John L. Cameron: A Historical Perspective.
: John Lemuel Cameron is one of the most influential surgeons of modern American surgery and has had a profound impact on shaping its current global landscape. He served as the president of 14 surgical societies and 4 of his presidential addresses focused on the lives of surgeons who he believed had made an exceptional contribution to the field of surgery: Harvey William Cushing, William Stewart Halsted, John Shaw Billings, and John Miller Turpin Finney. Many parallels can be drawn between these surgeons and Dr. Cameron. This paper describes Dr. Cameron's career in the context of the 4 legendary surgeons, about whom he chose to base his presidential addresses.


Hydrolysis of Irradiated Ovalbumin by Pepsin.
Solid ovalbumin has been irradiated at three doses, 6.5, 25, and 40 megarads, under high vacuum. The native and irradiated samples have been hydrolyzed at pH 1.4 by pepsin, after centrifugation of the aggregates, if necessary. The number of bonds broken per ovalbumin molecule has been estimated by comparing the rate of protein destruction with the rate of formation of NH(2) groups. Both rates increase very much with the dose, but the number of bonds broken decreases. Sedimentation measurements show a strong shape modification of the soluble fraction in the case of the 25 and 40 megarad samples. The increase in asymmetry is bound to the increase in the rate of attack on gamma-irradiated ovalbumin by pepsin. Infrared spectra of the aggregates show small difference from those of the native samples.


Renewed interest in centrally acting antihypertensive drugs.
Classic centrally acting drugs such as clonidine and alpha-methyldopa induce peripheral sympatho-inhibition via the stimulation of alpha(2)-adrenoceptors in the brainstem. From a haemodynamic point of view this appears to be a useful mechanism to lower elevated blood pressure in hypertensives. Although not known in full detail, a complex relationship exists between the sympathetic nervous system and hypertensive disease; sympathetic inhibition therefore appears to be a logical target of antihypertensive drug treatment. Numerous attempts have been made to improve the unfavourable side-effect profile of the centrally actingalpha(2)-adrenoceptor stimulants. None of these attempts were successful, since both the central antihypertensive activity and the major side-effects (sedation, dry mouth) are mediated by alpha(2)-adrenoceptors. A new approach in this field has been offered by the introduction of centrally acting antihypertensives which interact with central imidazoline (I(1))-receptors and thus cause peripheral sympatho-inhibition. As such they offer haemodynamic benefits similar to those of the classic alpha(2)-adrenoceptor agonists. However, it is hoped that their side-effect profile will be more favourable because of their much weaker affinity for alpha(2)-adrenoceptors. Moxonidine and rilmenidine are the prototypes of centrally acting I(1)-receptor stimulants. The antihypertensive activity of these agents is caused by vasodilatation and a reduction of peripheral vascular resistance. Left ventricular end-diastolic and end-systolic volumes are reduced, whereas heart rate, stroke volume, cardiac output and pulmonary artery pressures are largely unchanged. Left ventricular hypertrophy (LVH) is reduced in the long term. Both drugs when applied in a once-daily dosage schedule appear to control hypertension in most patients. Both drugs have been compared with representative agents from the major classes of antihypertensives in controlled trials and have been found to be equally effective with respect to blood pressure control. The incidence and severity of side-effects is lower than for clonidine, in particular with respect to sedation. A rebound (withdrawal) phenomenon has so far not been reported for moxonidine and rilmenidine. In conclusion, I(1)-receptor stimulants appear to offer the potential to be developed as centrally acting agents with a better side-effect profile than the classic alpha(2)-adrenoceptor stimulants, but with similar haemodynamic properties.


An evolutionary hypothesis of suicide: why it could be biologically adaptive and is so prevalent in certain occupations.
From an evolutionary perspective, suicide is a puzzle, because it has serious adverse effects, yet is remarkably common and heritable. An hypothesis is proposed to explain this puzzle, by explaining how suicide could be adaptive through reducing risk that individuals will transmit infections to kin. Empirical evidence supports four predictions from the hypothesis. There are well-established mechanisms by which infections and immune factors increase risk for mental disorders that contribute to suicide. Suicide is more prevalent in occupations with greater exposure to infection and immune-compromising factors and at higher latitudes, where key environmental factors increase vulnerability to infection. In several other highly social species, suicide-like behaviors have evolved to reduce transmission of infections. If the hypothesis is correct, detection and treatment of underlying infections and immune dysfunction should help predict and prevent suicidal behavior, while also combating spread of infectious diseases.


Evaluation of an RNAseq-Based Immunogenomic Liquid Biopsy Approach in Early-Stage Prostate Cancer.
The primary objective of this study is to detect biomarkers and develop models that enable the identification of clinically significant prostate cancer and to understand the biologic implications of the genes involved. Peripheral blood samples (1018 patients) were split chronologically into independent training (n = 713) and validation (n = 305) sets. Whole transcriptome RNA sequencing was performed on isolated phagocytic CD14+ and non-phagocytic CD2+ cells and their gene expression levels were used to develop predictive models that correlate to adverse pathologic features. The immune-transcriptomic model with the highest performance for predicting adverse pathology, based on a subtraction of the log-transformed expression signals of the two cell types, displayed an area under the curve (AUC) of the receiver operating characteristic of 0.70. The addition of biomarkers in combination with traditional clinical risk factors (age, serum prostate-specific antigen (PSA), PSA density, race, digital rectal examination (DRE), and family history) enhanced the AUC to 0.91 and 0.83 for the training and validation sets, respectively. The markers identified by this approach uncovered specific pathway associations relevant to (prostate) cancer biology. Increased phagocytic activity in conjunction with cancer-associated (mis-)regulation is also represented by these markers. Differential gene expression of circulating immune cells gives insight into the cellular immune response to early tumor development and immune surveillance.


Two cases of clear cell ovarian cancer in young patients.
Ovarian cancer is the most common cause of cancer death worldwide. Incidence of ovarian cancer is more common in postmenopausal women. Premenopausal onset is rare and the present study described two cases of ovarian clear cell tumors in young women. The patients presented with pelvic mass which was confirmed by sonography and laparotomy and final diagnosis was made according to histologic examination. Both patients showed a solid mass with cystic components in adnexal areas and explorative laparotomy demonstrated extension of tumors to abdomen in both patients. The level of CA 125 increased in both patients. For both tumors, immunohistochemical stainings were positive for CK7 and CD15, but CK20 was negative. Although ovarian clear cell tumor is usually diagnosed in postmenopausal women but its diagnosis should be suspected in young women with pelvic mass.


Development of Professionalism and Performance Accountability for Sustained FIM Accuracy Improvement.
Rehabilitation nurses have essential responsibility for detailed Functional Independence Measure (FIM) documentation and accurate scoring. The goal of this performance improvement project was to improve individual FIM accuracy and establish and maintain a FIM accuracy standard. Case Study. This performance improvement project was implemented in a 20-bed CARF-accredited Midwestern acute care hospital inpatient rehabilitation unit. Improved individual and overall RN FIM scoring accuracy, with increased length of stay and case mix index. Concurrent FIM scoring and documentation review holds staff accountable and allows for timely revisions following discrepancy identification. Leadership supports for concurrent FIM audits, utilization of available technology, transparent reporting of results, and individual accountability lead to sustained improvement in FIM scoring accuracy. Accurate FIM documentation and scoring drive appropriate length of stay and case mix group assignment. Assessment competence enhances nursing engagement, confidence, and process accountability.


Plasmid involvement in production of and immunity to the staphylococcin-like peptide Pep 5.
The staphylococcin-like peptide Pep 5 is produced by the penicillin resistant strain Staphylococcus epidermidis 5. This strain is immune to the peptide. Plasmid analysis of S. epidermidis 5 by agarose gel electrophoresis and electron microscopy demonstrated five plasmids with molecular weights ranging from 5.8 X 10(6) to 29 X 10(6). Variants of S. epidermidis 5 not producing Pep 5 or which had become penicillin sensitive were induced by various curing treatments. Strains lacking the 13.9 X 10(6) mol. wt plasmid (pED502) had lost penicillin resistance, and those lacking the 12.3 X 10(6) mol. wt plasmid (pED503) failed to produce Pep 5. pED503 is also responsible for the immunity of the producer cell to Pep 5. Plasmid pED502 could be transformed into S. aureus RN 981 which then became resistant to penicillin. pED503 could not be transformed into S. aureus RN 981, but could be transformed into S. epidermidis 5 variants previously cured of this plasmid; the transformants then regained the properties of Pep 5 production and immunity.


Epidemic Use of Benzodiazepines among Older Adults in Israel: Epidemiology and Leverage Points for Improvement.
Benzodiazepines and benzodiazepine-receptor agonists (BDZRAs, often known as "Z-drugs") are commonly used in older adults despite well-documented harms. To evaluate patterns of benzodiazepine and BDZRA use in Israel, focusing on potential leverage points where quality improvement initiatives might effectively curtail new use or the transition from intermittent to chronic use. We used national electronic medical data to assess a 10% random sample of adults receiving care in Clalit Health Services, which serves half of Israel's population. The sample included 267,221 adults, of whom 56,808 (21%) were age 65 and older. Medication use from 2013 to 2015 was ascertained using pharmacy dispensing data. In 2014, 7% of adults age 21-64 and 32% of adults age 65 and older received at least one benzodiazepine/BDZRA, including 49% of adults age 85 and older (P < 0.001). The majority of older users (59%) were long-term users of the drugs, and 21% of older adults who were short-term users in 2014 transitioned to medium- or long-term use in 2015. Older Arab Israelis were much less likely to receive benzodiazepine/BDZRAs than older Jewish Israelis (adjusted OR 0.28, 95% 0.25-0.31), but within each community there was no major variation in prescribing rates across clinics. Depression diagnosis was associated with particularly high rates of benzodiazepine/BDZRA use: 17% of older adults with depression received a benzodiazepine/BDZRA but no antidepressant, and 42% received both. Recent hospitalization increased the risk of new benzodiazepine/BDZRA use (adjusted OR 1.41, 95% CI 1.01-1.96), but the absolute risk increase was only 3%. Benzodiazepines/BDZRAs are used at exceptionally high rates by older Israeli adults, especially the oldest old. Important leverage points for quality improvement efforts include curtailing the transition from short-term to long-term use, reducing use in older adults with depression, and identifying reasons that explain large differences in benzodiazepine/BDZRA prescribing between different ethnic groups.


Biogenic methane production in formation waters from a large gas field in the North Sea.
Methanogenesis was investigated in formation waters from a North Sea oil rimmed gas accumulation containing biodegraded oil, which has not been subject to seawater injection. Activity and growth of hydrogenotrophic methanogens was measured but acetoclastic methanogenesis was not detected. Hydrogenotrophic methanogens showed activity between 40 and 80 degrees C with a temperature optimum (ca. 70 degrees C) consistent with in situ reservoir temperatures. They were also active over a broad salinity range, up to and consistent with the high salinity of the waters (90 g l(-1)). These findings suggest the methanogens are indigenous to the reservoir. The conversion of H(2) and CO(2) to CH(4) in methanogenic enrichments was enhanced by the addition of inorganic nutrients and was correlated with cell growth. Addition of yeast extract also stimulated methanogenesis. Archaeal 16S rRNA gene sequences recovered from enrichment cultures were closely related to Methanothermobacter spp. which have been identified in other high-temperature petroleum reservoirs. It has recently been suggested that methanogenic oil degradation may be a major factor in the development of the world's heavy oils and represent a significant and ongoing process in conventional deposits. Although an oil-degrading methanogenic consortium was not enriched from these samples the presence and activity of communities of fermentative bacteria and methanogenic archaea was demonstrated. Stimulation of methanogenesis by addition of nutrients suggests that in situ methanogenic biodegradation of oil could be harnessed to enhance recovery of stranded energy assets from such petroleum systems.


Laser induced fluorescence coupled to capillary electrophoresis for the determination of fluoroquinolones in foods of animal origin using molecularly imprinted polymers.
A method for the simultaneous determination of four fluoroquinolones of veterinary use (ciprofloxacin, danofloxacin, enrofloxacin and sarafloxacin) in two complex matrixes, such as bovine raw milk and pig kidney, has been established and validated. The method is based on the use of capillary electrophoresis (CE) coupled with a very sensitive detection mode, such as laser induced fluorescence (LIF) detection, due to the fact the all the compounds selected show native fluorescence. In order to achieve high selectivity in the sample treatment procedure, a commercially available molecularly imprinted polymer has been used for the solid phase extraction of the analytes. Once the retention and elution processes were optimized, the final extract was analyzed by CE-LIF using a 325 nm He-Cd laser. Optimum separation was obtained in a 70 cm x75 microm capillary using a 125 mM phosphoric acid solution at pH 2.8 with 36% methanol as background electrolyte. The method provided very low detection limits, ranging from 0.17 to 0.98 microg/kg for milk and 1.10 to 10.5 microg/kg for kidney, with acceptable precision and satisfactory recoveries.


Targeting critical kinases and anti-apoptotic molecules overcomes steroid resistance in MLL-rearranged leukaemia.
Acute lymphoblastic leukaemia with mixed lineage leukaemia gene rearrangement (MLL-ALL) frequently affects infants and is associated with a poor prognosis. Primary refractory and relapsed disease due to resistance to glucocorticoids (GCs) remains a substantial hurdle to improving clinical outcomes. In this study, we aimed to overcome GC resistance of MLL-ALL. Using leukaemia patient specimens, we performed bioinformatic analyses to identify target genes/pathways. To test inhibition of target pathways in vivo, we created pre-clinical therapeutic mouse patient-derived xenograft (PDX)-models by transplanting human MLL-ALL leukaemia initiating cells (LIC) into immune-deficient NSG mice. Finally, we conducted B-cell lymphoma-2 (BCL-2) homology domain 3 (BH3) profiling to identify BH3 peptides responsible for treatment resistance in MLL-leukaemia. Src family kinases (SFKs) and Fms-like tyrosine kinase 3 (FLT3) signaling pathway were over-represented in MLL-ALL cells. PDX-models of infant MLL- ALL recapitulated GC-resistance in vivo but RK-20449, an inhibitor of SFKs and FLT3 eliminated human MLL-ALL cells in vivo, overcoming GC-resistance. Further, we identified BCL-2 dependence as a mechanism of treatment resistance in MLL-ALL through BH3 profiling. Furthermore, MLL-ALL cells resistant to RK-20449 treatment were dependent on the anti-apoptotic BCL-2 protein for their survival. Combined inhibition of SFKs/FLT3 by RK-20449 and of BCL-2 by ABT-199 led to substantial elimination of MLL-ALL cells in vitro and in vivo. Triple treatment combining GCs, RK-20449 and ABT-199 resulted in complete elimination of MLL-ALL cells in vivo. SFKs/FLT3 signaling pathways are promising targets for treatment of treatment-resistant MLL-ALL. Combined inhibition of these kinase pathways and anti-apoptotic BCL-2 successfully eliminated highly resistant MLL-ALL and demonstrated a new treatment strategy for treatment-resistant poor-outcome MLL-ALL. This study was supported by RIKEN (RIKEN President's Discretionary Grant) for FI, Japan Agency for Medical Research and Development (the Basic Science and Platform Technology Program for Innovative Biological Medicine for FI and by NIH CA034196 for LDS. The funders had no role in the study design, data collection, data analysis, interpretation nor writing of the report.


Accelerated HF-rTMS Modifies SERT Availability in the Subgenual Anterior Cingulate Cortex: A Canine [11C]DASB Study on the Serotonergic System.
Repetitive transcranial magnetic stimulation (rTMS) is thought to partly exert its antidepressant action through the serotonergic system. Accelerated rTMS may have the potential to result in similar but faster onset of clinical improvement compared to the classical daily rTMS protocols, but given that delayed clinical responses have been reported, the neurobiological effects of accelerated paradigms remain to be elucidated including on this neurotransmitter system. This sham-controlled study aimed to evaluate the effects of accelerated high frequency rTMS (aHF-rTMS) over the left frontal cortex on the serotonin transporter (SERT) in healthy beagle dogs. A total of twenty-two dogs were randomly divided into three unequal groups: five active stimulation sessions (five sessions in one day, n = 10), 20 active stimulation sessions (five sessions/day for four days, n = 8), and 20 sham stimulation sessions (five sessions/day for four days, n = 4). The SERT binding index (BI) was obtained at baseline, 24 h post stimulation protocol, one month, and three months post stimulation by a [11C]DASB PET scan. It was found that one day of active aHF-rTMS (five sessions) did not result in significant SERT BI changes at any time point. For the 20 sessions of active aHF-rTMS, one month after stimulation the SERT BI attenuated in the sgACC. No significant SERT BI changes were found after 20 sessions of sham aHF-rTMS. A total of four days of active aHF-rTMS modified sgACC SERT BI one month post-stimulation, explaining to some extent the delayed clinical effects of accelerated rTMS paradigms found in human psychopathologies.


Circadian pattern of plasma 17-hydroxycorticosteroid: alteration by anticholinergic agents.
Atropine, administered to cats just prior to the time of the expected circadian rise in levels of 17-hydroxycorticosteroid in plasma, blocks this rise. Atropine does not alter this circadian pattern when administered at other times in the circadian cycle. Results similar to those obtained with atropine have been observed with short-acting barbiturates. Dibenzyline administered just prior to the time of the expected circadian rise is ineffective in blocking this rise. These findings support the hypothesis that the circadian pattern of plasma 17-hydroxycorticosteroid levels reflects activation, by the central nervous system, of the hypothalamicpituitary-adrenal axis during a "critical time period" in the circadian cycle.


Does presence of a VACTERL anomaly predict an associated gynecologic anomaly in females with anorectal malformations?: A Pediatric Colorectal and Pelvic Learning Consortium Study.
VACTERL association is defined by the presence of 3 or more anomalies in any of the following systems: vertebral, anorectal, cardiac, trachea-esophageal, renal, or limb. This study hypothesized that the presence of VACTERL association would correlate with an increased risk of gynecologic anomalies in patients with anorectal malformation (ARM). This study is a cross-sectional, retrospective analysis from the prospectively collected, multicenter registry of the Pediatric Colorectal and Pelvic Learning Consortium (PCPLC). The 834 female patients with ARM who were enrolled in the registry by January 1, 2020 were included in this study. The relationship of VACTERL association with presence of a gynecologic anomaly was evaluated with Fisher's exact test. The relationship of each VACTERL system with presence of a gynecologic anomaly was assessed in patients with cloaca, rectovestibular fistulas and rectoperineal fistulas. P-values reported were based on a 2-sided alternative and considered significant when less than 0.05. 834 patients with ARM underwent VACTERL screening and gynecologic evaluation with the three most common subtypes being cloaca (n = 215, 25.8%), rectovestibular fistula (n = 191, 22.9%) and rectoperineal fistula (n = 194, 23.3%). A total of 223 (26.7%) patients with ARM had gynecologic anomalies. VACTERL association was seen in 380 (45.6%) of patients with ARM. Gynecologic anomalies were present in 149 (39.1%) vs. 74 (16.3%) of subjects with vs. without VACTERL association (p < 0.001). VACTERL association did not significantly increase the risk of gynecologic anomaly in patients with cloaca and VACTERL (n = 88, 61.5%) vs. cloaca without VACTERL (n = 39, 54.2% p = 0.308). VACTERL association increased the risk of gynecologic anomalies in patients with rectoperineal fistulas (n = 7, 14.9% vs n = 9, 6.1% p = 0.014) and rectovestibular fistulas (n = 19, 31.1% vs. n = 13, 10.0% p<0.001). In patients with ARM who had a VACTERL association, when one of the associated anomalies was renal, there was an even higher risk of having an associated gynecologic anomaly (n = 138, 44.2% vs. n = 85, 16.3% p<0.001). VACTERL association in patients with rectoperineal and rectovestibular fistulas correlates with an increased risk of gynecologic anomalies. The presence of VACTERL associated findings, especially renal, should prompt a thorough evaluation of the gynecologic system. III. Retrospective comparative study.


The effect of parathyroidectomy compared to non-surgical surveillance on kidney function in primary hyperparathyroidism: a nationwide historic cohort study.
Patients with primary hyperparathyroidism (pHPT) and impaired kidney function (estimated glomerular filtration rate (eGFR) < 60 mL/min) are offered parathyroidectomy (PTX) to protect them from further complications. Surprisingly, two recent uncontrolled cohort studies have suggested a further decrease in kidney function following PTX. We aimed to examine the effects of PTX compared to non-surgical surveillance on kidney function in pHPT patients. Historic cohort study. From the Danish National Patient Registry (NPR) and major medical biochemistry laboratories in Denmark, we identified 3585 patients with biochemically confirmed pHPT among whom n = 1977 (55%) were treated with PTX (PTX-group) whereas n = 1608 (45%) were followed without surgery (non-PTX group). Baseline was defined as time of diagnosis and kidney function was re-assessed 9-15 months after PTX (PTX group) or 9-15 months after diagnosis (non-PTX group). At follow-up, eGFR had decreased significantly in the PTX- compared to the non-PTX-group (median - 4% vs. - 1%, p < 0.01). Stratification by baseline eGFR showed that the decrease was significant for those with a baseline eGFR value of 80-89 and > 90 mL/min, but not for those with lower eGFR values. Findings did not differ between patients with mild compared to moderate/severe hypercalcemia. However, after mutual adjustments, we identified baseline levels of calcium, PTH, and eGFR as well as age and treatment (PTX vs. no-PTX) as independent predictors for changes in kidney function. Compared to non-surgical surveillance, PTX is associated with a small but significant decrease in kidney function in pHPT patients with an initial normal kidney function.


Activity Intensity, Volume, and Norms: Utility and Interpretation of Accelerometer Metrics.
The physical activity profile can be described from accelerometer data using two population-independent metrics: average acceleration (ACC, volume) and intensity gradient (IG, intensity). This article aims 1) to demonstrate how these metrics can be used to investigate the relative contributions of volume and intensity of physical activity for a range of health markers across data sets and 2) to illustrate the future potential of the metrics for generation of age and sex-specific percentile norms. Secondary data analyses were conducted on five diverse data sets using wrist-worn accelerometers (ActiGraph/GENEActiv/Axivity): children (n = 145), adolescent girls (n = 1669), office workers (n = 114), premenopausal (n = 1218) and postmenopausal (n = 1316) women, and adults with type 2 diabetes (n = 475). Open-source software (GGIR) was used to generate ACC and IG. Health markers were (a) zBMI (children), (b) %fat (adolescent girls and adults), (c) bone health (pre- and postmenopausal women), and (d) physical function (adults with type 2 diabetes). Multiple regression analyses showed that IG, but not ACC, was independently associated with zBMI/%fat in children and adolescents. In adults, associations were stronger and the effects of ACC and IG were additive. For bone health and physical function, interactions showed associations were strongest if IG was high, largely irrespective of ACC. Exemplar illustrative percentile "norms" showed the expected age-related decline in physical activity, with greater drops in IG across age than ACC. The ACC and the IG accelerometer metrics facilitate the investigation of whether volume and intensity of physical activity have independent, additive, or interactive effects on health markers. In future studies, the adoption of data-driven metrics would facilitate the generation of age- and sex-specific norms that would be beneficial to researchers.


Blending Ionic and Coordinate Bonds in Hybrid Semiconductor Materials: A General Approach toward Robust and Solution-Processable Covalent/Coordinate Network Structures.
Inorganic semiconductor materials are best known for their superior physical properties, as well as their structural rigidity and stability. However, the poor solubility and solution-processability of these covalently bonded network structures has long been a serious drawback that limits their use in many important applications. Here, we present a unique and general approach to synthesize robust, solution-processable, and highly luminescent hybrid materials built on periodic and infinite inorganic modules. Structure analysis confirms that all compounds are composed of one-dimensional anionic chains of copper iodide (CumIm+22-) coordinated to cationic organic ligands via Cu-N bonds. The choice of ligands plays an important role in the coordination mode (μ1-MC or μ2-DC) and Cu-N bond strength. Greatly suppressed nonradiative decay is achieved for the μ2-DC structures. Record high quantum yields of 85% (λex = 360 nm) and 76% (λex = 450 nm) are obtained for an orange-emitting 1D-Cu4I6(L6). Temperature dependent PL measurements suggest that both phosphorescence and thermally activated delayed fluorescence contribute to the emission of these 1D-AIO compounds, and that the extent of nonradiative decay of the μ2-DC structures is much less than that of the μ1-DC structures. More significantly, all compounds are remarkably soluble in polar aprotic solvents, distinctly different from previously reported CuI based hybrid materials made of charge-neutral CumXm (X = Cl, Br, I), which are totally insoluble in all common solvents. The greatly enhanced solubility is a result of incorporation of ionic bonds into extended covalent/coordinate network structures, making it possible to fabricate large scale thin films by solution processes.


Cytoplasmic mislocalization of TDP-43 is toxic to neurons and enhanced by a mutation associated with familial amyotrophic lateral sclerosis.
Mutations in the gene encoding TDP-43-the major protein component of neuronal aggregates characteristic of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) with ubiquitin-positive inclusion bodies-have been linked to familial forms of both disorders. Aggregates of TDP-43 in cortical and spinal motorneurons in ALS, or in neurons of the frontal and temporal cortices in FTLD, are closely linked to neuron loss and atrophy in these areas. However, the mechanism by which TDP-43 mutations lead to neurodegeneration is unclear. To investigate the pathogenic role of TDP-43 mutations, we established a model of TDP-43 proteinopathies by expressing fluorescently tagged wild-type and mutant TDP-43 in primary rat cortical neurons. Expression of mutant TDP-43 was toxic to neurons, and mutant-specific toxicity was associated with increased cytoplasmic mislocalization of TDP-43. Inclusion bodies were not necessary for the toxicity and did not affect the risk of cell death. Cellular survival was unaffected by the total amount of exogenous TDP-43 in the nucleus, but the amount of cytoplasmic TDP-43 was a strong and independent predictor of neuronal death. These results suggest that mutant TDP-43 is mislocalized to the cytoplasm, where it exhibits a toxic gain-of-function and induces cell death.


Alternate light sources in sexual assault examinations: an evidence-based practice project.
The ability of sexual assault nurse examiners to correctly identify and collect DNA evidence improves patient outcomes and prosecution rates. The purpose of this paper is to present findings from a collaborative evidence-based practice (EBP) project between forensic nurses and baccalaureate nursing students. The goal of the project was to determine best practice using an alternate light source (ALS) to identify trace DNA evidence in sexual assault forensic examinations. Using the Johns Hopkins Nursing Evidence-based Practice model, the team searched several databases to summarize the limited amount of evidence available regarding this topic. Recommendations from the EBP project include: elimination of the Wood's lamp in sexual assault examinations; use of an ALS that provides appropriate wavelengths to detect DNA; education of forensic nurses about the advantages and limitations of an ALS; and additional research related to use of an ALS. By participating in similar collaborative efforts, practicing forensic nurses have the opportunity to collaborate with local colleges and universities to make complex projects more manageable while fulfilling the International Association of Forensic Nurses vision for ethical practice.


Lactic Acidosis and the Role of Sodium Bicarbonate: A Narrative Opinion.
Lactic acidosis occurs commonly and can be a marker of significant physiologic derangements. However what an elevated lactate level and acidemia connotes and what should be done about it is subject to inconsistent interpretations. This review examines the varied etiologies of lactic acidosis, the physiologic consequences, and the known effects of its treatment with sodium bicarbonate. Lactic acidosis is often assumed to be a marker of hypoperfusion, but it can also result from medications, organ dysfunction, and sepsis even in the absence of malperfusion. Acidemia causes deleterious effects in almost every organ system, but it can also have positive effects, increasing localized blood flow and oxygen delivery, as well as providing protection against hypoxic cellular injury. The use of sodium bicarbonate to correct severe acidemia may be tempting to clinicians, but previous studies have failed to show improved patient outcomes following bicarbonate administration. Bicarbonate use is known to decrease vasomotor tone, decrease myocardial contractility, and induce intracellular acidosis. This suggests that mild to moderate acidemia does not require correction. Most recently, a randomized control trial found a survival benefit in a subgroup of critically ill patients with serum pH levels <7.2 with concomitant acute kidney injury. There is no known benefit of correcting serum pH levels ≥ 7.2, and sparse evidence supports bicarbonate use <7.2. If administered, bicarbonate is best given as a slow IV infusion in the setting of adequate ventilation and calcium replacement to mitigate its untoward effects.


Sporulation, heat resistance, and biological properties of Clostridium perfringens.
A sporulation medium for 134 Clostridium perfringens strains, including types A, B, C, D, E, and F, was devised according to Grelet's observation that sporulation occurred when cultural environment became limited in any nutritional requirement indispensable for the growth of the organism. Sporulation took place most prominently when 10% cooked-meat broth (pH 7.2) containing 3% Proteose Peptone and 1% glucose was used for the preculture and 2% Poli Peptone medium (pH 7.8) was used for the subculture medium. Sometimes, terminal spores could be observed. A correlation between sporulation and heat resistance was examined by use of C. perfringens strains isolated from samples heated at different temperatures. Almost all strains isolated from unheated samples and from those heated at lower temperatures gave rise to spores in our sporulation medium, but the spores were weakly heat-resistant, whereas strains isolated from samples heated at 100 C for 60 min were highly heat-resistant but sporulated poorly. A majority of these heat-resistant strains were non-gelatinolytic and definitely salicin-fermenting.


Development of ⁶⁸Ga-labelled DTPA galactosyl human serum albumin for liver function imaging.
The hepatic asialoglycoprotein receptor is responsible for degradation of desialylated glycoproteins through receptor-mediated endocytosis. It has been shown that imaging of the receptor density using [(99m)Tc]diethylenetriamine pentaacetic acid (DTPA) galactosyl human serum albumin ([(99m)Tc]GSA) allows non-invasive determination of functional hepatocellular mass. Here we present the synthesis and evaluation of [(68)Ga]GSA for the potential use with positron emission tomography (PET). Labelling of GSA with (68)Ga was carried out using a fractionated elution protocol. For quality control thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) and size exclusion chromatography (SEC) techniques were evaluated. Stability of [(68)Ga]GSA was studied in phosphate-buffered saline (PBS) and human serum. For in vivo evaluation [(68)Ga]GSA distribution in Lewis rats was compared with [(99m)Tc]GSA by using a dual isotope protocol. PET and planar imaging studies were performed using the same scaled molar dose of [(68)Ga]GSA and [(99m)Tc]GSA. Time-activity curves (TAC) for heart and liver were generated and corresponding parameters calculated (t50, t90). [(68)Ga]GSA can be produced with high radiochemical purity. The best TLC methods for determining potential free (68)Ga include 0.1 M sodium citrate as eluent. None of the TLC methods tested were able to determine potential colloids. This can be achieved by SEC. HPLC confirmed high radiochemical purity (>98%). Stability after 120 min incubation at 37 °C was high in PBS (>95% intact tracer) and low in human serum (∼27% intact tracer). Biodistribution studies simultaneously injecting both tracers showed comparable liver uptake, whereas activity concentration in blood was higher for [(68)Ga]GSA compared to [(99m)Tc]GSA. The [(99m)Tc]GSA TACs exhibited a small degree of hepatic metabolism compared to the [(68)Ga]GSA curves. The mean [(68)Ga]GSA t90 was higher than the mean t90 for [(99m)Tc]GSA. The mean [(68)Ga]GSA t50 was not significantly different from the mean t50 for [(99m)Tc]GSA. This study provides a promising new (68)Ga-labelled compound based on a commercially used kit for imaging the functional hepatocellular mass.


Microglia complement astrocytes in neuromyelitis optica.
Neuromyelitis optica (NMO) is a central nervous system (CNS) inflammatory autoimmune disease caused by antibodies against aquaporin-4 (AQP4) expressed on astrocytes. Binding of AQP4-specific antibodies (NMO-IgG) triggers activation of the complement cascade, which is responsible for astrocyte loss and secondary demyelination. Although the role for the cytolytic complement proteins in astrocyte destruction in NMO is well established, little is known regarding the initial phase of astrocyte injury. In this issue of the JCI, Chen and colleagues evaluated the precytolytic phase when NMO-IgG binds astrocytes in vivo in the absence of exogenous complement. NMO-IgG alone caused astrocyte activation and AQP4 loss. Surprisingly, microglia, CNS-resident innate immune cells that produce endogenous complement, were required for clinical manifestations of disease, a finding that suggests microglia may serve as a therapeutic target in NMO.


A practical approach with drones, smartphones, and tracking tags for potential real-time animal tracking.
Drones are increasingly used for fauna monitoring and wildlife tracking; however, their application for wildlife tracking is restricted by developing such systems. Here we explore the potential of drones for wildlife tracking using an off-the-shelf system that is easy to use by non-specialists consisting of a multirotor drone, smartphones, and commercial tracking devices via Bluetooth and Ultra-Wide Band (UWB). We present the system configuration, explore the operational parameters that can affect detection capabilities, and test the effectiveness of the system for locating targets by simulating target animals in savanna and forest environments. The self-contained tracking system was built without hardware or software customization. In 40 tracking flights carried out in the Brazilian Cerrado, we obtained a detection rate of 90% in savanna and 40% in forest areas. Tests for targets in movement (N = 20), the detection rates were 90% in the savanna and 30% in the forest areas. The spatial accuracy obtained by the system was 14.61 m, being significantly more accurate in savanna ( x ¯ = 10.53) than in forest areas ( x ¯ = 13.06). This approach to wildlife tracking facilitates the use of drones by non-specialists at an affordable cost for conservation projects with limited resources. The reduced size of the tags, the long battery life, and the lower cost compared to GPS-tags open up a range of opportunities for animal tracking.


Factor H C-Terminal Domains Are Critical for Regulation of Platelet/Granulocyte Aggregate Formation.
Platelet/granulocyte aggregates (PGAs) increase thromboinflammation in the vasculature, and PGA formation is tightly controlled by the complement alternative pathway (AP) negative regulator, Factor H (FH). Mutations in FH are associated with the prothrombotic disease atypical hemolytic uremic syndrome (aHUS), yet it is unknown whether increased PGA formation contributes to the thrombosis seen in patients with aHUS. Here, flow cytometry assays were used to evaluate the effects of aHUS-related mutations on FH regulation of PGA formation and characterize the mechanism. Utilizing recombinant fragments of FH spanning the entire length of the protein, we mapped the regions of FH most critical for limiting AP activity on the surface of isolated human platelets and neutrophils, as well as the regions most critical for regulating PGA formation in human whole blood stimulated with thrombin receptor-activating peptide (TRAP). FH domains 19-20 were the most critical for limiting AP activity on platelets, neutrophils, and at the platelet/granulocyte interface. The role of FH in PGA formation was attributed to its ability to regulate AP-mediated C5a generation. AHUS-related mutations in domains 19-20 caused differential effects on control of PGA formation and AP activity on platelets and neutrophils. Our data indicate FH C-terminal domains are key for regulating PGA formation, thus increased FH protection may have a beneficial impact on diseases characterized by increased PGA formation, such as cardiovascular disease. Additionally, aHUS-related mutations in domains 19-20 have varying effects on control of TRAP-mediated PGA formation, suggesting that some, but not all, aHUS-related mutations may cause increased PGA formation that contributes to excessive thrombosis in patients with aHUS.


The Relationship Between Self-Esteem, Emotional Intelligence, and Empathy Among Students From Six Health Professional Programs.
Theory: Psychosocial skills such as communication, empathy, and emotional intelligence are now considered key attributes of health professionals. Self-esteem is another important construct that is less well studied. Self-esteem is important because low levels have been linked to depression, suicide, and eating disorders. Given that health professional students experience high levels of stress and are at increased risk for similar psychopathology, self-esteem may be an important variable in student well-being and performance after graduation. Hypotheses: This study sought to explore self-esteem during students' 1st year of training hypothesizing that several would demonstrate low self-esteem. It is also hypothesized that emotional intelligence and empathy would be associated with self-esteem. Method: A cross-sectional survey was conducted, and data were gathered from dental, medical, nursing, optometry, pharmacy and veterinary students. Self-report questionnaires assessing self-esteem, emotional intelligence, and empathy were completed and demographic information was collected. Scores were calculated and differences between groups analyzed with analysis of variance and chi-square testing. Pearson's correlation was used to assess associations between the constructs. Results: The mean self-esteem score was 26.2 ± 2.3 but 21% of the sample evidenced low self-esteem. There was no difference in the proportion of students demonstrating low self-esteem among programs. Gender did not have a significant effect on self-esteem scores, though ethnicity did. Emotional intelligence scores were higher among male individuals than among female. Emotional intelligence and empathy showed a small association with self-esteem. Conclusions: A significant proportion of health professional students suffer from low self-esteem during their 1st year of study. Such students may be more susceptible to the stresses associated with study and the development of psychopathology. More research needs to be conducted to explore the relationships between self-esteem, emotional intelligence, and empathy with a view to strengthening training in these areas and managing the challenges faced by health professional students.


Vision and visual plasticity in ageing mice.
Little is known about neuronal changes during ageing in the visual system of mice which are increasingly being used as animal models for human visual disorders. Measuring the optomotor response to moving gratings, visual acuity of C57BL/6-mice was 0.39 cycles/degree (cyc/deg) until 12 months of age and declined to 0.27 cyc/deg (by 30%) at 26 months. In the visual water task, a cortex-dependent task based on visual discrimination learning, visual acuity remained stable at 0.58 cyc/deg up to 21 months and then declined to 0.48 cyc/deg (by 19%) at 27 months. Visual cortical activity recorded by optical imaging declined by 33% between seven and 23 months of age. After monocular deprivation and daily testing of the optomotor response, visual acuity of the open eye increased by 29% in 4 to 7-month-old animals, while the increase was only 13% in 23-month-old mice. Interestingly, interindividual variability generally increased with age, so that some 23-month-old mice retained visual acuity and interocular plasticity like 4 or 7-month-old animals. In summary, reduced visual function was accompanied by a reduction of both visual cortical responses and interocular plasticity indicating a central nervous system component in age-related vision loss in mice.


Targeted Modulation of FAK/PI3K/PDK1/AKT and FAK/p53 Pathways by Cucurbitacin B for the Antiproliferation Effect Against Human Cholangiocarcinoma Cells.
Inadequate responses to traditional chemotherapeutic agents in cholangiocarcinoma (CCA) emphasize a requirement for new effective compounds for the treatment of this malignancy. This study aimed to investigate the antiproliferative property of cucurbitacin B on KKU-100 CCA cells. The determination of underlying molecular mechanisms was also carried out. The results revealed that cucurbitacin B suppressed growth and replicative ability to form colonies of CCA cells, suggesting the antiproliferative effect of this compound against the cells. Flow cytometry analysis demonstrated that the interfering effect of cucurbitacin B on the CCA cell cycle at the G2/M phase was accountable for its antiproliferation property. Accompanied with cell cycle disruption, cucurbitacin B altered the expression of proteins involved in the G2/M phase transition including downregulation of cyclin A, cyclin D1, and cdc25A, and upregulation of p21. Additional molecular studies demonstrated that cucurbitacin B suppressed the activation of focal adhesion kinase (FAK) which consequently resulted in inhibition of its kinase-dependent and kinase-independent downstream targets contributing to the regulation of cell proliferation including PI3K/PDK1/AKT and p53 proteins. In this study, the transient knockdown of FAK using siRNA was employed to ascertain the role of FAK in CCA cell proliferation. Finally, the effect of cucurbitacin B on upstream receptor tyrosine kinases regulating FAK activation was elucidated. The results showed that the inhibitory effect of cucurbitacin B on FAK activation in CCA cells is mediated via interference of EGFR and HER2 expression. Collectively, cucurbitacin B might be a promising drug for CCA treatment by targeting FAK protein.


Transfer of the virulence plasmid of Yersinia pestis to Yersinia pseudotuberculosis.
Transposon Tn5 insertion derivatives of the virulence plasmid pYV019 of Yersinia pestis were transferred by P1 transduction into a plasmid-free strain of Y. pseudotuberculosis. One of these plasmid derivatives conferred virulence upon the Y. pseudotuberculosis strain. This strain had the ability to express temperature-inducible plasmid-coded outer membrane proteins and was also found to be Ca2+ dependent.


[Radiation and combination treatment of maxillary cancer using metronidazole].
A study was made of the results of therapy of 66 patients with maxillary cancer, of them 31 (a control group) received routine therapy by a method of summary focal dose (SFD) fractionation--60-70 Gy. In the study group (35 patients) treatment was started with large fractions at a dose of 5 Gy 2 times a week up to SFD of 20 Gy using metronidazole 2.5-3 h before each of the first 4 large fractions and continued using a course with SFD up to 52 Gy by common fractionation. Staged assessment of the results of radiotherapy was done when the summary dose reached 40 Gy in the control group and 32 Gy in the study group. Complete regression of a tumor or a decrease in its volume by more than 50% at these doses were noted in 13 (41.9%) patients of the control group and in 23 (65.7%) patients of the study group. After combined treatment of a slightly decreased tumor preirradiated in a summary dose of 40 Gy and 32 Gy the 3-year recurrence-free survival was noted in 72.2% (13 patients) in the control group and in 91.7% (11 patients) in the study group. The results obtained are indicative of a marked radiosensitizing effect of metronidazole in radical radiation and combined radiation therapy of maxillary cancer improving long-term results.


Continuous monitoring in COVID-19 care: a retrospective study in time of crisis.
A new monitoring system was implemented to support nursing staff and physicians on the COVID-19 ward. This system was designed to remotely monitor vital signs, to calculate an automated Early Warning Score, and to help identify patients at risk of deterioration. Hospitalized patients who tested positive for SARS-CoV-2 were connected to 2 wireless sensors measuring vital signs. Patients were divided into 2 groups based on the occurrence of adverse events during hospitalization. Heart and respiratory rate were monitored continuously and an automated EWS was calculated every 5 minutes. Data were compared between groups. Prior to the occurrence of adverse events, significantly higher median heart and respiration rate and significantly lower median SPO2 values were observed. Mean and median automated EWS were significantly higher in patients with an adverse event. Continuous monitoring systems might help to detect clinical deterioration in COVID-19 patients at an earlier stage.


Salmon food matrix influences digestion and bioavailability of long-chain omega-3 polyunsaturated fatty acids.
The natural structure of whole food plays an important role in the physiological impact of bioactive compounds present within the food, also known as the "matrix effect". Long-chain omega-3 polyunsaturated fatty acids (LCn-3PUFAs) are one example of a food-derived nutrient, mostly found in fish, that is believed to be influenced by the food matrix. However, most previous studies have compared only the long-term bioavailability of fish versus fish oil and have used commercial sources of fish oil. The present study aimed to investigate whether fish (salmon) matrix influences the transit of LCn-3PUFAs during in vitro digestion and affects bioavailability in healthy females. Meals containing intact salmon (intact structure), minced salmon (some structure) and defatted salmon + oil (no structure) with identical macronutrient compositions were developed. Healthy female participants (n = 13) consumed the meals in a postprandial crossover study and blood was collected at regular time points for 6 h post meal consumption. In parallel, in vitro digestion of the meals was performed using a human gastric simulator (HGS) and digesta samples were collected at regular time points for 6 h. Results: showed that plasma concentration of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were significantly higher after participants consumed intact salmon compared to the other meals (covariate analysis p < 0.001). The in vitro digestion results showed defatted salmon + oil meal had a faster decrease in pH and faster fat emptying from the HGS than the other two meals. The defatted salmon + oil meal more closely followed fat emptying of a homogeneous unstructured meal, whereas the other meals exhibited phase separation with a delay in fat emptying. Conclusion: The fish matrix (salmon) plays an important role in the bioaccessibility and bioavailability of EPA and DHA. The differences are partly explained by fat digestion and emptying from the stomach. This study suggests that the natural structure of fish has a functional effect on the absorption and bioavailability of fish oil.


Long-term efficacy of a computer-tailored physical activity intervention for prostate and colorectal cancer patients and survivors: A randomized controlled trial.
Physical activity (PA) can improve the physical and psychological health of prostate and colorectal cancer survivors, but PA behavior change maintenance is necessary for long-term health benefits. OncoActive is a print- and web-based intervention in which prostate and colorectal cancer patients and survivors receive automatically generated, personalized feedback aimed at integrating PA into daily life to increase and maintain PA. We evaluated the long-term outcomes of OncoActive by examining the 12-month follow-up differences between OncoActive and a control group, and we explored whether PA was maintained during a 6-month non-intervention follow-up period. Prostate or colorectal cancer patients were randomly assigned to an OncoActive (n = 249) or a usual care waitlist control group (n = 229). OncoActive participants received PA advice and a pedometer. PA outcomes (i.e., ActiGraph and self-report moderate-to-vigorous intensity PA (MVPA) min/week and days with ≥30 min PA) and health-related outcomes (i.e., fatigue, depression, physical functioning) were assessed at baseline, 6 months, and 12 months. Differences between groups and changes over time were assessed with multilevel linear regressions for the primary outcome (ActiGraph MVPA min/week) and all additional outcomes. At 12 months, OncoActive participants did not perform better than control group participants at ActiGraph MVPA min/week, self-report MVPA min/week, or ActiGraph days with PA. Only self-report days with PA were significantly higher in OncoActive compared to the control group. For health-related outcomes only long-term fatigue was significantly lower in OncoActive. When exploratively examining PA within OncoActive, the previously found PA effects at the end of the intervention (6 months follow-up) were maintained at 12 months. Furthermore, all PA outcomes improved significantly from baseline to 12 months. The control group showed small but non-significant improvements from 6 months to 12 months (and from baseline to 12 months), resulting in a decline of differences between groups. The majority of previously reported significant between-group differences at 6 months follow-up were no longer present at long-term follow-up, possibly because of natural improvement in the control group. At long-term follow-up, fatigue was significantly lower in OncoActive compared to control group participants. Computer-tailored PA advice may give participants an early start toward recovery and potentially contributes to improving long-term health.


Are multiple cesarean sections safe?
To assess the maternal and neonatal risk associated with high-order cesarean sections, a case-control study was carried out in two university affiliated maternity wards. The outcome of 154 pregnancies of women undergoing cesarean section for the 4th time or more was compared with 148 women sectioned for the 2nd or 3rd time and 132 women of similar age and parity after spontaneous birth. The main outcome measures were maternal operative and postoperative morbidity and neonatal prematurity and its complications, Apgar scores, and the need for intensive care. Women undergoing multiple (> or = 4) cesarean sections had significantly more intra-abdominal adhesions (P < 0.0001) than women sectioned for the 2nd or 3rd time. However, the time interval from incision to delivery and the total duration of operation were similar. The postoperative course was not adversely affected by multiple cesarean sections. A high incidence (16.2%) of preterm cesarean deliveries was noted in the study group. This was due to non-elective repeat cesarean delivery rather than to poor timing of scheduled cesarean sections. The significantly increased (P < 0.05) need for neonatal intensive care was explained by the higher occurrence of prematurity. Low Apgar scores (< or = 7) at 1 and 5 min were significantly (P < 0.01) related to multiple cesarean sections, even after controlling for the effect of gestational age. We conclude that multiple cesarean sections pose little risk for the mother, but may be associated with increased neonatal risk, attributed mainly to preterm non-elective cesarean sections.


Tamoxifen therapy in breast cancer control worldwide.
In most developed and many developing countries, breast cancer is the most frequent cancer and the leading cause of cancer death among women. At least 50% of all breast cancer patients worldwide would survive longer, however, if public awareness about and early detection of the condition were increased and greater use were made of efficient treatment of proven value. With early-stage, localized breast cancer, local treatment combined with adjuvant hormonal therapy with tamoxifen, a synthetic estrogen, could save the lives of 6 women out of 100 compared with local treatment alone. Tamoxifen has anti-estrogenic effects not only on breast cancer cells but also on liver metabolism and bone, with concomitant decreases in risk factors for chronic skeletal and vascular system diseases. Long-term tamoxifen treatment causes major adverse clinical effects in < 5% of women; menopausal and vasomotor symptoms occur in the majority of treated women, but their severity lessens over time. Tamoxifen is being considered as a standard therapy and is included in the WHO list of essential drugs for the treatment of breast cancer patients in both developing and developed countries. For the control of breast cancer more successfully worldwide, one challenge is to make tamoxifen therapy available to greater numbers of women.


Association of circulating omentin-1 level with arterial stiffness and carotid plaque in type 2 diabetes.
Adipokines contribute directly to the atherosclerotic process, connecting metabolic disorders such as obesity and diabetes to cardiovascular disease. Omentin-1 is a recently discovered novel adipokine, so data about the relationship of this adipokine to vascular health in type 2 diabetes is limited. We enrolled 60 people with type 2 diabetes, with or without carotid plaque, and 30 participants with normal glucose tolerance. We measured serum omentin-1, high-sensitivity C-reactive protein (hsCRP) levels, and the homeostasis model assessment of insulin resistance (HOMA-IR), as well as other cardiovascular risk factors. Vascular health was assessed by brachial ankle pulse wave velocity (baPWV) and carotid intima-media thickness (IMT). Serum omentin-1 levels were significantly decreased in type 2 diabetes patients compared to normal glucose controls and was further reduced in type 2 diabetes patients with carotid plaque compared to those without carotid plaque. Multiple stepwise regression analysis showed that age, systolic blood pressure, history of use of statins, angiotensin receptor blockers or angiotensin-converting enzyme inhibitors, and serum omentin-1 level were independent factors determining baPWV in people with type 2 diabetes (r2 = 0.637). Furthermore, in multivariate logistic regression analysis, circulating omentin-1 level was an independent decisive factor for the presence of carotid plaque in type 2 diabetes patients, even after adjusting for age, gender, body mass index, systolic blood pressure, fasting blood glucose, low density lipoprotein cholesterol, and history of smoking and medication (odds ratio, 0.621; 95% confidence interval, 0.420-0.919; P = 0.017). Circulating omentin-1 level was independently correlated with arterial stiffness and carotid plaque in type 2 diabetes, even after adjusting for other cardiovascular risk factors and detailed medication history.


Altering MYC phosphorylation in the epidermis increases the stem cell population and contributes to the development, progression, and metastasis of squamous cell carcinoma.
cMYC (MYC) is a potent oncoprotein that is subject to post-translational modifications that affect its stability and activity. Here, we show that Serine 62 phosphorylation, which increases MYC stability and oncogenic activity, is elevated while Threonine 58 phosphorylation, which targets MYC for degradation, is decreased in squamous cell carcinoma (SCC). The oncogenic role of MYC in the development of SCC is unclear since studies have shown in normal skin that wild-type MYC overexpression can drive loss of stem cells and epidermal differentiation. To investigate whether and how altered MYC phosphorylation might affect SCC development, progression, and metastasis, we generated mice with inducible expression of MYCWT or MYCT58A in the basal layer of the skin epidermis. In the T58A mutant, MYC is stabilized with constitutive S62 phosphorylation. When challenged with DMBA/TPA-mediated carcinogenesis, MYCT58A mice had accelerated development of papillomas, increased conversion to malignant lesions, and increased metastasis as compared to MYCWT mice. In addition, MYCT58A-driven SCC displayed stem cell gene expression not observed with MYCWT, including increased expression of Lgr6, Sox2, and CD34. In support of MYCT58A enhancing stem cell phenotypes, its expression was associated with an increased number of BrdU long-term label-retaining cells, increased CD34 expression in hair follicles, and increased colony formation from neonatal keratinocytes. Together, these results indicate that altering MYC phosphorylation changes its oncogenic activity-instead of diminishing establishment and/or maintenance of epidermal stem cell populations like wild-type MYC, pS62-MYC enhances these populations and, under carcinogenic conditions, pS62-MYC expression results in aggressive tumor phenotypes.


An evaluation of least-squares fits to COSY spectra as a means of estimating proton-proton coupling constants. II. Applications to polypeptides.
A new computational method for simultaneously estimating all the proton-proton coupling constants in a molecule from COSY spectra [Yang, J.-X. and Havel, T.F. (1994) J. Biomol. NMR, 4, 807-826] is applied to experimental data from two polypeptides. The first of these is a cyclic hexapeptide denoted as VDA (-D-Ala1-Phe2-Trp3-Lys(Z)4-Val5-Phe6-), in deuterated DMSO, while the second is a 39-residue protein, called decorsin, in aqueous solution. The effect of different data processing strategies and different initial parameter values on the accuracy of the coupling constants was explored. In the case of VDA, most of the coupling constants did not depend strongly on the initial values chosen for the optimization or on how the data were processed. This, together with our previous experience using simulated data, implies strongly that these values are accurate estimates of the coupling constants. They also differ by an average of only 0.36 Hz from the values of the 14 coupling constants that could be measured independently by established methods. In the case of decorsin, many of the coupling constants exhibited a moderate dependence on their initial values and a strong dependence on how the data were processed. With the most successful data processing strategy, the amide-alpha coupling constants differed by an average of 1.11 Hz from the 21 values that could be measured by established methods, while two thirds of the three-bond coupling constants fell within 1.0 Hz of the ranges obtained by applying the Karplus relation to an independently computed ensemble of distance geometry structures. The averages of the coupling constants over multiple optimizations using random initial values were computed in order to obtain the best possible estimates of the coupling constants. Most clearly incorrect averages can be identified by large standard deviations in the coupling constants or the associated line widths and chemical shifts, and can be explained by strong coupling and/or overlap with the water signal, the diagonal peaks or other cross peaks.


Anti-CD25 antibody enhancement of vaccine-induced immunogenicity: increased durable cellular immunity with reduced immunodominance.
An efficacious vaccine strategy must be capable of inducing strong responses of an appropriate phenotype that are long lasting and sufficiently broad to prevent pathogen escape mechanisms. In the present study, we use anti-CD25 mAb to augment vaccine-induced immunity in mice. We demonstrate that coformulation of Ab and poxviral- or adenoviral-vectored vaccines induces significantly increased T cell responses to a malaria Ag; prior anti-CD25 Ab administration was not required for this effect. Furthermore, this vaccination approach subverts immunodominant epitope hierarchies by enhancing responses to subdominant epitopes induced by recombinant modified vaccinia virus Ankara immunization. Administration of anti-CD25 with a vaccine also induces more durable immunity compared with vaccine alone; significantly higher T cell responses were observed 100 days after the primary immunization. Enhanced immunogenicity is observed for multiple vaccine types with enhanced CD4+ and CD8+ T cell responses induced by bacillus Calmette-Guérin and a recombinant subunit protein vaccine to hepatitis B virus and with multiple Ags of tumor, viral, bacterial, and parasitic origin. Vaccine strategies incorporating anti-CD25 lead to improved protection against pre-erythrocytic malaria challenge. These data underpin new strategies for the design and development of more efficacious vaccines in clinical settings.


Complex wound care by a nurse practitioner in primary care: a quality improvement evaluation on healing rates, costs and patient satisfaction.
Complex wounds are a major burden for healthcare professionals. Patients with complex wounds are often referred to hospitals or wound expertise centres. Complex wound care could be organised in primary care, but very little published evidence for this is available. In this study, members of a primary healthcare organisation were interviewed to ascertain how the wound care was organised. Patient characteristics and data regarding wound care, healing rates and costs were collected from medical records. Patients filled in a questionnaire concerning accessibility, communication, medical counselling and the overall experience of their treatment. This study followed 25 patients with a total of 42 wounds. Some 82% of the wounds were healed within a mean treatment duration of 9.7 weeks. Mean treatment costs were calculated at €155 (£130) for healed patients. Patients were generally very satisfied with their treatment, illustrated by an overall 8.7 out of 10 scoring on an amended survey based on the Consumer Quality Index. Treating complex wounds in primary care seemed adequate, with high patient satisfaction and tolerable costs.


The cultural construction of childhood diarrhoea in rural Nicaragua: relevance for epidemiology and health promotion.
As a component of a series of studies of childhood diarrhoea in rural Nicaragua, lay knowledge regarding the condition and its appropriate management has been investigated through semistructured interviews with 70 mothers. These data have been combined with information from focus group discussions and observations from investigators who have been resident in the study area for many years. For any episode of childhood diarrhoea, the lay nosology influences the treatment path followed. Thus for some types of diarrhoea, treatment at a health centre or health post and the use of rehydration fluids is seen to be appropriate, while for other types the use of traditional healers or home-based treatment, often explicitly without the use of rehydration fluids, is applicable. The implications of lay nosological systems for the interpretation of epidemiological studies and for the implementation of health promotion programmes are discussed.


Which diagnostic procedures in the elderly? The case of late-onset Huntington's disease.
The typical adult-onset form of Huntington's disease (HD) is a clinical condition in which the latest advances of genetic research can be usefully applied during the course of the diagnostic process; not so clear are the guidelines for the much less frequent late-onset variant. We have recently seen three patients in their late sixties who had been misdiagnosed for up to 10 years due to the apparently isolated, mild, and slowly progressive nature of their hyperkinetic movements or cognitive disorders. Only after the results of DNA sequencing on a blood sample became available could the appropriate diagnosis of late-onset HD be reached. By contrast, neuroimaging studies lacked sufficient sensitivity and specificity. Appropriate neurogeriatric assessment in these cases should follow specific guidelines and should always include selected high-technology procedures.


[Evaluation of the use of caerulein (ceruletide) in the postoperative of abdominal surgery].
In order to evaluate the efficacy of Caerulein in the postoperative stage of abdominal surgery, 25 patients received 0.3 mcg/kg intravenously every 4 hours. Bowel Movements, passage of gases, first evacuation and general improvement were compared to 25 patients in similar conditions who didn't receive the drug. The study group showed an earlier appearance of intestinal motility as compared to the control group. This contributed in a relevant matter to an earlier discharge from the hospital. The differences with the control group were statistically significative. The benefits of the drug in the management of the postoperative stage of abdominal surgery are discussed.


[Some ecomorphologic aspects of nodular goiter].
Determine microcalcifications' rule in nodular goiter, to classify those calcifications and identify US features suspect for tumoral pathology. In Endocrine Surgery Unit we ultrasonographically detected 655 patients in a period of twelve months (Jan to Dec 2005). Patients aged from 22 to 75 years. Multinodular goiter were 42% and solitary nodular 33.8%, higher than other studies, as we checked selected patients. We determined microcalcification' incidence in nodular goiter and thyroiditis and studied their correlations with tumours. We also studied other US features suspect for malignant lesions. We divided microcalcifications in four groups. Thyroiditis prevalence was 55.8%. Microcalcifications' prevalence was 32.3%, divided as follow: Type I 18.1%, type II 4%, type III 81.8%, type IV 9%. In 9% of patients was present more than one kind of calcifications. Microcalcification rate was 28.9% in inflammatory disease, 55.17% in multinodular goiter and 18, 18% in single thyroid nodules. "Suspect" microcalcifications were present in 9% of whole thyroid pathology and in 12% of MNG. The majority of microcalcifications belonged to type III and their nature is questionable. Several Authors mentioned microcalcifications as a sensitive and highly specific feature for tumour diagnosis. Microcalcifications have a 59.2% sensitivity rate and a specificity rate of 85.8-95%. Their diagnostic accuracy is 77.4%. Other us suspect features are hypoecogenicity and irregular nodule outline. Ultrasonography remains an operator-dependent exam and it's necessary a correct interpretation of morphological data. Meticulous characterization of all ultrasonographic criteria enounced above is useful to reach satisfactory results and identify suspect lesions.


Evaluating Retention Index Score Assumptions to Refine GC-MS Metabolite Identification.
As metabolomics grows into a high-throughput and high demand research field, current metrics for the identification of small molecules in gas chromatography-mass spectrometry (GC-MS) still require manual verification. Though steps have been taken to improve scoring metrics by combining spectral similarity (SS) and retention index (RI), the problem persists. A large body of literature has analyzed and refined SS scores, but few studies have explicitly studied improvements to RI scores. Here, we examined whether uninvestigated assumptions of the RI score are valid and propose ways to improve them. Query RIs were matched to library RI with a generous window of ±35 to avoid unintentional removal of valid compound identifications. Each match was manually verified as a true positive (TP), true negative, or unknown. Metabolites with at least 30 TP identifications were included in downstream analyses, resulting in a total of 87 metabolites from samples of varying complexity and type (e.g., amino acid mixtures, human urine, fungal species, and so on.). Our results showed that the RI score assumptions of normality, consistent variance across metabolites, and a mean error centered at 0 are often violated. We demonstrated through a cross-validation analysis that modifying these underlying assumptions according to empirical metabolite-specific distributions improved the TP and negative rankings. Further, we statistically determined the minimum number of samples required to estimate distributional parameters for scoring metrics. Overall, this work proposes a robust statistical pipeline to reduce the time bottleneck of metabolite identification by improving RI scores and thus minimize the effort to complete manual verification.


Redefining clinical venous thromboembolism phenotypes: a novel approach using latent class analysis.
Patients with venous thromboembolism (VTE) are commonly classified by the presence or absence of provoking factors at the time of VTE to guide treatment decisions. This approach may not capture the heterogeneity of the disease and its prognosis. To evaluate clinically important novel phenotypic clusters among patients with VTE without cancer and to explore their association with anticoagulant treatment and clinical outcomes. Latent class analysis was performed with 18 baseline clinical variables in 3062 adult patients with VTE without active cancer participating in PREFER in VTE, a noninterventional disease registry. The derived latent classes were externally validated in a post hoc analysis of Hokusai-VTE (n = 6593), a randomized trial comparing edoxaban with warfarin. The associations between cluster membership and anticoagulant treatment, recurrent VTE, bleeding, and mortality after initial treatment were studied. The following 5 clusters were identified: young men cluster (n = 1126, 37%), young women cluster (n = 215, 7%), older people cluster (n = 1106, 36%), comorbidity cluster (n = 447, 15%), and history of venous thromboembolism cluster (n = 168, 5%). Patient characteristics varied by age, sex, medical history, and treatment patterns. Consistent clusters were evident on external validation. In Cox proportional hazard models, recurrence risk was lower in the young women cluster (hazard ratio [HR], 0.27; 95% CI, 0.12-0.61) compared with the comorbidity cluster, after adjusting for extended anticoagulation. The risk of bleeding was lower in young men, young women, and older people clusters (HR, 0.50; 95% CI, 0.38-0.66; HR, 0.23; 95% CI, 0.11-0.46; and HR, 0.55; 95% CI 0.41-0.73, respectively). The heterogeneity of VTE cases extends beyond the distinction between provoked and unprovoked VTE.


Equity in antenatal care visits among adolescent mothers: An analysis of 54 country levels trend and projection of coverage from 2000 to 2030.
Ensuring utilization of antenatal care (ANC) services by adolescent mothers (ages 10-19) is an enormous challenge in low-and middle-income countries (LMICs). This study provides the first comprehensive analysis of ANC visits among adolescent and adult mothers. Using all available Demographic and Health Survey and Multiple Indicator Cluster Surveys between 2000 and 2019 in 54 LMICs, we estimated proportion of ANC visits among women. Bayesian hierarchical regression models were used to estimate trend, projection, and determinants of single and four ANC visits (ANC1 and ANC4) independently. Equity analysis were performed to assess the magnitude of wealth-based and urban-rural inequalities in access to ANC visits. Compared to women aged 36-49 years, coverage of ANC1 and ANC4 are expected to increase significantly for adolescent mothers and women aged 20-35 years. This increase was observed at the national level, as well as both urban and rural areas in most countries between 2000 and 2030. By 2030, the coverage of ANC1 is predicted to reach 80% or more in all countries except Angola, Central African Republic and Togo, whereas only 16 countries are predicted to reach 80% or more for ANC4. According to wealth quintile, the lowest inequalities with highest coverage of 80% or more ANC4 will be observed in Armenia, Cambodia, Dominican Republic, Ghana, Maldives, Indonesia, and Sao Tome and Principe in 2030. Determinant analysis found increased odds of receiving ANC visits during pregnancy for adolescent mothers with higher educational levels, frequency of listening/watching mass media, and various household socio-economic status factors. This study calls for advanced, innovative and cost-effective approaches to increase ANC coverage among adolescent mothers, particularly in rural areas and/or in low socioeconomic groups.


15 years of litigation following laparoscopic cholecystectomy in England.
We aimed to analyze trends in litigation following laparoscopic cholecystectomy (LC) in England and compare our findings with data from the United States. Several studies from the United States have highlighted the medico-legal repercussions of complications following LC. In 2007-2008, litigation claims cost the National Health Service in England over 660 million Great British Pounds (GBP) (1.1 billion USD). Despite this, there has been little examination of litigation following LC in England. Data from the National Health Service Litigation Authority on clinical negligence claims between 1995 and 2009 following LC were obtained and analyzed. Four hundred eighteen claims were made of which 303 were settled. One hundred ninety-eight (65%) were found to be in the claimants favor for a total cost of 20.4 million GBP (33.4 million USD). Litigation claims have leveled since 2001. Operator error was the most likely cause to result in a claim and the only cause associated with a successful claim (P = 0.023). A delay in the recognition of complications was the second most common reason for initiation of a claim. Bile duct injury was the most frequent injury resulting in litigation and the most likely injury associated with a successful claim (P < 0.001). The average payout for a successful claim was 102,827 GBP/168,337 USD. Findings from US studies were similar, although the magnitude of payouts was 4 times higher. Strategies that minimize bile duct injury and speed up recognition of injuries should be adopted to reduce the litigation burden and improve patient care.


Spatio-temporal evolution of traditional soybean planting structure in Songnen Plain, China in 1996-2016.
Under the impacts of genetically modified soybean, the planting area of traditional soybean in China has fallen sharply. Understanding the traditional soybean planting structure and dynamics in the main soybean producing areas is of great significance to China's soybean import and export trade and safeguarding national food security. With the Landsat TM/OLI remote sensing images from 1996 to 2016, we extracted the spatial distribution information of soybean planting using random forest (RF) method based on remote sensing (RS) and geographic information system (GIS) techniques and analyzed the spatiotemporal evolution characteristics of soybean planting structure in the Songnen Plain using landscape pattern index. The results showed that the soybean planting area in the Songnen Plain was fluctuating during 1996-2016. Among the area conversion with other land-use types, the interchanges between soybean and other crop lands were the most obvious. The proportion of soybean patch area to the total crop area increased first and then decreased, while the patch density index and the resolution index decreased first and then increased. In conclusion, the spatiotemporal evolution of the traditional soybean planting structure in the Songnen Plain showed the characteristics of fluctuation in planting area, increase in fragmentation, and discrete spatial distribution. The domestic and foreign markets as well as policy guidance were the important factors afecting the traditional soybean planting structure.


Role of radiation therapy in patients with resectable pancreatic cancer.
The 5-year overall survival of patients with pancreatic cancer is approximately 5%, with potentially resectable disease representing the curable minority. Although surgical resection remains the cornerstone of treatment, local and distant failure rates are high after complete resection, and debate continues as to the appropriate adjuvant therapy. Many oncologists advocate for adjuvant chemotherapy alone, given that high rates of systemic metastases are the primary cause of patient mortality. Others, however, view locoregional failure as a significant contributor to morbidity and mortality, thereby justifying the use of adjuvant chemoradiation. As in other gastrointestinal malignancies, neoadjuvant chemoradiotherapy offers potential advantages in resectable patients, and clinical investigation of this approach has shown promising results; however, phase III data are lacking. Further therapeutic advances and prospective trials are needed to better define the optimal role of adjuvant and neoadjuvant treatment in patients with resectable pancreatic cancer.


Molecular and biochemical characterization of mannitol-1-phosphate dehydrogenase from the model brown alga Ectocarpus sp.
The sugar alcohol mannitol is important in the food, pharmaceutical, medical and chemical industries. It is one of the most commonly occurring polyols in nature, with the exception of Archaea and animals. It has a range of physiological roles, including as carbon storage, compatible solute, and osmolyte. Mannitol is present in large amounts in brown algae, where its synthesis involved two steps: a mannitol-1-phosphate dehydrogenase (M1PDH) catalyzes a reversible reaction between fructose-6-phosphate (F6P) and mannitol-1-phosphate (M1P) (EC 126.96.36.199), and a mannitol-1-phosphatase hydrolyzes M1P to mannitol (EC 22.214.171.124). Analysis of the model brown alga Ectocarpus sp. genome provided three candidate genes for M1PDH activities. We report here the sequence analysis of Ectocarpus M1PDHs (EsM1PDHs), and the biochemical characterization of the recombinant catalytic domain of EsM1PDH1 (EsM1PDH1cat). Ectocarpus M1PDHs are representatives of a new type of modular M1PDHs among the polyol-specific long-chain dehydrogenases/reductases (PSLDRs). The N-terminal domain of EsM1PDH1 was not necessary for enzymatic activity. Determination of kinetic parameters indicated that EsM1PDH1cat displayed higher catalytic efficiency for F6P reduction compared to M1P oxidation. Both activities were influenced by NaCl concentration and inhibited by the thioreactive compound pHMB. These observations were completed by measurement of endogenous M1PDH activity and of EsM1PDH gene expression during one diurnal cycle. No significant changes in enzyme activity were monitored between day and night, although transcription of two out of three genes was altered, suggesting different levels of regulation for this key metabolic pathway in brown algal physiology.


The effect of hypoglycemia on myocardial ischemic injury during acute experimental coronary artery occlusion.
To determine the effect of hypoglycemia on myocardial ischemic injury following coronary artery occlusion epicardial electrograms were recorded 15 minutes after two 20-minute coronary artery occlusions in seven anesthetized dogs. The first occlusion was a control (blood glucose 85 plus or minus 5(sd) mg per cent). Before the second occlusion hypoglycemia was induced (blood glucose 40 plus or minus 5 mg per cent) by the intravenous administration of insulin (2 units/kg). The average ST-segment elevation in leads during control was 3.5 plus or minus 1.0 mV which rose to 6.1 plus or minus 1.4 mV during the second occlusion (P smaller than 0.05). The number of sites showing ST-segment elevation exceeding 2 mV increased from 7.6 plus or minus 1.6 during control to 10.6 plus or minus 1.4 (P smaller than 0.05) during the occlusion with hypoglycemia. In other dogs, a coronary artery was occluded for 24 hours. Epicardial ST-segment elevations were compared to creatine phosphokinase (CPK) activity and histological appearance from the same sites. CPK activity in sites with normal ST segments (0-2 mV) was 33.1 plus or minus 6.0 IU/mg protein. Six additional dogs received insulin following the 15 minute epicardial map and blood sugar was maintained at a level of 46 plus or minus 6 mg per cent for the 24 hours. These dogs showed more myocardial necrosis than predicted by the ST-segment elevation prior to insulin administration. Forty-six percent of sites, which in control dogs would have been expected to have normal CPK and histological appearance, showed depressed CPK activity and histological evidence of early myocardial necrosis. Thus, hypoglycemia increases myocardial damage, as reflected by enzymatic and histological analyses.


Aging in attraction-driven colloidal glasses.
Aging in an attraction-driven colloidal glass is studied by computer simulations. The system is equilibrated without attraction and instantaneously "quenched," at constant colloid volume fraction, to one of two states beyond the glass transition; one is close to the transition, and the other one deep in the glass. The evolution of structural properties shows that bonds form in the system, increasing the local density, creating density deficits (holes) elsewhere. This process slows down with the time elapsed since the quench. As a consequence of bond formation, there is a slowing down of the dynamics, as measured by the mean-squared displacement and the density, bond, and environment correlation functions. The density correlations can be time rescaled to collapse their long time (structural) decay. The time scale for structural relaxation shows for both quenches a superlinear dependence on waiting time; it grows faster than the bond lifetime, showing the collective origin of the transition. At long waiting times and high attraction strength, we observe arrested dynamics for more than three decades in time, although individual bonds are not permanent on this time scale. The localization length decreases as the state moves deeper in the glass; the nonergodicity parameter oscillates in phase with the structure factor. Our main results are obtained for systems with a barrier in the pair potential that inhibits phase separation. However, when this barrier is removed for the case of a deep quench, we find changes in the static structure but almost none in the dynamics. Hence our results for the aging behavior remain relevant to experiments in which the glass transition competes with phase separation.


Objects are selected for attention based upon meaning during passive scene viewing.
While object meaning has been demonstrated to guide attention during active scene viewing and object salience guides attention during passive viewing, it is unknown whether object meaning predicts attention in passive viewing tasks and whether attention during passive viewing is more strongly related to meaning or salience. To answer this question, we used a mixed modeling approach where we computed the average meaning and physical salience of objects in scenes while statistically controlling for the roles of object size and eccentricity. Using eye-movement data from aesthetic judgment and memorization tasks, we then tested whether fixations are more likely to land on high-meaning objects than low-meaning objects while controlling for object salience, size, and eccentricity. The results demonstrated that fixations are more likely to be directed to high meaning objects than low meaning objects regardless of these other factors. Further analyses revealed that fixation durations were positively associated with object meaning irrespective of the other object properties. Overall, these findings provide the first evidence that objects are, in part, selected by meaning for attentional selection during passive scene viewing.


Measuring a frequency spectrum for single-molecule interactions with a confined nanopore.
Nanopore analysis is a powerful technique for single molecule analysis by virtue of its electrochemically confined effects. As a single molecule translocates through the nanopore, the featured ionic current pattern on the time scale contains single molecule characteristics including volume, charge, and conformational properties. Although the characteristics of a single molecule in a nanopore have been written to the featured ionic current, extracting the dynamic information from a complex current trace is still a big challenge. Here, we present an applicable nanopore analysis method employing the Hilbert-Huang Transform (HHT) to study the vibrational features and interactions of a single molecule during the dynamic translocation process through the confined space of a nanopore. The HHT method is specially developed for analyzing nonlinear and non-stationary data that is highly compatible with nanopore data with a high frequency resolution. To provide proof-of-concept, we applied HHT to measure the frequency response for the wild-type (WT) aerolysin and mutant K238E aerolysin nanopores with and without the presence of poly(dA)4, respectively. The energy-frequency-time distribution spectra demonstrate that the biological nanopore contributes greatly to the characteristics of the high frequency component (>2 kHz) in the current recording. Our results suggest that poly(dA)4 undergoes relatively more consistent and confined interactions with K238E than WT, leading to a prolonging of the duration time. Therefore, the characteristics in frequency analysis could be regarded as an "single-molecule ionic spectrum" inside the nanopore, which encodes the detailed behaviours of single-molecule weak interactions.


Modelling the erythroblastic island niche of dyserythropoietic anaemia type IV patients using induced pluripotent stem cells.
Introduction: Congenital dyserythropoietic anaemia (CDA) type IV has been associated with an amino acid substitution, Glu325Lys (E325K), in the transcription factor KLF1. These patients present with a range of symptoms, including the persistence of nucleated red blood cells (RBCs) in the peripheral blood which reflects the known role for KLF1 within the erythroid cell lineage. The final stages of RBCs maturation and enucleation take place within the erythroblastic island (EBI) niche in close association with EBI macrophages. It is not known whether the detrimental effects of the E325K mutation in KLF1 are restricted to the erythroid lineage or whether deficiencies in macrophages associated with their niche also contribute to the disease pathology. Methods: To address this question, we generated an in vitro model of the human EBI niche using induced pluripotent stem cells (iPSCs) derived from one CDA type IV patient as well as two iPSC lines genetically modified to express an KLF1-E325K-ERT2 protein that could be activated with 4OH-tamoxifen. The one patient iPSC line was compared to control lines from two healthy donors and the KLF1-E325K-ERT2 iPSC line to one inducible KLF1-ERT2 line generated from the same parental iPSCS. Results: The CDA patient-derived iPSCs and iPSCs expressing the activated KLF1-E325K-ERT2 protein showed significant deficiencies in the production of erythroid cells with associated disruption of some known KLF1 target genes. Macrophages could be generated from all iPSC lines but when the E325K-ERT2 fusion protein was activated, we noted the generation of a slightly less mature macrophage population marked by CD93. A subtle trend in their reduced ability to support RBC enucleation was also associated with macrophages carrying the E325K-ERT2 transgene. Discussion: Taken together these data support the notion that the clinically significant effects of the KLF1-E325K mutation are primarily associated with deficiencies in the erythroid lineage but it is possible that deficiencies in the niche might have the potential to exacerbate the condition. The strategy we describe provides a powerful approach to assess the effects of other mutations in KLF1 as well as other factors associated with the EBI niche.


[Fluid suspension co-culture of Dendrobium officinale protocorm and living fungus].
To set up a system for fluid suspension co-culture of Dendrobium officinale protocorm and living fungus MF24, so as to provide certain scientific evidence for industrial production of protocorm. Whether the protocorm culture system was suitable for the normal growth of MF24 fungus were studied, the growth of protocorm cultured alone and co-cultured with the fungus were researched under light and dark culture conditions, the biomass and proliferation times were determined, and HPLC method was used to analyze and compare the changes of 11 characteristic peak areas in D. officinale protocorm. The MF24 fungus could grow normally in the 6,7-V liquid medium used to culture the protocorm, and when it was cultured by 8-10 hours per day under 1 500 lx, the growth rate of the fungus was slowed. Protocorm could grow normally in light and dark culture conditions, and add the MF24 fungus in the early cultivation stages of protocorm, both inhibit the growth of each other. In the protocorm for the growth stability to add 5 diameter 9 mm fungi block, the protocorm growth and chemical composition type had no significant effect. However, under illumination, co-cultured for 5 days protocorm of which 10 compounds content decreased 13.64% to 138.47%, in dark conditions, co-cultured for 5 days protocorm of which 7 compounds increased by 0.71% to 12.82%, and 4 compounds slightly reduced by 3.03% to 14. 14% compared with the control. Under the appropriate condition, living fungus MF24 could co-culture with the D. officinale protocorm, and affected the latter's secondary metabolite levels.


Acetylation of Abelson interactor 1 at K416 regulates actin cytoskeleton and smooth muscle contraction.
Actin cytoskeletal reorganization plays an important role in regulating smooth muscle contraction, which is essential for the modulation of various physiological functions including airway tone. The adapter protein Abi1 (Abelson interactor 1) participates in the control of smooth muscle contraction. The mechanisms by which Abi1 coordinates smooth muscle function are not fully understood. Here, we found that contractile stimulation elicited Abi1 acetylation in human airway smooth muscle (HASM) cells. Mutagenesis analysis identified lysine-416 (K416) as a major acetylation site. Replacement of K416 with Q (glutamine) enhanced the interaction of Abi1 with neuronal Wiskott-Aldrich syndrome protein (N-WASP), an important actin-regulatory protein. Moreover, the expression of K416Q Abi1 promoted actin polymerization and smooth muscle contraction without affecting myosin light chain phosphorylation at Ser-19 and vimentin phosphorylation at Ser-56. Furthermore, p300 is a lysine acetyltransferase that catalyzes acetylation of histone and non-histone proteins in various cell types. Here, we discovered that a portion of p300 was localized in the cytoplasm of HASM cells. Knockdown of p300 reduced the agonist-induced Abi1 acetylation in HASM cells and in mouse airway smooth muscle tissues. Smooth muscle conditional knockout of p300 inhibited actin polymerization and the contraction of airway smooth muscle tissues without affecting myosin light chain phosphorylation and vimentin phosphorylation. Together, our results suggest that contractile stimulation induces Abi1 acetylation via p300 in smooth muscle. Acetylation at K416 promotes the coupling of Abi1 with N-WASP, which facilitates actin polymerization and smooth muscle contraction. This is a novel acetylation-dependent regulation of the actin cytoskeleton in smooth muscle.


[Changes in lipids of granulation fibrous tissue in rats at various doses and routes of melatonin administration].
The experimental data indicate that melatonin actively influences time-causes of changes of lipid content in rats tissue during the inflammation process. Its effect depends on a dose, modes of administration (intraperitonial, hypodermic or local) and duration of treatment. A single dose intraperitonial administration of melatonin (4 mg/kg) did not influence lipid content in the granular-fibrose tissue, while repeated injections of this hormone limited the increase in contents of lipids and phospholipids at the 5th and 8th days of regeneration. Long-term subcutaneous injections of melatonin caused distinct changes of lipids: at the dose of 0.3 mg/kg it prevented, and at the dose of 4 mg/kg it promoted the increase of lipid content in the granular-fibrose tissue. Local application of a melatonin solution (1.5 mg/ml) at early periods of regeneration caused insignificant changes of total lipids and total phospholipids in the granular-fibrose tissue. However, the higher concentration (15 mg/ml) of melatonin caused the decrease of total lipids due to reduced content of cholesterol and triglycerides and the increase of total phospholipids and some of their fractions.


Modification of motor cortex excitability during muscle relaxation in motor learning.
We postulated that gradual muscle relaxation during motor learning would dynamically change activity in the primary motor cortex (M1) and modify short-interval intracortical inhibition (SICI). Thus, we compared changes in M1 excitability both pre and post motor learning during gradual muscle relaxation. Thirteen healthy participants were asked to gradually relax their muscles from an isometric right wrist extension (30% maximum voluntary contraction; MVC) using a tracking task for motor learning. Single or paired transcranial magnetic stimulation (TMS) was applied at either 20% or 80% of the downward force output during muscle release from 30% MVC, and we compared the effects of motor learning immediately after the 1st and 10th blocks. Motor-evoked potentials (MEPs) from the extensor and flexor carpi radialis (ECR and FCR) were then measured and compared to evaluate their relationship before and after motor learning. In both muscles and each downward force output, motor cortex excitability during muscle relaxation was significantly increased following motor learning. In the ECR, the SICI in the 10th block was significantly increased during the 80% waveform decline compared to the SICI in the 1st block. In the FCR, the SICI also exhibited a greater inhibitory effect when muscle relaxation was terminated following motor learning. During motor training, acquisition of the ability to control muscle relaxation increased the SICI in both the ECR and FCR during motor termination. This finding aids in our understanding of the cortical mechanisms that underlie muscle relaxation during motor learning.