Many proteins necessary for sound transduction have been identified through positional cloning of genes that cause deafness. We report here that mutations of LRTOMT are associated with profound nonsyndromic hearing loss at the DFNB63 locus on human chromosome 11q13.3-q13.4. LRTOMT has two alternative reading frames and encodes two different proteins, LRTOMT1 and LRTOMT2, detected by protein blot analyses. LRTOMT2 is a putative methyltransferase. During evolution, new transcripts can arise through partial or complete coalescence of genes. We provide evidence that in the primate lineage LRTOMT evolved from the fusion of two neighboring ancestral genes, which exist as separate genes (Lrrc51 and Tomt) in rodents.


Taxonomy dimension reduction for colorectal cancer prediction.
A growing number of people suffer from colorectal cancer, which is one of the most common cancers. It is essential to diagnose and treat the cancer as early as possible. The disease may change the microorganism communities in the gut, and it could be an efficient method to employ gut microorganisms to predict colorectal cancer. In this study, we selected operational taxonomic units that include several kinds of microorganisms to predict colorectal cancer. To find the most important microorganisms and obtain the best prediction performance, we explore effective feature selection methods. We employ three main steps. First, we use a single method to reduce features. Next, to reduce the number of features, we integrate the dimension reduction methods correlation-based feature selection and maximum relevance-maximum distance (MRMD 1.0 and MRMD 2.0). Then, we selected the important features according to the taxonomy files. In this study, we created training and test sets to obtain a more objective evaluation. Random forest, naïve Bayes, and decision tree classifiers were evaluated. The results show that the methods proposed in this study are better than hierarchical feature engineering. The proposed method, which combines correlation-based feature selection with MRMD 2.0, performed the best on the CRC2 dataset. The dataset and methods can be found in http://lab.malab.cn/data/microdata/data.html.


Effect of pressure intensity of graduated elastic compression stocking on muscle fatigue following calf-raise exercise.
The purpose of the present study was to examine the effects of wearing a graduated elastic compression (EC) stocking, with different pressure profiles during a fatiguing calf-raise exercise session, on the torque generating capacity after exercise. Fourteen subjects performed 15 sets of 10 repetitions of calf-raise exercise, wearing one of three stockings of different compression pressure profiles: two kinds of EC stockings (one (EC30) with 30mmHg at the ankle, 21-25mmHg at the calf, and 10mmHg below the knee, and the other (EC18) with 18, 12-14, and 7mmHg at the same regions, respectively) and a non-EC sport stocking as a control (CON). Before and after the exercise, torque and electromyographic (EMG) signals of medial gastrocnemius and soleus muscles were recorded during an evoked triplet contraction and the maximal voluntary contraction (MVC). All stockings had no effect on the decline of the MVC torque whereas the reduction of the evoke triplet torque in EC30 stocking condition was significantly smaller compared with that in CON (-6.4±8.5% for EC30 and -16.5±9.0% for CON, P<0.05). The reduction of the mean power frequency of EMG during MVC in the EC30 condition was significantly smaller than that in CON (-4.4±10.9Hz for EC30 and -18.7±7.9Hz for CON, P<0.05). These results suggest that the EC stocking with adequate pressure at the calf region relieves muscle fatigue of the triceps surae induced by calf-raise exercise.


CT whole-body scanning in pediatric radiology.
In an 8 month period 57 scans in 28 children have been performed using an EMI-CT 5005 General Purpose Scanner. Thirteen primary malignant tumours were evaluated, and scanning was of great value in localizing and delineating the lesion, and hence predicting the operability. Furthermore, secondary spread of malignant tumours was easily detected. In the continuing evaluation of treated malignant tumours CT has been used to estimate regression and relapse.


Best practices for IACUCs in the evaluation of multiple major operative procedures.
Evaluation of multiple major operative procedures can be a daunting task for IACUCs. Committee members need to know what a major operative procedure is, when multiple major operative procedures are permitted under the Animal Welfare Act, when permission from the Administrator of the US Department of Agriculture's Animal and Plant Health Inspection Service is required for multiple major operative procedures, and what information must be included when requesting such permission. The author discusses the intent and requirements for multiple major operative procedures under the Animal Welfare Act and describes best practices that IACUCs may use in evaluating multiple major operative procedures both within and across protocols.


Validation of Chimeric Fusion Peptides Using Proteomics Data.
Chimeric RNAs as well as their fused protein products have therapeutic applications ranging from diagnostics to being used as therapeutic target. Many algorithms have been developed to identify chimeric RNAs, however, identification and validation of fused protein product of the chimeric RNA is still an emerging field. These chimeric proteins can be validated by searching and identifying them in publicly available proteomics datasets. Here we describe the detailed steps for (1) downloading and processing publicly available proteomics datasets, (2) developing fusion peptide database by performing in silico tryptic digestion of chimeric proteins, and (3) software used to identify chimeric peptides in the proteomics data.


Monocyte-mediated activation of endothelial cells occurs only after binding to extracellular vesicles from red blood cell products, a process mediated by β-integrin.
Red blood cell (RBC) transfusion is associated with organ failure. The mechanism remains unknown, but may include adherence of blood cells to the microvasculature. We hypothesized that RBC-derived extracellular vesicles (EVs) interact with monocytes to activate endothelial cells. Human umbilical vein endothelial cells were incubated with supernatant from fresh and stored RBC units either containing EVs or depleted from EVs, with or without the addition of immune cells. We measured expression of adhesion markers by flow cytometry and markers of coagulation and inflammation in the culture medium. We studied phagocytosis of EVs by monocytes by using confocal microscopy and flow cytometry. Incubation of endothelial cells with monocytes alone did not induce up regulation of adhesion markers. The addition of both monocytes and supernatant from RBCs containing EVs resulted in up regulation of endothelial expression of intercellular adhesion molecule 1 and E-selectin when compared to baseline. Up regulation was absent when stimulated with RBC supernatant depleted from EVs. EVs are phagocytosed by monocytes, which was partly abrogated after coincubation with two different complement receptor 3 (CR3)-blocking antibodies. Addition of RBC-derived EVs also increased levels of von Willebrand factor (VWF). There were no differences between groups related to storage time. EVs from RBC transfusion bags activate monocytes with subsequent up regulation of endothelial cell adhesion markers. EVs are phagocytosed by monocytes through CR3. Furthermore, these EVs proved to be a source of VWF. These effects are unrelated to storage time. Thereby, EVs from RBC transfusion bags induce a proinflammatory and procoagulant endothelial cell response.


[Patients' decision for aesthetic surgery].
Aesthetic surgery is a service which entails a high degree of trust. Service evaluation prior to provision is difficult for the patient. This leads to the question of how to manage the service successfully while still focusing on the medical needs. The decision to undergo an operation is not influenced by the operation itself, but by preoperative events which induce the patient to have the operation done. According to "buying decisions" for products or in service management, the decision for an aesthetic operation is extensive; the patient is highly involved and actively searching for information using different directed sources of information. The real "buying decision" consists of 5 phases: problem recognition, gathering of information, alternative education, purchase decision, and post purchase behaviour. A retrospective survey of 40 female patients who have already undergone an aesthetic operation assessed for problem recognition, which types of information were collected prior to the appointment with the surgeon, and why the patients have had the operation at our hospital. They were also asked how many alternative surgeons they had been seen before. Most of the patients had been thinking about undergoing an operation for several years. They mainly used the web for their research and were informed by other (non-aesthetic) physicians/general practitioners. Requested information was about the aesthetic results and possible problems and complications. Patients came based on web information and because of recommendations from other physicians. 60% of all interviewees did not see another surgeon and decided to have the operation because of positive patient-doctor communication and the surgeon's good reputation. Competence was considered to be the most important quality of the surgeon. However, the attribute was judged on subjective parameters. Environment, office rooms and staff were assessed as important but not very important. Costs of surgery were ranked second. Influence on patients' decision is only possible by high quality operation results, which in turn lead to good reputation with (non-aesthetic) physicians and patients. In contrast to print advertisements, the provision of information via the internet is of tremendous importance and must be both information and comprehensible. Not only the aesthetic result should be depicted, but also questions such as "when will I be fit after surgery", and possible problems and complications should be described honestly. Besides the described word-of-mouth recommendations web fora resemble a virtual extension of recommendation. Whether such web based fora will prevail as a significant source of information remains unclear. Board certification in plastic and aesthetic surgery ("Facharzt" qualification) was assessed as very important. Thus, this is relevant information which should be emphasized. Most important for the patients' decision, however, was patient-doctor communication. To develop a basis of trust, communication should be open and reliable. An adequate time span has to be planned for the first appointment with the patient. In addition to aesthetic results, risks and possible complications have to be discussed. An open and comprehensible conversation which does not sugar-coat the operation is assessed as very positive. Finally, based on this consultation the surgeon is suggestive of his surgical competence. This underlines that core competence in aesthetic surgery is a medical one.


Draft Genome Sequence of Methicillin-Resistant Staphylococcus aureus KT/Y21, a Sequence Type 772 (ST772) Strain Isolated from a Pediatric Blood Sample in Terengganu, Malaysia.
Here, we report the draft genome sequence of a methicillin-resistant Staphylococcus aureus (MRSA) strain, KT/Y21, isolated from a blood sample of a pediatric patient. This strain belongs to sequence type 772 (ST772), harbors the staphylococcal cassette chromosome mec element (SCCmec) type V, and is positive for the Panton-Valentine leukocidin (PVL) pathogenic determinant.